2015

VOLUME 57

Suppl. 1/2015

Published quarterly ISSN 0204-8043 (print) ISSN 1314-2143 (online)

C O L L E C T I O N O F RESEARCH WORK

EDITORIAL BOARD

Editor-in-chief Professor Stefan Kostianev

Deputy/ Managing Editor Professor Victoria Sarafian Professor Todorka Tsvetkova

Editorial Advisory Board Professor Blagovest Pehlivanov Professor Damianka Getova-Spasova Assoc. Professor Beniamin Anavi

Statistical Advisors Assoc. Professor Nonka Mateva, Kiril Simitchiev Assoc. Professor Maria Semerdjieva Technical Editors:Vanya Gjulina, Nina Atanasova

Members Professor Bernard Amour Paris, France Professor George Anastassov NYC, USA Professor Nikolai Boyadjiev Plovdiv, Bulgaria Professor Patrick Dhellemmes Lille, France Professor Christo Dobrev Plovdiv, Bulgaria Professor Ben Zion Garty Tel Aviv, Israel Professor Stefan Goranov Plovdiv, Bulgaria Professor Ivan Ivanov Plovdiv, Bulgaria Professor Margarita Kasarova Plovdiv, Bulgaria Professor Konstantinos Kazakos Alexandroupolis, Greece Professor Konstantinos Kouskoukis Alexandroupolis, Greece Professor Atanas Krastev Plovdiv, Bulgaria Professor Ingrid Kreissig Munheim, Germany Professor Maria Kukleva Plovdiv, Bulgaria Professor John Kyriopoulos Athens, Greece Professor Jukka Meurman Helsinki, Finland Professor Mikhail Mikhailovsky Novosibirsk, Russia Professor Andrew Miles London, UK Professor Vanyo Mitev Sofia, Bulgaria

Professor Mariana Murdjeva Plovdiv, Bulgaria Professor Fedia Nikolov Plovdiv, Bulgaria Professor Michael Oellerich Göttingen, Germany Professor Philippe Pellerin Lille, France Professor Yves Poumay Namur, Belgique Professor Emanuela Signori Roma, Italy Professor Constantinos Simopoulos Alexandroupolis, Greece Professor Stefan Sivkov Plovdiv, Bulgaria Professor Francisco Soriano Madrid, Spain Professor Chavdar Stefanov Plovdiv, Bulgaria Professor Rumen Stefanov Plovdiv, Bulgaria Professor Yordanka Stoilova Plovdiv, Bulgaria Professor Drozdstoy Stoyanov Plovdiv, Bulgaria Professor Mariana Stoycheva Plovdiv, Bulgaria Professor Dobrin Svinarov Sofia, Bulgaria Professor Georgi Todorov Plovdiv, Bulgaria Professor Angel Uchikov Plovdiv, Bulgaria Professor Kichka Velkova Plovdiv, Bulgaria Professor Eberhard Wieland Stuttgart, Germany

Language Editors (English): Assoc. Prof. O. Obretenov (Plovdiv, Bulgaria), J. Oldham (Plovdiv, Bulgaria) Language Editor (Russian): P. Kancheva (Plovdiv, Bulgaria), B. Philipov (Plovdiv, Bulgaria) Typeset by Medical University - Plovdiv, Scientific Department, Dipl. Eng. N. Atanasova Printed in Bulgaria, Plovdiv, Avtoprint LTD ISSN 0204-8043 (print) ISSN 1314-2143 (online) Number of copies: 1500

Editorial Correspondence FOLIA MEDICA, MU, Library and Information Center, V. Gjulina (e-mail: [email protected]) 15 A Vassil Aprilov St., 4002 Plovdiv, Bulgaria

Tel. +359 32/602 588; Fax: +359 32/602 534 http://www.degruyter.com/view/j/folmed www.foliamedica.bg e-mail: [email protected]

57, Suppl. 1 2015

C O N T E N T S Anniversary Scientific Conference - 70 Years Medical University of Plovdiv (20-23 May 2015) .........................................................................................5 Organizing Committee ...................................................................................................................... 6 Welcome Address of the Rector of Medical University - Plovdiv - Prof. Stefan Kostianev ............ 7 Keynote Lecture ................................................................................................................................ 9

Plenary Lectures Biomedicine ....................................................................................................................................... 9 Biopharmacy .................................................................................................................................... 15 Public Health................................................................................................................................... 17 Therapy ............................................................................................................................................ 20 Surgery ............................................................................................................................................. 25 Dental Medicine .............................................................................................................................. 29

Oral presentations Biomedicine ..................................................................................................................................... 32 Biopharmacy .................................................................................................................................... 36 Public Health................................................................................................................................... 39 Therapy ............................................................................................................................................ 41 Surgery ............................................................................................................................................. 44 Dental Medicine .............................................................................................................................. 48

Posters Biomedicine ..................................................................................................................................... 52 Biopharmacy .................................................................................................................................... 68 Public Health................................................................................................................................... 78 Therapy ............................................................................................................................................ 94 Surgery ........................................................................................................................................... 112 Dental Medicine ............................................................................................................................ 128 Science and Youth ........................................................................................................................ 140

XI National Conference of Medical Biology (15-17 May 2015) ......................................................................................................173 Organizing Committee .................................................................................................................. 174 Welcome Address of the Vice Rector of MU Plovdiv - Prof. V. Sarafian ............................ 175 Plenary Lectures............................................................................................................................ 177 Posters ............................................................................................................................................ 182

57, Suppl. 1 2015

Anniversary Scientific Conference “Science for Health” 70 Years Medical University of Plovdiv (20-22 May 2015)

The 70th Anniversary Scientific Conference is held under the auspices of the Rector of Medical University - Plovdiv

ORGANIZING COMMITTEE of the 70th Anniversary Scientific Conference “Science for Health” 70 Years Medical University of Plovdiv (20-22 May 2015) President: Corr.-Mem. Prof. Stefan Kostianev, MD, PhD, DMSc Vice President: Prof. Victoria Sarafian, MD, PhD, DMSc Scientific committee: Head: Prof. Blagovest Pehlivanov, MD, PhD, DMSc Members: Prof. Hristo Dobrev, MD, PhD Prof. Chavdar Stefanov, MD, PhD, DMSc Professor Ana Maneva, PhD, DBSc Assoc. Prof. Snejana Tzanova, DDM, PhD Assoc. Prof. Nonka Mateva, PhD Eng. Angelina Kirkova Secretariat: Head: Assoc. Prof. Blagoi Marinov, MD, PhD Members: Delyan Delev, MD, PhD Stoilka Mandadzhieva, MD, PhD Dimitar Penkov, MPharm Ivan Shekerev

RECTOR'S ADDRESS

Dear colleagues,

It is a great pleasure for the academic community of Medical University - Plovdiv to welcome you to the Scientific Conference dedicated to the 70th Anniversary of our Alma Mater. This forum is intended to be a cross point for eminent academics and practitioners from all fields of medical science. Over the years of its existence, Medical University – Plovdiv went through a long way of recognition, strengthening and development. I hope that this conference will be a venue for in-depth discussions, original ideas and memorable meetings. I believe all participants will take advantage of seven decades of traditions to present their achievements and to share experience in all aspects of medical science, education and practice. I wish you a very successful and enjoyable conference! Welcome to Plovdiv!

Corr.-mem. Prof. Stefan Kostianev, MD, PhD, DMSc Rector of Medical University - Plovdiv

Folia Medica 2015; 57; Suppl. 1 Copyright © 2015 Medical University, Plovdiv, Bulgaria

KEYNOTE LECTURE

WHAT ARE THE SOLUTIONS TODAY TO ERADICATE THE HIV/AIDS VIRUS, MORE THAN 30 YEARS AFTER ITS DISCOVERY ? Jean-Claude Chermann Pasteur Institute Medical School, Marseille, France HIV/AIDS virus was isolated in 1983 by my group at the Pasteur Institute. A great progress was made to use the combined therapy with different antiretroviral drugs to avoid the AIDS disease. Why such situation? 1. The complexity of the retroviral genomic structure with the structural genes appears for the first time in virology, additional regulatory as well as the auxiliary genes increasing the viral production and then numerous mutations. 2. These mutations explain the diversity of the virus; it changes from one individual to another, also with time in the same individual and also with the antiretroviral treatment becoming resistant to the drugs. 3. This variability at the level of viral envelope explains the failure of the previous vaccine trials. 4. The virus induces numerous associated diseases: by infecting and killing the CD4 cells it induces AIDS, by infecting the macrophage it is responsible for the dementia moving in the brain, atypical pneumonia in the lung... By infecting the intestinal cells it provokes malabsorption (explaining the weight loss) and clean diarrhea. By infecting the CD34 - anemia, lymphopenia and thrombocytopenia are often induced. To eradicate a virus a need of a vaccine is obligatory, then a new way should be developed. I propose to use a cellular antigen (R7V) taken by the retrovirus during the budding at the cell surface and common to all isolates. Protective antibodies to R7V are found in non progressor patients and neutralize all HIV isolates including resistant viruses. Key words: HIV/AIDS virus

PLENARY LECTURES

Biomedicine THE FUTURE OF THE BRAIN Stefan Sivkov Department of Anatomy, Histology and Embryology, Medical University - Plovdiv, Bulgaria The human brain contains about 100 billion neurons, each interacting with 10,000 other neurons through an intricate connectivity. As a result, there may be one quadrillion connections between neurons in the brain each firing 10x in a second. Presently however this incredible network is beyond the capacity of technologies to trace it. Advances in nanotechnology, microelectronics, optics, compression and data storage, cloud computing, information theory and synthetic biology can help to make possible studies that were unthinkable to conduct before. For example, scientists can extend the value of traditional brain scans by implantation of nanosensors, wireless fiberoptic probes or by genetically engineered living cells to browse brain tissue and identify the neurons activated in response to miscellaneous incentives. The goal in the near future is to develop new technologies in the field of data processing, nanotechnology, optogenetics and other abstruse Folia Medica 2015; 57; Suppl. 1 © 2015 Medical University, Plovdiv, Bulgaria

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Anniversary Scientific Conference “Science for Health” 70 Years Medical University of Plovdiv

areas and explore the interactivity of those billions of brain cells and complex neural circuits. Acquired knowledge will create models of the organization of neural circuits and determine their effect on behavior and cognition - one of the greatest mysteries. It would also enable researchers in the biomedical science to develop more precise ways of diagnosis and treatment of disabling brain diseases like depression, schizophrenia, dementia, autism, stroke, parkinsonism. Key words: brain, neurons, circuits, technologies, diseases

STUDYING PATHOLOGIES OF THE BARRIER IN RECONSTRUCTED HUMAN EPIDERMIS Yves Poumay1, Evelyne De Vuyst1, Severine Giltaire2, Aline Chretien3, Michel Salmon3, Catherine Lambert De Rouvroit1 1University of Namur, 2University of Namur and Straticell Inc, 3Straticell Inc, Belgium The etiology of atopic dermatitis (AD) is still puzzling. Interplay between abnormal epidermal barrier and T helper type 2 (Th2) immune response is suspected to create vicious circles between both components since they induce each other. Transcriptomic studies on keratinocytes after cholesterol depletion have shown strong similarities with AD deregulated gene expression. Th2specific interleukins (IL) -4, -13 and -25 also altered keratinocyte gene expression reminiscent of AD. This study combined endogenous epidermal activation with Th2-interleukins to analyze potential synergistic effects on epidermal barrier. Reconstructed human epidermis (RHE) was chosen to analyze keratinocytes in tissue-specific context. Its activation was initiated by two hours of cholesterol depletion, a condition capable of initiating EGFR and p38MAPK-dependent signalling and transiently alter their gene expression. Interestingly, cholesterol depletion in mature RHE induced the expression of thymic stromal lymphopoietin which alerts the immune system in AD. The effects produced by IL-4, -13 and -25 also resulted in specific deregulated gene expression of CA2 and NELL2. They also altered RHE morphology as granular layer disappeared, intercellular edema developed, and loricrin, filaggrin and transglutaminase-1 expressions were reduced. Barrier function was evaluated by trans-epidermal electrical resistance and Lucifer Yellow diffusion through RHE. Each treatment alone was unable to alter barrier functions. Conversely, cholesterol depletion followed by exposure to Th2-ILs resulted in significant alterations of barrier functions and further reduced expression of keratinization markers. Thus, epidermal or immune activation alters keratinocyte within RHE, but combined activation is required to modify the barrier and reproduce AD in vitro. Key words: epidermis, interleukins, barrier, atopic dermatitis

PERSPECTIVES ON PERSONALIZED MOLECULAR DIAGNOSTICS Michael Oellerich Institute for Clinical Pharmacology, University Medical Center Göttingen, Germany Technological advances in the use of novel molecular biomarkers have paved the way for personalized diagnostics. Transplantation biomarkers have especially attracted attention, because there are still unresolved problems (e.g. irreversible chronic allograft dysfunction, side effects of standard immunosuppression) that limit long-term outcome. There are limitations to how immunosuppressive drugs are currently monitored. Therapeutic drug monitoring is more useful to prevent toxicity than to predict efficacy. Biomarkers are needed that can be used to facilitate personalized immunosuppression. Various strategies are currently being evaluated, including biomarkers of immune response (IL-2 expression in CD8+ T cells, DSA), predictors of tolerance (e.g. nTregs,

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Folia Medica 2015; 57; Suppl. 1 © 2015 Medical University, Plovdiv, Bulgaria


specific gene expression signatures), and molecular markers of graft injury. A particularly promising new approach for early detection of graft rejection is based on the determination of graftderived circulating cell-free DNA, using droplet digital PCR. This assay has the advantage that it directly interrogates the health of the donor organ (“liquid biopsy”). It may also be useful to monitor immunosuppression minimization. New diagnostic platforms used for high-quality genomic analysis will also be critical for a personalized medicine approach to cancer patient management where genome sequencing can potentially identify the molecular abnormalities that predict either good or poor outcomes and to identify new targets for therapy. Tumor-specific rearrangements identified in cell-free DNA isolated from serial patient blood samples will be helpful in monitoring relapse or response to treatments. In the future, such personalized medicine approaches will shift emphasis from reaction to prevention and could improve outcomes at lower healthcare costs. Key words: biomarkers, transplantation, cancer, personalized diagnostics

TH17 CELLS IN INFECTIOUS IMMUNITY - BALANCE OR DAMAGE ? Marianna Murdjeva Department of Microbiology and Immunology, Faculty of Pharmacy, Medical University - Plovdiv, Bulgaria Three distinct subsets of T helper (Th) cells mediate immunity in infections - Th1 are responsible in cellular immune reactions against intracellular bacteria and viruses, Th2 are involved in humoral immunity against parasites, and regulatory T cells (T regs) keep check in inflammatory responses. Recently, a novel lineage of T helpers - Th17, bridging innate and adaptive immunity, was established with a critical role in mucosal homeostasis, especially against extracellular bacteria, colonizing airways, skin and gut lumen. These cells secrete many cytokines - interleukin-17 (IL-17), IL-17F, IL-22, IL-6 and TNF-α. It is questionable whether Th17 cells are associated with antimicrobial mechanisms or cause tissue destruction. Their protective function in infections is confirmed by the observation that IL-17 and IL-22 induce production of antimicrobial peptides (beta-defensin 2, lipocalin-2) from gut, lung and skin epithelia. IL-17 potentiates neutrophil recruitment, cytotoxicity and phagocytosis. Defective IL17 signaling leads to Klebsiella pneumoniae, Bordetella pertussis and Streptococcus pneumoniae infections. The potentially damaging nature of Th17 cells is demonstrated in pathological abscess formation in response to Bacteroides fragilis. In HIV/AIDS patients the defect of IL-17 results in higher susceptibility to opportunistic infections by Staphylococcus aureus, Haemophilus influenzae and Streptococcus pneumoniae. IL-17 deficiency in disseminated and oral candidiasis and pulmonary aspergillosis exacerbates pathology. A synergy between Th1 and Th17 lineage is required for better protection against intracellular bacteria (Mycobacterium tuberculosis, Listeria monocytogenes). Elucidation of mechanisms governing amplification and attenuation of Th17 cells in infections is important for the development of therapeutic strategies aiming at modulating the immune response. Key words: Th17 cells, IL-17, infectious immunity

IMMUNOLOGY OF AGING Elissaveta Naumova Department of Clinical Immunology and Stem Cell Bank, Alexandrovska University Hospital, Medical University, Sofia, Bulgaria Aging is a process of progressive loss of body functions, leading to a decreased ability to adapt to environmental changes and maintenance of homeostasis. Longevity is achieved by the preserva-


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Plenary Lectures


tion of good health, depending in 75% on lifestyle and environmental factors. However, genetic factors, including polymorphic immune genes, contribute in 25% to successful aging. The immune system is a reliable marker in determining the biological age, and in predicting longevity. The decline of physiological potential of immune system with age affects the optimal function of other systems (eg. nervous, endocrine). The clinical implications of the aging of the immune system are: increased susceptibility to infectious diseases and severity of infections leading to increased mortality, partial loss of efficacy of the vaccines, increased autoreactivity and tendency to develop tumors. Identifying immune biomarkers that indicate the beginning of aging, the level of immunocompetence, vaccine responses and susceptibility to infections will allow the development of strategies to restore the immune system, better selection approaches for prevention and treatment of elderly people, and thus contributing to successful aging. The findings of our collaborative study focused on the role of immune genes and changes in the immune system related to aging will be presented. In addition, in the light of our results and the data from the literature we will attempt to summarize and discuss the current knowledge on different aspects of immunology of aging. Key words: immune system, aging, longevity, immunosenescense

NOVEL INSIGHT FROM A COLORECTAL CANCER MOUSE MODEL: NOTUM AND GLYPICANS AS POSSIBLE TARGET FOR GENE THERAPY PROTOCOLS Emanuela Signori University Campus Bio-Medico and CNR Institute of Translational Pharmacology, Rome, Italy After some decades of extensive research efforts, many of the original fears associated with gene transfer have disappeared. Public perception has become positive in the hope of more effective treatments, cures, or preventive interventions. However, preclinical studies of effective therapeutic agents delivered in animal models to translate into the clinic remain a crucial feature. To select adequate genes for a target gene therapy, studies on the mechanisms of onset and development of cancer diseases in appropriate in vivo models are becoming of utmost importance, especially in case of tumors whose regulatory pathways result difficult to study in humans, due to ethical considerations which make impossible to recur to sampling specimens of patients, through repeated biopsies, from the early to the late stage of the disease. Colorectal cancer (CRC) represents the third most common neoplastic disease worldwide and the second leading cause of cancer death in the western world. To identify potential novel genes related to tumor progression, we performed a genome-wide expression profiling of AOM/DSS induced tumors. We found 2036 differentially expressed genes, 1092 upregulated and 944 downregulated genes, by comparing the colon mucosa in affected and control mice. The most altered genes were significantly involved in the activation of the Wnt/β-catenin pathway. Particularly, we demonstrated significant changes in NOTUM, GLYPICAN-1 and GLYPICAN-3 gene expression during CRC development. We propose these new genes as potential target for novel gene therapy strategies against CRC and we suggest the AOM/DSS mouse model as a valid translational platform for preventive/therapeutic protocols. Key words: animal model, colorectal carcinogenesis, gene expression profiling, gene therapy, glypicans, Notum

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THE PIONEERING OF E-LEARNING IN MEDICAL PHYSICS Slavik Tabakov Department of Medical Engineering and Physics, Clinic of Medical Physics, King’s College, London, UK Medical Physics was one of the first professions in the world to develop its own original e-learning materials. The projects EMERALD and EMIT (1994-2004) developed extensive e-Learning materials – training tasks, image databases (including 4000+ images), simulations, and one of the first e-learning web sites (www.emerald2.eu). Further, the project EMITEL (2005-2013) developed the first e-Encyclopaedia of Medical Physics (3100+ articles) with Multilingual Dictionary of terms (translated in 29 languages) – both major reference materials for the profession (www. emitel2.eu). These materials are used by thousands of colleagues all over the world. The success and impact of Medical Physics e-learning through these projects was recognised with the first educational award of the European Union – the Leonardo da Vinci Award. These projects, initially including specialists from UK, Sweden, Italy, Portugal, Ireland, France, Czech Republic and Bulgaria, expanded rapidly through the International Organization for Medical Physics (IOMP) to include more than 320 senior colleagues from 36 countries. Specialists from Medical University Plovdiv took an important role in the development of the web technology of these projects. Medical Physics e-learning was a stable support for the overall development of the profession. This was one of the important factors for the 40% global increase of medical physicists in the past 20 years. Currently almost 15,000 colleagues from all over the world are using these e-learning materials. Medical University - Plovdiv took an active part in these projects, what is now widely recognised. Key words: medical physics education, e-learning

MEDICAL PHYSICS EDUCATION IN HUNGARY Csilla Pesznyak, David Legrady, Attila Aszodi, Szabolcs Czifrus Budapest University of Technology and Economics, Institute of Nuclear Techniques, Budapest, Hungary The interdisciplinary education in medical physics at master level was organised by the Institute of Nuclear Techniques, Faculty of Natural Sciences, Budapest University of Technology and Economics in 2010. The medical physics specialisation aims at providing high level interdisciplinary theoretical and practical knowledge that can be applied to both clinical and research and development fields. The medical physics discipline was organised in order to educate qualified personnel to acquire the necessary skills to apply modern physical methods in medical diagnostics and therapy. The MSc programme is established on the basis of EFOMP, IPEM, AAPM and IAEA recommendations. The course curriculum comprises fundamental physical subjects (atomic and molecular physics, nuclear physics, particle physics) as well as fundamental medical knowledge (anatomy, physiology, radiobiology) required for subjects of clinical diagnostics and radiation therapy. Students may choose further subjects from the “compulsory optional” array concerning medical imaging, X-ray diagnostics, radiation therapy, magnetic resonance imaging, radiation protection, Monte Carlo calculation and its clinical applications, ultrasound diagnostics and nuclear medicine, respectively. The education provided by this programme is greatly sought-after on the labour market; therefore, it ensures the training of highly skilled professionals demanded in the region capable of equipment handling, operation, calibration, Q&A, trade, research and development. Key words: medical physics, medical diagnostics, radiation therapy

Folia Medica 2015; 57: Suppl. 1 © 2015 Medical University, Plovdiv, Bulgaria

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SIMULATION IN MEDICAL EDUCATION: QVO VADIS Evangelos Georgiou University Medical School, Athens, Greece Halsted’s training program, which has served medicine for almost a century, can be epitomized as ”See one, teach one, do one”. However, the extended introduction of advanced technology in medicine produces an exponential disruption of clinical practice, thus making urgent the need for a new education model and for validated assessment methodology. Today, the traditional teaching and skills assessment, previously responsibility of the supervising consultant, shifts into new standardized methodologies and has became a mandatory part of training and career development. The incorporation of the new technology into medical simulators allow instructors to deliver evidence-based training and assessment within a curriculum. This new paradigm shift can be summarized as “See one, practice many, do one”. One of the most important parts of the simulation curriculum (regardless of the simulation platform it is delivered on) is feedback. Through deliberate practice, simulation allows to augment feedback on performance. Metrics are an extrinsic augmented form of feedback. A simulator without metrics is really no better than an expensive video game. The most valuable metrics is identification of errors. Thus simulations should be a vehicle for the delivery of metric-based performance feedback rather than something that approximates the look and feel of performing the procedure on a patient. Evidence exists from multiple prospective randomized clinical studies that demonstrate unequivocally that simulation-based training improves skills performance and therefore convince the sceptical part of the medical community that this is a better way to prepare physicians for delivering better health services to patients. Key words: simulation, medical education

DISCREPANCY IN EXPRESSION OF MIRNAS AND MESSENGER RNAS FOR CARDIAC MYOSIN ISOFORMS IN HUMAN END-STAGE FAILING HEART Jan Kyselovic Department of Pharmacology, Faculty of Medicine, Faculty of Pharmacy, Comenius University, Bratislava, Slovakia In the heart, three distinct myosin heavy chain isoforms coexist in delicate balance and the shift from one myosin isoform to another may be one of the key factors causing heart failure. The aim of our study was to analyse the ‘cardiac myosin switching’ via gene expression of microRNAs in correlation with isoforms myosin MYH6, MYH7, MYH7B in human end-stage failing heart. The study included patients (n = 40) indicated for heart transplantation. The following clinical information about the patients was collected from hospital records: age, sex, BMI, blood pressure, type of heart failure (DCM, CAD, RCMP, AS), ECG, ECHO, catheterization data, serum biochemistry, coexisting disorders and pharmacotherapy. We examined samples from left ventricles of explanted hearts. In adult human explanted hearts, the slow-twitch myosin heavy chain MYH7 is expressed isoform (~98%) whereas fast-twitch MYH6 isoform constitutes only 1% of all myosin isoforms. Low expression of miR-208a and increased miR-208b has a significant correlation with expression cardiac MYH6 and MYH7 of the heart, with significant discrepancy between expression alpha-myosin and miR 208a vs control. There is a possibility of involving MYH7B and miR-499 with myosin switching. miR-1 and miR-133a seem to be in close relation with other cardiac microRNAs included in this study. In conclusion, dysregulated gene expression of myosin heavy chains resulting in MYH7 upregu-

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lation and MYH6 downregulation might be a pathological adaptation in heart failure regulated through a complex net of relationships between MHCs – transcription factors – epigenetic modulators, but probably no microRNAs 208a and 208b. Supported by VEGA 1/0905/14, VEGA 1/0949/15 Key words: heart failure, cardiac myosin switching, microRNAs

TRADITION AND INNOVATION IN MODERN AGE PATHOLOGY. GROSS ANATOMY AND MOLECULAR MEDICINE ARE COHABITERS Andrea Onetti Muda University Campus Bio-Medico, Rome, Italy Pathologists are physicians who specialize in the diagnosis and characterization of disease based on the examination of tissues removed from diseased body parts or biopsy samples. Pathology plays a vital role across all facets of medicine throughout our lives, from pre-conception to post mortem. In fact it has been said that “Medicine IS Pathology”. The pathologist’s work is performed by the use of several techniques, which have improved over the centuries. Interestingly, since the dawn of human history all medical activities were based solely on the resolving power of the human eye. A true revolution was brought about by the advent of the microscope, which allowed the study of the fine structure of living organisms. This also started a technological progress which led to today’s current molecular biology and digital pathology techniques, now part of the standard experience of modern pathologists. However, autopsy is still a fundamental activity in the practice of pathology. The purpose of an autopsy is to determine the exact clinical history of the patient, therefore identifying the cause of death as the last of the events which, linked together, lead to death. However, during the second half of the twentieth century there was a decline in the rate of autopsies performed, still persisting in present days. Post-mortem examination in modern, twenty-first century medical school should be considered always in intimate connection with the clinical study of the patient, and be perceived by educators as having a multifactorial role in providing the doctors of tomorrow with the appropriate knowledge and attitudes needed for the practice of medicine. Key words: pathology, gross anatomy, molecular medicine

PLENARY LECTURES

Biopharmacy MEDICATION ERRORS Ivanka Kostadinova Department of Pharmacology and Clinical Pharmacology, Medical University – Plovdiv, Bulgaria The causes for most of the common medication errors are reviewed in the lecture. A medication error is each preventable event causing or which may have caused harm to the patient while their treatment has been under the control of a medical specialist (a physician, a pharmacist or a nurse)


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Plenary Lectures


or under their own control. Such errors are related to the professional practice of medicine, communication related to this, labelling of medical products, leaflets, combination treatment, dosing, distribution, administration, education as well as application methods. Importance of the issue: according to US-derived statistics, more than 1,5 million people annually have been affected by inappropriate use of medication. Many drug therapy errors are never reported, meaning that the actual numbers may be even higher. Each year more than 100 000 people die from medication errors, and at least 400 000 people suffering from damage caused by the same are admitted into hospitals. That means that each day in the USA at least one patient experiences a medication error. In care homes, damage from medication errors has been done to 800 000 people, and is linked to incorrect use of drugs. With the available 30 billion prescriptions annually, it has been calculated that this leads to 51.5 million medication errors. The data from Europe are the following: 7.5% of medication errors are experienced when drugs are prescribed in the ambulatory; Worldwide the costs incurred due to medication errors amount to 4.5- 21.8 billion euro. Key words: medication errors, medical specialist

CHANGES IN CHOLINERGIC NEURONS OF NUCLEUS BASALIS DURING DEVELOPMENT OF ALZHEIMER’S DISEASE Damianka Getova1, S. Peres2, E. Muson2 1Department of Pharmacology and Clinical Pharmacology, Medical University – Plovdiv, Bulgaria, 2Rush University, Chicago, IL, USA Cholinergic neurons of the basal forebrain (CBF) of human brain provide the innervation to the cerebral cortex and hippocampus, play a key role in memory and attention. The neurons of nucleus basalis are preferably affected in patients with Alzheimer’s disease (AD). They are excellent model to study mechanisms of cell death and survival. The CBF projection system is plastic during the early stages of AD. The neurotrophin nerve growth factor (NGF) and its receptors in cholinergic neurons are required for CBF neuronal maintenance, survival and biological activity. There 2 main types of NGF receptors: TrkA – cell survival receptor and p75NTR – cell death receptor. CBF neurons express high TrkA and low p75NTR receptor levels in normal brain. In the early stage of AD the balance between these 2 receptors is disrupted. The cell cycle protein Rac1, plays a crucial role in the cascade of events leading to neuronal death during the progression of Alzheimer’s disease (AD). Rac1 interacts with p75NTR and induces neuronal cytoskeletal abnormalities in AD neurons and regulate the formation, reorganization, and maturation of neurons. Our study shows biochemical and morphologic evidence that Rac1, increases in a subpopulation of p75NTR-positive NB neurons that colocalize with pathologic tau epitopes during AD. Our data suggest that increased Rac1b expression acts as a modulator of various signaling pathways and leads to NFT formation and membrane dysfunction in a subgroup of p75NTR cholinergic NB neurons in AD. Key words: Alzheimer disease, cholinergic neurons, n. basalis

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PLENARY LECTURES

Public Health HEALTH POLICY FOR RARE DISEASES Rumen Stefanov Department of Social Medicine, Faculty of Public Health, Medical University - Plovdiv, Bulgaria Rare diseases represent a public health threat as they are life-threatening or chronically debilitating conditions with a low prevalence and a high level of complexity. Despite their rarity, there are so many different types of rare diseases that millions of people are affected. Rare diseases call for a global approach based on combined efforts to prevent significant morbidity or avoidable premature mortality, and to improve the quality of life and socioeconomic potential of affected persons. This is why the principles and overarching values of universality, access to good quality care, equity and solidarity are of paramount importance for rare disease health policy. Over the last decade rare diseases have been gradually identified as a health policy priority at both European and national level. The European Union health policy requires the adoption of national plans and strategies, the establishment of a system of European Reference Networks, union-wide information databases and registries for rare diseases. Key words: rare diseases

BIOTERRORISM: HISTORY, CURRENT STATE, ANALYSIS, COORDINATION AND ORGANIZATION OF ANTI-TERRORISTIC MEASURES, MANAGEMENT AND PREPAREDNESS OF CIVIL, MILITARY AND MEDICAL UNITS Krassimir Metodiev Department of Preclinical and Clinical Sciences, Medical University - Varna, Bulgaria Biological weapons used for bioterrorism are: microorganisms, bacteria, viruses, fungi or toxins. Their application must inflict harm on the opponent. The UN Convention on the Prohibition of Development, Production and Stockpiling of Biological and Toxin Weapons of 1972 banned the use of such agents. However, there are still many official and hidden laboratories world-wide where scientists develop either methods to defend the population against bioterrorism (the peacemakers) or new ways to affect human health (war-dreamers). A number of viral and bacterial agents such as small pox, Ebola, HIV, West-Nile, SARS, Marburg, hepatitis, influenza, plague, glanders, B. anthracis, Y. pestis, F. tularensis, Brucella species, V. cholerae, TBC, etc. are recognized as the etiologic reason for terrifying infections. The author makes a review of the current threats world-wide with possible attacks using bioagents. The history, present state and future possibilities are discussed, giving examples for the most often etiologic microorganisms applied for bioterrorism. Special attention has been paid to the organization, coordination, preparedness and management of civil and military units, hospitals and medical centers for active responses before, during and after a potential bioterrorist attack, as a corner-stone for a successful defence of the population. Key words: bioterrorism, risk infections, organization, coordination


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Plenary Lectures


REFLECTIONS ON “TEN PRINCIPLES OF THE PHYSICIAN AND THE MANAGER” Tsekomir Vodenicharov Faculty of Public Health, Medical University of Sofia, Bulgaria Revealing the basic steps towards success in medicine and health care, the book “The Ten Principles of the Medical Professional and the Manager” introduces the reader to the world of the integrated and associative thinking. The author of the book, Prof. Tzekomir Vodenitcharov shares his experience in the field of health management and identifies 10 principles which can promote the potential of the physician and the manager. In fact, these are universal principles, which application will lead to success in life and profession, whatever the surrounding. Prof. Vodenitcharov formulates the following principles: 1) Have an immortal goal. 2) Do not believe, but analyze. 3) Think in an associative and integrated way. 4) Follow the main goal, but do take care of the details, try to manipulate. 5) Be useful. 6) Plan. 7) Predict. 8) Organize. 9) Dream. 10) Act. “Anyone who aspires to become a Medical Professional with a capital M, and a person who strives to become a Manager with a capital M, will succeed if they are able to develop their potential to the full”. “The ambition of a Medical Professional is to reach beyond the limits in the struggle to save health and human life, and that of a Manager – in the efforts to reach greater efficiency. Their limit is – reaching perfection. Both the new health care managers, and the new generation of medical professionals should be ethical pragmatists”. The book tries to provoke an unconventional and creative style of thinking in the health management. Key words: health management

THE MEDICAL UNIVERSITY OF PLOVDIV, BULGARIA, AND THE EUROPEAN SOCIETY FOR PERSON CENTERED HEALTHCARE Andrew Miles European Society for Person Centered Healthcare, London, UK; Medical University - Plovdiv, Bulgaria Recent decades have seen a shift of the clinical and institutional ‘gaze’ away from the patient as a person towards a vision of practice that focuses preferentially on the technical application of government and payer-approved guidelines as the basis of medical care. Resulting unintended consequences have collectively led modern medicine into crisis: a crisis of knowledge (what is / is not ‘evidence’ for decision-making), care (deficits in empathy, compassion, dignity, autonomy), safety (neglect, iatrogenic injury, excess mortality), costs (which are increasingly unaffordable), governance (a failure of basic/advanced leadership) and of clinician wellbeing (burnout rates are at an unprecedented level) [1]. To deal with these contemporary problems a new way of ‘thinking’ and ‘doing’ has been advanced and termed person-centered healthcare´. To facilitate its progress, a major initiative has recently emerged, represented by the creation of a European Society for Person Centered Healthcare. MUP is playing a major role in establishing the work of the Society in Eastern Europe, illustrating one of the myriad ways in which the University is and remains, as per its Motto, “Committed to Humanity”. Key words: person-centered healthcare

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RADIOACTIVITY IN THE ENVIRONMENT – SOURCES AND EFFECTS Marios Anagnostakis School of Mechanical Engineering, Nuclear Engineering Department, National Technical University of Athens, Greece Radioactivity in the environment is being studied for more than a century. Initially, interest was limited to natural radionuclides, while the beginning of Atomic Era introduced a series of artificial radionuclides to the environment, which were studied as well. Nuclear accidents significantly contributed to the development radionuclides’ analysis techniques, dosimetric risk assessment and the field of radioecology. The study of natural radionuclides in the soil, originally limited to Ra-226, Th-232 and K-40 was then extended to include: Th-234, Pb-210, U-235 and Be-7, as a result of both, advances in detector technology and specific needs that had to be met, such as the study of radioactive equilibrium in the environment and Depleted Uranium contamination. At the same time research was extended to building materials, NORM (Naturally Occurring Radioactive Materials), TENORM (Technologically Enhanced Naturally Occurring Radioactive Materials) with potentially significant environmental impact and radon (Rn-222). Another source of exposure – potentially significant for some population groups – studied the last few years, is cosmic radiation. For the estimation of human risk due to the exposure to various sources of ionizing radiations, such as at: the ground or the built environment, dosimetric models have been developed, usually based on the activities of the radionuclides involved. This work is a short review of environmental radioactivity – mainly that of natural origin. The most significant sources of human exposure and simple models for the estimation of human risk due to the exposure to radiations emitted from radionuclides existing in the environment, are also presented. Key words: natural radioactivity, environment, dose risk assessment

FROM CONCEPT TO PRACTICE - DEVELOPING A PERSON CENTRED ACUTE HOSPITAL AT HOME SERVICE Jonathan Asbridge European Society for Person Centered Healthcare Headquarters, London, UK Care processes within hospitals are invariably inefficient often leading to frequent visits for patients to access investigations, consult physicians and receive treatment (Muijen 1999, Shepherd et al 1997, Ford et al 1998). Increasingly acute hospitals have been identified as places of risk rather than sanctuaries for recovery as antibiotic resistant infections, accidents and errors rise causing real harm to patients (Nursing Times 2013, Berwick 2013). They are also expensive to operate and slow to change (McKay NL, Deily ME 2008). Applying a person centred approach to the design of acute services leads to the change of the internal hospital process around the needs of the individual patient rather than the convenience of the institution (Clissett et al 2013). Evidence is clear that physician and patient satisfaction improves as does the level of safety and clinical outcome (Gladman et al 2010). Increasingly the evidence is emerging that these three factors increase further if the hospital stay is reduced to the bare minimum required for the patient to access typically 24hr nursing care and essential technologies (Carter and Oldman 2013).


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Plenary Lectures


Transferring patients to their own homes at the earliest possible time and replacing hospital care with domiciliary skilled nursing and rehabilitation teams, assisted by technological support has been found to reduce the time spent under the care of the acute medical team, improve clinical outcome and dramatically improve patient experience (Royal college of Physicians 2012). This paper describes the design of such a service and its mobilisation. It also provides evidence of outcomes after one year of operation. Key words: person centred healthcare, care process

PLENARY LECTURES

Therapy RE-EMERGING OF TULAREMIA IN BULGARIA – 2015 Todor Kantardziev National Center of Infectious and Parasitic Diseases, Sofia, Bulgaria Tularemia is severe zooanthroponosis which emerged in Bulgaria around the Sofia district in 1997 and gradually expanded its geographic range to the north and south. For a period of ten years approximately 400 clinical cases were laboratory confirmed. Analyses showed that this is a new focus for tularemia in the country. The emergence of epidemic zooanthroponosis is related to poor farming practices on agricultural land. In the last five years tularemia has become endemic along with the occurrence of single cases. Not harvesting of crops last year was followed by increase in the population of field rodents, which are important carriers in the spread of this disease. Since the beginning of 2015 seven cases of tularemia were confirmed in humans for a period less than one month. The area of the focus is expanding and affecting new territories in the central and southeastern part of the country. Sample investigation analyses are performed on patients, animals and environment. Molecular techniques are used for diagnosis and epidemic typing. Key words: tularemia

DIABETES EPIDEMIC, A CHALLENGE FOR OUR SOCIETY Paolo Pozzilli Department of Endocrinology & Diabetes, Faculty of Medicine, University Campus Bio-Medico, Rome, Italy Type 2 diabetes (T2DM) is a chronic metabolic disorder characterized by hyperglycemia due to an imbalance between insulin requirement and insulin production. T2DM is one of the most prevalent noncommunicable diseases claiming millions of victims around the world. The total number of people with diabetes in 2013 worldwide is estimated to be approximately 382 million, more than 5 million people die of diabetes and many more suffer from diabetes related complications. It is therefore worrisome that the pandemic of T2DM is so far from stopping: the global prevalence of T2DM is expected to rise from 8.3% to 10.1% by 2035. Sadly, despite the large number of available treatment strategies, the overall results of therapies for diabetes are not really satisfactory. T2DM is still the second cause of disability-adjusted life years lost and in most countries is the leading cause of death. In particular, subjects with T2DM continue to have excess cardiovascular morbidity and mortality compared with the general population. The risk for T2DM individuals to develop cardiovascular diseases is at least two fold than

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those without T2DM. Even multifactorial approaches including lowering of blood glucose, and smoking cessation do not completely eliminate the increased cardiovascular risk associated with T2DM. In this respect, new approaches to halt cardiovascular events in subjects with T2DM are needed. Key words: type 2 diabetes, cardiovascular complications, epidemiology and treatment of diabetes

METABOLIC DISTURBANCES IN POLYCYSTIC OVARY SYNDROME Maria Orbetzova Second Department of Internal Diseases, Clinic of Endocrinology and Metabolic Diseases, University Hospital St. George, Plovdiv, Medical University - Plovdiv, Bulgaria Polycystic ovary syndrome (PCOS) is one the most common endocrine disorders affecting about 5-14% of women of reproductive age and leading cause of infertility. The global medical importance of the disease is based on the fact that according to the new concepts PCOS is considered as typical representative of metabolic syndrome with insulin resistance and compensatory hyperinsulinaemia being the underlying mechanism. PCOS is associated with higher prevalence of visceral obesity, unfavorable ratio between atherogenic and antiatherogenic adipocytokines, impaired glucose tolerance, diabetes mellitus type 2. There is strong evidence that even in younger age women with PCOS carry on markedly atherogenic lipid profile and other cardiovascular risk factors that later in life (after 15-20 years) might result in higher incidence of cardiovascular morbidity and mortality. Premature development of atherosclerosis, more frequent occurrence of hypertension, gestational diabetes and hypertension during pregnancy are observed in women with this syndrome. A review of literature data and author’s experience on the prevalence and characteristics of overt metabolic syndrome and its separate components in PCOS women with different phenotypes are presented. The author’s results are based on several retrospective and prospective studies investigating anthropometric characteristics, carbohydrate and lipid disturbances, alterations in adipose tissue hormones and appetite regulators, circadian rhythms of blood pressure and effect of different therapeutic strategies on metabolic parameters and total cardiovascular risk in PCOS. Key words: PCOS, metabolic syndrome, cardiovascular risk

HEART FAILURE - PAST, PRESENT AND FUTURE Maria Tokmakova First Department of Internal Diseases, Division of Cardiology, Medical University - Plovdiv, St. George University Hospital, Plovdiv, Bulgaria Heart failure (HF) has a major public significance due to its growing prevalence, worsened quality of life, high mortality and huge economic burden. Co-morbidities frequently accompany HF thus contributing to increased morbidity, hospitalizations and mortality. Despite multiple evidence-based drug and device therapies outcomes of patients with chronic HF improve slowly and in-hospital and 1 year mortality of patients with acute HF remain disturbingly high. Prevention of HF and reduction of mortality due to HF are global health priority. Compliance with clinical practice guidelines is associated with improved outcome. Novel therapies aim to improve symptoms, survival as well as quality of life. Key words: heart failure, epidemiology, economic burden, prognosis


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Plenary Lectures


VALIDITY AND VALIDATION OF THE DIAGNOSTIC ASSESSMENT METHODS AND CLASSIFICATION IN PSYCHIATRY Drozdstoy Stoyanov Department of Psychiatry and Medical Psychology, Medical University - Plovdiv, Bulgaria Psychiatry as a hybrid discipline that embraces facets from both the natural sciences and humanities has adopted relevant heterogeneous approaches to validity and classification. On the one hand, psychiatry adopted from medicine the categorical stance which resembled typical medical nosological units with narrow borders constituted from exclusion and inclusion criteria. On the other hand, psychiatry adopted from psychology the more liberal and flexible approach of dimensional diagnosis, which is to some extent incorporated in the latest revisions of Diagnostic and Statistical Manual of Mental Disorders. Unfortunately, both models in itself turned out to be inconsistent as well. In effect, validity was erroneously displaced with statistical reliability and utility of diagnostic criteria, which are both subject to critical considerations. This crisis called for the development and implementation of alternative perspectives on psychiatric validity and validation, such as Research Domain Criteria (RDoc), prototype, cluster, narrative-based and person-centered diagnosis, four domains of mental illness (FDMI); translational validity, etc. Key words: psychiatry, validity, models

EFFECTIVENESS OF EXTRACORPOREAL SHOCKWAVE THERAPY AS A NEW METHOD IN THE TREATMENT OF MUSCULO-SKELETAL DISORDERS Elena Ilieva Division of Physical Medicine and Rehabilitation, Medical Faculty, Medical University - Plovdiv, Bulgaria Еxtracorporeal shock wave therapy (SWT) is a new method in the treatment of musculoskeletal disorders, which has become significant subject of research worldwide. The physical and biological effects of shock wave therapy are presented: pain relief and stimulation of regeneration due to: increased cell permeability, reduction of non-myelinated nerve fibres and selective neural inhibition, stimulation of microcirculation, release of substance P and neuropeptides, release of nitric oxide, induction of growth factors [eNOS, VEGF, PCNA, IGF – in blood vessels (neo-vascularization), epithelium, bones, collagen], stimulation of stem cells. The author discuses the evidence concerning the effect of SWT in chronic tendinopathies, resistant to other treatment modalities (calcifying tendinopathy of the rotator cuff, lateral epicondylitis of the elbow, tendinopathy of the Achilles tendon; plantar fasciitis); in bone non-unions and avascular necrosis; for reduction of muscle spasticity in central motor neuron disorders and in patients with diabetic ulcers. The author presents the results of own studies about the application of radial shock wave therapy in patients with chronic tendinopathies (tennis elbow, plantar fasciitis); for reduction of muscle spasticity in children with cerebral palsy and in patients with osteoarthritis of the knee as a new and not yet studied field of application. Conclusion: SWT is a treatment of choice in chronic musculoskeletal disorders, resistant to other treatment modalities with proved effectiveness and safety. The evidence in comparison with the vast majority of other conservative methods is above average. New fields of application are studied and give promising results. Key words: radial shock wave therapy, musculo-skeletal disorders, chronic tendinopathy, bone-nonunions, knee osteoarthritis

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HEREDITARY ANGIOEDEMA - 2015 Henriette Farkas National Angioedema Center, Third Department of Internal Medicine, Medical Faculty Semmelweis University, Budapest, Hungary Hereditary angioedema (HAE) is a rare, autosomal dominant disorder with recurrent edema formation in subcutaneous and/or submucosal tissues. In HAE type I and II, a mutation in the C1-inhibitor (C1-INH) gene causes deficiency of the C1-INH protein, which regulates the complement, contact, coagulation, and fibrinolytic systems. In HAE with normal C1-INH function, the Factor XII gene can be defective. The diagnosis is established by complement testing. The manifestations of the different HAE types are similar, but their individual presentation varies. HAE is a bradykinin-mediated form of angioedema and therefore, it does not respond to conventional treatment. Knowledge on the pathomechanism has increased lately. However, the relationship between geno- and phenotype has not been elucidated. The occurrence and frequency, as well as the likely location and possible triggers of edematous attacks remain unpredictable. New, safe, and effective drugs are available. Although these are characterized by diverse modes of action, targets, and posology, all relieve attacks in the first place. They include different types of C1-INH, ecallantide, and icatibant. No treatment capable of preventing edematous attacks with certainty is currently available. Innovative agents are developed for prophylaxis. Early diagnosis and appropriate treatment can improve the patients’ quality of life. Increasing disease awareness and patient education are all indispensable. The goal is to expand access to state-of-the art therapies and to administer these according to international guidelines; however, in an individualized fashion. As HAE is a rare disease, centralized management can improve patient care – this calls for dedicated HAE centers. Key words: hereditary angioedema complement C1-inhibitor, prophylaxis, management

ASSESSING ESOPHAGEAL MOTILITY: IS HIGH-RESOLUTION MANOMETRY WORTH ITS COST? Dan Dumitrascu Iuliu Hatieganu University of Medicine and Pharmacy, Cluj-Napoca, Romania BACKGROUND: Esophageal manometry (EM) is the gold standard in evaluating patients with esophageal symptoms, after upper digestive endoscopy. It is the first investigation of the esophageal motility disorders. Recently, the high-resolution manometry (HRM) arrived on the market and is spreading over. AIM: To check the usefulness of the most expensive high-resolution esophageal manometry (HREM) compared to standard EM. METHODS: A single center audit to compare the cost-efficiency of EM with perfusion catheter vs. HREM. The study was carried out in a lab for functional digestive investigations with 15 years of experience in manometry and recently equipped with HRM. R ESULTS: EM is a well standardized examination capable of diagnosing primary and secondary esophageal motility disorders. HREM is much more expensive, but faster, thus less time consuming, more elegant and more spectacular. The results are less well standardized, but using the latest version of the Chicago classification, we have been able to diagnose subtle changes in esophageal motility disorders. CONCLUSIONS: HREM is worth its cost. It should be used in tertiary centers. Standard EM with perfusion catheters is still very useful for didactic purposes and for diagnosis in secondary centers. Key words: esophageal motility, high-resolution manometry Folia Medica 2015; 57; Suppl. 1 © 2015 Medical University, Plovdiv, Bulgaria

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Plenary Lectures


SEMIOLGY OF EARLY CHILDHOOD DEVELOPMENT Ivan Ivanov Department of Pediatrics and Medical Genetics, Medical University - Plovdiv, Bulgaria Semiology of early childhood development studies the significance of particular signs and symptoms in history, оbjective examination and laboratory evaluation for previous and present neurodevelopmental pathology and for sequelae. In pediatric perspective, birth is the onset point. Almost any second and third trimester pathology of pregnancy may have negative impact on neurodevelopment, with preterm birth, intrauterine growth retardation, preeclampsia and abnormal fetal morphology being the most important. In the objective examination, head circumference, its trajectory and its relation to the width of the extracerebral CSF spaces have the greatest diagnostic and prognostic significance. Motor examination (posture, range of passive movements, spontaneous motor activity repertoire, and maturation of voluntary activity) needs detailed knowledge of its normal dynamics and variability, and of the diagnostic significance of the multiple combinations of signs. Language delay is diagnosed commonly after the first year of life, with major diagnostic factors being history of preverbal development, differential assessment of expressive and receptive language development, evaluation of the other domains of neurodevelopment and of the social milieu. Abnormalities in emotional-social development are most often missed by the parents. Therefore, early diagnosis of autism relies on examiner’s focused examination of this domain. Neuroimaging proves present and past lesions, while the range of brain atrophy and the site of the lesion have prognostic significance. Biochemical screening is indicated for etiological diagnosis of delayed neurodevelopment. Familial and social evaluations are also important, knowing that two major predictors of normal neonate’s development are maternal education and socio-economical status. Key words: neurodevelopment, risks, prediction, semiology

TARGET THERAPY – CURRENT STATE AND NEW PERSPECTIVE J. Grudeva-Popova NOVARTIS, Department of Clinical Oncology, Faculty of Medicine, Medical University - Plovdiv, Bulgaria The recent, rapid development of molecularly targeted therapy (ТТ) is best illustrated by advances in oncology. TТ of malignancies seems to be an effective and less toxic approach to the treatment of such disorders. A broad spectrum of targeted drugs affecting diverse set of targets is readily available in practice: antiCD20, antiCD33, antiCD52; antiVEGF(R), antiEGFR; ABL, Src/ABL-kinase and JAK2 inhibitors; m-TOR, FLT3, histone-deacetylase inhibitors and some others. Some of the resistance mechanisms have already been successfully identified. The authors underline that further studies are needed to determine where and when such management fits into a therapeutic regimen for any given disorder, whether upfront or as salvage therapy, alone or in combination with chemotherapy (concurrent or sequential), as well as what potential adverse effects to expect and their late toxicities. The new knowledge about ‘’targets’’ will hopefully help us choose personalized treatments for the patients, improving their survival with minimal impact on their quality of life. Key words: target therapy, current state, perspective

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PLENARY LECTURES

Surgery PATIENT SAFETY IN LAPAROSCOPIC CHOLECYSTECTOMY Rossen Dimov Department of Special Surgery, Medical University - Plovdiv, General Surgery Clinic, Kaspela University Hospital, Plovdiv, Bulgaria In the last decade laparoscopy has become the gold standard in the implementation of the majority of procedures in the abdominal cavity. Despite standardized protocols for education and training, practical schooling for all surgeons starts and for the most part continues with the implementation of laparoscopic cholecystectomy. Since the dawn of its implementation to the present time it has become a ‘best seller’, the most often practiced laparoscopic procedure in general surgery. All innovations in the improvement of laparoscopy as the technique of single-port surgery and NOTES technology have made their first steps with the performance of cholecystectomy. Huge in quantity and quality information has been accumulated on how to perform the operation. And amid this rapid development as “a thorn in the Achilles heel “possible errors, threats and complications of laparoscopic cholecystectomy remain, which invariably accompany each of us in the constant path of improvement. Unfortunately, neither modern technologies, nor team work principle have resulted in the elimination of Intraoperative complications. Their share remains at approximately the same range for the last five years. Learning and self-perfection oriented behaviour can protect to a large extent by the bitterness of “failure” and the complicated treatment of iatrogenic influenced patients. In this aspect, I consider right a sentence that crystallized during the accumulation of my personal experience in laparoscopic surgery. It reads: “A surgeon is ready to perform alone a surgery only when he is ready to meet and adequately treat potential complications.” Key words: patient safety, laparoscopic cholecystectomy

INTERNATIONAL COOPERATION IS A POWERFUL ENGINE FOR IMPROVING UNDERSTANDING, DIAGNOSIS AND CARE FOR RARE CRANIOFACIAL CLEFTS Philippe Pellerin Centre National de Reference des Malformations Cranio Maxilla Faciales Rares, Lille France; Plastic and Craniofacial Unit, Medical University - Plovdiv, Bulgaria In his 1976 publication (J Max Fac Surg 4, 1976, 69-92) completed in 1977 in the book “Chirurgie Orbito-Palpebrale” Paul Tessier gave a comprehensive, logical classification of rare craniofacial clefts. It was the result of the analysis of 336 cases that were referred to this genius. Tessier’s work has provided distinctive landmarks in diagnosing and understanding these malformations, yet there is still a lot to do in respect of the systematisation of their management. As these cases are so rare there are very few teams over the world that may have the chance to collect a sufficient number of cases so that proper surgical techniques can be developed. The Ministry of Health Care in France has organised Centres of Reference for rare diseases and our centre, given its wide experience over 40 years and thousands of craniofacial patients, has been nominated to be included in this system. The senior author of this article and the team based in Plovdiv have been collaborating for 25 years. This has allowed the physician in this centre to collect a large number of well documented cases of very rare malformations with long follow-ups which has helped improve both the process diagnosis and management. The partnership between the respective


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Plenary Lectures


centres in Lille and in Plovdiv bound duly by formal agreement has been a powerful force in boosting the progress in the treatment of these severe and very disfiguring malformations Key words: craniofacial clefts

WEAR OF COMPONENTS IN PROSTHETIC SURGERY Michele Attilio Rosa, Domenico Fenga Policlinico Universitario G. Martino, Messina, Italy Wear of prosthetic components is a cause of failure of an implant; it causes aseptic loosening and periprosthetic bone resorption. It’s important to study the mechanisms effecting the onset of wear. Mechanisms that lead to wear are four: sliding (the physiological motion of the artificial joint); from contact between a mating and non-scheduled surface for the coupling; by presence of a “third body” between two surfaces; from contact between two surfaces that do not include the coupling (McKellop, 2007). Risk factors are related to patient, to surgery and to characteristics of implant. Wear risk increases with obesity, with increasing of functional requirements and with tendency to implant a prosthesis in young patients. Among factors related to surgery we must mention the possible errors of cementation, the presence of interposed fragments and errors in the choice of size and rotation of implant; choice of materials plays a fundamental role in the onset of wear. The pathology of “debris” is often progressive and clinically “silent” and can mislead the patient and the surgeon. When clinical and radiographic evidence is clear, often debris damage is serious and treatment much more complex. Great interest and study was dedicated to the characteristics of implant system. Modularity undoubtedly represents a considerable advantage in recreating a more anatomical articular geometry, but results of first generation implants has led to a change of course. Increasing functional needs of patients are the greater risk of wear and a more increasing attention and effort must be obtained from research. Key words: wear, ceramic, metal, polyethylene

EVOLUTION OF FACIAL RECONSTRUCTIVE SURGERY George Anastassov Department of Maxillofacial Surgery, Mount Sinai School of Medicine, USA Facial reconstructive surgery has undergone remarkable evolution from simple camouflage with epitheses, through number of local, regional and free-vascularized flaps to allogeneic facial transplantations. This presentation will review the historical aspects of these developments, the specific indications and techniques for facial reconstruction depending on defects being reconstructed as well as present the author’ experience with these techniques. Key words: facial reconstruction, free flaps

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DIAGNOSIS AND TREATMENT OF COMPLICATED INTRA-ABDOMINAL INFECTIONS IN A WORLDWIDE CONTEXT – PRESENT AND PERSPECTIVES Boris Sakakushev Department of Propaedeutics of Surgical Diseases, Division of General and Operative Surgery with Coloproctology, Medical University - Plovdiv, Bulgaria Despite advances in medicine, the rates of morbidity, mortality and bad prognosis associated with complicated intra-abdominal infections (CIAI) remain very high. The disorders causing this severe pathology range from acute appendicitis to stercoral peritonitis. We are witnessing now a change of the epidemiological, clinical and pharmacological profile of peritonitis. With the introduction of advanced technologies in surgery and the more stringent patient requirements the rate of the noninvasive diagnostic procedures and mini-invasive surgical manipulations and operations increases. Many evidence based prospective randomized trials for successful application of percutaneous, laparo-endoscopic and robotic operative methods for the treatment of CIAI worldwide are a matter of controversies and discussions. The assembled international groups of experts and the conducted international trials over the last three years have allowed the medical community to formulate general guidelines and recommendations for the modern diagnosis and management of these infections and distinguish the fundamental research areas for further investigation of the problem. These facts logically suggest a stable tendency to achieve a definite decrease of lethality from 30% to 10% in the next decade, decline in morbidity, better prognosis and a better quality of life for patients. Key words: CIAI, diagnosis, treatment

MODERN SURGERY OF ADOLESCENT IDIOPATHIC SCOLIOSIS Mikhail Mikhaylovskiy Research Institute of Traumatology and Orthopaedics n.a. Y. L. Tsivyan, Novosibirsk, Russia Our concept of early detection and treatment of progressive adolescent idiopathic scoliosis (AIS) has been elaborated over the last 15 years. It contains a few major stages. 1. School screening for AIS performed using 3D analysis of the dorsal body surface. This method, which was developed at our institute and called computed topography, allows assessing the shape of the dorsal body surface in 1-2 instants without any radiation exposure. 2. The whole cohort (up to 300 children a day) is divided into subgroups as follows: 1) healthy children, 2) those who need observation only (Cobb angle of 0 to 20°), 3) the subgroup of conservative treatment (20-40°) and 4) surgical treatment subgroup (more than 40°). 3. Patients may migrate from one subgroup to another (for example, from the second group to the third one) during the observation period depending on progressive nature of the pathology. 4. Surgical correction of progressive AIS is performed according to the patient’s age, the curve type, magnitude, and mobility, comorbidities, and so on. 5. Postoperative rehabilitation. We treat the early onset scoliosis (before 10 years of age) using the vertical expandable prosthesis titanium rib (VEPTR) instrumentation. This type of surgical treatment includes initial distraction, staged corrections every 7-8 months, and definitive dorsal fusion at the age of skeletal maturity. Our department has had 70 cases, five of them completed. Adolescents aged between 11 and 13 years need one-stage surgical correction, and the purpose of this procedure is twofold: to correct the scoliotic curve and prevent further deterioration, since progression potentials still exist in this cohort of patients. We use third-generation spinal implants with total transpedicular fixation.


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Plenary Lectures


Our surgical strategy in patients aged 14 to 20 years depends on initial spinal mobility determined during preoperative assessment on lateral bending X-rays. Mobile deformities are treated using dorsal instrumentation and bone fusion only, and stiff ones (less than 30% correction in lateral bending) require anterior discectomy as a first step of the staged procedure. Adult patients (older than 20 years of age) need two-stage surgery in all cases unless contraindications are present. In two recent cohorts we preferred hybrid constructions including both transpedicular screws and hooks in order to diminish blood loss, operative time and complication rate. Key words: AIS, VEPTR

NEW INDICATIONS AND CHALLENGES IN ANTI-VEGF TREATMENT IN OPHTHALMOLOGY Petya Vasileva Academician Pashev Specialized Hospital and Clinic of Ophthalmology, Sofia, Bulgaria Anti-VEGF medications offered a paradigm shift in the treatment of pathological ocular neovascularization and associated complications in a number of diseases of the retina and the anterior eye segment. Anti-VEGF treatment offers a radical change in the prognosis of age-related macular degeneration (AMD) – a disease, until recently considered to be incurable, with escalating prevalence, mainly due to the increased life expectancy. Modern diagnostic methods, such as optical coherence tomography (OCT), which reveal the structural changes in the retina, in combination with routine imaging tests of the ocular vascular system, led to new concepts regarding disease classification and definition of stages of progression, as well as to the exceptional ability to monitor and document the treatment results. Research conducted by international scientists and findings from numerous randomized multicentre clinical studies provided us with new data on diagnosis, drug response and side effects of anti-VEGF treatment. Intravitreal application of anti-VEGF agents offers more targeted and effective drug delivery into the posterior segment of the eye. Indications for anti-VEGF application continue to increase beyond most common retinal pathology: AMD, diabetic macular edema, retinal vein occlusion (RVO), and include various rare retinal diseases associated with active retinal neovascularization, such as myopic degeneration, central serous retinopathy, dominant drusen, angiod striae. Anti-VEGF treatment is beneficial also in anterior segment pathology - vascularized leukoma, keratoplasty, rubeosis, antiglaucomatous surgery. We present our experience in anti-VEGF treatment for a period of 8 years - indications, treatment regimen, follow-up requirements, immediate and long-term results. A group of patients treated with a new type of anti-VEGF medication with considerably greater efficiency and less systemic safety is also analyzed. Possibilities of combined, personalized anti-VEGF treatment regimen are discussed. Key words: anti-VEGF

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PLENARY LECTURES

Dental Medicine LASER PHOTOCOAGULATION – A NEW APPROACH FOR NON SURGICAL TREATMENT OF BENIGN ORAL VASCULAR LESIONS Georgi Tomov Department of Operative Dentistry and Endodontics, Faculty of Dental Medicine, Medical University – Plovdiv, Bulgaria INTRODUCTION: The heterogeneous group of benign oral vascular lesions (BOVL) includes diverse disorders of congenital, inflammatory or neoplastic. BOVL are characterized by vascular proliferation, clinically manifested as soft red-bluish tumor. If traumatized BOVL run a significant risk of hemorrhage; their treatment usually requires caution to prevent bleeding. In the last decade lasers have changed the treatment options of managing BOVL permitting the application of non-surgical techniques that reduced the hemorrhage risks and unpleasant tissue deformities. AIM: The aim of this study was to evaluate the clinical outcome after application of laser assisted techniques for management of BOVL. MATERIALS AND METHODS: In this study ten BOVLs were treated by two different laser techniques: the transmucosal photocoagulation (TMP) and the intralesional photocoagulation (ILP). The laser used in this study was a diode laser (LiteMedics, Italy, 980nm) selected because of its great affinity to hemoglobin. R ESULTS: In each case, the laser permitted safe treatments of BOVL without hemorrhages, both during the intervention and in the post-operative period. The minimally invasive treatment (TMP and ILP) resulted in uncomplicated resolution of even big lesions without tissue loss. CONCLUSIONS: In this study diode lasers confirmed to be the gold standard in BOVL treatment, permitting the introduction of minimal invasive surgery principles and reducing the risks of hemorrhage, typical for these lesions. As usual in laser surgery to optimize the results and to reduce the risks and minimize the disadvantages, the specialists should be very familiar with laser technology and laser-tissue interaction. Key words: oral vascular lesions, laser, photocoagulation

THE CLINICAL USE OF LASERS IN ENDODONTIC THERAPY Adam Stabholz Faculty of Dental Medicine, The Hebrew University, Hadassah Faculty of Dental Medicine, Israel, Medical University - Plovdiv, Bulgaria Laser technology develops very quickly. New lasers with a wide range of characteristics are available today and are used in the various fields of dentistry. The search for new devices and technologies for dental procedures has always been a challenge in the last twenty years through which much experience and greater knowledge have been acquired. Our studies using SEM analysis following debridement and removal of pulp tissue and old gutta percha root canal fillings demonstrated heavy smear layers on the wall surfaces of the root canals as well as remnants of the old filling materials in cases of endodontic retreatments.


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Plenary Lectures


The use of laser irradiation in the root canal system enabled a thorough cleansing of the root canal and SEM photographs showed clean root canals and walls with open dentinal tubules. The walls of root canals lased with the Er: YAG laser after biomechanical preparation with NiTi rotary files were cleaner in comparison to root canals that were similarly prepared but not lased. Clinical cases with long follow-up will be presented and discussed. Key words: lasers, endodontic therapy

HOW ORAL INFECTIONS LINK TO SYSTEMIC HEALTH Jukka H. Meurman University of Helsinki, Finland Since the advent of medicine man has known that dental infections are detrimental to health, best illustrated by Egyptian mummies. When in the 19th century scientists started investigating these associations, the theory of focal infection was born. In the 1980s the first clinical studies were made showing a statistical association between poor dental health and heart infarction and stroke. Thereafter, periodontal disease in particular has been associated with a variety of systemic diseases and conditions such as obesity, metabolic syndrome and diabetes, kidney, liver, and rheumatic diseases, and lately even with malignancies. Poor oral health has also been shown to link to mortality. Frail elderly and immunosuppressed patients in general are in particular danger in this regard. The pathogenic mechanisms involved are the following: first, spread of oral microbes locally and to remote tissues and organs may cause abscesses and focal inflammations. Secondly, upregulation of cytokines and inflammatory mediators in the oral tissues generate systemic inflammatory conditions with even catastrophic consequences. The pathways include interleukins, tumor necrosis factor-alpha, radical oxygen metabolites, etc. These in turn trigger cascade reactions which may lead, for example, to atherosclerosis and also to malignant transformation. Hence, maintaining satisfactory daily oral hygiene is essential in order to avoid systemic complications. This seems to be the more important the more generally diseased patient we have in question. Key words: oral infection, periodontitis, systemic disease, malignancy, mortality

AC ELECTROKINETICS: THE DAWN OF A NEW ERA IN CONTROLLED DRUG DELIVERY FOR DENTISTRY Chris S. Ivanoff UTHSC College of Dentistry, TN, USA Several studies have shown both qualitatively and quantitatively that dielectrophoresis (DEP) and AC electroosmosis (ACEO) can enhance the absorption of fluoride (and other drugs) into human enamel by: (1) achieving greater penetration and (2) achieving greater concentration than diffusion. The studies suggest that DEP can deliver other drugs of variable molecular weight and structure. Current diffusion and electrochemical methods are inadequate for effective transport of antibiotics, anesthetics, analgesics and anti-inflammatory agents directly into intraoral targets. This pilot study explores the dielectrophoretic behavior of tetracycline, acetaminophen, lidocaine and benzocaine under the influence of non-uniform electrical fields at specific frequencies. By measuring the frequency response of approximately 1-10 μm sized drug particles in an aqueous

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media, appropriate frequencies for positive DEP, negative DEP and ACEO are identified. After optimizing the frequencies for particle translation under negative DEP, positive DEP and ACEO, an assembly composed of interdigitated electrode arrays (IDE), is used to trap, manipulate, and translate tetracycline, acetaminophen, benzocaine and lidocaine particles using single frequencies on the terminals of the IDE array. Drug particle movements at various frequencies are captured and analyzed with light microscopy in real time. The results show that drug flow can be controlled by manipulating electromechanical effects in the fluid due to drug particle-fluid and particle-particle interactions under DEP and ACEO. AC electrokinetics may potentially advance drug delivery in dentistry by: extending transport of tetracycline, acetaminophen, lidocaine and benzocaine to specific intraoral biological targets; eliminating reformulation; bypassing first-pass metabolism and improving the efficacy and safety of drug delivery. Key words: AC electrokinetics, AC electrosmosis, dielectrophoresis, diffusion, drug transport, antibiotics, analgesics, anti-inflammatory agents, anesthetics, intraoral targets

CROSS-NATIONAL CLERKSHIPS AND CULTURAL COMPETENCE TRAINING IN THE PREDOCTORAL DENTAL CURRICULUM - A COMPARISON OF BULGARIAN AND AMERICAN STUDENT PERCEPTIONS Chris S. Ivanoff 1, Bogomil Andonov2, Athena E. Ivanoff 3, Krassimira Borisova Yaneva-Ribagina4, Nina Musurlieva5 1Department Bioscience Research UTHSC College of Dentistry, Memphis, USA, 2Department of Operative Dentistry and Endodontics, Faculty of Dental Medicine, Medical University - Plovdiv, 3Department of Chemistry, Cleveland State University, USA, 4Faculty of Dental Medicine, Medical University, Sofia, 5Department of Social Medicine and Public Health, Faculty of Public Health, Medical University - Plovdiv, Bulgaria Seven hundred and nineteen American and Bulgarian dental students were surveyed at three colleges of dentistry to compare their perceptions about their dental school’s commitment to advance global dentistry and philanthropy. The study included questions about prior involvement in charitable dental missions both at home and abroad. Many respondents felt that their dental school does not advance global dentistry, nor adequately teaches students the virtues of philanthropy and volunteerism. The majority agreed, however, that dental schools have a moral obligation to raise global standards of oral health care and assist underserved communities gain equal access to dental care. They reported that an opportunity to spend a semester at a foreign dental school would enhance their dental education in ways that are not presently fulfilled; would help them better understand cultural diversity; and teach them about philanthropy and volunteerism. In their opinion, international exchange programs that provide clinical rotations and field experiences in economically challenged and underserved areas of the world would: (a) foster the global advancement of dentistry; (b) increase their cultural competence; (c) promote an appreciation for cultural diversity and socioeconomic disparity in the communities that graduates will be serving; (d) teach students the virtues of philanthropy and volunteerism. This study may contribute to understanding factors affecting student involvement in programs to advance global dentistry. Key words: student perceptions, curriculum, cultural competence


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Oral Presentations


ORAL PRESENTATIONS

Biomedicine KIR AND HLA GENETIC BACKGROUND INFLUENCE THE COURSE OF HBV INFECTION S. Mihaylova1, V. Varbanova2, A. Georgieva1, D. Dimitrova3, M. Ivanova-Shivarova1, E. Naumova1 1Department of Clinical Immunology, Alexandrovska University Hospital, Medical University, Sofia; 2National Hospital for Hematological Diseases, Sofia, 3Department of Gastroenterology, Alexandrovska University Hospital, Medical University, Sofia, Bulgaria Genes controlling innate and adaptive immunity might be responsible for variable outcome of hepatitis B virus (HBV) infection. The aim of this study was to evaluate the impact of two highly polymorphic immunogenetic systems - HLA and KIR on disease outcome. KIR and HLA genotyping was performed on 144 patients (124 with chronic hepatitis B and 20 spontaneously recovered from acute HBV infection). Development of chronic hepatitis B was significantly associated with increased frequency of KIR3DL1*004 allele (functionally inert) compared to controls (33.1% vs. 17.6%, p = 0.018). HLA alleles B*08 (OR-2.94; p = 0.0018), B*35 (OR-1.78; p = 0.03), DRB1*03 (OR-2.36; p = 0.002) were predisposing for chronic infection while DRB1*13 (OR-0.21; p = 0.006) was found to be protective. Patients recovered from HBV infection have lower frequency of KIR2DL5В (10.0% vs. 45.5%, р = 0.003; 10.0% vs. 33.9%%, р = 0.042) and higher frequency of HLA-A*25 and B*44 alleles compared to healthy individuals and chronic carriers. In addition KIR genotype consisting of 4 particular activating and 7 inhibitory KIRs was predisposing while a profile composed of 6 activating KIRs was protective for chronicity of the infection. However, the combination KIR3DL1(+)/3DS1(-)/HLA-B Bw4(-) which sets the pattern for immune inhibition was found in higher frequency in both patients groups. In conclusion our results showed the important role of KIR/HLA related immunoregulatory pathways for the outcome of a HBV infection. These genetic markers could be used as valuable predictive factors for the course and treatment outcome of the disease. Key words: HLA, KIR, HBV

ASSOCIATION OF -1082 A/G PROMOTER POLYMORPHISM OF IL-10 GENE WITH MULTIPLE SCLEROSIS (PRELIMINARY STUDY) Anastasia Trenova1, Boncho Grigorov2, Georgi Slavov1, Maria Manova1, Liuba Miteva2, Spaska Stanilova2 1Department of Neurology, Medical Faculty, Medical University - Plovdiv; 2Department of Molecular Biology, Immunology and Medical Genetics, Medical Faculty, Trakia University, Stara Zagora, Bulgaria Multiple sclerosis (MS) is a chronic disease with dysregulation of cytokines production involved in its pathogenesis. Treg cytokine IL-10 suppresses Th1-mediated immune response and inflammatory cytokine synthesis. Dependence of IL-10 production according to -1082A/G promoter polymorphism in IL-10 gene (rs1800896) has been reported. The role of this polymorphism in susceptibility and development to MS remains unclear. This study aims to compare -1082A/G IL-10 genotype distribution in 87 patients with MS with a group of 190 age and sex matched healthy controls in attempt to find a correlation with susceptibility to MS and the role of geno-

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types for the age of disease onset. Genotyping for -1082A/G polymorphism of IL-10 was performed using ARMS-PCR assay. Patients were divided according to the age of disease onset: under 29 years of age and above 30 years. We observed the following distribution of genotypes: in cases AA - 37%; AG - 47%; GG - 16%; in controls: AA - 27%; AG - 58%; GG - 14%. Regarding the genotype distribution in logistic regression analysis the presence of -1082AG was associated with decreased risk of developing MS (OR = 0.600, 95%CI = 0.327÷1.101; p = 0.077) with borderline significance. When MS patients were divided into two groups according to the age of disease onset and were separately compared to controls we observed lower risk of later onset for individuals with AG genotype (OR = 0.524, 95%CI = 0.237÷1.159; p = 0.08). In conclusion, this study demonstrated a tendency for association of promoter polymorphism -1082A/G in IL-10 gene with the age onset of MS. Carriers of heterozygous genotype-AG had two-fold lower risk for later-onset of MS in comparison with non-carriers. Key words: multiple sclerosis, cytokines, gene polymorphisms

ROLE OF MTHFR C677T SINGLE NUCLEOTIDE POLYMORPHISM IN PATIENTS WITH COPD Tatiana Vlaykova1, Georgi Golemanov2, Dimo Dimov3, Tania Tacheva1, Vania Ilieva3, Gospodinka Prakova3, Regina Komsapenkova2 1Department of Chemistry and Biochemistry, Medical Faculty, Trakia University, Stara Zagora, 2Department of Chemistry and Biochemistry, Medical University, Pleven, 3Department of Internal Medicine, Medical Faculty, Trakia University, Stara Zagora, Bulgaria Methylenetetrahydrofolate reductase (MTHFR) is a central regulatory enzyme in folate and methionine metabolism, which on turn play crucial role in DNA synthesis and methylation. MTHFR catalyzes the biologically irreversible reduction of 5,10-methylenetetrahydrofolate to 5-methyltetrahydrofolate, which supplies with the methyl group the process of remethylation of homocysteine to methionine. So far, 11 polymorphisms in the gene of MTHFR have been described as at least two of them (C677T and A1298C) have been shown to result in decreased enzyme activity. On turn, the reduced activity of MTHFR may lead to increased tissue and plasma homocysteine levels and DNA hypomethylation. Recent studies suggested that COPD is associated with hyperhomocysteinemia and hypomethylation of a large number of CpG sites. So far no study has been reported to explore the role of MTHFR C677T SNP in COPD. In this respect we aimed to determine the MTHFR C677T genotype frequency in patients with COPD and in control individuals and to evaluate its possible role for development of this disease. R ESULTS: We did not find significant difference between allele and genotype frequencies of COPD patients and controls (p = 0.870, p = 0.927). The polymorphism was not associated with the stage of the disease or with spirometric indexes. However, the carriers of TT genotype developed COPD significantly earlier (53.81 ± 13.02 yrs) than the heterozygous (62.00 ± 8.76 yrs, p = 0.044) and especially than carriers of CC genotype (64.22 ± 8.38 yrs, p = 0.012). CONCLUSIONS: These results suggest that MTHFR C677T polymorphism is not a risk factor for COPD but may contribute to early onset of the disease. Key words: COPD, MTHFR, polymorphism, risk


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Oral Presentations


NEUROMODULATORY EFFECTS OF TYR-MIF-1 FAMILY OF PEPTIDES ON CANNABINOID SYSTEM DURING HEAT STRESS-INDUCED ANALGESIA Adriana Bocheva, Hristina Nocheva Department of Pathophysiology, Faculty of Medicine, Medical University of Sofia, Bulgaria Stress is among the causes of different neuro-psychological diseases. Neuropeptides modulate many pathophysiological processes of the organism and this is where the need for studies on stress effects stems from. The Tyr-MIF-1 family of peptides comprises MIF-1, Tyr-MIF-1, Tyr-WMIF-1 and Tyr-К-MIF-1, which bind to opioid receptors as well as to non-opioid binding sites in the brain. Analgesia induced by stress is named stress-induced analgesia (SIA), and has an opioid and a non-opioid components. The non-opioid component comprises different neuro-mediatory systems, the cannabinoid one among them. The aim of the investigation was to evaluate the effects of the Tyr-MIF-1 family of peptides on the cannabinoid system during heat stress-induced analgesia (h-SIA). The experiments were conducted with male Wistar rats (180-200 g). Nociceptive tests were used with mechanical (paw-pressure test) and thermal (hot-plate test) nociceptive stimulus. Heat stress was induced by placing the animals in a thermal chamber. The peptides and the CB1 receptor agonist anandamide were injected intraperitoneally at a dose of 1 mg/kg. Results showed that Tyr-MIF-1 peptides interact with anandamide and influence h-SIA at a different degree. MIF-1 and Tyr-MIF-1’s analgesic effects are short-lasting, probably due to their interaction with their own binding sites that interact with the СВ1, but not with μ-opioid receptors. Tyr-W-MIF-1 and Tyr-К-MIF-1’s analgesic effects are due to binding both to their own binding sites (interacting with СВ1-receptors), as well as to μ-opioid receptors. In conclusion, peptidergic modulation of the cannabinoid system depends on the receptor binding of the peptides during heat stress-induced analgesia. Key words: Tyr-MIF-1 family of peptides, anandamide, heat stress-induced analgesia

EFFECT OF TRAINING AT LACTATE THRESHOLD INTENSITY ON MAXIMAL TIME TO EXHAUSTION AND ANXIETY BEHAVIOR OF SPONTANEOUSLY HYPERTENSIVE RATS AFTER KAINATE TREATMENT Katerina Georgieva1, M. Hadjieva1, M. Shishmanova-Doseva2, N. Georgiev3, G. Andreev3, Y. Chekalarova4 1Department of Physiology, 2Department of Pharmacology and Drug Toxicology, 3Faculty of Medicine, Medical University - Plovdiv; 4Institute of Neurobiology, BAS, Bulgaria The objectives of the study were to investigate the effects of exercise at lactate threshold intensity on maximal time of exhaustion (MTE) and heart rate (HR) during treadmill training and changes in anxiety level after kainate treatment of spontaneously hypertensive rats (SHRs). Male SHRs were divided in two groups. One group was sedentary and the other was exercised on treadmill at a speed of 20 m.min-1 and 0º elevation for 40 min.d-1, for 4 wk. At the beginning and at the end of the training period all rats were subjected to test for MTE and their HR was measured. Status epilepticus was evoked in half of the sedentary and trained rats by a repetitive intraperitoneal injection of kainate (KA) in low subconvulsive doses and other half of the groups received saline.

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All four groups were tested in the open field (OF) test and elevated plus maze (EPM) test a month after the KA/saline treatment. At the end of the training period MTE of trained rats was longer (P < 0.001) in comparison with baseline values, but had no effect on heart rate (P > 0.05). Kainate treatment after one month of training decreased the time in the corner in the OF test and increased the number of entries in the open arms in the EPM test and this effect was significant a month later (P < 0.05). Taken together, these results demonstrate beneficial influence of exercise on physical working capacity and anxiety behavior in a rodent model of essential hypertension and status epilepticus. Key words: training, epilepsy, anxiety, SHRs

APOPTOTIC BIOMARKERS IN NON-ALCOHOLIC FATTY LIVER DISEASE, INDUCED BY HIGH-FRUCTOSE DIET Kamelia Bratoeva1, Maria Radanova2, Ganka Bekiarova1, Maria Caneva3 1Division of Pathophysiology, Department of Physiology and Pathophysiology, Medical University - Varna, 2Department of Biochemistry, Molecular Medicine and Nutrigenomics, Medical University - Varna, 3Department of General and Clinical Pathology, Medical University - Varna, Bulgaria INTRODUCTION: The pathogenesis of non-alcoholic fatty liver disease as a component of metabolic syndrome (MS) is still unknown. Apoptosis may play an important role in pathophysiological mechanisms involved in liver damage and progression. Our aim was to detect early biomarkers of apoptosis in serum, as a cytokeratin-18 (CK-18) and its role in assessing the presence and severity of liver damage in high-fructose diet (HFD). METHODS: The study was performed on male Wistar rats divided into 2 groups (n = 7): control, high-fructose fed rats (35%, 16 weeks). Liver injury was evaluated immunohistochemically for presence of hepatic Bcl-2 family protein, biochemically for changes in aminotransferase levels and MDA (marker of lipid peroxidation) and histological (H&E) studies. Levels of CK-18 in hepatic and sera were measured by Western immunoblotting. R ESULTS: The results showed a significant increase of hepatic CK-18 levels, pro/anti apoptotic Bax/BCL-2 ratio by 92% (p < 0.01), elevation of liver MDA levels (p < 0.05) in HFD compared to the control group. HFD group showed microvesicular steatosis without changes in aminotransferase levels, inflammation and fibrosis. CK-18 serum levels correlated with changes in Bax/ BCL-2 proteins ratio, and the oxidative and histological changes in both groups. CONCLUSION: Increased serum CK-18 levels in the HFD group and its correlation with histological and biochemical changes in liver, suggested the key role of apoptosis in pathogenesis of the HDFinduced fatty liver in rats. The data shows that CK-18 may be a useful biomarker for non-invasive assessment of the liver damage and progression in MS. Key words: apoptosis, cytokeratin-18, non-alcoholic fatty liver disease, fructose


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Oral Presentations


ORAL PRESENTATIONS

Biopharmacy

EXPERIMENTAL STUDY ON THE DEVELOPMENT OF TOLERANCE AND WITHDRAWAL SYNDROME IN RATS TREATED WITH PREGABALIN TO PENTYLENETETRAZOLE-INDUCED SEIZURES Vesela Kokova, E. Apostolova, L. Peichev Department of Pharmacology and Drug Toxicology, Faculty of Pharmacy, Medical University - Plovdiv, Bulgaria INTRODUCTION: Pregabalin (PGB) was licensed in Europe as an add-on antiepileptic drug for the treatment of partial onset seizures for adults with uncontrolled seizures. AIM: The aim of this study is to investigate the development of tolerance and withdrawal syndrome in rats treated with pregabalin against the convulsant pentylenetetrazole (PTZ). MATERIALS AND METHODS: Male Wistar rats, divided into 12 groups (n=10), were treated as follows: groups 1 through 6 – with distilled water (controls) and groups 7 through 12 - with PGB in a dose of 150 mg/kg b.w. orally. The tolerance to the anticonvulsant effect to PTZ (120 mg/kg b.w.i.c.) was studied on days 1 and 14. The withdrawal syndrome to PTZ (60 mg/kg b.w. i.c.) was studied on day 14 of treatment and on days 1, 2, and 3 after abrupt termination of drug administration. R ESULTS: Data demonstrated slight decrease of the anticonvulsant activity of PGB on day 14 compared to day 1 of treatment, but with no significant difference. The seizure liability on days 1, 2 and 3 after the abrupt termination of drug administration didn’t show significant difference to the controls. CONCLUSION: Our data demonstrates that PGB does not lead to development of tolerance and withdrawal syndrome. Key words: pregabalin, tolerance, withdrawal syndrome, pentylenetetrazole, rats

STUDY ON THE BIOLOGICAL ACTIVITY OF FOLIA BETULAE EXTRACT Dimitar Penkov1, Margarita Kasarova1, Marianna Murdjeva2, Daniel Draganov3, Marian Draganov4 1Department of Pharmaceutical Sciences, Faculty of Pharmacy, Medical University - Plovdiv, 2Department of Microbiology and Immunology, Faculty of Pharmacy, Medical University - Plovdiv, 3Department of Biology, Molecular Biology and Plant Physiology, Faculty of Biology, Plovdiv University, 4Department of Medical Biology, Faculty of Medicine, Medical University - Plovdiv, Bulgaria AIM: To investigate the biological activity of birch leaf extract (Folia Betulae) on cell cultures in vitro. MATERIALS AND METHODS: The dry Folia Betulae extract was obtained by the method of percolation, and a subsequent removal of the solvent. It was standardized according to the content of rutin, quercetin, betulin and betulinic acid. The following cell lines were used in the experimental study: PC3 (human prostate cancer), HepG2 (human hepatocellular carcinoma) and SCP-1 (hTERT immortalized mesenchymal stem cells). Cells were treated for 24 hours with a solution of the extract in 10% DMSO. Proliferation, cytotoxicity and apoptosis assays were performed to determine the influence of the extract. R ESULTS: All cell lines showed reduction of cell numbers, changes in cellular morphology and integrity of the cell monolayer. The degree of changes and damages in cell cultures were dependent on the concentration of the extract. Cancer cells HepG2 and PC3 were more sensitive to treatment, compared to normal mesenchymal stem cells. It was confirmed that the extract of birch leaves induced apoptosis in the treated cells.

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CONCLUSIONS: Dry Folia Betulae extract has biological activity, demonstrated as changes in cell morphology and induction of cell death by apoptosis in PC3 and HepG2 cell lines. Stronger inhibition of proliferation and reduced cell vitality was observed in PC3 and HepG2 cell cultures, compared to normal mesenchymal stem cells SCP-1. Key words: Folia Betulae extract, biological activity

FLOWABILITY EVALUATION OF DRY POWDER FORMULATIONS FOR NASAL DRUG DELIVERY Bisera Pilicheva, Plamen Katcarov, Margarita Kasarova Department of Pharmaceutical Sciences, Faculty of Pharmacy, Medical University - Plovdiv, Bulgaria The development of effective and reliable nasal formulation is undoubtedly a serious challenge in drug delivery. The most acceptable form for nasal drug delivery is dry-powder inhalation formulation due to environmental sustainability, propellant-free design, little or no requirement of patient coordination and formulation stability. For optimum deposition in the nasal cavity drug particles’ size should be in the range of 1-25 microns. However, such particles are not free-flowing due to cohesion and static charge phenomena which interfere with handling during manufacture and could aggravate dose uniformity and cause retention within the device. Excipients can improve flowability and reduce particles aggregation. The aim of this study was to develop interactive mixtures of betahistine-loaded microspheres and different carriers for inhalation delivery with optimum flow properties. To study the effect of these carriers on microspheres flowability interactive (adhesion) mixtures at two different microsphere/carrier ratios were formulated and were studied for angle of repose and Hausner ratio (HR). Formulated mixtures showed values for the angle of repose within a broad range – from 23.86° to 52.53°, and for HR - from 1.09 to 1.95. Blending the cohesive microspheres with the carriers resulted in lower angle of repose and HR which indicates improved flowability. With high microsphere/carrier ratio improved flow properties were observed. The results show that good flowability depends on carrier properties and microspheres characteristics and confirm the thesis that choosing the proper carrier excipient is of great importance for effective nasal drug delivery. Key words: flowability, interactive mixtures, nasal delivery

TECHNOLOGICAL APPROACH FOR OPTIMIZATION OF BETAHISTINE RELEASE FROM BIOADHESIVE MICROSPHERES Plamen Katsarov, Bisera Pilicheva, Margarita Kasarova Department of Pharmaceutical Sciences, Faculty of Pharmacy, Medical University - Plovdiv, Bulgaria Polymer microspheres as drug delivery systems with sustained release of betahistine were already developed and characterized in a previous study. The in vitro drug release from the obtained microspheres was ‘biphasic’ with initial burst release of betahistine at a great rate - a phenomenon known as ‘burst effect’. Physiologically it could lead to toxic effects of the drug. In order to reduce this initial ‘burst effect’ and achieve more steady release of betahistine three new models of polymer microspheres were developed by modifying the production method through cross-linking the structure of the polymer matrix. Glutaraldehyde was used as a crosslinking agent. The obtained microsphere models were characterized by their shape and size, yield, drug content and drug loading efficiency. An in vitro test for the release of betahistine from the mod-


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Oral Presentations


els was performed, as well as a permeability test through an artificial membrane. The results indicate that the crosslinking of the polymer with glutaraldehyde during the preparation of the microspheres has reduced the rate of the initial release of incorporated betahistine, which is most probably due to the extended time for drug diffusion through the crosslinked polymer matrix. The elimination of the intense ‘burst effect’ permits a prolonged release of betahistine from the polymer carriers and a gradual increase of its concentration in the acceptor medium. This is important for maintaining serum levels of the drug within the therapeutic window for an extended period of time under in vivo conditions. Key words: polymer microspheres, cross-linking, sustained release

POLY(VINYL ACETATE) NANOCARRIERS IN EYE FORMULATIONS Velichka Andonova1, Stela Dimitrova2, Mariana Katsarova2, Georgi Georgiev3, Margarita Kasarova1 1Department of Pharmaceutical Sciences, 2Department of Chemistry and Biochemistry, Faculty of Pharmacy, Medical University - Plovdiv; 3Faculty of Chemistry and Pharmacy, St. Kliment Ohridski Sofia University, Bulgaria The focus of the present work was the investigation of the effect of polymer nanocarriers on the biopharmaceutical characteristics and chemical stability of drugs in aqueous medium when the nanoparticles are included in eye drops. The nanocarriers were obtained via one-stage emulsifier free radical polymerisation of vinyl acetate. For model drugs were selected indomethacin and acetylsalicylic acid due to their powerful anti-inflammatory activity and their inappropriate physical and chemical characteristics for inclusion in liquid dosage forms: low solubility in water and low chemical stability. Characterization of the nanocarriers was performed via transmission electron microscopy, infrared spectroscopy and dynamic light scattering. Their nanosized nature, the particle size distribution, -potential and also the degree of interaction of the drug with the polymer carrier we determined. The in vitro release studies at pH 7.4 showed that the rate and extent of the drug release from these nanocarriers could be adjusted by changing the composition of the copolymers, the polymer mixture, as well as the conditions for producing the nanoparticles. The stabilizing role of the polymer carrier on the included drugs in an aqueous medium was proved by accelerated tests and tests under ‘stress’ conditions using the technique of high performance liquid chromatography. The established biocompatibility of the polymer carriers, their stabilizing influence on the in-situ included drugs and the in vitro release studies at pH 7.4, give reasons to recommend these nanocarriers as drug delivery systems in eye drops. Key words: nanoparticles, drug delivery, eye drops

ANTI-NOCICEPTIVE EFFECT OF RHODIOLA ROSEA L. AND CURCUMA LONGA L. STANDARDIZED EXTRACTS IN RATS Lilia Vasileva, Damianka Getova 1Department of Pharmacology and Clinical Pharmacology, Medical Faculty, Medical University - Plovdiv, Bulgaria AIM: Rhodiola rosea L. is an adaptogen used to treat stress reactions. Curcuma longa L. with main active constituent curcumin has a range of beneficial effects. Our aim was to study the effects of standardized extracts of both plants on nociception in rats. METHODS: Male Wistar rats (6

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groups, n = 8) treated with: controls; Metamizole; Rhodiola 250 mg/kg p.o.; Rhodiola 500 mg/kg p.o.; Curcuma 250 mg/kg p.o.; Curcuma 500 mg/kg p.o. Four nociceptive tests were used. Criterion for hot-plate and tail flick tests was the latency of reaction, for analgesimeter – the paw pressure reaction and for formalin test - the time of licking the injected paw. The statistic evaluation was done by ANOVA. R ESULTS: In analgesimeter test rats treated with Rhodiola in the low dose significantly increased the paw reaction compared with ‘zero time’ of the group and with Metamizole. Groups treated with Curcuma significantly increased the paw reaction compared with ‘zero time’ of the group and with Metamizole. In hot-plate test rats treated with Rhodiola and with Curcuma in the low dose significantly increased the latency of reaction compared with the respective ‘zero time’. In tail flick test group treated with Curcuma in the high dose significantly increased the latency of reaction compared with “zero time” of the group and with controls. In formalin test rats treated with Rhodiola in the high dose and Curcuma significantly decreased the time of licking the injected paw in the late phase. CONCLUSIONS: Our data suggest that the studied extracts exert anti-nociceptive effect in all used pain models. Key words: adaptogen, plant extract, nociception, rats

ORAL PRESENTATIONS

Public Health IT IMPLEMENTATION IN DISASTER MEDICAL SUPPORT - REQUIREMENT AND CHALLENGE Rostislav Kostadinov, Nikolai Varshilov, Rumiana Etova Department of Epidemiology and Disaster Medicine, Faculty of Public Health, Medical University - Plovdiv, Bulgaria INTRODUCTION: Contemporary world frequently is described as global and wired, because of the Information Technology development and their impact on our daily life. On the other hand the last decades statistics reveal an exponential increase in both disasters frequency and severity. Recorded steady trend in disasters occurrence has raised the question about society preparedness to decrease population vulnerability and increase the community resilience. The AIM is to analyze how IT could enhance Disaster Medical Support capabilities. MATERIAL AND METHODS: Descriptive method is applied for presenting some of the recently achieved IT capacities in communication and Disaster Medical Support activities. Using a comparative method the Disaster Medical Support requirements for information and the new communications systems means and capabilities are analyzed. R ESULTS AND DISCUSSION: During all the disaster development phases Disaster Medical Support planners and managers are requiring constant information influx regarding observed changes in hazards, risk factors, hazardous sites and behaviour and health risk levels. The requirement for timely, accurate, precise and concise medical information is an imperative during prodromal, impact and response phases when information delay leads to lives lost. Implementing the new IT in medical information flow management in several disaster relief operations proved to benefit the Disaster Medical Support outcome. Key words: disaster medical support, IT, communication


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Oral Presentations


THE HTA GLOBAL ENVIRONMENT – POTENTIAL AS A TOOL FOR HEALTHCARE DECISION MAKING Ralitca Raycheva, Rumen Stefanov Faculty of Public Health, Medical University – Plovdiv, Bulgaria HTA activity – constantly growing over time at a global level – supports evidence based decisionmaking processes with sound knowledge and informs coverage and funding of new health technologies. The international survey on HTA was designed to attain enlightenment on the present status of HTA activities as well as to reveal some insights in HTA characteristics, processes and best practices. Between 2013 and 2015 an extensive international Web-based survey of HTA activities was conducted among 312 organizations with diversified profile located in 58 countries, including 11 international organizations. Data were obtained by semi-structured questionnaire, which contained 102 questions incorporated in six different panels, one specific section and two information segments with a total of 71 multiple choice and 31 open questions. A total number of 76 questionnaires from 23 countries on 5 continents were received, representing a response rate of 24.3%. The initial actions for HTA organization’s establishment were undertaken in 50.94% by government. They started in 98.11% as non-profit structures. The HTA agencies informed decision at the following levels: international – 7.55%; national – 39.62%; regional – 3.77%; international/national – 35.85%; international/national/regional – 7.55%; and can’t give an answer – 5.66%. Their key direction of activity is health technology assessment (81.13%). The final HTA product targets in 45.28% are decision-makers in health. HTA utilization is usually implemented at: top political and administrative level – 45.28%; institutional level – 26.42%; project level –18.87%; can’t give an answer - 9.43%. HTA prove to be a powerful tool for supporting and facilitating effective decision-making process on different levels of healthcare systems. Key words: health technology assessment, capacity building, decision support, evidenceinformed policymaking

IS DISEASE PREVENTION A PRIORITY FOR BULGARIAN PUBLIC HEALTH POLICY? Nadia Veleva, Makreta Draganova Faculty of Public Health, Medical University - Pleven, Bulgaria INTRODUCTION: Importance of prevention in achieving healthcare system aims is repeatedly proven and holds a central position in all WHO strategies. According to the Health Law and the National health strategy 2014-2020 prevention of diseases is set among the priorities of Bulgarian public health policy. The AIM is by analyzing the financial costs for preventive activities during the period 2003-2012 to assess the actual rank of prevention among the national public health priorities. MATERIAL AND METHODS: Data about current expenditures by basic financial sources from the System of Health Accounts of the National Statistical Institute database and financial reports of the National Health Insurance Fund (NHIF) for the period 2003-2012 was used. R ESULTS: reveal a prevalence of hospital services during the studied period. Financial costs for hospital treatment register continuously increasing rate which accounts for 5% in 2012 compared to 2011. The relative share of financial expenditures for primary care for 2012 is twice as low as the one for hospital care and the share of preventive medicine is significantly lower than the first one. CONCLUSIONS: Despite the legal reorientation towards preventive medicine “hospital healthcare” still dominates the Bulgarian health system. It is well known that this model is the most expensive

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one and is far away from both economic efficiency and medical effectiveness. Hence new public health policy measures are needed towards setting prevention on its due priority position. Key words: prevention, public health policy, priority

APPROACHES FOR EFFECTIVE TIME MANAGEMENT Makreta Draganova, Nadia Veleva, Silvia Aleksandrova-Yankulovska Department of Public Health Sciences, Faculty of Public Health, Medical University - Pleven, Bulgaria INTRODUCTION: Shortage of resources for health care is an inevitable fact. Provision of quality health services in times of financial crisis and scarcity of resources is a great challenge for healthcare managers. Time resource is becoming an object of increasing scientific research. AIM: The aim of this paper is to analyze different approaches for effective time management and the possibilities for their implementation in healthcare managers’ practice. MATERIAL AND METHODS: The study is based on systematic analysis of scientific publications in peer-reviewed conference proceedings and scientific journals. R ESULTS AND DISCUSSION: Description of six approaches for effective time management is presented. All of them are found to be applicable in healthcare managers’ everyday practice. An algorithm for effective time management is elaborated and specific tools are suggested for use in particular practical situations. The system of strategies, approaches and tools for effective time management provided is a real option for all healthcare managers and its practical implementation is a matter of personal choice. CONCLUSIONS: Time management is part of organizational management and is recognized as a key factor for success. Benefits of effective time management are proven both on organizational and on personal level. Effective time management skills are precondition for easier achievement of profits. Key words: management, time, healthcare, approaches

ORAL PRESENTATIONS

Therapy R ESTLESS LEGS SYNDROME (WILLIS-EKBOM DISEASE) AND SLEEP DISORDERS - DIAGNOSTIC AND THERAPEUTIC CHALLENGES Mario Milkov, Georgi Marinov, Plamen Nedev, Cvetan Tonchev, Hristiana Madjova Department of Prosthetic Dentistry and Orthodontics, Faculty of Dental Medicine, Prof. Paraskev Stoyanov Medical University - Varna, Bulgaria Restless legs syndrome (RLS), also known as Willis-Ekbom disease (WED), is a common neurological and sensorimotor disorder. It was first described by Thomas Willis in 1685 and the diagnostic criteria occurred in the monograph ‘Restless legs’ by Karl-Axel Ekbom in 1945. About 2-3% of adults in Europe and America suffer from clinically significant symptoms which exert a


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substantial negative impact on their sleep, quality of life, and health. RLS/WED is characterized by an uncomfortable urge to move the legs before sleep (periodic leg movements, PLMs), often accompanied by abnormal sensations in the legs, appearance of symptoms during periods of rest or inactivity, partial or complete improvement by walking or moving legs, and evening onset or worsening of symptoms. The International Restless Legs Syndrome Study Group develops updated diagnostic criteria enabling rapid development of research and treatment for RLS/WED. Surface electromyogram of the anterior tibialis muscle is the standard for counting PLM. Timely recognition, referral to polysomnography, and longitudinal patient’s follow-up are imperatives for neurologists in providing quality comprehensive patient care. Treatment strategies are divided into pharmacotherapy and non-pharmacological methods. Pramipexole, ropinirole and rotigotine are effective (Level A evidence) in treating RLS/WED for up to six months while pregabalin is effective for up to one year. On the other hand, probably effective (Level B evidence) in treating RLS/WED are gabapentin enacarbil (for up to one year), levodopa (for up to two years), and rotigotine (for up to five years). Key words: restless legs syndrome, periodic leg movements, sleep disorders, diagnosis, pharmacotherapy

CORRELATION BETWEEN END PRODUCTS OF OXIDATION OF LIPIDS AND PROTEINS WITH SISTEMIC INFLAMMATION IN PATIENTS WITH BRONCHIAL ASTHMA Vania Ilieva1, Vania Coneva2, Galina Nikolova3, Antoaneta Jeleva3, Yanica Jeliazkova4, Veselina Gadjeva3 1Department of Propaedeutics of Internal Medicine, Medical Faculty, Trakia University, Stara Zagora, 2Cenral Clinic Laboratory, Medical Faculty, Trakia University, Stara Zagora, 3Department of Chemistry and Biochemistry, Medical Faculty, Trakia University, Stara Zagora, 4Department of Propaedeutics of Internal Diseases, Faculty of Medicine, Trakia University, Stara Zagora, Bulgaria Bronchial asthma (BA) is a chronic inflammatory lung disease involving complex interactions between numerous cell types and mediators that results in airway hyperreactivity and variable airflow limitation. Oxidative stress (OS) is related to the etiopathogenesis of a lot of chronic diseases, including bronchial asthma and plays an essential role in their progress and therapy. OS can be diagnosed by measuring some final products of oxidation of lipids and proteins such as malondialdehyde (MDA) and protein carbonil concystence (PCC). Previous studies have reported inverse relationship between pulmonary function and C-reactive proteins (CRP), as well as the effect of asthma exacerbations on the level of CRP. The AIM of the present study was to clarify the correlation between the final products MDA and PCC with some markers of systemic inflammation CRP and fibrinogen in sera of BA patients. METHODS: 22 bronchial asthma patients and 20 controls were included in the study. MDA was found spectrophotometrically, PCC - using ELISA, CRP – by immunoturbidimetrical and fibrinogen – by coagulation chronotest. R ESULTS: Statistically significant higher values of MDA were found in BA patients compared to controls (mean 2.63 ± 0.35 vs. 1.76 ± 0.56; p < 0.0000). When PCC was measured significant difference was found between BA and controls (8.84 ± 3.19 vs. 5.53 ± 4.06, р < 0.05). Moreover, positive correlations were found between MDA with CRP and fibrinogen (r = 0.542 and r = 0.29, respectively). Conversely, negative correlations were found between PCC with CRP and fibrinogen (r = -0.13 and r = -0.26). CONCLUSION: The increased oxidative stress found in BA patients influences the systemic inflammatory markers. Key words: bronchial asthma, oxidative stress, fibrinogen, CRP

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GLOBAL CHALLENGES IN EPIDEMIOLOGY OF INFECTIOUS DISEASES Yordanka Stoilova Department of Epidemiology and Disaster Medicine, Faculty of Public Health, Medical University - Plovdiv, Bulgaria Nowadays, in the 21st century worldwide distribution of infectious diseases still remains a formidable challenge for the medical scientific communities. In this regard, our aim is to analyze the global challenges in epidemiology and to provide data on some of our own home research. Based on analytic and quantitative epidemiological methods global issues are assessed in the epidemiology of infectious diseases such as: high potential for epidemic spread; constant priority for healthcare systems of underdeveloped and developed countries; the lower mortality rate from infectious diseases during the last 30 years, today demonstrates a reverse trend; global population growth rate; migratory waves affecting almost the whole world result in epidemics and outbreaks; nosocomial infections associated with healthcare practices are among the leading problems in the world along with flu and viral hepatitis; unknown pathogens emerge; the appearance of new and newly emerging infections; climatic changes; antibiotic resistance; changes in the demographic structure; increased lifespan; innovations in medicine; variation in immune resistance of humans; organ and tissue transplantation; metabolic diseases; bio-terrorism etc. Conclusion: Globalization, the rise of old and newly emerging infectious diseases, changing epidemiology of infectious diseases, altering patterns of morbidity and mortality in the past 30 years require a reassessment of the strategy for prevention and control, based on professional expertise, laboratory support and scientific capacity. Molecular epidemiology designs, programs overcoming social, religious and regional barriers, scientific cooperation and business organizations ensure high result in the efforts to achieve disease control and prevention. Key words: globalization, prevention and control, newly emerge infections

INFECTIOUS DISEASES - QUO VADIS? Radka Komitova Department of Infectious Diseases and Parasitology, Medical University - Plovdiv, Bulgaria The specialty of infectious diseases is a broad discipline that is almost unique in modern medical science: it allows physicians to cure the diseases within its reference range, to prevent these diseases, to identify the specific causes (microbes) of these diseases, and to manage diverse, sometimes massive outbreaks of diseases. The value of infectious disease specialist is now getting higher all because of the current antibiotic resistance problem, the consequences of the expansion of international travel and the introduction of completely new pathogens diagnostics. In the future the approaches to conservation of antibiotics will include short-course regimens, molecularbased diagnosis designed to make the pathogen-target antibiotic choice better and more accurate, and optimal use of currently available agents. Clinical practice is now enriched by molecular diagnostic techniques of detecting epidemic pathogens or resistance genes with the salient features of speed, precision and great sensitivity. Studying the microbiome in different anatomical sites would afford the opportunity to treat or prevent multiple pathologic conditions such as obesity, allergy, autoimmune diseases, or cancer. Another important area is the development of bundles that prevent the complications of the infectious diseases. They are highly unpredictable, often making it necessary to include specialists in more than one field of medicine in their management. Despite all this, their significance is very often disregarded by healthcare systems and healthcare reforms. Key words: antibiotic stewardship, epidemics, infection control, microbiome


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ORAL PRESENTATIONS

Surgery NATIONAL REGISTER FOR FACIAL ANOMALIES Youri Anastassov1, Kostadin Gigov1, Regina Hatar1, Atanas Gyulev1, Radost Velikova1, Mariya Kazakova2, Neli Hashova3, Petya Tsarvulanova2 1Department of Propaedeutic of Surgical Diseases, Section of Plastic and Craniofacial Surgery, Medical University - Plovdiv, 2Association of Patients with Facial Anomalies and Their Parents – ALA, 3St. George University Hospital, Plovdiv, Bulgaria Our web based register for facial anomalies in Bulgaria (https://dfa-bg.org) was created thanks to a project of the Ministry of Education. The register allows also to monitor the follow-ups of cases performed by the specialists included in the national networks (feeding specialists, speech therapists, ENT doctors, orthodontists). An original, secured and protected software platform was created to document the outcomes from the treatment of the patients. The information shared by the specialists could be text, pictures and video files. In 2013, out of 61700 newborns in the country, we performed surgeries in 71 children with cleft lip and palate born in this year with this pathology, which shows that we dealt with almost 90% of the cases in the country. Twenty-seven of the cases had a cleft palate only, 20 - unilateral cleft lip and palate, 2 - bilateral cleft lip and palate and 23 - cleft lip. For a year and a half of operation 480 patients and 51 specialists have been registered. We did not have permission for inclusion in the register in only 3 (4%) cases of children raised in institutions. 80% of the users have active electronic addresses, 30% of them are active in posting to the system. The register has several advantages over the existing registers worldwide of being patient oriented, of having the capacity to process all types of files, and being able to be accessed in throughout the country. Although it is a hospital based register, it can be used in future as epidemiological database because it can clearly include almost 90% of the cases with this pathology in Bulgaria. Key words: facial anomalies, cleft lip and palate, register

PROBLEMATIC INFECTIONS IN SURGICAL PATIENTS – A 3-YEAR ANALYSIS Emma Keuleyan I. Batashki, B. Moshev, Sh. Teteva, D. Todorov, D. Panaiotova Medical Institute - Ministry of the Interior, Sofia, Bulgaria The AIM of the study was to review infections in patients of the Clinic of Surgery, Medical Institute, Ministry of the Interior, 2012 – 2014, and to analyze the etiological spectrum, antibiotic resistance and the administered therapy. MATERIALS AND METHODS: Retrospective analysis of surgical procedures, monitoring of antibiotic resistance and consumption, and audit of therapy. R ESULTS: For the 3-year period, 4727 patients were treated, 30% received antibiotic therapy. Among 3591 operations, 464 were referred for soft tissue infections, 38 for acute peritonitis, 75 for intra-abdominal infections and different organ infections; 22 patients with sepsis were treated. The leading pathogens were: 309 Staphylococcus aureus, 101 Enterococcus spp, 510 Enterobacteriaceae, 130 Gram negative non-fermenters, 65 anaerobes. The relative rate of problematic antibiotic resistant microorganisms was: MRSA 0%, 6%, 7%; ESBL-Enterobacteriaceae 17%, 20%, 25%, carbapenem – R: Acinetobacter spp – 14%, 25%, 67%, Pseudomonas aeruginosa – 25 %, 7 %, 15 %. Pathogens in sepsis were more resistant (from 15 bacteremiae with Enterobacteriaceae, 10 produced ESBL; from 5 with Acinetobacter spp, 4 were poly-resistant, icl. carbapenem – R),

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which necessitated a rapid diagnosis and targeted antibiotic treatment with reserve antibiotics (while cephalosporins of the I and III generation, aminoglycosides and metronidazole were the ones most commonly used (respectively, 47, 33, 16, 10 DDD/100 bed-day). CONCLUSIONS: This study found an increase of number of patients with severe infections together with an increase of the number of poly-resistant strains, which demands improvement of therapeutic approaches. Key words: surgical infections, antibiotic resistance, therapy

MAMMOGRAPHIC DENSITY AND RISK OF HISTOLOGICAL PRECURSORS OF BREAST CANCER IN POSTMENOPAUSAL WOMEN G. Baytchev, I. Inkov, N. Kyuchukov, A. Dimitrova, A. Goncharov Thoracic Surgery Clinic, Military Medical Academy, Sofia, Bulgaria Mammographic density has been proven to be an independent risk factor for breast cancer development, currently associated with a 4-6 fold increased risk of breast cancer. The aim of the present study is to analyze the relationship between different histological lesions and the mammographic density of the breast. The subjects in this prospective study were 150 post-menopausal women, treated at the Thoracic Surgery Clinic of the Military Medical Academy in Sofia, Bulgaria between 2010 and 2014. The patients were divided into 3 groups depending on the underlying pathology as follows: non-proliferative fibrocystic disease, atypical hyperplasia and invasive breast carcinoma. The radiographic density of breast was determined using the ACR BI-RADS Assessment Categories respectively, D1 < 25% - 82 cases, D2 = 20-50% - 44 cases, D3 = 51-75% - 17cases, D4 > 75% -7 cases. The results showed that 24 (16%) of the patients had a D3-D4 density of the breast, corresponding respectively to 5, 8, and 11 cases corresponding respectively to the above 3 groups. Women with higher fibro-glandular density of the breast have a higher risk for atypical hyperplasia and carcinoma. However, the sensitivity of a mammographic study and consequently the ability to detect pathological lesions is reduced. Therefore increased attention and additional diagnostic methods are needed for improving the diagnosis of breast pathology. Key words: mammographic density, breast cancer

ALGORITHM FOR TREATMENT OF GYNECOMASTY IN ADOLESCENTS PATIENTS Krasi Kalinova1, D. Dinkov1, Y. Dimcheva2, K. Georgiev1 1Department of Pediatric Surgery, Trakia University & University Hospital, Stara Zagora, 2Virgin Mary Hospital, Burgas, Bulgaria Gynecomastia is a benign, unilateral or bilateral enlargement of male breast due to the imbalance between the androgens and estrogens at the breast tissue level. This clinical condition is particularly common in boys during puberty. Gynecomastia may cause considerable psychological distress, anxiety, discomfort and fear of breast cancer, it is also commonly associated with breast pain, ranging in intensity from mild tenderness or sensitivity to constant pain and tension. AIM: The aim of this review was to present up-to-date information concerning the prevalence, pathogenesis, diagnosis and treatment of gynecomastia. At present, there are no generally accepted procedures for evaluation and treatment of adolescents patients. MATERIAL AND METHODS: After obtaining institutional review board approval, a retrospective review was conducted on all patients operated on for gynecomastia between January, 2000 and January, 20015. Data recorded


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included patient demographics, diagnostic evaluation, medical and surgical treatment, complications, and outcome in Department of Pediatric surgery, University Hospital Stara Zagora and Virgin Mary Hospital, Burgas. R ESULTS: Twenty-two patients underwent an operation for gynecomastia. Six patients had bilateral inferior reduction mammoplasty, and 16 patients underwent either unilateral or bilateral subcutaneous mastectomy. The mean age at operation was 15.6 years (range, 13-18 years). In all cases, the histopathology feature was consistent with gynecomastia. In two patient postoperative wound infections. Two patients developed a seroma after subcutaneous mastectomy requiring drainage. Two patients had subcutaneous asymmetry after a reduction mammoplasty that required no further management. 90% of patients subjected to reduction mammoplasty had a successful surgery. CONCLUSIONS: Milder grades of gynecomastia can be managed with subcutaneous mastectomy. Key words: gynecomastia, adolescents, surgical treatment

CLINICAL SIGNIFICANCE OF MONOCYTE HLA DR EXPRESSION AND SERUM LEVELS OF CRP IN THE PROGNOSIS OF PATIENTS WITH ACUTE PANCREATITIS Georgi Minkov1, Krasimira Halacheva2, Alen Petrov1, Tatiana Vlaikova3, Angel Popharitov1, Yovcho Yovchev1 1Department of Surgery, University Hospital, Stara Zagora, 2Department of Immunology, Medical Faculty of Trakia University, Stara Zagora, 3Department of Chemistry and Biochemistry, Medical Faculty of Trakia University, Stara Zagora, Bulgaria BACKGROUND: Early detection of severe forms is essential for reduction of morbidity and mortality in patients with acute pancreatitis (АP). MATERIALS AND METHODS: We analyzed the role of serum levels of CRP and monocyte expression of HLA DR on admission (CRP 1 / HLA DR 1) and at 48 hours (CRP 2 / HLA DR 2) as prognostic markers in patients with AP. R ESULTS: 39 (54.2%) patients had the disease in a mild form, 19 (26.4%) were with moderate AP and 14 (19.4%) had a severe form AP. 15 (20.8%) developed organ failure (OF) and local complications were found in 32 (44.5%). We found significant difference between levels of CRP 1, CRP 2, HLA DR 1 and HLA DR 2 in patients with severe AP (p = 0.009, p = 0,000, p = 0,000 and p = 0,000, respectively). The cut-off values of HLA DR 1 and HLA DR 2 for severe AP were 50.8% (AUC – 0.700) and 43.35% (AUC – 0.800), respectively. Patients who developed OF had significantly lower levels of HLA DR 1 and 2 (p = 0.005 and p = 0.007, respectively). There was a significant association between levels of CRP 2, HLA DR 1, HLA DR 2 and the development of pancreatic necrosis (p = 0.000). CONCLUSION: Levels of HLA DR’s monocyte expression and CRP, determined on admission and at 48 hours have an important role in the prognosis of patients with AP. Key words: acute pancreatitis, prognosis, HLA-DR, complications

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METAMORPHOSIS (TRANSFIGURATION) OF SPONTANEUS BACTERIAL PERITONITIS Nikolai Krastev1, Ventseslav Djurkov2, Penka Akrabova 2, Tanka Karparova2, Krasimir Asenov2, Georgi Kiprin2 1Second Department of Internal Medicine, Division of Gastroenterology, Medical University - Plovdiv; 2Department of Internal Medicine, Eurohospital, Plovdiv, Bulgaria Although currently mortality of spontaneous bacterial peritonitis (SBP) has decreased from 80% to 20%, this disease is now the main cause of death in patients with cirrhosis. SBP is detected in 10-15% of patients with cirrhosis and ascites, but its frequency is even higher (19%), the addition of the cases of culture negative neutrophilic ascites (a variant of the SBP). A diagnosis of SBP is made when polymorphonuclear leukocytes in ascites > 250 mm3, regardless of the outcome of the bacterial cultures. SBP occurs most often in the third year of occurrence of ascites (7-30% per year), but can also occur much earlier, especially during hospitalization, without regard to the age, gender and the etiology of cirrhosis. SBP may occur at any cirrhosis with ascites, regardless of the stage in Child-Pugh (Class C or B). Bleeding from esophageal varices is no longer a major predisposing factor for SBP, but in total protein in ascites < 10 g/l risk increases ten-fold. More frequent are the latent forms of oligosymptomatic SBP and the fulminant forms are rare. Deteriorated general condition, fever, encephalopathy and impaired liver or kidney function may be the only manifestations of SBP. Only rarely is the causative agent of SBP isolated even if hemoculture is used. The gram-positive microorganisms prevail especially after quinolone treatment of SBP. These microorganisms occur more rarely with polymorphonuclear leukocytes > 250 mm3. Gram-positive microorganisms, especially in nosocomial infections, are often resistant and multidrug-resistant, including to vancomycin. Serious diagnostic and therapeutic problems in SBP are expected in future. Key words: cirrhosis, ascites, SBP

INTRAVITREAL APPLICATION OF DEXAMETHASONE IMPLANT (OZURDEX): FIRST RESULTS Nelly Sivkova 1Department of Ophthalmology, Medical University - Plovdiv; Clinic of Ophthalmology, St. George University Hospital, Plovdiv, Bulgaria AIM: To present first own results from the application of Ozurdex – innovative method for treatment of macular edema (ME). PATIENTS AND METHODS: The study included the first 10 patients, 7 men and 3 women, aged 33 - 78 years who underwent intravitreal application of 0.7 mg dexamethasone. ME was due to central retinal vein occlusion. At 1, 3, and 6 follow-up months after the intravitreal application we examined visual acuity (VA) and average central macular thickness (ACMT) using spectral-domain optical coherence tomography. R ESULTS: Pretreatment data were: VA from 0.001 to 0.5 and ACMT from 478 to 712 μm; one month after Ozurdex treatment: VA from 0.01 to 0.6, ACMT from 302 to 657 μm; at 3 months: VA from 0.02 to 0.7, ACMT from 212 to 534 μm; at 6 months: VA from 0.15 to 1.0, ACMT from 194 to 312 μm. А significant decrease of retinal thickness was found, which corresponds to increase in visual acuity. CONCLUSION: The first results from the intravitreal application of dexamethasone implant in macular edema due to vein occlusion are encouraging. Multicenter randomized studies are needed to follow-up the late results of the treatment. Key words: Ozurdex, macular edema, venous occlusion


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OPTICAL COHERENCE TOMOGRAPHY IN PATIENTS WITH OPTIC NEURITIS DUE TO MULTIPLE SCLEROSIS Marin Atanasov, Marieta Konareva-Kostianeva, Snezhina Kostianeva, Ralica Kermedchieva, Aleksandar Gerdjikov Department of Ophthalmology, Medical University - Plovdiv; University Clinic of Ophthalmology, St. George University Hospital - Plovdiv, Bulgaria The aim of this study was to establish the type of damage to structures of the optic nerve detectable by optical coherence tomography (OCT) in patients with optic neuritis, due to multiple sclerosis (MS). PATIENTS AND METHODS: OCT was performed in 32 patients with optic neuritis due to MS with full functional recovery. The structures taken into account were the thickness of the retinal nerve fiber layer (RNFL) and the retinal ganglion cell complex (GCC). For comparison we used the fellow non-affected eyes. Except OCT, all patients received complete eye examination, including tests of visual acuity and visual field. R ESULTS: Even after full clinically detectable functional recovery of the affected eyes (return to pre-disease visual acuity and visual field), OCT establishes diffuse thinning RNFL and GCC. CONCLUSIONS: OCT can identify morphological changes in optic neuritis associated with MS, which until now could only be inferred. The method contributes to differential diagnosis of the disease and its confirmation. Key words: optical coherence tomography, multiple sclerosis

ORAL PRESENTATIONS

Dental Medicine INTERCANINE DISTANCE CHANGES FROM EARLY MIXED DENTITION TO PERMANENT DENTITION Laura Andreeva Department of Orthodontics; Faculty of Dental Medicine, Medical University - Sofia, Bulgaria AIM: To study the changes in the intercanine width in the upper and lower arches from early mixed dentition to permanent dentition in correlation to growth and development of the dental arch and the conducted orthodontic treatment. Material: A total of 14 patients in early mixed dentition were followed up until the final formation of permanent dentition. An original measurement protocol of intercanine widths was implemented. The patients had the first phase of orthodontic treatment commenced with removable or fixed appliance. After the eruption of the permanent teeth in the “support zone” the patients continued their treatment with comprehensive fixed technique treatment until malocclusion was corrected completely. Results: Frequency distribution of the maxillary and mandibular intercanine widths in early mixed dentition, as well as in completed permanent dentition was evaluated. The occurring changes in intercanine distances in the upper and lower arch were analyzed. Conclusion: After patients were evaluated for an average period of 6.2 years, the following conclusions were made: intercanine widths in the maxillary arch in the early mixed dentition varies from 14.81 mm to 28.40 mm. In the permanent dentition

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the values ranged from 15.56 mm to 27.60 mm, thus showing no significant differences in the maxillary intercanine width in the both investigated dentitions. Mandibular intercanine width at baseline was in the range 11.91 mm - 21.70 mm. At the end of treatment it was 10.78 mm to 23.74 mm, demonstrating that the mandibular intercanine width decreased during the permanent dentition. Key words: intercanine distance; early mixed dentition

COMPLEX FOCAL DIAGNOSTICS IN DENTAL MEDICINE - A METHODIC WHICH DOESN’T KNOW LIMITS Maria Dencheva1, Ivan Chakalov2, Julia Kamenova2, Angelina Kisselova1 1Department of Imaging and Oral Diagnostics, 2Department of Prosthetic Dentistry, Faculty of Dental Medicine, Medical University - Sofia, Bulgaria The complex focal diagnostics (CFD) in dental medicine is a sequence of procedures that is in full accordance with the development of modern scientific technology. This algorithm gives priority to dental health and its role in the patient’s health status. The diagnostics includes well-known diagnostic techniques such as pulp vitality test, corrosion potential measurement, Gehlen’s electro-skin test as well as other innovative methods such as thermovision and electroacupuncture measurement of energy in the body with Acugraph and T-scan. The patients we usually examine today present very often with chronic diseases with incomplete or delayed diagnosis. The CFD supports process of the energetically misbalance of the body and helps to develop an individual treatment plan. CFD can be improved with the advance of technology; the dentist in the focal team is responsible of the introduction of this new technique in clinical practice. Positive professional relations in the focal team and motivation of the patients to have all tests and examinations are crucial for the success of the complex diagnostics and dental treatment. Key words: complex focal diagnostics, thermovision, T-scan, electroacupuncture

LASER DOPPLER ASSESSMENT OF PULP BLOOD FLOW IN YOUNG INDIVIDUALS’ VITAL TEETH Vesela Stefanova, Snejana Tsanova, Neshka Manchorova, Maria Manolova Faculty of Dental Medicine, Medical University - Plovdiv, Bulgaria INTRODUCTION: Thermal and electrical pulp tests for pulp status determining are based on the sensitivity of neural transmission and also patient dependent. The laser Doppler flowmetry is a non invasive method with direct and objective registration of blood circulation, which allows the true vitality of the pulp to be measured. AIM: To measure and assess the pulp blood flow of vital teeth in young individuals. MATERIAL AND METHODS: Monitoring of blood perfusion of vital mandible and maxilla incisors, canines, premolars and molars of young individuals for three minutes by needle probes of moorVMS-LDF2, USA device was carried out. The obtained data were statistically studied by descriptive statistics and correlation analyses using specialized software (moorVMS-PCV3.1 for Vascular Monitor System; SPSS 13.0). Parametrical and nonparametric tests for comparison of means and percentage distribution were used. All means are given with their standard errors. R ESULTS: The mean age of the individuals was 24.2+/-0.22 yrs. The total number of monitored teeth was 50. The mean value of blood perfusion was 20.38 PU+/- 1.68. Mandible teeth showed significantly higher levels (23.38 PU+/- 2.52) compared to maxilla teeth


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(17.62 PU+/- 2.15), p < 0.05. In this study no significant differences of blood flow ranges in men and women, left or right teeth, frontal and distal and teeth types were found (p > 0.05). CONCLUSIONS: The established difference between mandible and maxilla teeth blood perfusion should be considered in laser doppler flowmetry as a diagnostic quantity method for pulp status assessment. Key words: laser doppler flowmetry, pulp vitality

INTERDISCIPLINARY APPROACH IN ORTHODONTIC TREATMENT PLANNING OF IMPACTED CANINE - A CASE REPORT K. Yovcheva1, E. Boyadzhieva2, P. Kanazirska3, N. Kanazirski3 1Department of Orthodontics, 2Department of Surgical Dentistry and Endodontics, 3Department of Radiology, Faculty of Dental Medicine, Medical University - Plovdiv, Bulgaria The case study aims to demonstrate the collaboration between dental specialists in planning an orthodontic treatment of a palatally positioned impacted upper canine. MT, 22 years old, was referred to the Department of Orthodontics with the esthetic complaint of persisted temporary upper canine. To make the orthodontic diagnosis, the following examinations were conducted: facial and intraoral examination, facial and intraoral photographs, dental casts, panoramic radiograph, lateral cephalometric radiograph. The examinations found palatal localization of the impacted tooth 13 and periapical lesions due to inadequate endodontic treatment in 15 and 26. The biometric analysis established unilateral crossbite on the left, retrusion of the maxillary incisors and overbite. To make an adequate comprehensive treatment plan, cone-beam computed tomography and segment parallel radiography of teeth 15 and 26 were conducted. Achieving realistic occlusal relationships in patients with complex dental problems depends not only on the correct treatment plan, but also on the exact sequence of dental procedures. Conducted diagnostic tests and collaboration between various dental specialties formed an interdisciplinary treatment approach. Key words: interdisciplinary approach, treatment plan, diagnostics, impacted canine

FIRST STEPS WITH CAD/CAM: A SINGLE CROWN DESIGN BY 3SHAPE DENTAL SYSTEM Angelina Vlahova, Rada Kazakova, Tania Bozhkova, Viktor Hadjigaev, Stefan Zlatev, Hristo Kisov, Georgi Todorov Department of Prosthetic Dentistry, Faculty of Dental Medicine, Medical University - Plovdiv, Bulgaria INTRODUCTION: Computer aided design / computer aided manufacturing (CAD/CAM) is a technology for prosthetic restorations design using software and fabrication by milling from a suitable material (ceramics, laboratory composite, plastic, metal alloy). The aim of this presentation is to acquaint the audience with the technology for taking an optical impression - with TRIOS (intraoral) scanner, optical working cast - with 3Shape WIELAND Dental D 800 (laboratory) scanner and the design possibilities with 3Shape Dental System software by presenting a clinical case finished with a single ceramic crown.

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MATERIALS AND METHODS: The clinical protocol for taking an optical impression and the laboratory scanning of plaster models were considered. The application possibilities of 3Shape Dental System for construction of various prosthetic restorations were summarized - single crowns and bridges, post-and-core restorations, splints, partial and complete dentures, individual impression trays, implant surgical guides, implant prostheses, orthodontic appliances, temporary restorations, prognostic wax-up. The materials and working tools were considered. The process of a single crown design with 3Shape Dental System was presented. R ESULTS AND DISCUSSION: All steps of the design were described: insertion of direction, outlining preparation junction, providing a cement gap, choice of an optimal shape of the crown, initially positioning and shaping, final shaping, selecting distance between adjacent teeth and antagonists, smoothing and completion. The computer software 3Shape Dental System provides extremely precise and accurate design and fabrication of the prosthetic restoration. CONCLUSION: 3Shape Dental System is an optimal solution for modern and efficient clinical and laboratory practice. Key words: CAD/CAM, design, a single crown

PROBLEMS ASSOCIATED WITH THE USE OF THE PENDULUM APPLIANCE IN MIXED DENTITION Greta Yordanova Department of Orthodontics, Faculty of Dental Medicine, Medical University - Sofia, Bulgaria The Pendulum appliance can be used in treatment of patients with mixed and permanent dentition if there is a stable premolar and molar anchorage zone. The clinical decision which teeth are going to be the anchorage, in case of a presence of primary molars depends on the level of their root resorption and the location of the germs of the permanent premolars and canines. The distalization of the first molars is always accompanied with a loss of anchorage. When the anchorage teeth are the primary molars, their medicalization can lead to interchange with their successor teeth and disturbance their natural root resorption. If the primary molars move mesially with their successors, this can lead to abnormal eruption of the canines. These problems can be overcome by proper prognosis of the dental developing, analyzing X-rays, clinical examination and designing the appliance. We suggest actual clinical decisions by modifying the Pendulum which we use. The changes of the appliance we suggest include enlargement or horseshoe-like modifying of the palatal Nance button, preforming of the occlusal stops or including the permanent molars to the anchorage, in case of unilateral distalization and other clinical tricks. Most often the use of the appliance in mixed dentition is associated with impacted premolars or canines, and lack of space for their eruption in the dental arch. We analyzed our decisions in each of the presented cases. The good results and comfort of the patient during treatment with this appliance are the grounds for our recommendation to use it in the orthodontic procedures. Key words: pendulum, mixed dentition, anchorage


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Posters


POSTERS

Biomedicine BM 1.

SPERM ANALYSIS OF MEN WITH A HISTORY OF INFERTILITY

Mina Pencheva1, Petia Darmonska1, Aleksandar Aleksandrov2, Denis Milkov1 1Department of Anatomy, Histology and Embryology, Medical University - Plovdiv, 2Medical Center Salmanida, Plovdiv, Bulgaria Infertility affects about 14% of couples of childbearing age. The first step to elucidate the etiology of infertility is through semen analysis. The evaluation of the quantitative and qualitative indicators of ejaculate was made according to the criteria of the World Health Organization. The aim of our study was to perform a quantitative and qualitative analysis of sperm indicators of men of Bulgarian origin with an anamnesis of infertility, compare them to the WHO criteria and seek reasons for their infertility. The study included 74 men (mean age 34.9 yrs). The quantitative and qualitative research was performed using an optical microscope. The data was processed using SPSS 17.0. Our study found that the average ejaculate volume is 2.89 ml, and according to WHO criteria, the minimum quantity of ejaculate should be 2 ml or more. The concentration of spermatozoids in 1 ml ejaculate was 17.72 million, which is below the WHO minimum concentration of 20 million/ml. Evaluating the sperm motility, we found that 41.81% were akinetic, 13.95% - hypokinetic and only 36.36% were normokinetic. WHO recommends a minimum of 50% of the sperm to be progressively motile. After a detailed assessment of the spermatozoid morphology we found defects: in the head - 6.65%, in the neck - 3.36%, in the body - 3.19% and in the tail - 6.07%. The men in our study suffer from infertility with a multifactor etiology due to a decreased sperm concentration and low motility. Key words: men infertility, semen analysis

BM 2.

ANGIOTENSIN II RECEPTOR BLOCKADE AND ITS IMPORTANCE FOR

INTESTINAL MOTILITY Petia Hadzhibozheva1, A. Tolekova, G. Ilieva1, R. Kalfin2, Ts. Georgiev1 1Department of Physiology, Pathophysiology and Pharmacology, Medical Faculty, Trakia University; 2Institute of Neurobiology, Bulgarian Academy of Sciences, Sofia, Bulgaria The localization and role of the different components of the renin-angiotensin system in the gastrointestinal tract are not sufficiently studied and defined. This study aims at investigating the presence and role of the two basic types of angiotensin II (AngII) receptors for the intestinal motility by application of losartan (AngII receptors type 1 blocker) and PD123319 (AngII receptors type 2 blocker). Smooth muscle preparations from different segments of Wistar rats’ intestinal tract were prepared and recording of the contractile activity after administration of AngII alone and in combination with the receptor blockers was performed. The analysis of power and kinetic parameters was performed with KORELIA-Dynamics Software. The speed of ileal contraction and relaxation after administration of PD123319 was significantly higher than those obtained after the application of AngII and Losartan+AngII. The speed parameters of colonic contraction and relaxation induced by AngII and PD123319+AngII did not display significant differences. The presence of losartan markedly delayed the speed of the colonic contractile process, and completely abolished AngII-induced jejunal contraction. The application of Losartan+AngII on rectum preparations revealed significantly rapid speed of relaxation.

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The obtained results demonstrate: 1) the presence of both AngII receptors types in all segments of the intestinal tract; 2) The specific importance for the change of power and kinetic parameters of the two phases of contractile activity in the different intestinal segments. Acknowledgments: This work is supported by Grant 22/2014 from Trakia University, Stara Zagora, Bulgaria. Key words: angiotensin II receptors, contraction, kinetic parameters

BM 3.

EFFECT OF NITRATES, THIOCYANATES AND SELENIUM ON IRON AND

IODINE STATUS IN BREASTFEEDING MOTHERS Anelia Bivolarska1, Ana Maneva1, Penka Gatseva2 1Department of Chemistry and Biochemistry, Faculty of Pharmacy, 2Department of Hygiene and Ecomedicine, Faculty of Public Health, Medical University - Plovdiv, Bulgaria The study included 41 new mothers (mean age 26.4 ± 5.9 yrs) from the town of Asenovgrad in whom we studied the effect of thiocyanates, nitrates and selenium on their iodine and iron status parameters. Iodine in the urine was determined using Sandell-Kolthoff reaction, thiocyanates – using the interaction of these ions with an acidic solution of KmnO, the nitrate levels in blood serum – using the colorimetric method, serum selenium – using electro-atomic absorption spectrophotometry. ELISA was used to determine FT4, TSH, SF, sTfR, and hematology analyzer to measure hemoglobin levels. With respect to the iodine status we found a statistically significant negative correlation between the levels of iodine and thiocyanates in the urine (R = -0.717, p 0.05). The body mass of the group subjected to combined nonpharmacological treatment was reduced (p < 0.05). The three nonpharmacological programs for treatment of the MetS such as dietary regimen, increased physical activity and combined program with diet and training resulted in decrease of the BMI of the animals. The physical training itself could not provide permanent reduction of the body mass. Key words: metabolic syndrome, diet, training; rats

BM 9.

ENDURANCE TRAINING AFFECTS THE EXPRESSION OF BRAIN-

DERIVED NEUROTROPHIC FACTOR IN SATELLITE CELLS IN SOLEUS MUSCLE OF RATS Katerina Georgieva1, Slavi Delchev2, Yveta Koeva2, F. Gerginska2, M. Shishmanowa-Doseva3 1Department of Physiology, 2Department of Anatomy, Histology and Embryology, Faculty of Medicine; 3Department of Pharmacology and Drug Toxicology, Faculty of Pharmacy, Medical University - Plovdiv, Bulgaria Neurothrophic factors are expressed not only by neurons, but also by muscle cells. Experimental data have shown that prolonged exercise training leads to increased expression of some neurotrophins in the rat skeletal muscles. It is not known whether this expression is specific to the muscle fibers only, or satellite cells are also involved in these adaptational processes of skeletal muscles to the endurance training. The aim of the study is to investigate the expression of brain-derived neurotrophic factor (BDNF) in soleus muscle of endurance trained rats. Male Wistar rats were randomly divided into two groups (n = 6 each) – controls and trained. Trained rats were exercised on a treadmill with submaximal loading (70-75%VO2max) for 8 weeks. At the end of the trial soleus muscles (85% type I myofibers) of all rats were taken. On paraffin sections immunohistochemical reactions for BDNF were applied. The intensity of the reaction was measured by specialized software. We found BDNF immunoreactivity in muscle fibers and satellite cells of soleus of the trained rats. The expression of the neurothrophic factor was mainly in the satellite cells and it was stronger than that of the controls. Some satellite cells contained more than one nucleus.

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Our results demonstrate that the increased BDNF expression in the satellite cells of soleus is one of the adaptational processes of skeletal muscles to endurance training. Key words: submaximal training, BDNF, muscles

BM 10.

HIGH LEAD CONCENTRATIONS IN BLOOD, CHANGES IN THE MOTILITY

OF THE GASTROINTESTINAL TRACT, AND PROSTAGLANDIN SYNTHESIS Vladimir Sirakov1, Natalia Prissadova2, Atanas Krastev2, H. Badakov2 1Department of Imaging Diagnostics, 2Department of Medical Physics and Biophysics, Medical University - Plovdiv, Bulgaria Lead is a toxic element found naturally in the environment. If lead concentration in the human body exceeds certain level, it can cause a wide range of health issues, including problems of the gastrointestinal tract (GIT), such as abdominal discomfort, pain, and vomiting–effects generally associated with motility disorders of the smooth muscle (SM) tissue. The relatively few investigations of this issue, conducted with isolated SM tissues, as well as the existing clinical data related to the motility of the GIT, make this study appropriate. Contrast X-ray investigation of patients with high levels of lead in the blood, show specific changes in the images of organs of the GIT. Elevated tonus, and strong segmenting peristaltic activity have been observed. A higher incidence of gastroduodenitis, colitis, and inflammation processes of the appendix have also been registered. In vitro investigation with Pb2+ of concentration range 1.10-7 – 1.10-5mol/l shows contractions of SM tissue in concentration-dependant manner. Pb2+induced contractions are not influenced by TTX, but are reduced significantly by the blockers of prostaglandin synthesis, indomethacin and acetysal. Analysis of the results leads to the conclusion that the effects of lead on the motility of GIT are related to the activation of prostaglandin synthesis. Key words: Pb2+, smooth muscle, prostaglandin synthesis

BM 11.

INFLUENCE OF BREASTFEEDING ON THE PARAMETERS OF IRON

METABOLISM IN CHILDREN WITH IRON DEFICIENCY ANAEMIA UP TO TWO YEARS OF AGE Tsvetelina Petkova-Marinova, Vania Nedkova, Boriana Ruseva 1Department of Physiology and Pathological Physiology, Medical University - Pleven; 2Department of Pediatrics, Medical University - Pleven, Bulgaria Anaemia is a serious problem now because of its high prevalence and adverse health consequences for the affected groups. Between the ages of 6 months and 2 years, there is a physiological susceptibility to iron deficiency states due to intensive physical development and depleted foetal stores of iron. The aim of the present study is to evaluate the influence of breastfeeding on the parameters of iron metabolism in children with iron deficiency anaemia (IDA) up to two years of age. 172 children aged from 2 to 24 months with IDA were examined. IDA was diagnosed according to criteria recommended by the World Health Organization: haemoglobin concentration below 110 g/l, MCV below the respective age-related reference values, serum iron (SFe) - below 8.95 μmol/l, and transferrin saturation (SatTf) - below 16%. Children were divided into two age groups: group G1 (from 0 to 12 mo) and group G2 (from 13 to 24 mos). Parameters: SFe, total iron-binding capacity, SatTf, haemoglobin, red blood cell count, Folia Medica 2015; 57; Suppl. 1 © 2015 Medical University, Plovdiv, Bulgaria

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erythrocyte indices - MCV, MCH, MCHC, and RDW, were compared between breastfed and non-breastfed children in each group. Data were processed by STATGRAPHICS Centurion XVI. Statistical analysis was performed by Student’s t-test. Statistically significant differences for SFe, SatTf, haemoglobin, MCH, MCHC, and RDW were observed between breastfed and non-breastfed children in both age groups. We found that in children up to 2 years of age, breastfeeding is a factor which does not prevent development of IDA, but improves parameters of iron metabolism. Key words: breastfeeding, iron, anaemia

BM 12.

MORPHOLOGICAL CHANGES IN RAT LEYDIG CELLS AND THEIR

CORRELATION WITH THE ACTIVITY OF KEY STEROIDOGENIC ENZYMES DURING AGING Darina Barburska, Yveta Koeva, M. Koutsogiannis Department of Anatomy, Histology and Embryology, Medical University - Plovdiv, Bulgaria Leydig cells (LC) are the main source of male sex hormones testosterone. It is now well established that testosterone reduced with aging both in human and in rodents, suggesting that there must be deficit in the steroidogenic capacity of aging Leydig cells. Experimental models of aging in rodents are suitable for clarifying the changes in LC that determine their reduced steroidogenic capacity. By using light microscopic observations our work demonstrate that the aging in the rat testis is accompanied by morphological changes – reduction of the diameter of the seminiferous tubules and atrophic changes of the LC. These observations reflect the ultrastructural changes of the cytoplasmic organelles – reduction in the smooth endoplasmic reticulum and apoptotic events in the nuclei of LC. The aim of study was to find a correlation between the morphological changes in rat LC and the activity of steroidogenic enzymes 3β HSD and NADH2-cytochrome-c reductase in aging. In the study, we used rat testis at 7, 18 and 24 months. The histological analysis revealed atrophic changes of the LC, reduction in their number, especially in 24-month-old animals. Enzymohistochemical analysis revealed decrease of the activity of steroidogenic enzymes by 18-month-old and 24-month-old rats compared with the control animals (7 months old). The observed decrease in enzymohistochemical activity of 3β -HSD and NADH2-cytochrome -c -reductase in the LC during aging correlated with the observed morphological changes in testis and reveals the correlation between suppressed activity of steroidogenic enzymes in LC and agerelated decreased steroidogenesis. Key words: Leydig cells, aging, steroidogenesis

BM 13.

SALIVA AND NITRIC OXIDE

M. Hadjieva, M. Koutsogiannis, Julia Nikolova Department of Physiology, Faculty of Medicine, Medical University - Plovdiv, Bulgaria Saliva is a multi-constituent fluid that could be considered the mirror of the body. Its components could be explored as diagnostic markers of different systemic and local changes in homeostasis. Nowadays an interesting direction of investigation is saliva nitric oxide (NO) monitoring. NO, identified as endothelium derived relaxing factor (EDRF) has strong reactivity with oxygen, superoxide and iron-containing compounds. It is believed to play not only a major role in blood flow and blood pressure regulation, but to play additional roles - synthesized by activated macrophages influences cells’ ability to kill bacteria, viruses and tumor cells being in relationship with

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cytokines, interleukins and TNF. In addition, it has damaging effects against cellular proteins, DNA and lipids leading to cell death and tissue injury. Its very short half life (1 to 10s) makes the assessment technically difficult to quantify in solution and NO production is attempted to be measured by the levels of nitrate and nitrite. NO registration is carried out in blood and urine. It is recently reported that freshly released human saliva contains measurable and even in oral mucosal diseases high levels of NO. Nitrate-nitrite-NO way is an alternative for NO formation. It is noted that there is a correlation between blood and saliva nitrite levels. The ability saliva to produce NO is discussed too. In diets, rich in leaf vegetables, it is noted that plasma and saliva nitrite are increased. The monitoring of saliva NO would be reliable noninvasive biomarker of systemic and oral disorders. Key words: saliva, nitric oxide

BM 14.

VITAMIN K2 IMPROVES ANXIETY AND DEPRESSION BUT NOT COGNITION

IN RATS WITH METABOLIC SYNDROME: INTERFERENCE WITH BLOOD GLUCOSE? Silvia Gancheva, Maria Zhelyazkova - Savova Department of Preclinical and Clinical Pharmacology, Medical University - Varna, Bulgaria In the present study we examined the metabolic and behavioral effects of vitamin K2 in rats with metabolic syndrome and looked for relationships between them. Male Wistar rats were divided in four groups: control (on a regular rat chow diet), metabolic syndrome (MS group, on a high fat high fructose diet), control group treated with vitamin K2 and MS group treated with vitamin K2. Vitamin K2 was administered orally throughout the experiment (11 weeks). At the end of the study fasting blood glucose was taken and behavioral tests were performed. The social interaction test was used to measure the level of anxiety. The Porsolt test was used to assess depression. The object recognition test was performed to estimate the effect on memory. The correlations between blood glucose and behavioral performance were analyzed. The rats from the MS group had elevated blood glucose. They displayed anxiety, depression and memory deficit. Anxiety (time of social interaction) and memory (index of recognition), but not depression (immobility time), were negatively correlated with blood glucose in the untreated rats. Vitamin K2 normalized blood glucose, exerted anxiolytic and antidepressant-like effects, but did not improve memory. When vitamin K2-treated rats were included in the analysis, the correlation of blood glucose with time of social interaction was preserved, but the one with the index of recognition was lost. It is possible that the anxiolytic effect of vitamin K2 in this setting be at least partly due to its effects on blood glucose, while the antidepressant effect is glucose-independent. Key words: vitamin K2, metabolic syndrome, rats, anxiety, depression, correlation with blood glucose

BM 15.

CEPHALIC AND MORPHOLOGICAL FACIAL INDEXES IN YOUNG GROWN-

UP BULGARIAN GIRLS Denis Chervanskiy Department of Anatomy, Histology and Embryology, Medical University - Plovdiv, Bulgaria We made a cephalometric characteristic of young grown-up Bulgarian girls to find correlations between some anthropological indexes. The study included 35 girls (mean age 19.00 yrs), in whom we measured the maximum head length (g-op), the maximum cranial breadth (eu-eu), the


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maximum facial breadth (zy-zy) and the morphological height of face (n-gn). We used the classical methods of Martin-Saller (1957), using sliding and spreading calipers accurate to 0.1 cm. Statistical analysis was performed using SPSS 17.0. The cephalic and morphological facial indexes were calculated by these measurements. The distribution of the subjects by the cephalic index was as follows: very long heads had 8.57% (hyperdolichocephalic), long skulls - 40% (dolichocephalic), medium skulls (mesocephalic) - 42.85%, 11 9% short skulls (hyperbrachycephalic) and 2.85% were determined to have very short skulls (ultrabrachycephalic). The distribution by facial index was: very short faces (hyper euryprosopic) - 2.85%, short faces (euryprosopic) - 8.57%, medium faces (mesoprosopic) -17.4%, long faces (leptoprosopic) - 22.85% and 48.57% - very long faces (hyper leptoprosopic). In conclusion young grown-up Bulgarian girls are characterized by long and medium skulls, while in their facial morphology the very long and long faces prevail. Key words: morphological face index, cephalic index

BM 16.

COMPARATIVE EVALUATION OF SOME PARAMETERS OF IRON

HOMEOSTASIS IN DIABETES AND RHEUMATOID ARTHRITIS Katia Stefanova1, G. Delcheva1, Ana Maneva1, Anastas Batalov2, Mariela Geneva-Popova2, R. Karalilova2, Dimitar Iliev3, Maria Orbetzova3 1Department of Chemistry and Biochemistry, Faculty of Pharmacy, Medical University - Plovdiv; 2Department of Propaedeutics of Internal Diseases, Medical University - Plovdiv; 3Second Department of Internal Medicine, Division of Endocrinology, Medical University - Plovdiv, Bulgaria Diabetes and rheumatoid arthritis (RA) are accompanied with a chronic inflammatory process. Hyperglycemia results in complications which activate cellular signals, causing the expression of inflammatory mediators. Hyposideremia is an adaptive response to the chronic inflammatory processes. It is associated with pathological deviation of serum iron in the ferritin depot and rise of iron deficiency. The latter has increased tissue expression of transferrin receptors, as well as increased serum concentration of soluble transferrin receptor (sTfR). The aim of the present study was to investigate and compare serum levels of some parameters of iron homeostasis and inflammation (serum ferritin, serum transferrin receptor, lactoferrin and CRP) in patients with diabetes (n = 81) and RA (n = 114). The serum levels of ferritin and lactoferrin were higher in diabetes compared with RA and the differences were statistically significant (p = 0.007 and p < 0.0001, respectively). We think that these results show preserved adaptive abilities for control of iron homeostasis in diabetes where unlike RA there is a negative significant correlation between sTfR and ferritin. The preserved mechanisms of regulation in diabetes are confirmed also by the positive correlations between ferritin and HbA1c (r = 0.265, p = 0.018) and between ferritin and CRP (r = 0.285, p = 0.025). The lower lactoferrin level in RA compared with diabetes probably concerns its continuous secretion from the specific granules of the neutrophils during phagocytosis which depletes the adaptive abilities of the cell for protein biosynthesis in patients with RA. Key words: ferritin, STfR, diabetes, rheumatoid arthritis

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BM 17.

THE TUMOR NECROSIS FACTOR ALPHA – A PROBABLE PROGNOSTIC

TARGET FOR CARDIOVASCULAR DISEASES Velichka Popova1, Anastas Batalov1, Zaprin Vazhev2, Yovko Ronchev4, Liudmil Peytchev5 1Department of Rheumatology, Medical University – Plovdiv, Kaspela University Hospital, 2Department of Cardiac Surgery, Medical University - Plovdiv, Kaspela University Hospital, 3 Kaspela University Hospital, Department of Pharmacology and Drug Pharmacology, Faculty of Pharmacy, Medical University - Plovdiv, Bulgaria INTRODUCTION: Cardiovascular diseases such as coronary heart disease, congestive heart failure, etc. interact with members of the tumor necrosis factor alpha (TNF) superfamily and their receptors, which results in several biological effects, that may be involved in the pathogenesis of cardiovascular diseases. TNF-α is inducer of apoptosis, regulates various cellular functions such as proliferation, migration, differentiation, cell death, inflammation, and angiogenesis. MATERIALS AND METHODS: We studied the serum levels of TNF-α in 20 patients with inflammatory joint diseases with high cardiovascular risk assessed by Reynolds risk score scale and carotid intima-media complex (CIMT). Control groups of patients - group 1 with high cardiovascular risk without inflammatory joint disease and group 2 - healthy controls. TNF-α was analyzed using ELISA Biomasters kits. The study of the CIMT was performed with a sonographic machine Essaote. R ESULTS: We found high serum levels of TNF-α in the group of inflammatory joint disease – the levels of TNF-α by (5.15 ± 2.6 pg/ml) compared to the group without the inflammatory joint diseases - (4.87 ± 2.6 pg/ml) and the serum levels in healthy controls are reference values (3.15 ± 2.3 pg/ml; p < 0,05). CIMT analysis indicates persistence over 1 mm in the groups at high cardiovascular risk. CONCLUSION: High serum levels of TNF-α correlate with disease activity, vascular risk in highrisk groups. Perhaps serum TNF-α could serve as a prognostic factor for assessing the high-risk groups. Key words: tumor necrosis factor alpha (TNF)

BM 18. EVALUATION

OF ANTIMICROBIAL RESISTANCE OF SALMONELLA AND SHI-

GELLA ISOLATES IN ST. GEORGE UNIVERSITY HOSPITAL - PLOVDIV, BULGARIA Mihail Petrov1, Atanaska Petrova1, Irina Stanimirova1, Marina Mircheva1, Lalka Koycheva1, Raina Velcheva2, Mariana Stoycheva2, Marianna Murdjeva1 1Department of Microbiology and Immunology, Faculty of Pharmacy, 2Department of Infectious Diseases and Parasitology, Medical Faculty, Medical University - Plovdiv, St. George University Hospital - Plovdiv, Bulgaria AIM: To study antimicrobial resistance of Salmonella and Shigella as causative agents of acute gastroenteritis to the most common antibiotics used for treatment at the St. George University Hospital, Plovdiv between 2009 and 2013. MATERIALS AND METHODS: 290 strains isolated in this period were tested in vitro for resistance to 15 antimicrobial agents including azithromycin. The presence of Extended-spectrum betalactamases (ESBLs) was demonstrated by a variety of specialized tests. Minimum inhibitory concentrations against the most common antibiotics were determined on 56 Salmonella and Shigella strains, isolated in 2013 from children under 8 years of age. A collection of 28 strains from both genera, submitted by the National Reference Laboratory for intestinal infections at the NCIPD, Sofia, were also tested for antibiotic susceptibility and production of ESBLs. A PCR detection of bla-CTX-M, bla-SHV and bla-TEM genes, responsissble for resistance to beta-lactams,


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was performed in isolates, phenotypically demonstrated as ESBLs producers. R ESULTS: Two hospital isolates - Salmonella Livingstone and Shigella flexneri, and 4 out of 28, submitted by NCIPD, Sofia, were phenotypically proved as ESBLs producers. The presence of bla-CTX–M gene was detected in all 6 of them, bla - SHV gene - only in Salmonella Livingstone strain from Plovdiv and bla-TEM gene was established in two Salmonella isolates from Sofia and one (Salmonella Livingstone, again) from Plovdiv. An interesting fact is the coproduction of the three tested ESBL enzymes in Salmonella Livingstone strain. CONCLUSION: Salmonella and Shigella strains from in-patients with acute gastroenteritis at St. George University Hospital, Plovdiv demonstrate good susceptibility to the most common antibiotic agents used for treatment. Key words: Salmonella, Shigella, enterocolitis, ESBLs, antimicrobial resistance, PCR

BM 19.

GAMMA-RADIATION INDUCED DNA DAMAGE ATTENUATION BY

NANO-SILYMARIN: AN IN VITRO APPROACH Manish Adhikari1, Radjesh Arora1, Yana Karamalakova2, Rach Kumar1, Veselin Ivanov2, Antoaneta Zheleva2, Veselina Gadjeva2, Stoiko Stoev3 1Radiation Biotechnology Group, Institute of Nuclear Medicine and Allied Sciences, Brig. S. K. Mazumdarmarg, Defence Research and Development Organization, Delhi-110054, India; 2Department of Chemistry and Biochemistry, Medical Faculty Trakia University, Stara Zagora, Bulgaria Radiation exposure caused formation of free radicals through ionizing reactions that are proficient of destroying healthy tissues. Hence we must continue to improve approaches to protect normal tissues from radiation-induced damage. Nanoparticles as a radioprotector are a vanguard development in scientific interest regarding the protection of normal cells and tissues from radiation. Nanoemulsions are submicron sized emulsion that is under extensive investigation as drug carriers for improving the delivery of therapeutic agents. Silymarin because of its low and poor oral bioavailability, was formulated as a nanoemulsion to increase its solubility (and so its oral bioavailability) as well as therapeutic activity. Results showed the protection of DNA by nanosilymarin against γ-radiation-induced toxicity. DNA damage revealed by plasmid relaxation in which maximum protection seen at 10μg/ml in which supercoiled DNA retention was >70% as compared to radiation group. Micronuclei count showed reduction in micronuclei frequency at different time-points in Human embryonic kidney (HEK) cell lines. When 10μg/ml of nanosilymarin was given 0.5h prior to 2Gy γ-radiation, decrease in MN level with maximum reduction (61%) at 72h time interval was observed. Microscopic analysis performed using Differential interference contrast (DIC) microscopy images in which pre-treated nano-silymarin irradiated group showed less morphological alterations as compared to radiation alone group indicating its protective potential against radiation induced morphological damage. Overall, nano-silymarin showed radioprotection by protecting DNA damage during radiation exposure which will encourage the development of innovative and new approaches to radiation protection, using nanotechnology. Key words: radiation, nano-silymarin,radioprotection, nanotechnology

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BM 20.

GEOGRAPHICAL DISTRIBUTION AND EPIDEMIOLOGICAL FEATURES OF

VISCERAL LEISHMANIASIS IN BULGARIA FOR THE PERIOD 2010 - 2014 Rumen Harizanov1, I. Kaftandjiev1, I. Rainova1, D. Yordanova1, N. Tsvetkova1, I. Mineva1, E. Kaneva1, T. Kostova2 1National Center of Infectious and Parasitic Diseases, Department of Parasitology and Tropical Medicine, 2National Diagnostic Veterinary Research Institute, Sofia, Bulgaria Visceral leishmaniasis (VL) caused by Leishmania infantum is an endemic illness in Bulgaria. Cases in humans are registered almost every year. The aim of this study was to determine some epidemiological characteristics of VL, and some specificities in the geographical distribution of the disease in this country for the last 5 years (2010-2014). Thirty-six cases of human disease were registered including 34 autochthonous and 2 imported cases. Out of these 36, 15 (41.7%) were children and 21 (58.3%) adults. Over 60% of cases (n = 22) were registered in Blagoevgrad region, mainly in Petrich city and municipality (n = 20). The average annual incidence in the region is over 25 times higher than the average annual incidence in the country for the studied period. A pilot seroepidemiological study of 51 healthy subjects from Petrich was carried out and found 3.9% seroprevalence. Clinical samples from 12 dogs from the region were studied with Real Time PCR and yielded one positive result. In the last 5 years, the region of Petrich municipality may be classified as permanently active endemic focus of VL, which calls for a large-scale research activities, both in terms of people, but also in terms of possible reservoir sources and vectors of the disease Key words: visceral leishmaniasis, epidemiology, geographical distribution

BM 21.

MOLECULAR EPIDEMIOLOGICAL ANALYSIS OF HIV-1 NON B STRAINS IN

BULGARIA (PRELIMINARY ANALYSIS) Reneta Dimitrova1, Anna Gancheva1, Asia Kostadinova1, Danail Beshkov1, Vania Dafova2, Ivailo Elenkov3, Mariana Stoycheva4, Ivailo Alexiev1 1National Center of Infectious and Parasitic Diseases, 2University of Sofia “St. Kl. Ohridski”, 3Hospital for Infectious and Parasitic Diseases, Sofia, 4Department of Infectious Diseases, Faculty of Medicine, Medical University - Plovdiv, Bulgaria OBJECTIVES: Subtype B is the most prevalent in North America, West Europe and in most Balkan countries. From 1986 to 2014 in Bulgaria 2019 HIV/AIDS cases were registered in three main transmission groups (Hetero, MSM and IDUs). The aim of this study was to analyze the diversity of non-B subtypes in different transmission groups of HIV-1 epidemic in Bulgaria. MARERIALS AND METHODS: 681/2019 (33.7%) pol sequences were generated with TruGene DNA Sequencing System during the patients follow-up. Preliminary subtype analysis was evaluated using REGA3 and COMET1.0. Phylogenetic analysis was conducted by Maximum Likelihood (ML) method implemented in FastTree2 program. Recombinations of the HIV-1 genome were analyzed with Bootscan analysis using SimPlot 3.5.1. R ESULTS: Phylogenetic analysis reviewed that subtype B is the most common HIV-1 subtype in Bulgaria (38.5%), followed by two major recombinant forms CRF01_AE (28.6%) and CRF02_ AG (23,4%). We also found a bouquet of more than 15 different subtypes, complex and unique recombinant forms. In addition, subtype B was more prevalent among MSM, while CRF01_AE and CRF02_AG were common among IDUs. Furthermore, HIV-1 subtype B and CRF01_AE was predominant in Sofia, while CRF02_AG in Plovdiv. Several clusters were found on the phylogenetic tree representing rapid development of local HIV-1 sub-epidemics.


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CONCLUSIONS: We found a wide HIV-1 diversity with unequal distribution among different transmission groups of the population. Our findings indicate that providing of detailed molecular epidemiological surveillance of HIV-1 epidemic in Bulgaria is of importance to better control the epidemic in the country. Key words: HIV-1, subtype, Bulgaria

BM 22.

MOLECULAR-EPIDEMIOLOGICAL ANALYSIS OF HIV-1 STRAINS INTRO-

DUCED FROM ABROAD (PRELIMINARY ANALYSIS) Asia Kostadinova1, Anna Gancheva1, Reneta Dimitrova1, Danail Beshkov1, Vania Dafova2, Ivailo Elenkov3, Mariana Stoycheva4, Ivailo Alexiev1 1National Center of Infectious and Parasitic Diseases; 2University of Sofia “St. Kl. Ohridski”; 3Hospital for Infectious and Parasitic Diseases, Sofia; 4Department of Infectious Diseases, Faculty of Medicine, Medical University - Plovdiv, Bulgaria OBJECTIVES: From 1986 to 2014, 343 (16 %) migrants with HIV/AIDS were diagnosed in Bulgaria out of a total of 2019 cases. The aim of this study was to investigate viral characteristics introduced in the country by genotyping and phylogenetic analysis. MATERIALS AND METHODS: In this study we analyzed 68 (19.8%) out of 343 migrants, 258 (73%) of which were Bulgarian citizens infected abroad and 85 (25%) foreigners diagnosed in Bulgaria. HIV-1 pol sequences were obtained using TruGene DNA Sequencing System. Preliminary HIV-1 subtypes were analyzed with REGA3 and COMET1.0. The phylogenetic analysis was performed with ML method using FastTree2 program. R ESULTS: We found wide HIV-1 genetic diversity in migrants diagnosed in Bulgaria. Phylogenetic analysis showed that most prevalent subtype among this population is subtype B (44.1%) followed by unique recombinant forms (25%), subtype C (8.8%), CRF02_AG (4.4%), CRF01_AE (2.9%), CRF14_BG (2.9%) and 1.5 % each: CRF05_DF, CRF12_BF, CRF36_cpx, subtype A1, recombinant B/D, subtype F1, subtype G and recombinant J/C. Our epidemiological data showed that migrants with HIV-1 registered in Bulgaria originated from at least 30 countries around the world, mostly from Nigeria, Cameroon and Germany. CONCLUSIONS: Significant number of HIV-1 cases in Bulgaria is caused by viral strains introduced from abroad by migrants. Our analysis showed that introduced viruses have great genetic diversity and in combination with a significant number of migrants defines these populations as of importance to Bulgarian HIV-1 epidemic. Further investigation of this target population would be of benefit to control the epidemic. Key words: HIV-1, migrants, subtype, Bulgaria

POLYMERASE-CHAIN REACTION ANALYSIS OF CARBAPENEM RESISTANCE IN GRAM NEGATIVE NON-FERMENTING ISOLATES FROM ST. GEORGE UNIVERSITY HOSPITAL IN PLOVDIV BM 23.

Atanaska Petrova1, Irina Stanimirova1, Marina Mircheva2, Mihail Petrov1, V. Kardjeva3, Marianna Mirdjeva1 1Department of Microbiology and Immunology, Faculty of Pharmacy, Medical University - Plovdiv; 2Laboratory of Microbiology , St. George University Hospital - Plovdiv, 3Aquachim JSC, Sofia, Bulgaria BACKGROUND: BlaOxa23, Oxa58 and the hyperexpression of Oxa51 enzymes due to Isaba1 insertion sequence are the leading cause of carbapenem resistance in Acinetobacter baumannii, a non-fermenting Gram negative bacterium. The main factor for carbapenem resistance in another harmful Gram negative non-fermenter - Pseudomonas aeruginosa is the loss of OprD transmem-

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brane protein whereas BlaVIM and blaIMP metaloenzymes’ production has a secondary role. The aim of this study was to determine the mechanisms of resistance to carbapenems in clinically significant isolates of A.baumannii and P.aeruginosa from the St. George University Hospital in Plovdiv by screening the presence of coding genes and carbapenemase production. MATERIALS AND METHODS: Bacterial DNA was extracted from 25 A.baumannii and 23 P. aeruginosa clinical isolates from in-patients, admitted to the hospital between 2010 and 2012. The amplification was performed using RT-PCR system 7300 (Applied-Biosystem) with subsequent gel-electrophoresis. R ESULTS: Oxa23 genes were established in all A.baumannii isolates. All of them were negative for the presence of Oxa58. All P. aeruginosa were negative for blaVIM genes. A. baumannii demonstrated phenotype of carbapenemase production by modified Hodge test, whereas P. aeruginosa did not show this phenomenon. The lack of metaloenzymes was affirmed by negative combined E-test with imipenem/imipenem+EDTA. CONCLUSIONS: The leading cause of carbapenem resistance in A. baumannii from our hospital is the presence of Oxa23 genes and carbapenemase production whereas in P. aeruginosa we consider the lost of transmembrane OprD protein and the efflux pumps’ hyperexpression as main mechanisms. Further screening of other carbapenemases with subsequent sequencing is planning. Key words: PCR, non-fermenting bacteria, carbapenem resistance

BM 24.

PRIMARY CULTURES FROM TUMOR-BEARING ANIMALS AS MODEL

SYSTEMS IN EXPERIMENTAL ONCOPHARMACOLOGY Radostina Alexandrova1, Abdulkadir Abudalleh1, Lora Dyakova2, Dimitar Ivanov1, Delka Salkova1, Ivailo Dankov3, Orlin Alexandrov4, Luminica Patron5 1Institute of Experimental Morphology, Pathology and Anthropology with Museum, Bulgarian Academy of Sciences, Sofia, Bulgaria; 2Institute of Neurobiology, Bulgarian Academy of Sciences; 3Faculty of Medicine, St. Kl. Ohridski Sofia University; 4Health Service in Gorna Malina, Bulgaria; 5Institute of Physical Chemistry Ilie Murgulescu, Timisoara, Romania Demand for new, highly efficient and biocompatible antitumor agents requires the development and application of appropriate model systems to assess their biological activity and potential toxicity. The aim of this study was to evaluate the influence of metal complexes with bile acids and mixed ligands on viability and proliferation of cultured cancer and healthy cells from one the same tumor-bearing animals. Ascitic tumor of Ehrlich in mouse and hepatoma of Zajdela in rat were used as experimental models. Primary cultures were obtained from ascitic and solid forms of the tumors as well as from bone marrow cells, peritoneal macrophages and spleen lymphocytes. The investigations were performed by thiazolyl blue tetrazolium bromide (MTT) test, neutral red uptake cytotoxicity assay and trypan blue dye exclusion technique. The results obtained demonstrate time- and concentration-dependent cytotoxic/cytostatic activity of the compounds examined. Bone marrow cells were found to be highly sensitive to the cytotoxic effects of metal complexes. Such comparative studies on primary cultures from tumors in laboratory animals and non-tumor cells (bone marrow cells, lymphocytes, macrophages) from the same tumor-bearing animals deserve special attention because this model system largely reflects the situation in chemotherapy patients. Acknowledgement: Supported by Grant DFNI - Б02/30 from 12.12.2014, Bulgaria, and a bilateral project between Bulgarian Academy of Sciences and Romanian Academy. Key words: primary cell culture, antitumor activity Folia Medica 2015; 57; Suppl. 1 © 2015 Medical University, Plovdiv, Bulgaria

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BM 25.

SEROLOGICAL AND MOLECULAR BIOLOGICAL STUDIES OF PARVOVIRUS

B19, COXSACKIE B VIRUSES AND ADENOVIRUSES AS POTENTIAL CARDIOTROPIC VIRUSES IN BULGARIA Stefka Ivanova1, Svetla Georgieva2, Asia Stoyanova3, Irina Georgieva3, Liubomira Nikolaeva-Glomb3, Zafira Mihneva1, Nelly Korsun2 1National Reference Laboratory of Measles, Mumps, Rubella, National Center of Infectious and Parasitic Diseases (NCIPD), 2National Reference Laboratory of Influenza and Acute Respiratory Diseases, NCIPD, 3National Reference Laboratory of Enteroviruses, NCIPD, Sofia, Bulgaria AIM: To determine the causative role of parvovirus B19, Coxsackie B viruses and adenoviruses in the development of myocarditis, pericarditis and dilated cardiomyopathy by demonstrating the presence of specific antiviral antibodies or viral DNA in patients serum samples. MATERIALS AND METHODS: Serum samples gathered in 2010-2014 from 235 patients with myocarditis (n = 108), pericarditis (n = 79), myopericarditis (n = 19), dilated cardiomyopathy (n = 7) and fever of unknown origin accompanied by cardiac complaints (n = 22), were tested. Serological (ELISA for specific IgM/IgG antibodies to parvovirus B19, Coxsackie B and adenoviruses) and molecular (PCR for parvovirus B19) methods were used. R ESULTS: A total of 40 of the 235 serum samples (17%) were Coxsackie B virus IgM positive. They were found in 17% (18/108) of the patients with myocarditis, in 15%(12/79) with pericarditis, in 16%(3/19) with myopericarditis and in 32%(7/22) with fever of unknown origin. The serum samples of the 63 Coxsackie B virus IgM negative patients were tested for adenovirus IgM antibodies. Positive results were obtained in 4 patients with pericarditis and in 2 patients with fever of unknown origin. Every IgM negative sample (n = 189) for Coxsackie B and adenovirus was tested for parvovirus B19 IgM/IgG. B19-IgM antibodies were confirmed in 14 (7.4%) of the samples. They were 8% (7/90), 5% (3/63) and 31% (4/13) among the patients affected with myocarditis, pericarditis and fever of unknown origin, respectively. Protective B19-IgG antibodies were found in 108 (57%) and positive B19-PCR signal in 15 (8%) of the samples. Key words: parvovirus B19, coxsackie B, adenoviruses, myocarditis, pericarditis, ELISA, PCR

BM 26.

CHOLINERGIC, TACHYKININERGIC AND NITRERGIC NEUROTRANS-

MISSION IN MОTОR RЕSPОNSЕS ОF LОNGITUDINAL MUSCLЕS IN DISTAL CОLОN Galia Stavreva1, Zornitca Gorcheva2, Radomir Radomirov3 1Department of Experimental and Clinical Pharmacology, Medical University - Pleven, Department of Nursing Care, Medical University - Pleven, 3Department of Experimental and Clinical Pharmacology, Medical University - Pleven, Institute of Neurobiology, Bulgarian Academy of Sciences, Sofia, Bulgaria The aim of this study was to investigate elеctrically-induced lоcal and dеscеnding mоtоr activity of lоngitudinal muscle in isоlatеd cоlоnic sеgmеnt as a display оf еxcitatiоn оf thе lоcal nеrvе nеtwоrks оr thе dеscеnding rеflеx pathways. Еlеctrical fiеld stimulatiоn (0.8 ms, 40 V, 5 Hz, 20 s) appliеd tо thе distal оr tо thе prоximal part оf sеgmеnt-prеparatiоns inducеd tеtrоdоtоxin

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(0.1 μM)-sеnsitivе lоcal оr dеscеnding cоntractiоns indicating thе nеrvе-mеdiatеd оrigin оf thе rеspоnsеs. Thе lоcal cоntractiоns wеrе mоrе еxprеssеd by amplitudе. Thе dеscеnding cоntractilе rеspоnsеs dеcrеasеd by amplitudе and a rеlaxatiоn rеvеalеd оn thе backgrоund оf atrоpinе (0.3 μM). Thе cоntractiоns wеrе dеcrеasеd mоrе, but nоt еliminatеd, and thе rеlaxatiоn wеrе еnhancеd whеn spantidе was (0.1 μM), anantagоnist оf NK1 rеcеptоrs, additiоnally was addеd tо thе bath sоlutiоn. L-NNA (0.5 mM), an inhibitоr оf nitric оxidе synthasе, incrеasеd thе dеscеnding cоntractilе rеspоnsеs during atrоpinе trеatmеnt and dеcrеasеd thе rеlaxatiоn, whilе L-Argininе (0.5 mM), a substratе оf nitric оxidе synthеsis, dеcrеasеd thе cоntractilе activity and dееpеn thе rеlaxatiоn. Immunоhistоchеmical and histоchеmical studiеs fоr substancе P and NADPH-d confirmed mechanographical results. Thеsе rеsults indicatе that in thе rat-mоdеl еxcitatоry chоlinеrgic and tachykininеrgic and inhibitоry nitrеrgic nеurоtransmittеr systеms arе invоlvеd in thе rеflеx pathways cоntrоlling thе cоlоnic dеscеnding mоtоr activity. Key words: lоngitudinal muscle, dеscеnding mоtоr activity

BM 27.

SENSITIVITY AND SPECIFICITY OF TTF1, NAPSIN A, CK 7, P63, CK34ΒE12

AND MUTATIONS IN THE RECEPTOR OF EPIDERMAL GROWTH FACTOR IN THE RESECTION MATERIALS IN NON-SMALL CELL LUNG CARCINOMAS Silvia Genova1, Stoian Bichev2, Veselin Belovezhdov3, Angel Uchikov4 1Department of General and Clinical Pathology and Forensic Medicine, Medical Faculty, Medical University Plovdiv; 2National Genetic Laboratory: Maternity Hospital, Medical University, Sofia; 3Department of General and Clinical Pathology and Forensic Medicine, Medical Faculty, Medical University, Plovdiv; 4Department of Special Surgery, Medical Faculty, Medical University - Plovdiv, Bulgaria INTRODUCTION: Personalized therapy in non-small cell lung carcinomas, as well as advances in targeted therapy with tyrosine kinase inhibitors require clarification of tumor subtypes and specific reference to the type of lung cancer. MATERIALS AND METHODS: From 2012 to 2014, we studied prospectively 72 RM of lung carcinomas derived from the Surgery Division in St. George Hospital, Plovdiv. Male / female: 59/13, ratio 4.5: 1, age: 28-84. Groups: Group I included 34 AC and 3 adenosquamous carcinomas (AdSCC); n = 37. Group II: n = 35 RM, initially considered a routine microscopy 22 SCC and 13-NOS. Biopsies were tested with Napsin A, TTF1, CK7, p63, CK34βE12 and were tested for EGFR mutations with real-time PCR technology. Fivefold negative undifferentiated carcinomas typified with Synaptophisin, Vimentin, CDX2, CK20. Results: After IHH, distribution of histology is: ACs - 31/72, AdSCC - 3/72, SCC - 22/72, large cell neuroendocrine carcinomas - 9/72, sarcomatoid - 3/72, 1/72 NOS. Sensitivity and specificity of the test markers was: Napsin A (76.5% and 100% ); TTF1 (82.4% and 80.6%); CK7 (94.3% and 57.9%); p63 (84.0% and 85.1%); CK34βE12 (88.0% and 51.06%). EGFR mutations were found in 1/22 SCC - S7681 and 1/3 AdSCC- L858R. All ACs were negative for EGFR mutations. Conclusion: TTF1 and Napsin A for ACs, p63 for the SCCs showed moderate sensitivity, at 100 % specificity for Napsin A and it should be used in combination. Although rare, EGFR mutations in the SCC and AdSCC justify their research and chance these patients receive targeted therapy. Key words: non-small cell lung cancer, epidermal growth factor, mutations, Napsin A, TTF1, CK7, p63, CK34βE12


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POSTERS

Biopharmacy BP 1.

ADHERENCE TO LIPID-LOWERING THERAPY IN ELDERLY –

A PHARMACY DATABASE STUDY IN BULGARIA Hristina Lebanova1, Konstantin Kalaidjiev2, Evgeni Grigorov3, Ilko Getov2 1Medical University - Pleven, 2Medical University - Sofia, 3Medical University - Varna, Bulgaria Patients with dyslipidemia are at high risk for cardiovascular diseases. Adherence with prescribed therapy can considerably mitigate this risk and is important in these patients. The aim of this study is to evaluate adherence to lipid-lowering therapy in patients over 65 years old using pharmacy dispensing records and define predictors of non-adherence. This retrospective study examined the refill patterns of 341 patients diagnosed with hyperlipidemia (ICD code E75.8) from their automatic pharmacy dispensing records for a period of 24 months. Adherence was measured as proportion of days covered by each one month interval following enrolment in study. Patients were considered adherent if they had filled prescriptions no later than 5 days after due date. Age, sex, comorbidities, generic/brand medicines were evaluated as potential predictors of adherence. Two-year adherence rates were 70% for women and 64% for men. Lower copayments were associated with higher levels of adherence for both groups. After adjustment for age, sex, comorbidities and other potential predictors, patients were more likely to be adherent if they were female, age 65-75 years old and with a history of coronary heart disease. Elderly patients have low rates of adherence to lipid-lowering therapy. This suggests that they receive limited benefit from their therapy. Key words: adherence, elderly, lipid-lowering therapy, database

BP 2.

MELATONIN PROTECTS BURN INDUCED HEPATOCYTE OXIDATIVE INJURY

BY INDUCING HEME OXYGENASE-1 VIA NRF2 PATHWAYS Ganka Bekyarova, M. Hristova, M. Tzaneva Department of Physiology and Pathophysiology, Medical University - Varna, Bulgaria Melatonin has beneficial effects against early liver injury by modulating hepatic oxidative stress. To understand the protective effect of melatonin against burn-induced hepatic injury we investigated the expression of 4-hydroxinonenal (4HNE), a major product of lipid peroxidation and mediator of oxidative response, the inducible heme-oxygenase-1 (HO-1), antioxidant enzyme and anti-oxidative stress regulator erythroid 2-related factor 2 (Nrf2) in burn rat model. Liver injury was evaluated immunohistochemically for presence of hepatic 4-HNE protein, biochemically for changes in serum aminotransferase levels and MDA (marker of lipid peroxidation) and histological (H&E) studies. Expression and location of HO-1 and Nrf2 in liver were investigated using light immunochemistry. Thermal skin injury caused significant elevation of hepatic 4-HNE, MDA and transaminase levels and degenerative changes in the liver. Concurrently, overexpression of HO-1, rate-limiting enzyme for heme degradation and an oxidative stress marker in SEC and hepatocytes without changes in expression of Nrf2 in the liver. Melatonin (20 mg/kg), augmented the increase the HO-1 expression, upregulated Nrf2 expression as well as decreased serum transaminase levels and histopathological changes in the liver in rats.

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In conclusion, our results show that melatonin ameliorates burn-induced liver injury through inhibiting oxidative stress injury and through upregulation of antioxidant enzymes, such as HO-1 and expression of the antioxidant transcription factor Nrf2 pathway. Stimulation of protective mechanisms of the cells by activating transcription antioxidant factors such as NRF2 is a new mechanism for protection against liver damage in burns. Key words: Nrf2, HO-1, 4-HNE, oxidative stress, liver, melatonin

BP 3.

IDENTIFICATION AND ANTIOXIDANT ACTIVITY OF ISOPRENOIDS IN

BIOMASS BY PSYCHROPHILIC YEASTS Stela Dimitrova1, Lyudmil Lukanov1, Konstanca Pavlova2, Mariana Katcarova1 1Department of Chemistry and Biochemistry, Faculty of Pharmacy, Medical University - Plovdiv; 2Institute of Microbiology, Bulgarian Academy of Sciences, Sofia, Bulgaria Psychrophilic yeasts are interesting as new and insufficiently investigated microorganisms with a potential to synthesize biologically active substances. Yeasts from different genera are able to synthesize carotenoids, coenzyme Q10 while ergosterol is one of the most important components in their cell membranes. They are lipophilic substances with isoprenoid structure and pronounced biological activity. The main task of this study is identification of isoprenoids extracted from the psychrophilic yeast biomass, estimation of their antioxidant activity and research of the aspects of their applications as ingredients in sunscreen cosmetics. Important scientific and practical results were achieved in the process of the research. Several methods were developed for extraction and HPLC-identification of β-carotene, coenzyme Q10 and ergosterol from the Antarctic yeasts. Torularodin and torulen were identified. Correlations were found between the conditions in which the cultures of Antarctic yeasts were developed and the biosynthesis of β-carotene, coenzyme Q10, ergosterol and unsaturated higher fatty acids. Positive effect on the biosynthesis of β-carotene, coenzyme Q10 was proven at combination radiation with ultraviolet and visible spectrum lights at the time of cultivation of the Antarctic yeasts. Oxygen Radical Absorbance Capacity (ORAC) method for lipophilic substances and an electrochemical method were both used to determine the antioxidant activity of β-carotene, torularodin, torulen, coenzyme Q10 and ergosterol extracted from the biomass of Antarctic strain S. salmonicolor AL1. Versatile biotechnological potential was proved for strain S. salmonicolor AL1 to produce exopolysaccharide with emulsifying qualities and endogen metabolites with antioxidant activity and prospective use as components in photoprotective cosmetics. Key words: carotenoids, coenzyme Q10, antioxidant activity

BP 4.

COMPARATIVE EFFECTIVENESS OF HORMONE STIMULATION MEDICINES,

USED IN ASSISTED REPRODUCTION THERAPY – PRELIMINARY RESULTS FROM A RETROSPECTIVE DATABASE STUDY Svetoslav Stoev1, Hristina Lebanova2, Ilko Getov1 1Faculty of Pharmacy, Medical University - Sofia, 2Medical University - Pleven, Bulgaria The AIM of the study is to compare IVF/ICSI parameters and the rate of pregnancy in patients who were stimulated with several different follicle-stimulating hormones /FSHs/, FSHs plus recombinant human luteinizing hormone or menopausal gonadotropins and gonadotropin-releasing hor-


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mones /GnRH/ antagonists. Another task is to identify whether there is any correlation between anthropologic characteristics of patients in the study cohort and the rate of pregnancy. METHODS: A retrospective analysis of a hospital based IVF database for one year period PATIENTS: patients undergoing IVF with/without ICSI. R ESULTS: The participants were 665 women who received assisted reproduction treatment between January and December 2013. The overall clinical pregnancy rate was 21%. The average age of the patients is 35 yrs (range 23-45 yrs). The comparison of stimulation characteristics shows that human follicle-stimulating hormones are administerеd in the highest total dose during stimulation /7500 IU/. The stimulation period is between 3 and 19 days. Stimulation protocols that include recombinant Follitropin alfa, Follitropin beta and Collifolitropin lead to higher number of follicles produced. There is no statistically significant difference between the number of explanted oocytes and the number of transferred embryos in accordance to the type of the ovarian stimulation. Most of the patients with a positive clinical outcome (pregnant) had been treated by a stimulation protocol, including a combination of a recombinant and a human FSH, and a GnRH antagonist. These results are preliminary, and need to be confirmed by detailed further research and analysis. Key words: comparative effectiveness, pregnancy rate, IVF

BP 5.

EFFECTS OF KYOTORPHIN AND D-KYOTORPHIN ON PAIN PERCEPTION

AFTER HOT STRESS IN RATS Department of Pathophysiology, Faculty of Medicine, Medical University -Sofia, Bulgaria Mimoza Tzvetkova, Radka Hadjiolova, Hristina Nocheva Kyotorphin (Кyо) and D-kyotorphin (D-Kуo) are dipeptides which modulate pain perception. Кyо and D-Kуo indirectly influence μ- and δ-opioid receptors, exerting naloxone-reversible and prolonged analgesia due to release of met-enkephalin and β-endorphin. Analgesia induced after stress is a common phenomenon in many animal species and could be triggered by different stressors – immobilization, low or high temperatures. The aim of the present study was to evaluate the effects of Kyo and D-Kyo on analgesia after heat stress. The experiments were carried out on male Wistar rats (180-200 g). The peptides were administered intraperitoneally at a dose of 5 mg/kg after the heat stress. Nociceptive tests: Antinociceptive effects were evaluated using paw pressure (PP) and hot plate (HP) tests. Stress model - 1 hour of heat stress (1hHS) was provoked by placing the animals for 1 hour in thermal chamber (38 ± 1°C). The results of our study showed that hot stress increased pain threshold and HP-latency during the whole investigated period. Kyo increased pain threshold at 15 and 30 minutes of the experiment, while D-Kyo increased pain during the whole investigated period compared to the control animals. Куо prolonged HP-latency during the whole investigated period, while D-Куо prolonged HPlatency at 15 and 30 min of the evaluated time compared to the controls. After 1hHS Куо and D-Куо decreased the pain threshold and НР-latency of the animals for the same time of investigation compared to animals after 1hHS without the peptide. Key words: kyotorphin, D-kyotorphin, stress, nociception

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BP 6.

EFFECTS OF ATYPICAL ANTIPSYCHOTICS ARIPIPRAZOLE AND

OLANZAPINE ON A MODEL OF CLONIDINE RETROGRADE AMNESIA Marian Topolov1, Damianka Getova2 1Department of Pharmacology and Drug Toxicology, Faculty of Pharmacy, Medical University - Plovdiv; 2Department of Pharmacology and Clinical Pharmacology, Medical Faculty, Medical University - Plovdiv, Bulgaria INTRODUCTION: Aripiprazole is antipsychotic drug, alternative to conventional antipsychotics that impair memory. Atypical antipsychotic olanzapine acts as dopamine, serotonin and alpha adrenergic antagonist and these neuromediators could account for memory impairment, psychomotor skills and locomotor activities. The AIM of the study was to assess the impact of the drugs on locomotor activity of rats with clonidine amnesia with particular emphasis on their memory reactions after repeated application. MATERIAL AND METHODS: 32 male Wistar rats were used, divided in 4 groups (n = 8). Passive avoidance tests, step through and step down were used. They were performed in 2-day learning session, short (3rd day) and long (9th day) memory retention tests. Criteria for step through were latency of reactions 180s staying in the light chamber and for step down test was the maximal latency of reaction 60s staying on the platform. Locomotor activity was studied in an open field chamber for 5 minutes, horizontal and vertical movements were observed. R ESULTS: In step-through test rats treated with aripiprazole and clonidine increased the latency time during the learning session and long memory retention test. Rats treated with olanzapine and clonidine decreased latency time only during the learning session. In step-down test only the animals treated with aripiprazole and clonidine increased latency time on learning session. In locomotor activity test only rats treated with aripiprazol and clonidine decreased the number of horizontal and vertical movements. CONCLUSION: The study reveals that both antipsychotics influenced locomotor activity, learning and memory in rats with clonidine amnesia. Key words: aripiprazole, olanzapine, clonidine retrograde amnesia

BP 7.

EVALUATION OF THE EFFECTS OF CHOLECYSTOKININ ON PROTEIN

KINASE B (AKT) PHOSPHORYLATION IN A MODEL OF DIET-INDUCED OBESITY IN RATS Teodora Stankova1, G. Valchev2, B. Merino3, B. Cano3, M. Ruiz-Gayo3, Delian Delev4 1Department of Chemistry and Biochemistry, Faculty of Pharmacy, Medical University- Plovdiv; 2Regulatory Affairs Expert Actavis; 3Departamento De Farmacología, Tecnología Y Desarrollo Farmacéutico, Universidad Ceu-San Pablo, Madrid, Spain; 4Department of Pharmacology and Clinical Pharmacology, Faculty of Medicine, Medical University - Plovdiv, Bulgaria In the last decades, a large number of metabolic disturbances like insulin-independent diabetes and cardiovascular diseases have been related to the consumption of high fat diet. Obesity is defined as a chronic positive energy imbalance resulting in an excessive fat accumulation in adipose tissue. An important role in the regulation of energy homeostasis plays the satiety signal cholecystokinin (CCK), a peptide that is distributed widely throughout the gastrointestinal tract and the central nervous system. At cellular level protein kinase B (Akt) is a key regulatory molecule integrating several different signals important in cell growth and metabolism. Activation of Akt signaling has great implications in obesity and insulin resistance. Therefore this study aimed to evaluate the possible effect of cholecystokinin administration on phosphorylation of Akt in adipose tissue in a high fat diet (HF) induced obesity model. The results demonstrated that the consumption of HF differentiated the rats into two groups according to the gain of body weight, obesity resistant (OR) and obesity prone (OP) animals. The Folia Medica 2015; 57; Suppl. 1 © 2015 Medical University, Plovdiv, Bulgaria

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administration of CCK had a tendency to increase the level of phosphorylation of Akt in both OP and the control low fat diet (LF) treated rats, but to decrease it in OR group. This effect did not reach statistical significance and suggested that longer dietary treatment is necessary to be sufficiently affected. Key words: cholecystokinin, obesity

BP 8.

HIPPOCAMPAL ASYMMETRY IN ANGIOTENSIN II MODULATORY EFFECTS

ON LEARNING AND MEMORY IN RATS Roman Tashev1, Margarita Stefanova2, Stiliana Belcheva3, Iren Belcheva4 1Department of Pathophysiology, Medical University - Sofia; 2Department of Physiology and Physiology, Medical University - Varna; 3Faculty of Pre-School and Primary School Education, SU Sv. Kl. Ohridsky; 4Institute of Neurobiology, Bulgarian Academy of Sciences, Sofia, Bulgaria Learning and memory effects of angiotensin II (Ang II) microinjected unilaterally (left or right) and bilaterally into hippocampal CA1 area on the background of the losartan inhibited hippocampal angiotensin 1 receptors type (AT1) of male Wistar rats were studied. It was found that the combination (losartan 100 μg + Ang II 0.5μg) microinjected bilaterally or into the left CA1 area improved learning and memory in shuttle-box and step through behavioral tests as compared to the respective controls. The effects were more pronounced after injection into the left CA1 area as compared to the right-side. These findings suggest that Ang II infused on the background of the inhibited CA1 hippocampal AT1 receptors ameliorated the cognitive processes. The data show also an asymmetric effect of Ang II on learning and memory processes in the hippocampus. The stronger modulating effect after microinjection of the combination (losartan + Ang II) into the left CA1 hippocampal area suggests a leftward bias in the rat. The results point to a differential distribution of angiotensin II receptors modulating the learning and memory processes in the left and right hippocampal CA1 area. Key words: losartan, AT1 receptors, hippocampus, learning, memory, rat

BP 9.

IMPORTANCE OF PATIENTS REPORTS OF ADVERSE DRUG EVENTS

Hristina Lebanova1, Svetoslav Stoev2, Ilko Getov2 1Medical University - Pleven, Medical College, 2Medical University - Sofia, Bulgaria Patient reports of adverse drug events could contribute to the pharmacovigilance system qualitatively as well as quantitatively. There are a lot of factors that influence patients’ participation such as personal characteristics, disease perception, previous experience and education. Patients in the European Union are direct and active participants in the ADR reporting systems since July 2012 due to changes in the European pharmaceutical legislation. Their inclusion as a source of information will help to change the perspective and coverage of local and community pharmacovigilance systems. The factors that affect the decision of the patient’s report of an adverse drug reaction and possible external influences that would shape their opinion and could affect the quality of reports are not well studied. The need of educational materials is well recognized in the literature. They should meet certain criteria in order to maximize their usefulness. Their aim is achieved through different strategies to improve readability, graphic design and patients’ understanding. In this presentation we summarize and discuss the possible options and approaches to improve patients role in ADRs reporting. Key words: pharmacovigilance, patients, adverse drug reactions

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BP 10.

INFLUENCE OF ROSUVASTATIN ON ACUTE INFLAMMATION AND IL-6

LEVELS IN RATS Maria Georgieva-Kotetarova1, Ivanka Kostadinova1, Marianna Murdjeva2, Delian Delev1, Ilia Kostadinov1 1Department of Pharmacology and Clinical Pharmacology, Faculty of Medicine, Medical University - Plovdiv; 2Department of Microbiology and Immunology, Faculty of Pharmacy, Medical University - Plovdiv, Bulgaria Recent evidence indicates that statins possess pleiotropic effects apart from their lipid-lowering activity. The AIM of the present study was to investigate the anti-inflammatory effect of rosuvastatin in a model of acute inflammation. MATERIALS AND METHODS: Male Wistar rats were treated orally for 90 days with rosuvastatin 10 and 20 mg/kg bw. Control group was treated orally with saline 1 ml/100 g bw. At the day of experiment a fourth group received orally indomethacin 9 mg/kg bw. After the 90-day period paw edema was induced by carrageenan injection and the footpad volume was measured at the 4th hour using plethysmometer. Blood was collected and plasma concentrations of IL-6 were measured. R ESULTS: Indomethacin significantly reduced the extent of the paw edema 4 hours after the carrageenan injection compared to the saline treated group. Administration of rosuvastatin in both doses produced significant inhibition of the edema compared to the saline treated group. There was no significant difference in plasma levels of IL-6 in rosuvastatin-treated animals compared to the animals, receiving saline. CONCLUSION: Rosuvastatin exerts anti-inflammatory effect in a model of carrageenan-induced paw edema, but does not reduce IL-6 plasma concentrations. Key words: statins, anti-inflammatory, IL-6, carrageenan-induced paw edema

BP 11.

INTERACTION BETWEEN NALOXONE AND THE CANNABINOID SYSTEM

AFTER THREE MODELS OF STRESS Hristina Nocheva, Adriana Bocheva Department of Pathophysiology, Faculty of Medicine, Medical University - Sofia, Bulgaria Several physiological processes change in the body during stress, pain perception among them. Decreased pain sensitivity is known as stress-induced analgesia (SIA), which has an opioid and a non-opioid component. The opioid component comprises opioid receptors (μ-, δ-, κ-, λ-, σ-) and their endogenous ligands (β-endorphins, enkephalins, and dynorphin). The non-opioid one comprises the adrenergic, the serotoninergic, the nitric oxide, and the cannabinoid systems. The latter consists of two types of cannabinoid receptors (СВ1 and СВ2), their endogenous ligands, and the enzyme systems involved in their synthesis and degradation. It is known that the opioidergic and the endocannabinoid systems interact with each other. The aim of the study was to evaluate the interaction between the two systems targeting the cannabinoid and the opioid receptors. The opioid and the non-opioid components are differently involved in cold, immobilization and heat stress. Antagonizing the opioid receptors with naloxone would differently influence SIA after each of the three stresses. The experiments were conducted on sexually mature male Wistar rats. SIA was induced through cold, immobilization, and heat stress. The substances were administered intraperitoneally. СВ1 agonist anandamide and the non-selective opioid receptors antagonist naloxone were injected at a dose of 1 mg/kg. Results showed that antagonizing the opioid receptors influenced at a different degree the effect of exogenous anandamide on SIA after cold, immobilization, and heat stress. Key words: stress-induced analgesia, anandamide, naloxone Folia Medica 2015; 57; Suppl. 1 © 2015 Medical University, Plovdiv, Bulgaria

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BP 12.

INTERDEPENDENCE BETWEEN THE USE OF DRUGS, REIMBURSEMENT

LEVELS AND HOSPITALIZATION RATES IN THE TREATMENT OF PEDIATRIC ASTHMA Anna Todorova1, Antoaneta Tzvetkova2 1Faculty of Pharmacy, Medical University - Varna; 2Assistant Pharmacist - Education and Research Center, Medical College of Varna, Bulgaria The results showing an increased use of drugs in outpatient care facility and a decrease of the number of children hospitalized between 2008 and 2013 were the reason we set out to explore the interdependence between the following variables: drug use, level of reimbursement and hospitalization rates of pediatric patients with asthma. The survey was carried out in the town of Varna. For the purposes of our study, we conducted a data analysis of the asthma medications prescribed to children in two age groups (6-12 years and 13-16 years) and the number of hospitalizations of the pediatric patients within these age groups. We found a statistically significant (p < 0.05) positive correlation between the drug use and the level of reimbursement (R = 0.91 for the age group 6-12 yrs and R = 0.89 for the age group 13-16 yrs). The results showed a very strong negative correlation (p 0.05) between the outpatient drug use and the hospitalization rates (R = 0.99 and R = 0.88), and between the reimbursement level and hospitalization rates (R = 0.94 and R = 0.88) within the two age groups alike. Better access of patients to prophylaxis and the increased use of medications in ambulatory care provide better asthma control and reduce the hospitalization rates. Key words: asthma, children, drug use, reimbursement, hospitalization rates

BP 13.

EFFECTS OF SELECTIVE BLOCKADE OF THE PURINERGIC RECEPTOR

P2X7 ON THE ELECTRORETINOGRAPHIC (ERG) RESPONSES Petia Kupenova, E. Popova, L. Vitanova Department of Physiology, Medical University - Sofia, Bulgaria The ionotropic purinergic receptor P2X7 is widely expressed in the retinal neurons as well as in the Müller glial cells. However, its contribution to physiological retinal responses is not clarified. In order to elucidate this question, we investigated the effects of blockade of the P2X7 receptors in the retina by the selective P2X7 blocker A438079 on the amplitude of the electroretinographic (ERG) responses. We studied the effects on the responses of the frog (Rana ridibunda) retina to mesopic stimuli under dark adapted conditions. The effect of the selective P2X7 blocker was dose-dependent. After application of A438079 in saturating concentration (200 μM), the amplitude of the ERG response to stimulus onset (ONresponse; the ERG b-wave) was increased, while the amplitude of the response to stimulus offset (OFF-response; the ERG d-wave) was decreased. The effect on the ON-response had a fast time course and was fully reversible. The effect on the OFF-response was stronger as compared to the effect on the ON-response. However, it followed a slower time course and the OFF-response was not fully recovered during the recovery period. The results obtained show that the purinergic receptor P2X7 participates in the control of the physiological retinal responses and that its effects on the ON- and OFF-responses are asymmetric. The differences in the direction, degree, time course and reversibility of the effects of the selective P2X7 blockade on the ERG ON- and OFF-responses indicate that these responses are controlled by different P2X7 receptor-mediated purinergic mechanisms. Key words: retina, purinergic, P2X7, ERG, frog

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BP 14.

NON-ALCOHOLIC FATTY LIVER DISEASE (NAFLD) IN RATS WITH

METABOLIC SYNDROME AFTER SUCROSE OR FRUCTOSE CONSUMPTION Rositca Sandeva1, Gergana Sandeva2 1Department of Physiology, Pathophysiology and Pharmacology, Medical Faculty, Trakia University, Stara Zagora; 2Department Hygiene, Infectious Diseases and Epidemiology, Medical Faculty, Trakia University, Stara Zagora, Bulgaria The increased incidence of metabolic syndrome and type 2 diabetes mellitus in recent decades is associated with overconsumption of sugar, fructose and artificial sweeteners. Indulging in simple carbohydrates can also lead to inflammatory changes in the liver, resulting in development of non-alcoholic fatty liver disease (NAFLD). Objective: To model the metabolic syndrome in rats by increased intake of fructose or sucrose and to track changes in the liver of the control and experimental groups. Methods: The present study compared the effects of 8-week treatment of male Wistar rats with a 15% fructose solution (Group F, n = 7), 10% sucrose solution (Group S, n = 7) and control group (n = 7). Changes in body weight, basic metabolic parameters (glucose, triglycerides, total, LDL and HDL cholesterol) and urea levels, ASAT, ALAT and C-reactive protein were traced. In the seventh week ultrasound examination was performed on the livers of all animals. Results: Statistically significant difference (p < 0.05) was found between the experimental and control groups in terms of mean body weight: sucrose group > fructose group> controls. Significant increase of serum glucose, triglycerides, total and LDL cholesterol, hepatic transaminases and C-reactive protein was registered in the experimental groups compared to the controls. Urea in the experimental groups was also increased, but not significantly. Ultrasound scans showed the development of fatty liver in animals from the experimental groups. Conclusions: The development of metabolic syndrome and fatty liver in rats after 8-week overconsumption of fructose or sucrose showed the need to limit the intake Key words: metabolic syndrome, fructose, sucrose, non-alcoholic fatty liver disease, Wistar rats

BP 15.

EFFECTS OF LOSARTAN INFUSED INTO HIPPOCAMPAL CA1 AREA ON

EXPLORATORY AND LOCOMOTOR ACTIVITY OF RATS Roman Tashev1, Margarita Stefanova2, Stiliana Belcheva3, Iren Belcheva4 1Department of Pathophysiology, Medical University - Sofia; 2Department of Physiology and Pathophysiology, Medical University - Varna; 3Faculty of Pre-School and Primary School Education, St. Kliment Ohridsky Sofia University; 4Institute of Neurobiology, Bulgarian Academy of Sciences, Sofia, Bulgaria We examined the effects of losartan, a specific antagonist of angiotensin II type I (AT1) receptors, infused uni- or bilaterally into the hippocampal CA1 area of male Wistar rats, on exploratory and locomotor activity. The changes in exploratory behavior and locomotor activity were registered in an OptoVarimex apparatus. Microinjected bilaterally at a dose of 100 μg, losartan significantly decreased the horizontal and vertical movements. Microinjections of losartan into the left hippocampal CA1 area suppressed the exploratory and locomotor activity, while right-side losartan administration did not affect them, as compared to the respective controls. The effect was more pronounced when losartan was infused into the left-side as compared to the right hippocampal CA. The main finding was the presence of hippocampal asymmetry in exploratory and locomotor activity to unilateral microinjections of losartan depending on the microinjected hemisphere, suggesting different distribution of AT1 receptors in left and right CA1 hippocampal area. Key words: losartan, AT1 receptors, exploratory activity, locomotor activity, hippocampus, rat


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BP 16.


THE ROLE OF THE PHARMACIST IN EXTEMPORANEOUS DISPENSING

Valentina Petkova1, Shahnaz Husain1, Milen Dimirov1, Kalina Andreevska2, Elina Petkova1, Stanislav Georgiev2, Daniela Grekova2, Vasil Madjarov2 1Faculty of Pharmacy, Medical Universty - Sofia; 2Faculty of Pharmacy, Medical Universty - Plovdiv, Bulgaria INTRODUCTION: Access to a special dosage form of medication is essential when administration to infants and children and selected other populations is required. Extemporaneous preparation describes the work involved in supplying the drug in a form or dose that is not otherwise available. MATERIALS AND METHODS: The aim of the study was to compare the most prescribed extemporaneous preparations and to outline the disease for which they are prescribed. Comparative analysis is performed together with statistical processing of the data. The questionnaire was applied in Bulgaria and Greece for 6 months during the period March – September 2014. Statistical analysis was performed using SPS ver. 22. Data from the questionnaires were entered on an ongoing basis into a database (SPSS ver. 22.0) after each interview in order to perform the interim analyses. CONCLUSIONS: Analysis of the results suggests that there is no difference between the main problems for which extemporaneous preparations are prescribed. The main diseases are dermatological such as acne, dermatitis, etc. The main population that uses extemporaneous medication includes infants, elderly patients and patients with dermatologic chronic disorders. Despite the great number of commercially manufactured drugs extemporaneous preparations are still prescribed and dispensed both in Greece and Bulgaria. Key words: extemporaneous preparations, pharmacist

BP 17.

CARDIOPROTECTIVE PROPERTIES OF ARONIA MELANOCARPA –

SOMATOMETRIC AND SERUM INDICES Elena Daskalova1, Slavi Delchev1, Julia Peeva2, Liudmila Vladimirova-Kitova3, Maria Kratchanova4 1Deparment of Anatomy, Histology and Embryology, Faculty of Medicine; 2Deparment of Social Medicine and Public Health, Faculty of Public Health; 3Department of Internal Diseases, Division of Cardiology, Medical University Plovdiv; 4Laboratory of Biologically Active Substances, Bulgarian Academy of Sciences, Sofia, Bulgaria One of the potential strategies to counteract the age-dependant oxydation damages is the delivery of antioxidants with food. Aronia melanocarpa (A.m.) has been rated first in antioxidative potential among anthocyanin fruits. Certain polyphenolic ingredients in A.m. have revealed cardioprotective effects according to the literature data. AIM: To determine the cardioprotective properties of juices from A.m. by studying somatometric and serum parameters as well as correlations between them in rats fed with such juices. MATERIAL AND METHODS: An experimental study has been carried out on 24 Wistar rats fed with A.m. juices for 3 months. R ESULTS AND DISCUSSION: The Lee’s obesity index remained within the norm in all groups of animals (p > 0.05). Statistically significant differences were found in weights of the heart but not in the heart weight index. An optimization of the lipid profile was found in cardioprotective direction with both groups of juice-fed animals whereas the combination of both constituents (A.m. and pectin) resulted in an increase of the effect. The clues were confirmed by the atherogenetic and cardioprotective indices. The RESULTS of the study indicated that the juice from A.m., and especially its combination with pectin, possesses organ-preserving and cardioprotective properties and can be recommended as prophylactic agent for conversion of the unsuccessful into successful vascular aging. Key words: successful aging, Aronia melanocarpa

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BP 18

CASTRATION-INDUCED TESTOSTERONE DEFICIENCY INCREASES FASTING

GLUCOSE IN MALE RATS Delian Delev, Ilia Kostadinov, Ivanka Kostadinova, Maria Georgieva-Kotetarova Department of Pharmacology and Clinical Pharmacology, Faculty of Medicine, Medical University Plovdiv, Bulgaria INTRODUCTION: Low testosterone levels have been shown to be predictive for the development of the metabolic syndrome in men. Castration is a way of studying the consequences of extreme testosterone deficiency in animal models. The AIM of this study was to investigate the effect of castration on the dynamics in the concentrations of serum testosterone and fasting blood glucose during replacement therapy with testosterone propionate (TP) in a dose of 4 and 8 mg/kg bw in rat model of androgen deficiency. MATERIAL AND METHODS: 80 male Wistar rats were included in the study, distributed in the following groups - orchiectomized controls, sham operated controls and treated with testosterone propionate in a dose of 4 and 8 mg/kg bw for 2 weeks (acute trial) and 3 months (chronic trial). R ESULTS: Orchiectomy lowered significantly the levels of serum T in the acute trial and insignificantly in the chronic trial. The supplementation with testosterone propionate raised its levels at the higher dose. Fasting blood glucose levels were significantly increased in castrated and sham operated rats 2 weeks after the intervention, compared with groups treated with TP in dose of 4 and 8 mg/kg bw. The decrease of blood glucose in chronic treated rats did not reach statistical significance. Conclusions: 1. Eight mg/kg b.w. of TP, applied in rat model of androgen deficiency restores the physical T levels. 2. Fasting blood glucose decline is observed in 2-week TP-treated castrated rats. Key words: testosterone propionate, castration, serum glucose

BP 19.

EFFECT OF ARONIA MELANOCARPA FRUIT JUICE ON BEHAVIOR AND

OXIDATIVE STRESS IN RATS EXPOSED TO DISRUPTION OF DIURNAL RHYTHM Stefka Valtcheva-Kuzmanova, Miroslav Eftimov, Krasimir Kuzmanov Department of Pre-clinical and Clinical Pharmacology, Medical University - Varna, Bulgaria Constant light leads to arrhythmicity of circadian locomotor activity, depressive-like behavior and excessive generation of free radicals. The present study investigated the effects of Aronia melanocarpa fruit juice (AMFJ) on the behavior and oxidative stress in male Wistar rats exposed to disruption of diurnal rhythm (DDR) via exposition to constant light for 14 days. Rats (n = 40) were divided into four groups: control, DDR, DDR+AMFJ5, and DDR+AMFJ10. Control and DDR animals were treated daily orally with water while DDR+AMFJ5 and DDR+AMFJ10 rats received AMFJ at doses of 5 ml/kg and 10 ml/kg, respectively. After the treatment period, the open field test (OFT), social interaction test (SIT) and forced swim test (FST) were carried out to evaluate behavioral changes. Oxidative stress was assessed by the concentration of thiobarbituric acid reactive substances (TBARS) in rat brains. Neither DDR, nor AMFJ affected significantly the locomotor activity in the OFT. In the SIT, DDR caused a non-significant reduction of the interaction between the test partners while the treatment with the two AMFJ doses significantly increased (p < 0.01 vs. DDR) the interaction time. In the FST, DDR significantly increased (p < 0.01 vs. controls) the immobility time. The immobility time of AMFJ-treated animals was not significantly different from that of the controls. TBARS in the brains of DDR rats were significantly higher (p < 0.01) than the control levels. AMFJ dose-dependently reduced TBARS which in DDR+AMFJ10 rats were not significantly different from the control values. AMFJ showed an anxiolytic and antidepressant-like effect, and decreased oxidative stress in rats exposed to constant light. Key words: Aronia melanocarpa, disruption of diurnal rhythm, behavior, oxidative stress, rats Folia Medica 2015; 57; Suppl. 1 © 2015 Medical University, Plovdiv, Bulgaria

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POSTERS

Public Health PH 1.

A CONCEPT OF NEW RELATIONSHIP BETWEEN THE COMPONENTS OF

COMMUNICATIVE COMPETENCE IN THE MODERN LANGUAGE TRAINING FOR MEDICAL PURPOSES Aneta Tosheva1, Victoria Sarafian2, Ana Coiug, Sofi Le Gal3 1Medical University - Plovdiv, Bulgaria, 2UMF Cluj-Napoca, Romania, 3University of Nantes, France The European language policy promotes task-oriented training in foreign languages and communication-oriented language training programmes, including those for professional purposes. Communicative competence is a key objective and the main methodological task in this training. Traditionally the ratio of its three core components (linguistic, sociolinguistic and pragmaticdiscursive competences) in foreign language textbooks for medical specialists has always been in favor of linguistic competence, especially its lexical component - medical terminology. A team of university authors – medical professionals and lecturers of medical French from Romania, Bulgaria, Moldova and France - suggests a concept of change in this relationship by putting the emphasis on the other two competences: pragmatic-discursive (related to situational functional speech behavior and text genres) and sociolinguistic (with characteristic descriptors for social, cultural and professional status of the communicators, for inter- and intra-professional registers and speech strategies). The textbook ‘Manuel de langue française pour les sciences et les métiers de la santé+CD’ Cluj-Napoca 2014 issued under the project AUF/BECO/PI/01/46125FT, prepares learners from different medical specialties for adequate speech behavior in situations of academic and/or professional mobility in a French-speaking environment. An appendix containing 40 authentic documents introduces all the basic text genres of medical records. The CD and the numerous web links provide audio and video materials and dozens of interviews of students and lecturers in real situations, in academic and professional environment. Key words: language policy, French, medicine

PH 2.

ADOLESCENTS’ ATTITUDE TOWARDS THE PHYSICAL ACTIVITY IN

SCHOOL Elena Merdjanova, Gergana Petrova Department of Midwifery Care, Faculty of Public Health, Medical University - Plovdiv , Bulgaria INTRODUCTION: Educating young people to have positive attitude to physical activity and sport is a constant process which includes the free time as well as the time spent at school. AIM: To survey adolescents’ opinion of physical activity and their attitude to sport classes at school. MATERIALS AND METHODS: A pilot study was performed among 12-year-old students in Hristo Danov secondary school, Plovdiv in February for the 2013/2014 school year. R ESULTS: Approximately half (56%) of the respondent students said the sport classes at school were sufficient to increase their physical culture and efficiency. It is notable that the percentage of those that did not answer this item is quite big (31%). Probably this is the reason why 88% consider the possible increase of sport classes in school curriculum as the right thing to do. The majority (88%) of the examined students regard the classes as interesting. The question: “Do they want rates in sport classes at school?”, almost half of them (44%) gave an affirmative answer and the same percentage do not

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think it is correct to fulfill rates. CONCLUSION: The bigger part of the examined students prefer games and entertainment in sport classes rather than systematical, planned and productive classes to increase their physical efficiency and young people’s physical development. Key words: physical education, physical activity, students

PH 3.

ADOLESCENTS HEALTHY EATING – AN INDICATOR OF GOOD HEALTH

Elena Merdjanova Department of Midwifery Care, Faculty of Public Health, Medical University - Plovdiv, Bulgaria INTRODUCTION: Childhood and adolescence are crucial periods in people’s lives - during these periods the body structure and the physiological changes define the variability of food rettquirements and behavioral physical activity. AIM: To study the food habits of adolescents and compare them with the recommendations for healthy eating. MATERIALS AND METHODS: A pilot study was performed among 12-year-old students in Hristo Danov secondary school, Plovdiv in February for the 2013/2014 school year. Results: Only 44% of the examined students have breakfast in the morning, the same number of students say they have it “sometimes”. Basic food products as meat and fish are eaten several times a week by 63% of the examined, while 24% have this food every day. The daily consumption of milk and dairy food is only for 37% of the students but 63% of the respondents eat baked products and pastry every day. CONCLUSION: The promotion of healthy eating among the children and adolescents is getting more important for public health because of the increase and prevalence of overweight and obesity among children and adolescents. Formation of healthy eating habits is of great importance for preventing obesity and other food disorders. Key words: healthy eating, healthy behavior, obesity, public health

PH 4.

AN ALGORITHM FOR KINESITHERAPEUTIC PROGRAM FOR CEREBRO-

VASCULAR DISEASE APPLIED BY STUDENTS IN CLINICAL PRACTICE Petia Kasnakova, I. Gatevski, N. Nikolova Division of Rehabilitation, Medical College, Medical University - Plovdiv, Bulgaria In their clinical internship, the students majoring in Rehabilitation at the Medical College are trained to work with patients diagnosed with cerebrovascular disease in the training facilities, to acquire professional skills and competencies. The kinesitherapeutic programs are an integral part of the overall recovery process in this disease. Use of algorithms in the preparation of students is an effective method in their training. The AIM of this study is to develop and implement an algorithm for a kinesitherapeutic program in cerebrovascular diseases geared to match the severity of the motor deficit, and apply it in clinical practice in the training of students. MATERIAL AND METHODS: We studied patients with cerebrovascular disease treated in Department of Physical Therapy and Rehabilitation at St. George University Hospital, at St. Panteleimon Hospital, and the Diagnostic and Consultative Centers 1, 2, and 5 in Plovdiv. An algorithm for medical rehabilitation was applied to the patients, which includes physiotherapy, massage and occupational therapy in the early and late periods. R ESULTS AND DISCUSSION: At the beginning and end of the rehabilitation procedures we performed the necessary tests and measurements. As a result of the applied complex kinesitherapeutic program an algorithm was developed to improve the func-


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tional status of patients. CONCLUSION: The application of this algorithm during clinical internship aided students in the rehabilitation activities and achieved one of the main tasks of recovery of locomotion, early ambulation, and prevention of complications. Key words: algorithm, kinesitherapeutic program

PH 5.

DETERMINING THE LEARNING STYLE OF E-LEARNING STUDENTS

Angelina Kirkova1, Nonka Mateva1, Daniela Taneva2, Yordanka Tzokova2 1Department of Medical Informatics, Biostatistics and E-Learning, Faculty of Public Health; 2Department of Nursing Care, Faculty of Public Health, 1Medical University - Plovdiv, Bulgaria INTRODUCTION: Achieving educational targets depends to a great extent on the cognitive and emotional characteristics of the audience. Such an important characteristic, informative for the planning and organization of training resources is the e-learning user’s learning style. AIM: The aim of the present study is to determine the learning styles of healthcare students for development of a relevant structure of the e-learning content. METHODS AND PARTICIPANTS: A momentary study was carried out which included 161 students from healthcare major at the Medical University in Plovdiv in December 2014. The study used anonymous questionnaire. The style of learning was determined according to Kolb’s Learning Style Inventory. Alternative, correlation, nonparametric statistical methods were used to analyse the results statistically. R ESULTS AND DISCUSSION: First year students demonstrate the following styles: 42% - assimilative, 35% - convergent, 14% - accommodative and 9% - divergent. Among third year students learning styles are distributed as follows: 49% - assimilative, 40% - convergent, 8% - accommodative and 3% - divergent. Assimilating students have a logical approach, they prefer lectures and theory. Converging way of learning implies practical work, case studies and simulations. CONCLUSION: Two styles stand out as typical for healthcare students – the assimilative and convergent styles. Preference for these styles is growing in the third year of study. E-learning resources and content structure relevant to these styles should be leading in e-courses design. Key words: learning style, students, e-learning

PH 6.

E-LEARNING – AN INNOVATIVE APPROACH TO IMPROVING THE QUALITY

OF STUDENTS EDUCATION Ruska Paskaleva Department of Medical Rehabilitation and Occupational Therapy, Physical Medicine and Sport, Medical Faculty, Trakia University, Stara Zagora, Bulgaria Training of students studying in the major Rehabilitation therapist at the Medical College and Medical Rehabilitation and Ergotherapy is designed to master relevant skills and competencies that meet the modern requirements for employment in the healthcare system and in specialized rehabilitation centres. The AIM of this study was to investigate the effect of innovative training methods on the quality of education and its application in clinical practice. MATERIAL AND METHODS: We included 85 students majoring in Rehabilitation therapist and Medical Rehabilitation and Ergotherapy majors in Trakia University, Stara Zagora in the 2014-2015 academic year, using a specially designed questionnaire. The results were analysed statistically. R ESULTS AND DISCUSSION: We found that students from both majors were satisfied with the e-learning modality, the organization and the conduct of clinical practice, but they recommended the introduction of new clinical facilities and rehabilitation centres in the region. CONCLUSIONS: The introduction of

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e-training is an innovative approach to improving the quality of education and leads to improvement of professional skills of students in clinical practice. Key word: innovation, e-learning, students, educational and practical facilities, practical training

PH 7.

EFFECT OF DIETARY INTAKE OF CHOLINE DURING PREGNANCY ON

OFFSPRINGS BRAIN DEVELOPMENT Rositca Stancheva Department of Hygiene, Medical University - Varna, Bulgaria A complete and balanced diet of a pregnant woman is essential for the fetus development and the health of future offspring. Deficiency of macronutrients and some micronutrients and biologically active substances in the diet of the mother has an unfavorable impact. The purpose of this review is to focus on the importance of choline for fetal brain development. The results of clinical studies published in Science Direct, Scopus and Bio Med Central are summarized. Pregnancy is a physiological condition which requires a high dietary intake of choline. It can affect the development of memory, cognitive function and mental activity of the offspring. During early pregnancy, choline is needed for the normal closing of the fetal neural tube. Key words: choline, pregnancy, brain development

PH 8.

HEALTH RELATED QUALITY OF LIFE AND PSYCHOLOGICAL WELL-BEING

IN PATIENTS WITH CHRONIC CORONARY ARTERY DISEASE Marieta Todorova, Gergana Petrova Faculty of Public Health, Medical University - Plovdiv, Bulgaria The association between the quality of life and health is displayed in the contemporary concepts of holistic approach to the individual and patient-centered medicine. One of the important objectives of healthcare worldwide is improvement of the quality of life of patients, their psycho-social adaptation and independence from the disease. The PURPOSE of the survey is to analyze the quality of life of patients with ischemic heart disease and its importance for their psycho-emotional adaptation to life with a chronic disease. METHODS: A pilot survey was conducted among patients with chronic coronary artery disease. The methodology employed two parts: an adapted Bulgarian version of the questionnaire EuroQoL EQ-5D, a generic tool for assessment of the quality of life and Spielberger Trait-Anxiety Inventory. R ESULTS: The dimensions most affected by coronary artery disease were pain/discomfort, followed by usual activities and mobility. Self-assessment of personal anxiety showed values mainly in the moderate scale. There was a correlation between emotional status and energy (r = 0.65). Anxiety was found in 23.40% of the respondents leading to shortening professional and daily tasks, feeling of tiredness and reduced vitality. CONCLUSION: Anxiety is a sign of emotional resistance of the personality and very often its importance is neglected by medical specialists. For patients with chronic cardiac diseases the quality of life is mainly influenced by psycho-social factors and the physical condition. Key words: quality of life, anxiety, psychological well-being, coronary artery disease


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PH 9.


HEALTH RISK FROM SOME HARMFUL LIVING HABITS AT THE WORKING

PLACE OF WORKERS IN SEWAGE WATER TREATMENT PLANTS (PRELIMINARY RESULTS) Elka Toseva1, Tania Turnovska1, Rositca Stoyanova2 1Department of Hygiene and Ecomedicine, Faculty of Public Health, Medical University - Plovdiv, Bulgaria; 2Department of Health Management, Health Economics and General Medicine, Faculty of Public Health, Medical University - Plovdiv, Bulgaria Harmful living habits at the working place of workers in the wastewater treatment plants (WWTP) adversely affect their health due to exposure to biological agents even if requirements for safe working conditions and instruction by the employer are good. The AIM of this study is to determine compliance with the rules of non-harmful living habits in the work environment among employees in the WWTP. MATERIALS AND METHODS: A study was carried out between November and December 2014 among 48 workers in two companies “Water-supply and Sewerage” of East Aegean region: 32 people (66.67%) of 2 WWTP (studied contingent) and 16 (33.33 %) of the treatment and delivery of drinking water (control contingent). Smoking and eating at the workplace, use the mobile phones and personal dressing style duirng work, hand washing and others were analyzed. R ESULTS: Smoking in the workplace: do not smoke 50.0% of WWTP vs. 46.7% of “Drinking water” (р > 0.05), rarely smoke - 21.9% vs. 26.7% (р > 0.05), often - 28.1% vs. 26.7% (р > 0.05), Eating at the workplace: often - 15.6% vs. 25.0% (р > 0.05), rarely 9.4% vs. 56.3% (р < 0,001), never 75.0% vs. 18.8% (р < 0,001), use of personal clothing during work - never 53.1% vs. 62.5% (p > 0.05), hands washing during work - often 96.9% vs. 93.8% (p > 0.05). CONCLUSION: It was found that there were violations of the instructions for behavior at the workplace among the two groups. The danger of exposure to biological agents was higher in the control group at Eating on the workplace. Key words: domestic waste water, daily habits, health risk, workers

PH 10.

INDIVIDUAL WORK AND FURTHER TRAINING AS ACTIVATING FACTOR

IN THE PROFESSIONAL COMPETENCE OF STUDENTS OF MEDICAL COLLEGE Petia Kasnakova Medical College, Medical University - Plovdiv, Bulgaria Self-employment and further training are critical factors for the active role of the students in the learning process. The aim of the study is to investigate the opinion of students studying in medical colleges on the use of self-employment as one of the most commonly used active methods and opportunities of further training. MATERIALS AND METHODS: The survey was conducted with 624 students I, II and III year on various aspects of the learning process through a questionnaire containing 25 questions. For our purpose we focused on issues related to self-employment and further training. R ESULTS AND DISCUSSION: According to 76.12% ± 1.1% of the surveyed students the amount of self-study was sufficient. The aid which students received in the development of their coursework in preparation of educational material, participation in scientific conferences (reports, announcements and posters), and scientific information was appropriate for promoting self-work. Most often students were assisted by teachers in their self-activity in development of assignments - 50.80 ± 2.00%. Teachers encouraged independent work in 69.07% ± 1.85% of all cases. 79.01% ± 1.63% of all students assessed positively their preparation and believed that the acquired theoretical knowledge and practical skills were sufficient. CONCLUSION: In terms of the continuous growth of scientific information and its considerable complexity, training should give students skills for independent orientation in knowledge so that they could acquire new knowledge and develop cognitive abilities for successful realization. Key words: individual work and further training 82



INTERACTIVE EDUCATION OF PHARMACEUTICAL STUDENTS IN

PH 11.

MATHEMATICS AND INFORMATION TECHNOLOGIES Hristo Manev Department of Pharmaceutical Sciences, Faculty of Pharmacy, Medical University - Plovdiv, Bulgaria The increased usage of information technologies in education and everyday life allows for conducting training courses through educational sites. In this particular case we developed a platform for the university course “Information Technologies” in the curriculum of students of Pharmacy major. This work aims to enable constructing of a course using modern e-learning environments. A hybrid system has been developed, i.e. a compulsory attendance of lectures in combination with interactive conduction of the test. Distance learning is an organization of the learning process where the teacher and the students are separated by location but not necessarily in time. The distance is compensated by technological resources – a web-based system “Interactive education in Information Technologies for pharmaceutical students”. The online courses have difficulties in controlling cheating on tests and exams precisely because of the lack of teaching supervision. The setting of the duration of the exam contributes to the objectivity of the evaluation. So, even if students try to check the answers of the questions in a textbook or online, they are going to fail to end on time if the gained knowledge of the students in their preparation is not enough. We designed a model for an educational platform of a university major which can be applied in other teaching courses. The relevance of e-learning nowadays and the need of modernization of the educational courses are the main reasons for the usage and the integration of this form of teaching more and more. Key words: interactivity, e-learning, educational course, Moodle

PH 12.

QUATERNARY PREVENTION - AN INNOVATIVE IDEA IN THE FIELD OF

PREVENTION Mariela Kamburova, Stela Georgieva, Silvia Aleksandrova-Yankulovska, Yoana Simeonova Department of Public Health Sciences, Faculty of Public Health, Medical University - Pleven, Bulgaria INTRODUCTION: Traditionally prevention as a combination of medical and non-medical activities conducted to achieve better health and quality of life is categorized in primary, secondary and tertiary prevention. The concept of quaternary prevention have appeared and gained recognition in the last decades. AIM: the aim of this report is to present this innovative preventive approach in medical practice and analyze the development of the concept of quaternary prevention. MATERIAL AND METHODS: Extensive literature review of over 20 sources, analyzing the place of prevention in the overall healing process and in particular the place and role of quaternary prevention was carried out. Publications focusing on current approaches for quaternary prevention of leading noncommunicable diseases were reviewed. R ESULTS AND DISCUSSION: “Primum non nocere (first, do not harm)” is the fundamental principle in medical practice. It supports the idea of quaternary prevention, whose focus is on the prevention of “unnecessary medicine” or prevention of overtreatment. It is subject to a number of studies mainly in the field of primary health care. Quaternary prevention is important not only in primary care, but also in the specialized out-patient medical care and in Bulgaria also in the hospital care due to the constantly increasing number of unjustified hospitalizations. CONCLUSION: Implementation of the quaternary prevention would increase the quality of life of Folia Medica 2015; 57; Suppl. 1 © 2015 Medical University, Plovdiv, Bulgaria

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people, would lead to a significant reduction in the unnecessary diagnostic and therapeutic procedures and unnecessary hospitalizations. As a result, healthcare resources will be spent more efficiently. Key words: quaternary prevention, quality of life, resources

PH 13.

MEDICAL EPONYMS NAMED AFTER SAINTS

Irina Mitarcheva, Zlatina Zheleva, Jana Delipavlova – Mladenova Department of Language and Specialized Education, Section of Foreign Languages, Medical University Plovdiv, Bulgaria The poster presents medical diseases and disorders named after saints. Eponyms are quite common not only in medicine but in other fields of study - they usually use the name of their creator, of the person who has invented or discovered the phenomenon in question. Eponyms in medicine are most frequently the names of the physicians who discovered a new disease, a function or a method but a variety of disorders have the names of saints attached to them. A saint is a person who has been canonized by the Church. Historically, many saints have become traditionally viewed as patrons of various illnesses, people, and places. From St. Agatha disease to the mysterious St. Vituss Dance, the authors try to introduce several conditions and diseases underlying these interesting names and attempt to reveal why exactly these saints are used to name a disease, a disorder or a syndrome. Key words: medical eponyms, patron saints, disease

PH 14.

METHODS OF CONTROL AND ASSESSMENT OF KNOWLEDGE IN

THE SPECIALIZATION IN GENERAL MEDICINE Gergana Petrova1, Gergana Foreva2, Radost Asenova2 1Department of Midwifery Care, Faculty of Public Health, Medical University - Plovdiv; 2Department of Health Management, Health Economics and General Medicine, Faculty of Public Health, Medical University - Plovdiv, Bulgaria INTRODUCTION: The specialization in General Medicine includes theoretical and practical training in 12 modules of different duration. PURPOSE OF THE TRAINING: to study the opinion of the General Practitioners (GP) regarding the applied methods of control and assessment of their knowledge during specialization in General Medicine. MATERIAL AND METHODS: The type of this sociological survey is anonymous direct group questionnaire carried out in two stages within a period of five years. During the first stage (2007-2008 academic year) 79 general practitioners from the Medical University of Plovdiv were questioned and during the second stage (2012-2013 academic year) 82 general practitioners from the Medical University of Plovdiv and the Medical University of Varna were questioned. R ESULTS: The data from the survey shows that the three most frequently applied methods of control and assessment of the knowledge of general practitioners during their specialization in General Medicine are written, oral and practical examination. The most preferred form of final control after each module is test examination 36.71 ± 5.42% for the respondents during the first stage and 47.06 ± 5.41% for the respondents during the second stage of the survey. CONCLUSION: The data obtained shows that in the process of specialization in General Medicine

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the most frequently used forms of control and assessment are the classical forms - written and oral examination, and the most preferred one is test examination. Key words: assessment, general medicine, postgraduate studies

PH 15.

MORAL CHALLENGES OF THREE-DIMENSIONAL BIOPRINTIG

TECHNOLOGY Atanas Anov, Silvia Aleksandrova-Yankulovska Department of Public Health Sciences, Section of Medical Ethics, Faculty of Public Health, Medical University - Pleven, Bulgaria INTRODUCTION: Bioprinting is a revolutionary technology for tissue engineering that seeks to solve the shortage of organs for transplantation through creating such from human cells. Like any new technology 3D-printing raises a number of ethical issues. The AIM of this report is to present and discuss emerging ethical issues of bioprinting. METHODOLOGY: Review, analysis and philosophical reflection on literature. DISCUSSION: In three-dimensional bioprinting human cells are printed or deposited on a surface layer by layer, instead of ink. Thus a whole organ or tissue can be produced for transplantation or cosmetic correction. Time in the waiting list is shortened to the «printing time». Ethical problems posed by 3D-printing are in two aspects: economic and social. The first aspect discusses the issue of equal access to the technology on macro- and micro-level. At the free market there are already companies that produce 3D-printed tissues and organs. Is this liberal approach appropriate when it comes to technology with vital benefits to patients? The social aspect of the discussion concerns the loss of personal identity related to the use of engineered organs. Does a patient remain a person after transplantation or do we create a robot piece by piece? CONCLUSION: In light of proactive ethics, ethical debates in connection with the newly developed technology must precede its mass introduction. Now is the exact time to reflect how we are going to answer the questions of patients who fear 3D-printed-organ robotization and those that cannot afford the price of the technology. Key words: bioprinting, ethics, identity, economic, social problem

PH 16.

NEW ASPECTS IN THE MIDWIFE’S EDUCATION IN RELATION TO

SEXOLOGICAL CONSULTING Elena Mihaylova, Teodora Momchilova, Ivanka Dimitrova, Diruhi Amudjyan, Sofka Kutryanska, Boris Kuzmanov Department of Midwifery Care, Faculty of Public Health, Medical University - Plovdiv, Bulgaria Sexual counselling of young people is an issue of increasing popularity and interest, for the number of young people that encounter problems in their sexual development is on the rise. The problems are mostly related to the young people’s inability to define their sexual identity and orientation. The present study included 77 students in the major specialty of Nurse Midwifery using a direct anonymous survey. Purpose: The aim of the study was to determine the level of knowledge of these future professionals about the problems in sexual counselling and their willingness to work as counseling specialists. METHODS: Sociological survey, statistical analysis.


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R ESULTS: The survey showed that 66.23% of the respondents are not familiar with the term “sexual counselling”. Similar percentage of respondents was ignorant of the tasks and procedures applied in sexual counselling. Fifty-five per cent of the respondents declared their willingness to works as counsellors in a team, but felt they were insufficiently prepared for the work. Almost 100% of the respondents believe that the special training should be done within the current study schedule in family planning and sexology, or medical psychology or may be included as an elective course. CONCLUSION: We propose inclusion of a sexual counselling training course within the frames of the major of sexology and family planning on the grounds that there is a high percentage of students that are ignorant of these problems. Key words: sexual counselling, education

PH 17.

RESEARCHING ELDERLY PEOPLE’S NEEDS OF MEDICAL ASSISTANCE

Diana Paskaleva1, Maria Semerdjieva2, Stoika Tufkova3 1Department of Nursing, 2Department of Health Care, Medical University - Plovdiv, 3Department of Toxicology, St George University Hospital, Plovdiv, Bulgaria One of the most important features of the past 20th century was an increase in the absolute number and proportion of older and elderly people. This requires society to pay attention at the needs of medical assistance to them. AIM: The need of medical assistance for older and elderly people. MATERIALS AND METHODS: The object of the study were 94 patients. The information is collected from Surgical Department, Neurology Department and Internal Care Department of HAT* “Plovdiv” between 20.04.2014 and 25.07.2014. The study was conducted as an anonymous questionnaire. Variance and non-parametric analyses were used to analyse data statistically. R ESULTS AND DISCUSSION: 93.6% of respondents were looking for ambulatory medical assistance over the last year (mean 3.36 ± 0.23 times). 70.2% needed specialized medical care and 80.9% were hospitalized (mean 1.42 ± 0.07 times). The least wanted were home visits by a doctor (17%) and by a nurse (6.4%). Adults over 71 were hospitalized at average 1.64 times, while those over 60, only 1.0 times. The same holds true for the demand for Outpatient care. CONCLUSION: While getting older, the need of medical care increased due to the high incidence of chronic and degenerative diseases. This requires providing a quality nursing care for the elderly people. Key words: elderly people, medical assistance, needs

PH 18.

THE ROLE OF NURSES FOR LIMITING TOBACCO SMOKING

Liliana Mancheva, I. Mirtcheva Department of Social Medicine and Organization of Health, Faculty of Public Health, Prof. Paraskev Stoyanov Medical University, Varna, Bulgaria On a global scale more than one billion people are smoking, one of 10 young people are smoking, while 50% of the young people are exposed to tobacco smoke in public places. Nurses from around the world assist the patients in their attempts to quit smoking, participate in promoting the creation of an environment, free from tobacco smoke and in support of effective policies for control of the tobacco smoking. The study aims to clarify the attitudes of the nurses in the country for active participation in the fight to reduce the tobacco smoking among them and among the society as a whole. A double sociological survey was conducted (2007 and 2013), as in this

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summary are presented the results, concerning the health behavior regarding tobacco smoking in medical professionals, working in St. Marina University Hospital, Varna. The results the following: Despite the high result of regular smokers - 42.0% in 2007 and 40.4% in 2013 there is a decrease of 2%, which we consider a result of the ongoing government policy concerning the restriction of tobacco smoking. Nurses have all the insruments to influence patients during the hospital stay, therefore attracting them to the side of the global fight against the tobacco smoking will be a big success for our health system. Prevention of damage from tobacco smoking should be included in the training syllabuses for educational programs for the nurses. Key words: restriction of tobacco smoking, nurses, basic competences, strategy

SOCIAL CHARACTERISTICS AND THEIR EFFECTS ON SELF-RATED

PH 19.

HEALTH IN PERSONS OVER 18 YEARS OF AGE Joana Simeonova, Angelika Velkova, Silvia Tsvetkova, Penka Kostadinova, Stela Georgieva, Mariela Kamburova Medical University - Pleven, Bulgaria Introduction: One of the basic mechanisms through which social inequalities influence health is stratification of people in social classes according to their control over resources. This social stratification results from differences in education, occupation and income, as people from lower social classes are exposed to different health risks and are deprived of ability to control their health. Aim: To study the effect of individual social status on self-rated health. Material and methods: A qualitative study was performed among 90 persons over 18 years of age in 2013. Self-rated health (SRH) and social status (education, income, property ownership, qualification) of the respondents were studied by an interview. Data were processed by SPSS v.19. Non-parametric statistical methods were applied. Results: Most of the participants (47.8%) assessed their health as good. Stratification by some basic social characteristics showed that 5.6% of the respondents had elementary education, 12.4% were unemployed. One third of them existed on monthly income less than 310 BGN, 30% defined their financial resources as insufficient, and 12.2% lived in rented accommodation. Every third person reported a disparity between the current job position and the professional competences they had. Education was the only variable significantly associated with self-rated health – those with elementary education perceived their health as poor more often (р = 0.001). Symmetrical distribution of poor SRH among the groups with lower incomes explained partly the lack of significant difference between groups (p = 0.469). Conclusion: Education was the sole variable significantly associated with self-rated health among all studied social factors. Key words: self-rated health, education, property ownership

PH 20.

STRESS MODERATORS AMONGST MIDWIFE STUDENTS THROUGH

THE PROCESS OF EDUCATION AT THE MEDICAL UNIVERSITY OF PLOVDIV Ivanka Dimitrova, Diruhi Amudjyan, Elena Mihaylova, Sofka Kutryanska, Teodora Momchilova, Boris Kuzmanov Department of Midwifery Care, Faculty of Public Health, Medical University - Plovdiv, Bulgaria Under conditions of restructuring higher education system, the evaluation of stress factors


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and situations through the process of education provide the possibility to create strategies for control and relaxation of stress. PURPOSE: To clarify the specific character of the stress effects amongst the students from the Medical University of Plovdiv through the process of their education and to offer concrete measures for a reduction and prevention of stress. PRESENTATION: The rapid development of higher education, the introduction of new student’s plans and programs into the educational process require that students should adapt rapidly to the specific conditions of education. Not with a lesser significance is the physiological part of the process of education because under a pressure of the adaptation mechanisms, it is possible to get unfavourale results. The performance of special manipulations through the patient’s caring process, the procedure of delivery, as well as during the initial cares for the newborn are manipulations which have turned into routine ones through the course of time but when they take place for the first time, they are related to stress reactions for 69.5% of freshmen. In the indicated situations there was also conducted an open and direct observation in a natural professional environment. METHODS: Sociological; method of observation; a statistic one; a graphical one. CONCLUSION: it becomes clear that medical profession which the students study to get generates stress-genetic factors different in character and impact. Key words: stress, moderators, stress situations, reactions

PH 21.

STUDENTS’ OPINION SURVEY ON REQUIRED PERSONAL SKILLS AND

COMPETENCES OF TEACHERS AT THE MEDICAL COLLEGE Petia Kasnakova Medical College, Medical University - Plovdiv, Bulgaria In relation to improving the quality of training students, opinions are periodically examined and the use of student ratings is expanded, which makes it possible to hear the opinions of students consumers of educational services. The AIM of the study is to examine students opinion on the assessment of the personality of the teacher and the latter’s professional activity. MATERIAL AND METHODS: The survey was conducted with 230 first-year students in the specialties of the Plovdiv Medical College R ESULTS AND DISCUSSION: All interviewed students put professionalism as a primary quality of the teacher (first year - 80.43 ± 2.01%; third year - 94.78 ± 2.72%). The interest in innovations is typical of students, as indicated by the high percentage in the survey (first year - 78.26 ± 2.71%; third year - 89.57 ± 2.02%), thus reflecting the requirement for the teacher to be an erudite, to acquaint the students with the latest innovations and achievements of medical science needed to practice. Communicative skills, as one of the personal qualities required of the teacher, are referred to by the students in the third place (first year - 74.78 ± 2.86%; third year - 90.87 ± 1.90%). All interviewed students indicated that teachers need to constantly nurture love for the profession. CONCLUSION: Students scored the highest the following qualities: professionalism, which spells out as the ability to dispense new practically oriented knowledge, to possess communicative skills so as to show compassion and to know the modern ways of teaching and presentation. Key words: personal skills, competences, teachers

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PH 22.

TECHNIQUES OF HOMEOPATHIC TREATMENT OF INJURIES IN EXPERT

SYSTEMS Zhivko Peychev Department of Medical Informatics, Biostatistics and E-Learning, Faculty of Public Health, Medical University - Plovdiv, Bulgaria Homeopathy enters more widely in the everyday live of people. It is used to treat any kind of trauma: household, sports, surgery, transport, mental trauma, etc. The AIM of this study is to develop an expert system module which provides the physician homeopathic rational choice of medicine according to the type of trauma. MATERIAL AND METHODS: Analysis of drugs of a specialized homeopathic reference book in traumatic conditions was carried out. Database was created with the programming language SQL and Java language was applied to write an algorithm to search for a medication. R ESULTS: According to the location of trauma the following groups were differentiated: medications for the soft tissue injuries (Arnica Montana, Ledum Palustre, for the trauma of bones, teeth and locomotor system (Ruta Graviolens, Rhus Toxicodendron, Symphytum), CNS and psycho trauma (Aconitum Napeus, Natrum Sulfuricum). To enable the programming module to customize the selection of P-drug according to the presence of external bleeding, bruises, broken bones, hyperextension of tendons and ligaments or damage to the phyche. CONCLUSIONS: The module developed by us helps to find quickly symptomatic homeopathic medication in traumatic conditions and would improve the work of the homeopathic doctor in outpatient care. Key words: trauma, homeopathy, expert systems

PH 23.

THE ATTITUDES OF NURSES TO THE SELECTION OF WORKPLACE -

HOME OR ABROAD Liliana Mancheva, I. Mirtcheva Department of Social Medicine and Organization of Health, Faculty of Public Health, Prof. Paraskev Stoyanov Medical University, Varna, Bulgaria At the beginning of the transformation in our country many young nurses left our hospitals and settled in European Union countries. After the massive outflow of medical doctor in the past years the issue that there is shortage of nurses in our hospitals became slightly forgotten. The AIM of our study is to find what the prevailing attitudes are now to the problem of selecting workplace by nurses. The analysis and the presentation are based on the results from the double sociological survey, conducted in 2007 and 2013 in St. Marina Hospital, Varna. The results suggested that the number of people who are willing to leave the country continues to increase (from 52.0% in 2013 to 56.5% now) which is particularly disturbing. These were more than half of the participants in the survey. From the beginning of the healthcare reform the number of nurses has decreased almost by half. In 2006 the availability of nurses is 2 times lower than the average for the EU. CONCLUSION: The issue of labor remuneration should immediatly be tabled for discussion as this is the main reason for leaving the country. As other reasons people give the opportunities for improving their qualification, improvement of working conditions and the organization of the work in order to reduce the tension in their daily activity. Key words: nurses, leaving the country, labor remuneration, qualification


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PH 24.

THE INTERACTIVITY - NEW HORIZON IN DISASTER MEDICINE

TUTORIAL PROCESS Rostislav Kostadinov, Rumiana Etova, Nikolai Varshilov Department of Epidemiology and Disaster Medicine, Faculty of Public Health, Medical Unviersity - Plovdiv, Bulgaria INTRODUCTION: The complexity of Disaster Medicine subject proved to be a challenge for almost half of the students in Medical University, Plovdiv. Discussing structure and organization of the Medical Support to affected population by various damaging factors with third-year-students is a challenge, because of their scarce medical background. Organizing triage, stabilization and medical evacuation for casualties in ‘exotic’ for the audience conditions results in incomprehensiveness. Contemporary Information Technologies development is creating incredible opportunities for enhancing the comprehensiveness of the Disaster Medicine. The AIM of this study is to present some of the possibilities for Disaster Medicine Tutorial Process improvement by implementation of the newest IT products. MATERIAL AND METHODS: Challenges observed in students comprehension of the Disaster Medicine along with possible solutions are analyzed by applying descriptive and comparative methods. By deductive analysis solutions are linked to the capabilities of the interactive teaching. R ESULTS AND DISCUSSION: One of most frequently revealed shortfall in the Disaster Medicine understanding of the interconnectivity between entire Disaster Medical Support chain activities. By implementing the interactivity in the tutorial process we could demonstrate to the students the results of the wrong assessment impact on the following decision making and activities execution. Giving students the possibility to enter in the disasters harsh atmosphere, looking for medical data, planning and managing medical activities in limited time frame surrounded by dying people, for sure, will increase their comprehensiveness. Key words: disaster medicine, students teaching, interactivity

PH 25.

THE MARKET FOR HEALTH CARE SERVICES - MONOPOLY OR

MONOPOLISTIC COMPETITION Reneta Roseva Department of Finance and Credit, Faculty of Finances, D. A. Tsenov Academy of Economics, Svishtov, Bulgaria In modern economy, an important factor for economic growth is human capital. Health is an integral part of quality of life and has an important role in the economic development of the country. It reveals the need for special demands on the health sector, respectively, the need to increase public satisfaction of medical care. Health is a national priority, but also in times of crisis, a decline in living standards, economic disruption, there is a shortage of public funding, which affects the system of health care. Healthcare system is different from other sectors and because of this the market forces of supply and demand do not have the same impact as in other markets. The health services as a product is fundamentally a public good. It is provided by the state, funding of the costs of the state budget, but is paid by the relevant consumer of the goods. Health care is a product which is marketed, it means that it has a certain value. Value is the price that consumers pay to get the service. In this report, the object of research is health care as a commodity. The goal, which the author sets is to answer the question: ‘Is it pure monopoly the market of health care services or it is a complex combination of different market structures?. Key words: healthcare, pricing policy, market, finance

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PH 26.

THE ROLE OF REHABILITATION FOR IMPROVEMENT OF THE HEALTH

STATUS OF PATIENTS WITH CHRONIC KIDNEY DISEASES Desislava Bakova1, Nonka Mateva2, Deliana Davcheva3, Teodora Dimcheva2 1Department of Health Care Management, Faculty of Public Health, Medical University-Plovdiv; 2Department of Medical Informatics, Biostatistics and E-Learning, Faculty of Public Health, Medical University - Plovdiv; 3Department of Clinical Laboratory, Faculty of Pharmacy, Medical University - Plovdiv, Bulgaria INTRODUCTION: Chronic kidney diseases occur as a huge social and economic problem based on its increasing incidence and frequency. As part of the therapeutic approach rehabilitation contributes to improvement and maintaining of the physical and social status in patients with chronic kidney diseases, for the timely reintegration into the family and work environment, keeping their social role and achieving a high quality of life. The AIM of the present study was to analyse and evaluate the health status of patients with chronic kidney diseases after rehabilitation. MATERIALS AND METHODS: The present study was conducted in the Specialized hospitals for rehabilitation - National complex EAD, in Hissar. The study included 83 patients aged from 20 to 80 years (mean age 58.17 ± 1.28 years). Questionnaire IRES-3 validated for Bulgarian population was used to rate the rehabilitation effect. Assessments were performed at the beginning, at the end and three months after rehabilitation. R ESULTS: We found statistically significant better score for satisfaction with health status at the end and three months after rehabilitation compared to the beginning of rehabilitation. CONCLUSIONS: Rehabilitation has a positive impact on the health status of patients with chronic kidney diseases. Key words: rehabilitation, assessment, chronic kidney diseases

THE ROLE OF ROYAL JELLY IN HUMAN HEALTH

PH 27.

Rositca Stancheva, St. Porozhanova Department of Hygiene, Medical University - Varna, Bulgaria Royal jelly is a bee product, unique in its composition and biological activity. Its proven biochemical properties attract the attention of researchers in recent years. Bee products including royal jelly are valuable components of modern pharmaceutical industry. Either used as food or food supplements, they promote health and longevity, increasing the chances for adaptation. The purpose of this review is to focus on the biological activity of royal jelly and its role in human health. The review is made referring to publications found in Science Direct and Bio Med Central. Key words: royal jelly, biological activity, human health

PH 28.

THE SIGNIFICANCE OF SCHOOL HEALTH SERVICES AS AN ELEMENT OF

SCHOOL HEALTH PROMOTION Stela Georgieva, Marieta Kamburova, Yoana Simeonova, Silvia Aleksandrova-Yankulovska Department of Public-Health Sciences, Faculty of Public Health, Medical University - Pleven, Bulgaria INTRODUCTION: School health services are an essential element of school health promotion. The school-based model has the greatest potential to achieve best health outcomes because various functions of school health professionals, mostly nurses, are fully targeted to the school environ-


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ment and its influence on health. The aim of this paper is to examine the organization of school health services in Pleven region and their importance in the overall process of school health promotion. MATERIALS AND METHODS: A comprehensive study of health promotion at school in Pleven region was conducted. Data were collected by self-administered questionnaire of school health professionals and health promotion experts, interview with school principals and documents review. R ESULTS: In ¾ of schools there are school health offices served by nurse and equipped according to the requirements. Over 80% of the respondents believe that nurses have competencies to deal with their diverse responsibilities. Difficulties in studying risk factors distribution among students were pointed out by 64% of the respondents. Special attention for children with high medico-social risks was reported by 56.3% of nurses. Parents discuss existing health problems of children mainly with the class teacher, only in 3.8% with school nurse. More than a half of medical staff is not satisfied with family-school collaboration but doesn’t take actions to improve it. CONCLUSION: School-based model of health services for students and school staff is leading in our country. This is favorable to achieve the health promotion goals if efforts are made to overcome the pointed difficulties and limitations. Key words: school health services, health promotion

THE PROBLEM OF PROFESSIONAL STRESS IN MEDICAL SPECIALISTS

PH 29.

FROM ST. ANNA MULTIPROFILE HOSPITAL FOR ACTIVE TREATMENT, VARNA AND SPECIALIZED HOSPITAL OF OBSTETRICS AND GYNECOLOGY, VARNA Liliana Mancheva, S. Toncheva Department of Social Medicine and Organization of Health, Faculty of Public Health, Prof. Paraskev Stoyanov Medical University, Varna, Bulgaria Occupational stress is in the focus of researchers as the subjects of the research are the workers in different areas. Modern requirements for safety in the workplace oblige the employers to detect and eliminate any harmful factors in the working environment, which include stress. The purpose of this study is determining the effect of new labor conditions on medical specialists working at both hospitals, the level of occupational stress and a comparison of its parameters. A questionnaire survey was conducted of the health status of nurses in St. Anna Hospital in Varna and the maternity nurses in Obstetrics and Gymecology Hospital, Varna, divided in two groups - surgery nurses and general nurses. Conclusions were made about the causes for the tension and stress in the daily routine. The profile of the department, where the medical specialists work exerts a negative impact on the health of the nurses to a greater extent than the effect of stress on the health of the maternity nurses. The negative assessment according to the self-assessment of the mental and physical health of the nurses is in a higher extent than the self-assessment of the maternity nurses. Key words: nurse, maternity nurse, working environment, stress, health

PH 30.

USE OF ARTIFICIAL NEURAL NETWORKS IN OUTCOME PREDICTION OF

PATIENTS WITH CHRONIC LYMPHOCYTIC LEUKEMIA Nonka Mateva Department of Medical Informatics, Biostatistics and E-Learning, Faculty of Public Health, Medical University - Plovdiv, Bulgaria Chronic lymphocytic leukemia (CLL) is characterized by remarkable variability in clinical

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presentation, course and prognosis. Many clinical and pathological variables affect patient prognosis, which in turn influences the treatment and surveillance of patients. This underlines the importance of prognostic modeling and outcome prediction. Recently, techniques such as artificial neural networks (ANNs) have become popular for medical classification decisions. This is largely due to their modeling flexibility in that they can detect certain predictor variable relationships (e.g. interactions, nonlinearities). The AIM of the present study is to construct an ANNs model for outcome prediction in patient with CLL. MATERIALS AND METHODS: We used a database of 131 patients with CLL followed up and treated in Clinic of Haemathology in Medical University, Plovdiv. In our study we built a Multylayer perception (MLP) network by using IBM SPSS Neural Networks software. As a measure of predictive ability the area under the receiver operating characteristic curve is computed. R ESULTS: The ANNs were trained using the data from 87 randomly selected patients (2/3 of original data base). As the input layer we used a set of six characteristics (ECOG performance status, Rai clinical stage, extranodal involvement, mediastinal lymphadenopaty, FAB morphological type, abdominal lymph nodes) and the outcome as an output layer. Overall prediction accuracy for the training and testing samples was 76.9 % and 83.3 %, respectively. CONCLUSION: This study demonstrates that ANNs is a potentially useful mathematical tool for predicting outcome of patients with CLL with high prediction accuracy. Key words: ANNs, CLL, outcome prediction

PH 31.

WATER BIRTH - RISK OR ALTERNATIVE

Milena Sandeva, Violeta Paskaleva Department of Obstetrics and Gynecology, St. George University Hospital, Medical University - Plovdiv, Bulgaria INTRODUCTION: Birth in water is widely practiced throughout the world and is quite popular in this country. Previous criticism of giving birth in water is based on assumptions that have no bearing to today’s level of knowledge of fetal physiology and newborn adaptation. AIM: The aim of the study was to eliminate the risks for mother and child during birth in water. MATERIALS AND METHODS: This prospective comparative case-control study included 140 mothers (age range 26-33 yrs) and newborns divided into two groups: water birth group - 70 women who gave birth in a bathtub, control group - 70 women that gave birth in the conventional way. R ESULTS: 31.5% of the women giving birth in water stayed in the bathtub less than an hour, and 33.9% were there between 1 and 2 hours, and two thirds of the women in this group stayed in water for more than 2 hours. Pain relief was demanded differently by the two groups – 1.1% of water-birth women and 20.1% of the women with spontaneous delivery. Episiotomy was performed in 19% of women giving birth in water versus 38% for women with conventional delivery. Concerns that placental bleeding may occur more frequently because of the warm water were not justified. The fetal parameters in both groups were similar. No infant or woman in labor died in either groups. CONCLUSION: The results give a good ground to support the hypothesis that water birth is a good alternative to the spontaneous birth delivery out of water; it poses no additional risks for mother and infant. Key words: birth, risk, adaptation, fetal parameters


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Therapy TH 1.

ISCHEMIC HEART DISEASE AS A RISK FACTOR FOR ISCHEMIC STROKE

Daniela Arabadzhieva1, Zhaneta Georgieva1, Ara Kaprelyan1, Zdravko Slavov2 1Department of Neurology, Professor Paraskev Stoyanov Medical University, Varna, 2Chernorizets Hrabar Free University, Varna, Bulgaria Recent research worldwide demonstrates a mutual relationship between the cardiovascular and cerebrovascular risk. The purpose of this investigation was to reveal the association between ischemic heart disease (IHD) as a particularly common cardiovascular disease (CVD) and acute ischemic stroke (AIS). A total of 258 AIS patients at a mean age of 70.59 ± 7.22 yrs were followed up. They were hospitalized in the Department of Neurology, Medical University of Varna, between 2007 and 2013. The comorbidity of eight CVDs and pathological conditions such as IHD, myocardial infarction (MI), coronary atherosclerosis (CA), heart failure (HF), effort angina pectoris (AP), ventricular tachyarrhythmia (VTA), atrial fibrillation (AF), arterial hypertension and hypertensive heart was analyzed. Data were statistically processed by means of variance, correlation (Pearson’s coefficient) and cluster analyses. IHD was diagnosed in 154 AIS patients (in 59.59% of the cases). There was predominantly weak or moderately positive correlation between these diseases among AIS patients such as r = 0.367 - between IDH and CA; r = 0.246 - between IDH and HF; r = 0.243 - between IDH and MI, etc. There was a considerable correlation between IHD and AP (r = 0.667). The cluster analysis revealed the closest proximity between the following three CVDs among AIS patients: AF, VTA, and MI. Single risk CVDs on their own account were not the only one and reliable criterion for the extent of risk for AIS. Their presence should comprehensively be interpreted with the concrete patient and be taken into consideration during the individualized diagnostic and therapeutic behaviour. Key words: ischemic stroke, cluster analysis

TH 2.

RISK PROFILE IN YOUNG SUBJECTS WITH HIGH NORMAL ARTERIAL

BLOOD PRESSURE Julia Nikolova, M. Bacelova, P. Hrischev, M. Hadjieva, P. Nikolov, D. Koleva Department of Physiology, Medical University - Plovdiv, Bulgaria According to the classification of JNC-7 (Joint National Committee) – USA and ESC-2013 prehypertension (PH) and high normal arterial blood pressure (ABP) are categories between normal and pathological values. PH is not only a state of increased ABP, but endothelial activation and metabolic alterations. Aim: To be followed up the risk profile and some hemodynamic indices in young subjects with high-normal ABP. Material and methods: The study was carried up with 51 individuals at mean age of 23 ± 1.3 yrs: 28 with optimal ABP (systolic BP ≤ 120 mm Hg and diastolic BP ≤ 80 mmHg) and 23 with high-normal ABP (systolic 130-139 mm Hg and diastolic 8589 mm Hg). ABP was measured considering the rules using CNAP®”HD” technology. Inquiry method was applied to check the prevailing risk factors. Body mass index (BMI) was followed by TANITA BC – 420. The data were processed by paired t-test. Results: The group with optimal ABP was presented by: systolic (S) ABP: 115.76 ± 3.7 mm Hg, diastolic (D) ABP: 72.80 ± 6.24 mm Hg, pulse pressure (PP): 42.96 ± 4.35 mm Hg, mean (M) ABP: 87.12 ± 3.13 mm Hg, heart rate (HR): 90.7 ± 11. In high-normal ABP individuals the data were: SABP: 132.84 ± 4.7 mm Hg,

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DABP: 86.44 ± 2.5 mm Hg, PP: 46.4 ± 2.2 mm Hg, MABP: 101.9 ± 2.34 mm Hg, HR: 93.7 ± 16.4. Conclusion: 42.6% from the followed-up individuals had high-normal ABP and hypertension I degree and 14.8% - with overweight. In smokers: 61.5% were with high normal ABP and 15.3% - with I degree obesity. 36.5% were the non smokers with high normal ABP and 14.3% – with increased BMI - I degree obesity. The possibility hypertension to be delayed would be concerned with preventive non medical means application. Key words: high-normal ABP, smoking, BMI

TH 3.

AMBULATORY BLOOD PRESSURE MONITORING: ADDITIONAL ANALYSIS

Nina Belova Department of Physiology, Medical Faculty, Medical University - Sofia, Bulgaria AIM:. This study was aimed at utilizating ambulatory blood pressure monitoring for evaluation of arterial stiffness and blood pressure variability. MATERIALS AND METHODS: The 25 studied young and normotensive individuals were divided into two groups: Н(+) – subjects with family history based elevated cardiovascular risk and Н(-) – their age matched controls. Ambulatory blood pressure was recorded by means of the Schiller MT-300 Holter-BPR device. In addition to the conventional parameters, the ambulatory index of arterial stiffness (AASI), based on the correlation between the diastolic and systolic values was calculated. The 24-hour blood pressure variability was assessed using the standard deviation SD and the weighted SD. The double product as a measure of myocardial stress was calculated as well. R ESULTS: According to the current guidelines all studied individuals were normotensive. However, the Н(+) individuals showed significantly higher blood pressure values as compared to Н(-), especially during the active part of the day (the statistical significance varied between P < 0.05 and 0.001). AASI was also higher in Н(+) – 0.74 as compared to 0.54 in Н(-) (P < 0.04). The weighted SD and the double product were also significantly higher in Н(+). All these differences could be interpreted as a manifestation of the higher sympathetic outflow to the cardiovascular system in the individuals with family history based elevated cardiovascular risk. CONCLUSION: The additional analysis of the data from the ambulatory blood pressure monitoring (ABPM) is informative and might be utilized for evaluation of the arterial wall state and the autonomic cardiac function. Key words: ABPM, arterial stiffness, arterial pressure variability

TH 4.

APNEAGRAPH - A NEW MODERN DIAGNOSTIC TOOL FOR OBSTRUCTIVE

SLEEP APNEA AND SNORING Mario Milkov, Plamen Nedev, Tzvetan Tonchev, Hristijan Madzhova, Georgi Iliev, G. Marinov Department of Prosthetic Dentistry and Orthodontics, Faculty of Dental Medicine, Prof. Paraskev Stoyanov Medical University, Varna, Bulgaria ApneaGraph provides an ambulatory sleep study with localization of obstruction for snoring, obstructive sleep apnea (OSA) and OSA-related breathing disorders. It is an innovative sleep analysis tool and the only investigation that combines polysomnography with nasal-pharyngeal pressure recording. It efficiently identifies the site of obstruction and detects the OSA type (obstructive or central) indicating the most appropriate surgical procedure or conservative therapy modalities. The principle features of ApneaGraph include: sleep study validated against full poly-


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somnography; localization of obstruction level during natural sleep to improve diagnosis and surgical outcomes; it does not require a sleep technician for set up; it does not require a sleep physician for data interpretation, it is performed under ambulatory conditions and is more cost effective than conventional polysomnography. Our experience with ApneaGraph was based on the examination of seven OSA patients. They were six males and one female aged between 43 and 62 years (mean age 48 ± 9.2 years). The computerized analysis of the constellation of physiological parameters allowed the diagnosis of a central OSA type in one patient and of an obstructive one - in the remaining six patients. The precise identification of the localization of the reason for OSA enabled the most correct choice of conservative management of these patients. Key words: ApneaGraph, polysomnography, obstructive sleep apnea, snoring, diagnosis

TH 5.

STATINS AS THE MAIN THERAPY IN PREVENTION OF CARDIOVASCULAR

DISEASE IN THE ELDERLY Idowu Adebayo, Dobrin Vasilev, Jivka Stoykova, Kiril Karamfiloff Clinic of Cardiology, Aleksandrovska University Hospital, Sofia, Bulgaria OBJECTIVES: The purpose of the paper is to assess if Statins reduce mortality and cardiovascular events in elderly people without established cardiovascular disease BACKGROUND: Because of the progressive aging of the population and the fact that cardiovascular diseases rank top as the leading cause of death in the elderly. It is apparent that prevention of cardiovascular diseases in the elderly is really important. According to the guidelines on the treatment of cardiovascular diseases, the use of statins is strongly recommended. Whereas the recommendation on the use of statins to prevent cardiovascular diseases in the elderly without previous cardiovascular events is ambiguous. METHODS: Randomized trials comparing statins versus placebo and reporting all cause and CV mortality, myocardial infarction and stroke onset in elderly subjects (age ≥ 65 years without previous CV diseases were included in the trials). R ESULTS: 5 trials enrolling 100 subjects (45% females; mean age 67 ± 2.9 years, mean follow-up 1.5 ± 0.5 years) were included in the analyses. Statins compared with placebo, significantly reduced the risk of MI by 39.4% (relative risk [RR] 0.606 [95% confidence interval (CI): 0.434 to 0.847] P = 0.003) and the risk of stroke by 23.8% (RR: 0.762 [95% CI: 0.626 to 0.926]: P = 0.006) In contrast, the risk of all cause death (RR: 0.941 [95% CI: 0.856 to 1.035]: P = 0.210 and of CV death (RR: 0.907 [95% CI: 0.686 to 1.199]: P = 0.493) were not significantly reduced. CONCLUSION: statins significantly reduce the incidence of MI and stroke. Key words: statins, cardiovascular disease

TH 6.

SHOULD WE INCLUDE BRAIN IN THE SCANNED FIELD OF THE WHOLE-

BODY 18F-FDG PET/CT? Dora Zlatareva1, I. Kostadinova2, M. Garcheva2, T. Petrov2, V. Hadjiiska2, V. Hadjidekov1 1Department of Diagnostic Imaging, Medical University - Sofia, Alexandrovska University Hospital, Bulgaria; 2Department of Nuclear Medicine, Alexandrovska University Hospital, Sofia, Bulgaria PURPOSE: Whole-body 18F-FDG PET/CT is widely used in oncology for diagnosis and staging. No standardized field of view (FOV) currently exists for whole-body FDG PET/CT. The aim of this study is to assess the utility of including brain in the scanned field of the whole-body 18FFDG PET/CT. We demonstrate the brain findings in patients with non-central nervous system malignancy.

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METHODS AND MATERIALS: A total of 4739 patients with non-central nervous system malignancy were studied between October 2011 and November 2014. The patients were scanned on GE Discovery 16 slices scanner from the top of the head to mid-thigh one hour after administration of 5MBq/kg 18F-FDG. All images were interpreted by nuclear medicine physician and radiologist with experience in oncologic imaging. Fifty-seven patients out of 4739 patients (1.2%) had pathological brain PET/CT findings which were retrospectively analyzed. Thirty-one patients out of 57 were female and 26 were male. The mean age was 58.5 years (range 35-86 yrs). R ESULTS: The most common abnormalities are brain metastases (14 patients), postoperative findings without metabolic activity (12 patients), residual formation or recurrence (5 patients), meningeoma (2 patients), extra-axial metastases (5 patients), cerebrovascular accidents in chronic stage (16 patients) and arachnoid cyst (3 patients). Detecting new metastatic lesions changed management in 19 patients. The most common primary malignancy was lung carcinoma (26 patients), breast carcinoma (14) and melanoma (7). CONCLUSION: Including the brain in the scanned field of the whole-body PET/CT may provide crucial information which is important for patient’s management. Key words: PET/CT, oncology, brain findings

TH 7.

ТHE SINGLE NUCLEOTIDE POLYMORPHISM IN CYP2C8 IS ASSOCIATED

WITH MYOCARDIAL INFARCTION IN BULGARIAN POPULATION WITH CARDIOVASCULAR RISK PROFILE Galya Atanasova1, Penka Stefanova2, Anna Atanasova3, Stefan Kostadinov4 1Department of Propaedeutics of Internal Diseases, University Hospital; 2Department of Social and Preventive Medicine, Medical Statistics, Pedagogy and Psychology, Medical University - Pleven, Regional Health Insurance Fund - Pleven; 3Medical University - Pleven, Bulgaria In the present study we analyzed the impact of a genetic variant in CYP2C8*3 and on myocardial infarction (MI) in Bulgarian population. We conducted a case-control study to determine whether the common genetic variation rs890293 (CYP2J2*7) in CYP2J2 gene was associated with the risk of MI. The study included 99 patients with MI and 377 population control subjects. To determine the genotypes of the samples real time PCR with predesigned. TaqMan SNP Genotyping Assays (Applied Biosystem) was used. The deviation of allele polymorphism CYP2C8 * 3 on the balance of Hardy-Weinberg and the frequency of the T allele with χ2 test was studied. The rare allele CYP2C8*3 was found in 16% of the affected and 10% of the non affected cases and it showed statistical significance [OR (95% CI): 1.64 (1.00-2.56), P = 0.03]. The CYP2C8*1/*1 genotype occured more often in the control group compared with the patients with MI [OR (95% CI): 0.58 (0.35-0.96), P = 0.039]. The heterozygous genotype of CYP2C8 was found to be significantly associated with a risk of myocardial infarction [OR (95% CI): 2.25 (1.06-4.75), P = 0.036] in women. Possession of the rare genetic variant rs1050968 in CYP2C8 gene is associated with a modestly increased risk of MI in Bulgarian population. Key words: genotype, risk, myocardial infarction


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TH 8.


ANTHRAX IN BULGARIA - AN OVERVIEW OF THE PROBLEM FOR THE

PAST 70 YEARS Iskra Tomova, Rumiana Nenova, Vladimir Tolchkov, Todor Kantardjiev Department of Microbiology, National Center of Infectious and Parasitic Diseases, Sofia, Bulgaria Anthrax is a zoonotic infection known since antiquity and still actual in the 21st century. The continuing importance of this disease is due to: the severity of some of its classic clinical forms; the recent description of outbreaks with a new clinical presentation and high mortality among injection heroin users; the importance of Bacullus anthracis as a bioterrorist agent. On this background, the evolution of anthrax in Bulgaria for the past 70 years will be presented. The review focuses on the scientific and practical achievements which led to overpower and limitation of the disease in humans and animals and removal of the majority natural foci of the disease. As a result only single cases in certain regions were registered in the country over the last decade. Key words: anthrax, epizootic, epidemy

TH 9.

LEGIONELLA CONTAMINATION IN THREE HOSPITAL DEPARTMENTS

IN ONE YEAR Iskra Tomova1, V. Stoeva2, R. Nenova1, Y. Stoilova2 1NRL High Medical Risk Infections, National Centre of Infectious and Parasitic Diseases, Sofia; 2Department of Epidemiology and Disaster Medicine, Medical University - Plovdiv, Bulgaria INTRODUCTION: In Europe, mortality from hospital-acquired Legionnaires disease (HALD) for 2013 is 32%, which significantly exceeds that from the community-acquired form (10%). In Bulgaria there are scarce, snapshot reports for the presence of Legionella in hospitals. AIM: Investigation of contamination with legionellae in three hospital departments over 1 year. METHODS: Water samples from various sources, related to care for patients at risk from inhalation and/or aspiration, were taken quarterly and cultured for legionellae. R ESULTS: A total of 60 samples were examined. Thirty-nine were positive (+ 65%), distributed as follows: 16 samples from 4 taps (+ 68.75%, with concentration 101-103 cfu/ L-1), 16 samples from 4 showers in patients’ rooms (+ 87.5%, 101-103 cfu/ L-1), 12 samples from 2 therapeutic tubs (+ 66.6%, 102- >105 cfu/ L-1), 17 samples from dH2O for medical use (+ 35%, 101-102 cfu/ L-1). Variations in the level of contamination can not be connected with the seasons, but rather with the practices of maintenance and use of the water devices. The highest legionella concentrations were found in samples from “dead legs”. There were no positive results from dH2O transport/storage containers, but the investigation found that medical equipment was occasionally rinsed under the tap before recharging with dH2O, and though exceptionally - filled with tap water. CONCLUSION: There is a need for staff training and implementation of HALD prevention program by applying Legionella contamination control methods, appropriate for the different water devices and medical equipment. Key words: Legionella, hospital-acquired Legionnaires

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TH 10.

MORTALITY IN HIV/AIDS PATIENTS MONITORED AT THE CLINIC OF

INFECTIOUS DISEASES, ST GEORGE UNIVERSITY HOSPITAL, PLOVDIV Mariana Stoycheva1, A. Dineva2, V. Georgieva1, T. Popova2, M. Atanassova3, N. Vatev4 1Department of Infectious Diseases and Parasitology, Medical University - Plovdiv,2 Clinic of Infectious Diseases, St. George University Hospital, Plovdiv, Bulgaria, 3Department of Microbiology and Immunology, Medical University - Plovdiv, 4Department of Epidemiology and Disaster Medicine, Faculty of Public Health, Medical University - Plovdiv, Bulgaria The introduction of combined antiretroviral therapy (cART) led to dramatical decrease in the incidence of death in HIV positive patients. However, it continues to be high, especially in some patients’ groups. The AIM of the study was to investigate mortality rate in HIV/AIDS patients, the most important causes and predictors of death, with a view to earlier diagnosis and adequate therapy. MATERIALS AND METHODS: The study involved 53 deceased patients with AIDS, who had been treated at the Clinic of Infectious Diseases, St George University Hospital, Plovdiv, between 1st Jan 2010 and 31st Dec 2014. Methods of clinical observation, laboratory, microbiological, serological tests (HCV, HBV, toxoplasmosis), ELISA, PCR were used. R ESULTS: There were 53 deaths among 316 HIV+ patients monitored at the Clinic, during the study period. Lethality rate was 16.7% in the entire cohort. It was significantly higher in patients with coinfections, as follows HIV + M tuberculosis - 46%, and HIV+ HCV - 17.8%. Lethality rate was compared between different patients groups, according to age, gender, level of immunodeficiency (CD4+, viral load-VL), and cART. CONCLUSIONS: The lethal outcome in our HIV+ patients was associated with coinfections with M tuberculosis and HCV. Predictors of fatal outcome were pulmonary tuberculosis and advanced immune deficiency with VL > 500 000 c/μl and CD4+ < 100/mm3, and absence or irregular cАРТ. Key words: HIV/AIDS, mortality, coinfections

TH 11.

PLACE OF THE NATIONAL REFERENCE LABORATORY OF PARASITOLOGY

IN SURVEILLANCE AND CONTROL OF PARASITIC DISEASES Diana Jordanova, I. Rainova, R. Harizanov, I. Marinova, I. Kaftanfjiev, I. Bikov Department of Parasitology and Tropical Medicine, National Center of Infectious and Parasitic Diseases, Sofia, Bulgaria Parasitic diseases are subject to the general epidemiological patterns, but also have a number of features such as mass distribution, steady level of long-lasting prevalence and ability of selfregulation in the endemic foci. Surveillance is aimed not only at the diseases but also at the abiotic and biotic environmental factors involved in the transmission. With some parasitic diseases the absence of exact specific etiological diagnosis and therapy may lead to a lethal outcome. In the supervision and control of parasitic diseases in the country participate the structures of RHI at local level and the National Reference Laboratory of Parasitology at national level. After the healthcare reform in the country were established many private diagnostic facilities and the diagnostic work of RHI was greatly reduced. This event motivated the need to create a National Reference Laboratory of Parasitology. Its status was established by Order 09-415 / 04.06.2001 of the Ministry of Health. Activity of the NRL is directly related to the quality assurance of parasitological diagnosis in the country, ensuring the implementation of inter-laboratory comparison tests in private diagnostic facilities, Municipality Hospitals and University Hospitals. The aim of this study was to systematize the activities of the NRL in the system of surveillance


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and control of parasitic diseases in terms of diagnostic abilities, analysis of their incidence, training of graduate and doctoral students, preparation of strategies for reduction of the local parasitic diseases and prevention of disease import from endemic countries. Key words: parasitic diseases, surveillance, control

TH 12.

SEROLOGICAL DATA FOR THE SPREAD OF FOUR HEPATOTROPIC

VIRUSES (A, B, C, E) IN THE PLOVDIV REGION Ani Kevorkyan1, Pavel Teoharov2, Ralitsa Raycheva3, Yordanka Stoilova1, Nedyalka Petrova3 1Department of Epidemiology and Disaster Medicine, Faculty of Public Health, Medical University - Plovdiv; 2National Reference Laboratory of Viral Hepatitis, National Center of Infectious and Parasitic Diseases, Sofia, 3Department of Social Medicine and Public Health, Faculty of Public Health, Medical University - Plovdiv, Bulgaria INTRODUCTION: The globally distributed viral hepatitis A, B, C and E are distinguished by a number of epidemiological characteristics (sources, mechanisms and routes of transmission, risk groups, demographic structure of the infected, etc.). OBJECTIVE: To determine the epidemic size distribution by conducting seroprevalence study on the spread of the four main hepatotropic viruses in the second largest administrative region of the country. MATERIALS AND METHODS: Based on a wide age range the estimation of the overall and age specific prevalence of hepatitis A, B, C and E was performed by testing 705 serum samples for antiHAV-IgG, 865 for HBsAg and anti-HCV and 741 for anti-HEV-IgG. The study was conducted at the National Reference laboratory of Viral Hepatitis, NCIPD Sofia. R ESULTS: Anti-HAV-IgG was proved in 46.5%; HBsAg in 3.9%; anti-HCV in 0.7%; and antiHEV-IgG in 9.04% of all respondents. Hepatitis A and hepatitis E established a clear trend of increasing effectiveness of the immune response with age particularly in the latter stages of life. CONCLUSION: Population-based seroprevalence studies provide important epidemiological data on: susceptible groups; potential for future outbreaks; surveillance, prevention, and control activities, as well as a basis for comparison in future studies. Key words: viral hepatitis, distribution, age-specific prevalence

TH 13.

RESULTS OF THE EUROMELANOMA SKIN CANCER PREVENTION

CAMPAIGN IN THE UNIVERSITY CLINIC OF DERMATOLOGY – PLOVDIV (2011-2014) Hristo Dobrev, Desislava Nocheva Department of Dermatology and Venereology, Medical University - Plovdiv, Bulgaia BACKGROUND: Euromelanoma is a skin cancer screening campaign that started in 1999 in Belgium. Since 2000, it is called ‘Euromelanoma Day’ and is organized annually by the European Academy of Dermatology and Venereology in many European countries. In the year 2006, the Bulgarian Dermatological Society initiated the campaign in Bulgaria. OBJECTIVES: To analyze the results of the ‘Euromelanoma Day’ in the Clinic of Dermatology of Plovdiv during the period 2011-2014. MATERIAL AND METHODS: Everyone who requested an appointment has been examined free of charge. Euromelanoma questionnaire, clinical examination, clinical photos and dermatoscopical investigation were used in order to collect demographic, epidemiological and clinical data, and to detect suspicious lesions. R ESULTS: A total of 126 subjects (47 male, 79 fe-

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male, average age 49 years) was examined. Most of them were of skin phototype II and III (80%). Sunburn during childhood was reported in 51%, while family history of melanoma was reported in 3.4% of participants. A total of 345 target lesions was determined and photo documented. 314 lesions were additionally studied using videodermatoscopy. There were 44% non-pigment, 54% pigment lesions, and 2% skin lesions suspected of being melanoma (2) and basal cell carcinoma (5). The most frequent non-pigment lesion was seborrheic keratosis (24%), while the most frequent pigment lesion was dermal nevus (21%). The incidence of atypical nevi was 6%. CONCLUSION: The campaign ‘Euromelanoma Day’ showed relatively low detection rate of melanoma and non-melanoma skin cancers, however, this rate is comparable with the established incidence of these disorders among Bulgarian population. Key words: skin cancer, dermatoscopy

TH 14.

SEVERE VIRAL HEPATITIS E IN PATIENTS WITH PRIOR LIVER DAMAGE

(ACUTE-ON-CHRONIC LIVER FAILURE) Andrey Petrov1, M. Stoycheva1, N. Vatev2, I. Boev1, M. Atanasova3 1Department of Infectious Diseases, Parasitology and Tropical Medicine, Faculty of Medicine, 2Department of Epidemiology and Disaster Medicine, Faculty of Public Health, 3Department of Microbiology and Immunology, Medical University, Plovdiv, Bulgaria INTRODUCTION: The course of hepatitis E viral infection varies widely from asymptomatic to moderate and to severe forms. A number of patients develop hepatitis with a risk of acute liver failure. A chronic clinical course is possible in immunosuppressed individuals. AIM: The aim of this study was to investigate cases of severe viral hepatitis E (VHE) in patients with prior damage of the liver. MATERIALS AND METHODS: The study included all patients hospitalized between January 1, 2013 and December 31 2015 with acute viral hepatitis. Various methods of clinical monitoring, laboratory, biochemical tests, sonography and CT were used. ELISA was performed in the Department of Immunology and Microbiology. R ESULTS: Over the study period 255 patients with acute viral hepatitis were hospitalized: 86 (33.7%) with HAV, 41 (16.1%) with HBV, 30 (11.8%) with HCV, and 77 patients (33.2%) with unspecified viral hepatitis. The cases of VHE were 21 (8.2%, 17 men and 4 women, mean age 54 yrs, range 30 - 77 yrs). Acute-on-chronic liver failure was found in five patients who showed signs of liver failure with low prothrombin time, hypoalbuminemia, extreme hyperbilirubinemia, ascites, ultrasound evidence, etc. CONCLUSION: Due to the improved genotyping we found that in Bulgaria there is viral hepatitis that is different from the Asian viral hepatitis. A particularly severe course of the disease was observed in cases with prior liver damage. Key words: VHE, liver failure, acute viral hepatitis

TH 15.

INTRODUCTION AND DISTRIBUTION OF HIV-1 SUBTYPES AMONG DRUG

USERS IN BULGARIA (PRELIMINARY ANALYSIS) Anna Gancheva1, Reneta Dimitrova1, Asya Kostadinova1, Danail Beshkov1, Vanya Dafova2, Ivaylo Elenkov3, Mariyana Stoycheva4, Ivailo Alexiev1 1National Center of Infectious and Parasitic Diseases, Sofia, 2 St. Kl. Ohridski University of Sofia, 3Hospital for Infectious and Parasitic Diseases, Sofia, 4Department of Infectious Diseases, Parasitology and Tropical Medicine, Medical University - Plovdiv, Bulgaria OBJECTIVES: The aim of the present study was to analyze the genetic variety of НІV-1 among IDUs in Bulgaria and to evaluate the evolutionary relationship (possible phylogenetic clusters) of Folia Medica 2015; 57; Suppl. 1 © 2015 Medical University, Plovdiv, Bulgaria

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sequences from drug users (IDUs). MATERIALS AND METHODS: The genotype analysis of НІV-1 was performed in 118 (32.9%) out of 359 HIV infected IDUs in Bulgaria. HIV-1 pol sequences were obtained using TruGene DNA Sequencing System. HIV-1 subtypes were analyzed with REGA3 and COMET1.0. The dataset included Bulgarian sequences, reference sequences from Los Alamos and all available CRF01_AE and CRF02_AG sequences from Gene Bank. The phylogenetic analysis was performed using ML method using FastTree2 program. R ESULTS: Phylogenetic analysis reviewed wide diversity of HIV-1 strains found in IDUs. The most prevalent subtypes were two circulating recombinant forms (CRF), CRF02_AG (42.7%) and CRF01_AE (35%), followed by unique recombinant forms, subtypes B and F1 and CRF14_BG. In both CRF01_AE and CRF02_AG trees Bulgarian IDU sequences cluster together representing short evolutionary history comparing to the other non-IDU sequences which were distributed across the tree. The geographic distribution demonstrates two separate independent epidemics among IDUs in Sofia and Plovdiv caused by two different HIV-1 CRFs. CONCLUSIONS: The rapid increase of HIV-1 cases among IDUs has been caused by the introduction of two major CRFs in two different geographical regions in Bulgaria. Evolutionary related clusters demonstrated development of independent local outbreaks with HIV-1 among IDUs. Our results showed that the molecular epidemiological surveillance is of major importance to better understand and control HIV-1 epidemic in Bulgaria. Key words: HIV-1, intravenous drug users, Bulgaria

TH 16.

THE ROLE OF SPECIFIC IGG ANTIBODIES AVIDITY IN THE DIAGNOSIS

OF TOXOPLASMOSIS Iskra Rainova, D. Jordanova, N. Tsvetkova, R. Harizanov, I. Marinova Department of Parasitology and Tropical Medicine, National Center of Infectious and Parasitic Diseases, Sofia, Bulgaria By determining the avidity of specific IgG antibodies in toxoplasmosis we distinguish a recently acquired from a chronic infection both in pregnant women and in suspected patients. In NRL for diagnosis of parasitoses in NCIPD for the period from 2013 to 2014, avidity of specific IgG antibodies was studied by ELISA in 54 patients from a total of 424 tested by clinical indications for the presence of IgG, IgM and IgA toxoplasma antibodies. The results showed a low avidity, or evidence of acute infection in 23 studied patients, most of them positive after serological testing for the presence of IgG, IgM and IgA toxoplasma antibodies. Only in two patients with positive results for IgG and IgM and negative for IgA antibodies we found low avidity. In patients, positive only for the presence of IgG toxoplasma antibodies we found predominantly high avidity or data for latent infection, and only in two of them IgG avidity was borderline. The data collected from our study showed that in most cases determination of the avidity of specific IgG antibodies in toxoplasmosis is a useful additional method for the distinction between acute from chronic infection. Key words: toxoplasmosis, IgG avidity

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ON CONSERVATIVE TREATMENT OF HUMAN HYDATIDOSIS AS

TH 17.

A COMBINATION OF ALBENDAZOLE AND AN IMMUNOMODULATOR (ISOPRINOSIN, RESPIVAX) AND CLINICAL FOLLOW-UP Dimitar Vuchev, Galya Popova-Daskalova Department of Infectious Diseases, Parasitology and Tropical Medicine, Medical University - Plovdiv, Bulgaria The morbidity rate of hydatid diseases showed an increase over the last two decades. The most important clinical aspects are adequate treatment and follow-up after making an acurate diagnosis. Due to introduction of benzimidazole carbamates as a concervative treatment the therapeutic options have expanded. However, immunosuppression also presents in humans with hydatidosis. Combined treatment with an antihelmintic drug and an immunomodulator was used in patients with liver or lung hydatidosis to achieve better clinical effects. The immunomodulators (isoprinosin in liver hydatidosis, respivax in lung hydatidosis) was administered with an antihelmintic drug, albendazole, to three different groups of patients with a hydatid disease: (1) after surgery – to prevent recurrence of liver and lung hydatidosis; (2) accompanying PAIR – to prevent recurrence of liver hydatidosis; (3) as a concervative treatment of inoperable cases, reccurant disease, multiple organ involvement, cyst rupture. After treatment 215 patients were followed up using imaging (US of the liver and X-rays of the lungs) and serological testing over a period of 2-10 years. No side effects of immunomodulators were noted. No recurrences of the disease were detected during the follow-up and clinical symptoms resolved. Most of the patients with lung echinococcosis eliminated the cyst content and the the residual membranes through the sputum, but in some patients cyst degeneration was noted on X-ray. In all patients with liver hydatidosis degeneration signs in the cyst were observed, incl. significant deformation, size reduction and disappearance of hydatid cyst. CONCLUSION: On the basis of the accumulated data it might be concluded that combined with conservative treatment, albendazol and immunomodulators (isoprinosin, respivax) in echinococcosis lead to faster and effective results - for 2-3 months for pulmonary and 6-8 months for liver hydatid disease, and prevention of recurrence after surgery and PAIR (1-2 months). Key words: human hydatidosis, conservative treatment, immunomodulation

TH 18.

COMBINED METHOD FOR DIAGNOSTICS OF ABNORMAL PARATHYROID

GLANDS COMBINING SINGLE-ISOTOPE DUAL-PHASE SCINTIGRAPHY WITH 99MTC – TETROFOSMIN WITH SPECT TECHIQUE Albena Botushanova1, Marianna Yaneva1, Nikolay Botushanov2 1Department of Radiology, St. George University Hospital, Plovdiv, 2Clinic of Endocrinology, St. George University Hospital, Plovdiv, Medical University - Plovdiv, Bulgaria Preoperative parathyroid scintigraphy is highly beneficial for reduction of the duration and extent of surgery in minimal invasive parathyroidectomy which is the commonly used modality nowadays. AIM: To evaluate the diagnostic value of combining single isotope dual-phase scintigraphy with 99mTc -tetrofosmin with SPECT technique.


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MATERIALS AND METHODS: 15 patients (range 38 to 62 yrs) were included in the study. Of these 14 were women and 1 man. 12 patients were with primary hyperparathyroidism and 3 with secondary hyperparathyroidism. Elevated levels of serum calcium and parathyroid hormone were registered in all of them. The examination was conducted on a dual-headed SPECT gamma camera SIMBIA using a protocol for single-isotope dual-phase technique with 99mTc - tetrofosmin. Patients were injected with 740 MBq 99mTc - tetrofosmin, after which planar images were taken at an early phase (20 min) and late phase (120 min). SPECT registration of the neck and upper thorax was performed after the early phase. R ESULTS: Hyperfunctioning parathyroid glands were registered in 10 patients in both phases and on the SPECT images as well. Areas suspicious for abnormal parathyroid glands were visualised in the early SPECT images in 5 patients, and these were not found in the late phase. This may be due to early washout of the radionucleotide. CONCLUSIONS: By combining both techniques we get complementary information for the localization of abnormal parathyroid glands, and do not expose patients to additional radiation. The technique is easy to perform technically. Key words: parathyroid scintigraphy, 99mTc -tetrofosmin, SPECT

TH 19.

SIGNIFICANCE OF ULTRASOUND PHENOMENA - DORSAL ACOUSTIC

ENHANCEMENT AND SHADOWING TO ESTIMATE THE RISK OF MALIGNANCY IN THYROID NODULES Boyan Nonchev1, Nonka Mateva2, Vladimir Danev3, Antoaneta Argatska1 1Division of Endocrinology, Faculty of Medicine, Medical University - Plovdiv, 2Department of Medical Informatics, Biostatistics and E-Learning, Faculty of Public Health, Medical University - Plovdiv, 3Department of General and Clinical Pathology, Faculty of Medicine, Medical University - Plovdiv, Bulgaria INTRODUCTION: The modern clinical approach to the diagnosis of thyroid nodules is based on the results of the ultrasound evaluation in combination with the results of fine-needle aspiration biopsy of the thyroid (FNAB). Ultrasound characteristics associated with malignancy are widely discussed in the scientific literature in order to refine the risk stratification of the nodular formations. Dorsal enhancement and shadowing are acoustic phenomena occurring behind the focal thyroid lesions. They are associated with amplification or attenuation of the ultrasound depending on the nodular tissue characteristics. OBJECTIVE: To evaluate the significance of ultrasound phenomena - dorsal acoustic enhancement and shadowing for the estimation of the risk of malignancy in thyroid nodules. PATIENTS AND METHODS: 270 patients with thyroid nodules were included in the study (males – 21, females – 249, mean age – 49y.). An ultrasound evaluation and FNAB of a total of 329 nodules were performed. Cytological evidence of malignancy was identified in 28 of the studied nodules (in 28 patients) and the diagnosis differentiated thyroid cancer was confirmed histologically. R ESULTS: Dorsal acoustic enhancement was found in 70 nodules and acoustic shadowing showed 32 nodules. The analysis found that the dorsal acoustic shadowing is a predictive sign of malignancy (OR = 8.627; 95% CI - [3.584; 20.770]). It was detected in 11 of 28 malignant nodules. It seems that dorsal acoustic enhancement is a protective characteristics, but due to the small number of malignant nodules possessing this trait (n = 2) definitive conclusions cannot be drawn. CONCLUSION: dorsal acoustic shadowing is an ultrasound feature indicating an increased risk of malignancy and it should be taken into account in the complex ultrasound assessment of the risk profile of thyroid nodules. Key words: thyroid nodules, ultrasound examination

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TH 20.

HIGH FREQUENCY ULTRASONOGRAPHY OF THE SALIVARY GLANDS IN

SJOGREN’S SYNDROME Krasimir Kraev1, Rositsa Karalilova1, Zguro Batalov1, Stefka Peitcheva2, Plamen Todorov1, 1Department of Propaedeutics of Internal Diseases, 2Division of Craniofacial Surgery, 1Medical University - Plovdiv, Bulgaria AIM: Evaluation of the diagnostic and prognostic value of ultrasonography of the salivary glands in the diagnosis of pSS. PATIENTS AND METHODS: 34 patients with pSS (32 women и 2 males, age 52.8 ± 10.4 years) and 46 healthy controls were included in the study. US assessment of the parotid and submandibular salivary glands was done with Esaote MyLab 40 (7-14 MHz linear transducer). The echogene structure and volume of salivary glands were evaluated. The US changes were compared with clinical and histological findings. R ESULTS: US changes were found in 24 of the patients with pSS and in 10 of the control group (р < 0.0001), thus proving sensitivity of 72%, specifity of 81% of this method. CONCLUSION: US of the salivary glands is a reliable, cheap, accessible and fast instrumental method for diagnose of pSS. Matching it with Elastography (another technique which we developed for the evaluation of the density and elasticity of the gland) may replace in future the invasive biopsy and the less used nowadays sialometric methods. Key words: Sjogren`s syndrome, high frequency ultrasonography

TH 21.

VAGINOSES AND ANTECEDENT ANTIMICROBIAL THERAPY

T. Dermendzhiev1, K. Hadzhieva1, S. Stanev1, G. Totev2 1Department of Microbiology and Immunology, Medical University - Plovdiv, 2Clinic of Obstetrics and Gynecology, Pre-Hospital Medical Care, Plovdiv, Bulgaria INTRODUCTION: Vaginal complaints are often a reason to visit the specialised clinics at the prehospital medical care. The interest to the cytolytic vaginosis (CV) and vaginal lactobacillosis (VLB) has increased. Little is known about the causes that lead to overgrowth of vaginal lactobacilli and to appearance of long lactobacilli formations. AIM: To study the relation between vaginoses and the antecedent antimicrobial therapy in women with chronic vaginal complaints. MATERIALS AND METHODS: A total of 280 patients have been studied. The patients have been divided into two groups: 1. experimental - with PAMT (n = 170), and 2. control - without PAMT (n = 110). Research methods used: clinical, microbiological and statistical (variation analysis, logistic regression, graphical analysis). R ESULTS AND DISCUSSION: A study of 63 patients diagnosed with CV established a statistically significant difference between the examined group – 46 (73.02%) and the control group - 17 (26.98%) (p < 0.05). Statistically significant difference between the examined and the control group was found also in cases of VLB. A statistically significant difference between PAMT and womens with bacterial vaginosis, vuvlovaginal candidiasis and aerobic vaginitis has not been established (p > 0.05). CONCLUSION: The study found a statistically significant difference between the patients with cytolytic vaginosis and vaginal lactobacillosis with an antecedent antimicrobial therapy and the control group. Key words: vaginoses, vaginitides, antimicrobial therapy


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TH 22.

ACUTE MEGAKARYOCYTIC LEUKEMIA IN ASSOCIATION WITH

PULMONARY FIBROSIS IN 71-YEAR-OLD WOMAN Katya Sapunarova1, Pencho Georgiev1, Vasko Graklanov2 , Emil Spassov1 , Stefan Goranov1, 1First Department of Internal Diseases, Division of Hematology, Medical University - Plovdiv, 2 St. George University Hospital, Plovdiv, Bulgaria Acute megakaryocytic leukemia (AMKL) was incorporated into the French- American- British (FAB) classification in 1985. The low disease frequency (2% of acute leukemias) reflects the experience of describing clinical spectrum of the disease. Although it is classified as acute leukemia, more than 60% of cases are reported to have previous history of myeloproliferative or myelodysplastic disorder. Literature reports focuse attention mainly on the peripheral blood characteristics, bone marrow findings and genetics of the disease, while data of non-hematopoietic organ involvement are scarce. Clinical case: 71- year-old woman was hospitalized in March 2014 in the division of Clinical Hematology for diagnostic evaluation of a documented peripheral pancytopenia. Bone marrow examination by biopsy shows AMKL with marrow fibrosis (MF) = 2-3, probably with preceding myelodysplastic syndrome. Since 2011 our patient is diagnosed with idiopathic pulmonary fibrosis and was treated with systemic corticosteroids and bronchodilators, without any improvement. Induction chemotherapy for AMKL was started on May 2014. Monthly courses of very low doses alexan (20 mg/m 2 i.v. 5-7 days per month), were the mainstay of therapy, together with supportive and prophylactic treatment. After four months of treatment, the patient’s hematological parameters remained stable, while clinical and imaging techniques documented improvement of pulmonary disease was surprisingly achieved. There are well documented cases in the literature of a relationship between pulmonary fibrosis and idiopathic myelofibrosis, acceleration phase or blast crysis of chronic myelogeneous leukemia and AMKL in Down syndrome. Based on these data, we suggest that in this case there is a correlation between the two disorders in the case reported herein. We discuss the possible hypotheses to explain the common molecular genetic mechanisms. Key words: acute megakaryocytic leukemia pulmonary fibrosis

TH 23.

EXTRAMEDULLARY FORMS OF MULTIPLE MYELOMA - CLINICAL

FEATURES, THERAPEUTIC OUTCOME AND SURVIVAL. A 30-YEAR ANALYSIS Veselina Goranova-Marinova, K. Sotirova, E. Spassov, K. Sapunarova, M. Dimitrova, P. Georgiev, V. Graklanov, V. Babacheva, I. Hristova, St. Goranov First Department of Internal Medicine, Division of Hematology, Medical University - Plovdiv, Bulgaria BACKGROUND: Extramedullar disease (EMD) in multiple myeloma presents in two different forms: ectopic proliferation of plasma cells through destruction of cortical bone and hematogenous dissemination. AIM: to analyse the incidence, therapeutic outcome and survival of patients with EMD over a 30-year period. PATIENTS AND METHODS: 629 patients with multiple myeloma (MM) were analysed for the period 1980 – 2013. EMD was assessed at the diagnosis and in the course of MM. EMD were compared to the rest of MM in the major disease characteristics, therapeutic regimens and survival. Statistics were performed with SPSS v. 18.0. R ESULTS: In 108 (17.2%) patients ЕМD was found. In 67 - at the diagnosis and in 41 - in the evolution of MM. The incidence of EMD significantly increases after year 2000 ( p < 0,01), and after the introduction of novel agents. The mean age was similar in the EMD and classic MM, male predominance was found in EMD (20.6% vs 13.4%). EMD at diagnosis is associated with an early clinical stage, solitary plasmocytomas, non-secreting and light chain variants; EMD in the progression of MM is predominantly at multiple sites, characterized by massive bone marrow infiltration, resistance

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to therapy and early relapses; MS of the whole group is 37 months, 31 month for EMD (NS). DISCUSSION AND CONCLUSION: EMD is associated with many unsolved problems in their biology , unpredictable therapeutic outcome, poor prognosis. Key words: extramedullary disease, multiple myeloma

TH 24.

HAIRY CELL LEUKEMIA - CLINICAL FEATURES, DIAGNOSIS AND

THERAPEUTIC APPROACHES, THERAPEUTIC OUTCOME AND PROGNOSTIC FACTORS Katya Sapunarova, Vesselina Goranova-Marinova, Pencho Georgiev, Emil Spassov, Katya Sotirova, Maria Dimitrova, Stefka Tuleva, Stefan Goranov First Department of Internal Medicine, Division of Hematology, Medical University - Plovdiv, Bulgaria AIM: To analyse clinical characteristics, diagnostic and therapeutic approaches, therapeutic response, event-free survival and overall survival and the prognostic factors in patients with hairy cell leukemia (HCL). PATIENTS AND METHODS: between 2005 and 2014, 37 patients with HCL were diagnosed, treated and followed-up, 4 (11.8%) with HCL-variant. Male 31 (83.8%), female 6 (16.2%), mean age 57.9 ± 13.6 yrs. Diagnosis was made by flowcytometry proving expression of CD 11c+, CD25+ and CD103+ and by histological and immune hystotochemical examination with additional assessement of TRAP+ and the degree of reticuline fibrosis. Patients were treated with interferon-alfa, purine analogues and splenectomy. Response rate and type of response were assessed. EFS, OS, MS and prognostic factors were analysed by Kaplan Maier method with long rank-test (SPSSv22.0). R ESULTS: The initial manifestation was massive splenomegaly in 37 (100%) in combination with anemia in 21 (56.8%), leukopenia in 19 (51.4%) or thrombocytopenia 18 (48.6%). First line therapy with cladribin was administered to 16 (43.2%), fludarabin 12 (32.4%), interferon-alfa - 8 (21.6%), and splenectomy 1 (2.7%). Complete therapeutic response was registered in 7 (19.0%), partial - in 20 (57.1%), no response in 8 (22.9%). EFS was 37 mos (CI 95% 30.2-43.8). Relapses occurred in 17 (54.8%). OS at 5year was 82%, OS at 10 year - 73%. MS is not reached by the end of analysis. DISCUSSION AND CONCLUSION: modern diagnostics and treatment in HCL transformed this rare disease into highly curable form of leukemia and patients have near-normal life expectancy. Key words: hairy-cell leukemia, therapeutic response

TH 25.

PHILADELPHIA POSITIVE (PH +) ACUTE LYMPHOBLASTIC LEUKEMIA IN

ADULTS - THE REVISED PROGNOSIS. DESCRIPTION OF 2 CASES AND REVIEW OF LITERATURE Pencho Georgiev, V. Goranova-Marinova, V. Gryklanov, E. Spasov, St. Goranov Department of Clinical Hematology, St. George University Hospital, Medical University - Plovdiv, Bulgaria INTRODUCTION: Acute lymphoblastic Ph+ leukemia (ALL) in adults is a rare malignant hemopathy with a very high unfavorable prognostic risk. Until the introduction of tyrosin kinase inhibitors (TKI) the five year survival rate was below 10%. AIM: To present and discuss two clinical cases of patients with ALL (Ph+). Clinical cases: First case: a 35-year-old woman, WBC, 290 x G/l, 95% blasts in DBC, ‘Common preВ’ phenotype, RT-PCR – М-BCR-ABL+(р210). Treatment


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regimen was hyper–CVAD/ MTX-AraC+Imatinib. After the first induction block and continuous TKI, complete remission was achieved. Second case: a 33-year-old man, WBC, 73 xG/l, 90% blasts in DBC, ‘Common preВ’ phenotype, RT-PCR – m-BCR-ABL+(р190). Since 12.2014 he has been treated with hyper–CVAD/ MTX-AraC+Imatinib. DISCUSSION: The introduction of I and II generation TKI (imatinib, nilotinib, dasatinib) unprecedentently improved the prognosis and survival rate of patients with ALL (Ph+), and has transformed the disease into a chronic and controllable one. The appearance of additional mutations, resistant to II generation TKI is still a problem. This implies the necessity of search for compatible donor soon after complete remission is achieved. Unfortunately, less than 50% from the patients are suitable for stem-cell transplantation (SCT). Treatment with hyper-CVAD and TKI with consequent supportive treatment with TKI is a good alternative. CONCLUSION: Despite improved prognosis, patients with ALL (Ph+) remain unfavorable prognostic group. In view of the presented cases and literature review, best results are achieved with polychemotherapy + TKI and consolidation of the remission with allogenic SCT. Key words: tyrosine kinase inhibitors; acute lymphoblastic leukemia

TH 26.

SIX CASES OF RARE HEREDITARY ANGIOEDEMA

Svetlan Dermendzhiev1, Petya Deleva1, Zlatka Stoyneva-Paskaleva1, Iliya Arolski1, Velcho Velev2, Atanaska Todorova1, Bogomil Penchev1 1Second Department of Internal Diseases, Division of Occupational Diseases and Toxicology, 2St. George University Hospital, Plovdiv, Bulgaria INTRODUCTION: Hereditary angioedema (HAE) is an autosomal dominant disease that afflicts 1 in 10,000 to 1 in 150,000 persons. It manifests as recurrent attacks of intense, massive, localized edema without concomitant pruritus. Due to the rare character of the disease the presentation of any case of the kind contributes to its better understanding. OBJECTIVE: The primary objective of our report is to present the rare disease to more physicians. Thus it will help the efficient diagnosis and treatment of HAE. MATERIALS AND METHODS: We used the medical documentation of all six patients along with laboratory test results representing the specific changes in the immune system as well as commonly used tests depicting the overall state of the patients. R ESULTS: As a result of the immune testing we found that all of the patients had decreased levels of antigenic and functional C1-esterase inhibitor with the exception of one of the patients in which the level of antigenic C1INH was close to the lower range. None of the patients had positive results from testing of anti-rabbit epithelium IgE. All of the patients with the exception of one had family history of the disease. CONCLUSIONS: HAE is a rare disease that is often difficult for GPs to diagnose quickly. It has specific treatment that differs completely from other forms of angioedema and needs to be diagnosed correctly and before the presentation of lethal exacerbations that include laryngeal localization of the swelling. Key words: hereditary angioedema, immune testing, C1-esterase inhibitor

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TH 27.

A RARE CASE OF ANGIOEDEMA AND URTICARIA IN A PATIENT WITH

ESSENTIAL THROMBOCYTHEMIA Svetlin Dermendziev1, Petya Deleva1, Zlatka Stoineva-Paskaleva1, Iliya Arolski1, Velcho Velev2, Atanaska Todorova1, Bogomil Penchev1 1Second Department of Internal Diseases, Division of Occupational Diseases and Toxicology, Medical University - Plovdiv, 2St. George University Hospital, Plovdiv, Bulgaria Urticaria (hives) is a common disorder that often presents with angioedema. Angioedema can occur in the absence of urticaria, with angiotensin-converting enzyme (ACE) inhibitor-induced angioedema and idiopathic angioedema being the more common causes. Urticaria is an outbreak of swollen, pale red bumps, patches, or welts on the skin that appear suddenly. Angioedema is similar to hives, but the swelling occurs beneath the skin instead of on the surface. Both disorders involve the same pathologic processes but with different localization. The etiological factor for urticaria and angioedema could be one and the same especially when the exacerbations happen simultaneously. However, often such a factor can’t be found and there is a possibility of different etiology of both conditions in one and the same patient. In most cases the pathogenic mechanisms include IgE mediation but sometimes pseudoallergic non-immune mechanisms are involved. The presence of possible autoimmune processes along with long-term therapeutic administration of medications makes it even more difficult to differentiate the cause of the urticarial rash and angioedema. Hereby we present a rare case, which according to our literature review, has not been described – chronic urticaria with concomitant angioedema and essential thrombocythemia. Key words: angioedema, urticarial, essential thrombocythemia

TH 28.

THERAPEUTIC EXPERIENCE WITH BORTEZOMIB IN PATIENTS WITH

MULTIPLE MYELOMA V. Goranova-Marinova1, A. Nedeva2, P. Georgiev1, K. Sotirova1, E. Spassov1, I. Hristova1, M. Dimitrova1, K. Sapunarova1, V. Graklanov1, V. Babacheva1, J. Raynov2, St. Goranov1 1First Department of Internal Medicine, Division of Hematology, Medical University - Plovdiv, 2Clinic of Hematology, Military Medical Academy, Sofia, Bulgaria INTRODUCTION: Therapy with bortezomib is the golden standard in the treatment of patients with multiple myeloma (ММ). PATIENTS AND METHODS: Between January 2006 and June 2014, 203 patients were studied, and treated in two university clinics of hematology – in St. George University Hospital - 148 patients and in Military Medical Academy - 55. M/F ratio =1.05/1.0, mean age 59.8±9.1y, ISS-I- 39 ( 19.2%); ISS-II- 77(37.9%) and ISS-III - 87 (42.9%); with renal failure (RF) 72 (29.6%), extramedullar forms 47 (23.2%). The therapeutic response (OR) was assessed according to EBMT. Statistical analysis was performed using SPSS v 22.0, survival was assessed by Kaplan-Maier method with log-rank test. R ESULTS: ORR was 148 (74.0%): CR 45 (22.5%), VGPR 24 (12.0%), PR - 79 (39.5%). EFS for patients with OR is 14 mos (95% CI 10.2-17.8) vs 5 mos (95% CI 2.08-7.92) p0.05) and significantly reduced microvascular dilator capacity of the patients during heat test was established (р < 0.01). Sympathetic vasoconstrictor responses to cold water immersion stimulation revealed delayed recovery time of the superficial skin temperature and skin perfusion in patients compared to controls (р < 0.01). Paradoxical venoarteriolar responses were found in 80% of patients. CONCLUSIONS: CTS with occupational etiology is associated with neurovascular microcirculatory thermoregulatory and venoarteriolar dysfunctions as a result of autonomic and sensory nerve fiber damage of the median nerve. Investigations of neurovascular reactivity by LDF contributes to the diagnosis and expertise evaluation of the disease. Key words: carpal tunnel syndrome, autonomic neurovascular reactivity, thermoregulation, occupational disease


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Surgery SU 1.

TECHNIQUES APPLIED IN COMPROMISED SKIN SURFACE IN

THE TRAUMATOLOGIC PRACTICE Stanislav Karamitev Department of Orthopedics and Traumatology, St. George University Hospital, Medical Faculty, Medical University - Plovdiv, Bulgaria The aim of the present study was to share our extended experience and strategy in the plastic reconstruction of open bone surfaces after heavy traumatic injuries of the locomotion apparatus. Obligatory in all cases of naked bone surface after injury the bone must be covered with soft tissues and skin in order to avoid any necrosis of the bone. In order to achieve our goals we used different techniques: split skin, rotational flaps, crossfinger flaps, crossleg flaps – primary and using the Tichinkina method, free skin and musculo-dermal flaps as well as transplants microvessel sutures. In secondary exposure of the bone with substantial bone necrosis after necrectomy we wait for fresh granulations and after that we initiate the non free skin flapping by tailoring the skin-dermal flap, or shaping the Filatov’s stem. The results in all operated 45 patients with different operative techniques for covering of the naked bone surfaces were rated as very good. Key words: skin surface, split skin, rotational flaps

SU 2.

A CASE OF PARTIAL TISSUE REPLANTATION IN THE SCALP AREA

Elean Zanzov, Ivan Traikov, Krasimir Yanev, Elena Krasteva, Parvan Voinov Department of Propaedeutics of Surgical Diseases, Division of Plastic, Reconstructive and Aesthetic Surgery, Medical University - Plovdiv, Bulgaria The replantation is a method with undeniable advantages. It offers tissues which are of the same anatomical structure and at the same time give a better cosmetic result, because of the same color, size, shape, figuration and physiology. We are representing a case with replantation of tissue in the temporal area, the necessity of which came as a consequence after a mechanical trauma. Our attention is directed to the pretreatment of the donor tissue, as a prerequisite for a successful replantation. Key words: replantation, scalp, defect

SU 3.

COMBINED METHODS FOR RECONSTRUCTION OF TISSUE DEFECTS

IN THE SCALP Elean Zanzov, Ivan Traikov, Krasimir Yanev, Vanya Anastasova, Stancho Prodanov Department of Propaedeutics of Surgical Diseases, Division of Plastic, Reconstructive and Aesthetic Surgery, Medical University - Plovdiv, Bulgaria The defects of the scalp are a challenge for reconstructive surgery and are associated with many limitations that have to be overcome in order to achieve a successful outcome. The intelligent reconstruction goes through a detailed assessment of the physiology and the anatomy of the area, a careful analysis of the defect and the general condition of the patient, an assessment of the po-

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tential for cosmetic compromise. The choice of surgical techniques is crucial. In this study we report 27 cases treated at the Cauterization, Plastic, Reconstructive and Aesthetic Surgery Clinic at Saint George University Hospital in Plovdiv for the past ten years (from 2003 to 2012), where we used multiple methods of reconstruction for covering tissue defects in the scalp area. Lately there has been a view that in the reconstruction of the scalp one has to avoid excessive tension at the edges of the wound, as the local ischemia may lead to dehiscence and/or partial loss of the flap and the involution of hair folicles. This fact mentioned above as well as the reconstructive solutions for large or dispersed tissue defects of the scalp are the main reasons for us to use the combined reconstruction methods. Key words: scalp, reconstructiоn, defects

SU 4.

EARLY ARTHROSCOPIC DIAGNOSIS AND TREATMENT OF

GONARTHROSIS Stanislav Karamitev, Krasin Ivanov Department of Orthopedics and Traumatology, St. George University Hospital, Medical Faculty, Medical University - Plovdiv, Bulgaria The goal of this report is to focus on the diagnosis and treatment of primary lesions in osteoarthritis of the knee joint, while in a stage of cartilage changes, through minimally invasive (arthroscopic) surgery. MATERIAL AND METHODS: Between June 2003 and July 2014 in the Clinic of Orthopedics and Traumatology of St. George University Hospital, Plovdiv we performed arthroscopic intervention in 326 patients. We operated 180 women (55.21%) and 146 men (44.79%). For early diagnosis and arthroscopic treatment of cartilage changes occurring in the knee joint we were guided by the classification of Joe Tippett. We implementated primarily the technique of abrasive arthroplasty. R ESULTS: In 256 patients (78.53%) we diagnosed I, II, III, IV, V, VI degree of cartilage damage and the function of the affected joint was fully restored. In other 52 (15.95%) we identified VII and VIII degree. The general condition of these latter patients improved, there was relief of pain and increased range of motion and disappearance of the chronic synovitis. In 10 operated patients (3.07%) there was no change in the condition of the joint, while in another 8 (2.45%) the condition worsened. CONCLUSION: Early arthroscopic diagnosis and abrasion arthroplasty are adequate prevention for the disease progression and when irreversible bone changes have come to the point to require serious knee surgery such as high tibial osteotomy and joint replacement. Key words: knee arthroscopy, abrasive arthroplasty, sclerotic subchondral bone

SU 5.

EARLY TREATMENT OF MUTILATING BURNS TO ORBITAL UNIT OF THE

FACE K. Kalinova, St. Prodanov, I. Traikov Department of Propaedeutics of Surgical Diseases, Division of Burns, Plastic, Reconstructive and Aesthetic Surgery, St. George University Hospital, Medical University - Plovdiv, Bulgaria Mutilating burns to the orbital unit requiring reconstructive surgery are an uncommon entity, their treatment is difficult and long and the relevant literature - scarce. AIM: To systematize early treatment of mutilating burns to orbital part of face. MATERIAL AND METHOD: A retrospective analyses is made of mutilating burns to the orbital third


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of face treated in the department over a 10-year period. R ESULTS AND CONCLUSIONS: The three major issues to be addressed by the surgeon managing mutilating burns of the orbital unit are: 1. To downsize at the maximum the initial damages; 2. To save vision by all possible means; 3. To restore as much as possible the person’s anatomical, functional and aesthetic entity of the face. Eyelids are particularly vulnerable and their subsequent reconstruction – dubious. Preservation of vision is in a close correlation with immediate provision of stable protection of eyeball by functioning eyelids. Priority is given to early aggressive surgical approach aimed at rapid healing of initial lesions thus reducing the risk of infection and extensive fibrosing. A long term reconstructive strategy should be built up immediately after the burn has taken place. K EY

SU 6.

WORDS:

early treatment, mutilating burn, orbital unit

EXTRACORPOREAL LITHOTRIPSY FOR TREATMENT OF RENAL AND

URETERIC STONES. PREDICTORS FOR SUCCESS Atanas Ivanov, I. Dechev, P. Antonov, I. Bakurdzhiev Department of Urology, Medical Faculty, Medical University - Plovdiv, Bulgaria INTRODUCTION: Extracorporeal shock-wave lithotripsy (ESWL) is defined as a first-line treatment in all patients with renal and ureteric stones. Several factors have been reported to influence the “stone-free clearance”. OBJECTIVE: To define the predictive value of six prognostic factors for success after ESWL- age, sex of the patient, BMI, size and site of the stone, presence of hydronephrosis. PATIENTS AND METHODS: The study included 443 patients, diagnosed and treated with ESWL in the Department of Urology at St. George University Hospital, Plovdiv between August 2013 and August 2014. The patients were divided into groups by prognostic factors. R ESULTS: 286 of 378 patients (75.66%) were presented with total fragmentation with the first session. The success rate was significantly higher (p < 0.05) for the stones up to 10 mm (75.77%), these between 10 and 20 mm (72.56%), pelvical (83.69%), upper calyceal stones (82.55%), ureteric stones in the proximal ureter (63.63%), in his pelvical part (78.20%) and in patients with a BMI within normal limits. The results obtained on the other investigated factors - age, sex and presence of hydronephrosis do not present statistically significant information about their role as predictors of success of ESWL . CONCLUSION: Stones’ size, site and BMI are the most important predictors for the success in ESWL of renal and ureteric stones. Key words: extracorporeal lithotripsy, urolithiasis, factors, success

SU 7.

FEMALE URETHRAL CARCINOMA – A BRIEF SURVEY OF LITERATURE

AND ANALYSIS OF THREE CASES A. Shopov, S. Zdravtchev, D. Delkov, A. Ivanov Department of Urology, Medical University - Plovdiv, Bulgaria AIM: We present three cases of primary carcinoma of female urethra as this type of tumors is very rare. MATERIALS AND METHODS: The cases were diagnosed by clinical investigations and by urethrocystoscopy, CT scan and pathohistological studies while examining the resectional line for residual tumors. R ESULTS: Two of the women were not subjected to diagnostic inguinal lymph dissection. The third female patient was diagnosed with inguinal and iliac lymphadenomegaly

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- the tumor was localised in a diverticulum in the middle part of the urethra (staging Т2N2M0). The patient had previously undergone hysterectomy because of myoma in the uterus. We performed urethrectomy with resection of the urinary bladder, the colon and cystostomy, inguinal and pelvic lymph dissection. The histological samples were studied by histochemistry: clear cell carcinoma of the urethra, inguinal metastases were the findings. The patients received distant radiotherapy and chemotherapy and survived for two years after therapy. CONCLUSION: In the early clinical stages, tumors of the anterior female urethra can be treated by surgery while tumors in advanced stages can be managed by extended surgical treatment. Key words: carcinoma, female urethra

SU 8.

FIBROUS DYSPLASIA OF FACIAL BONES – A PRESENTATION OF THREE

CASE REPORTS Regina Khater, Kostadin Gigov, Yourii Anastassov Department of Propaedeutics of Surgical Diseases, Division of Plastic and Craniofacial Surgery, Medical University - Plovdiv, Bulgaria Fibrous dysplasia is a benign disorder of bones, presenting as excessive growth which affects both axial and craniomaxilofacial skeleton. Depending on the number of affected bones, it is categorized as monostotic (monocystic), polystotic (polycystic) and the triad of polystotic fibrous dysplasia, precocious puberty with different hormonal disorders and “café au lait” spots, known as McCune-Albright syndrome. Hereby, the authors are presenting three cases of fibrous dysplasia, diagnosed and treated in the Division of Plastic and Craniofacial Surgery in St. George University Hospital. In all three patients: 1) a 20-year-old woman, 2) a 35-year-old man and 3) a 5-year-old child, the disorder affected the midfacial skeleton, as in cases 2) and 3) it was a monostotic form and in 1) polystotic, more apparently manifested in the maxilla region. The adults were treated by plastics of contours with a satisfactory result and the child – by total extirpation and closure of the defect. The present study discusses the stages of medical diagnosis and the advantages and disadvantages of the chosen therapeutic methods. Key words: fibrous dysplasia

SU 9.

NEURONAVIGATED RESECTION OF GLIAL TUMOR LOCATED IN

THE PARAHYPOCAMPAL GYRUS AND THE UNCUS OF THE RIGHT TEMPORAL LOBE - INITIAL EXPERIENCE Ivo Kehayov, Hristo Zhelyazkov, Stefan Raykov, Atanas Davarski, Borislav Kitov Department of Neurosurgery, Medical Faculty, Medical University - Plovdiv, Bulgaria We report a 26-year-old male who has been suffering from paroxysmal attacks of disorientation and confusion for 5 years. Some of these attacks lasted only couple of minutes while others lasted 10 to 15 minutes. His relatives reported that the patient used to lose consciousness during these attacks and his extremities were shaking. He was unable to recall these accidents. He was prescribed the anticonvulsant finlepsin 2x200 mg/day. Gradually, he became aggressive and his behavior changed over time. Several weeks prior to hospital admission, the attacks had become more frequent and taken place once a day. Despite the suggested epileptic origin of his complaints, the electroencephalography that was conducted failed to demonstrate any abnormal elec-


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trophysiological activity. The computed tomography and magnetic resonance imaging showed parenchymal brain tumor with diffuse calcifications infiltrating the parahypocampal gyrus and the uncus of the right temporal lobe. Antero-medial temporal lobe resection, amygdalohippocampectomy and brain tumor removal was performed guided by the neuronavigational system Medtronic StealthStation S7®. The histological examination demonstrated gemistiocytic astrocytoma grade II according to the WHO classification of CNS tumors. The patient was seizure-free at the third postoperative month. Key words: neuronavigation, gliomа, surgery

SU 10.

OUR EXPERIENCE OF IMPLEMENTING GIL-VERNET METHOD ABOUT

URINE INCONTINATION AFTER RADICAL RETROPUBIC PROSTATECTOMY A. Shopov1, D. Delkov1, A. Ivanov1, V. Sirakov2, N. Sirakov2 1Department of Urology, 2Department of Medical Imaging, Medical University - Plovdiv, Bulgaria This short study of the problems connected with urine incontination after radical prostatectomy shows that these problems have not been solved yet and the examinations are still actual. AIM: To implement and investigate a method of Gil-Vernet for reducing the urine incontination after radical prostatectomy MATERIALS AND METHODS: Betwenn March 2011 to February 2012 we made retropubic prostatectomyusing a novel approach for our practice in 14 patients with local advanced prostate adenocarcinoma (staging Т2 N0M0) and Glisson score of 7. From 2011 we began to use intraoperative investigation of the urinary bladder continence before making bladder-urethral anastomosis, the same as Gil-Vernet’s. The main part of the manipulation consists of filling the urinary bladder with liquid by Nelaton catheter. The orifice of the urinary bladder we create around supported anatomic pincet till there was no flow from the bladder after removing the pincet. After that we make bladder urethral anastomosis. R ESULTS: In 2 cases (14.3%) we had urine incontinence. But we had obstruction if the level of bladder urethral anastomosis in 4 cases /28,6%/ - it was solved by transurethral resection. CONCLUSIONS: Manipulating the urinary bladder orifice to hold liquid inside the bladder before making bladder urethral anastomosis prevents incontinence after prostatectomy. Key words: incontinence, Gil-Vernet method

SU 11.

PENILE OLEOGRANULOMA – SURGICAL TREATMENT

Ivan Bakardzhiev1, Ivan Dechev1, Atanas Ivanov1, Peter Antonov1, Mladena Mladenova2, Svetlana Bachurska3 1Department of Urology, 2Department of Obstetrics and Gynaecology, 3Department of General and Clinical Pathology, Medical University - Plovdiv, Bulgaria INTRODUCTION: Oleogranuloma also known as paraffinoma of the penis originate as a result of subcutaneous injection of foreign substances (liquid paraffin or vaseline) in the penile body in order to enlarge the penis and to achieve better sex. AIM: The aim of the study is to share our experience in surgical treatment of penile oleogogranuloma. PATIENTS AND METHODS: We present 176 cases of oleogogranuloma - 65 patients with light form, 88 patients with more severe form of oleogogranuloma and 24 with severe form. Patients were aged between 14-46 years (29.78 ± 2.87). Radical excision of skin defect is the only option in

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treatment of oleogogranuloma, as the reconstruction of the penis can be achieved by using several surgical techniques. R ESULTS: In 87 of our patients single stage surgery was performed, the other 89 cases were treated with two stage procedure. Dehiscence was observed in 12 patients, suppuration in 5, and 3 of our patients had haemorrhagia. No lethal outcome was registered. Postoperative inability for sufficient intercourse was observed in only 4 patients. CONCLUSION: The postoperative complications were a few. There are good cosmetic and functional results of the reconstructed penises. Key words: penile oleogranuloma, paraffinoma

SU 12.

POSTOPERATIVE PAIN AND PRESENCE OF COMPLICATIONS AND

DISADVANTAGES AFTER APPLICATION OF LIPOFILLING – A REVIEW OF THE LITERATURE AND PERSONAL RESEARCH Regina Khater, Kostadin Gigov, Yourii Anastassov Department of Propaedeutics of Surgical Diseases, Division of Plastic and Craniofacial Surgery, Medical University - Plovdiv, Bulgaria Nowadays, despite of the growing number of aloproducts the autologous fat grafting is recognized as one of the basic techniques for correction of soft tissue defects. In the recent years an object of broad discussion in the literature are the different technical aspects of the method, the need of transplant “enriching”, its indications and postoperative care. In the present material the authors provide a review of the literature and the results of their own research, concerning the postoperative pain, the disadvantages and complications after application of autologous fat grafting – poorly covered subject in literature, despite the method’s popularity. 148 protocols of lipofilling applied in 108 patients were investigated for the aims of the study. The postoperative pain, staged from 1 to 10 and the presence of edema and ecchymoses and influence on social life and work were assessed by a questionnaire. Our investigation found out that the postoperative period is characterized with low intensity of pain (3.95 ± 0.29 in recipient and 4.22 ± 0.26 in donor site according to subjective pain assessment from 1 to 10) and low rates of complications 4.05% in the donor and 12.16% in the recipient site. Edema and ecchymoses were observed respectively 65.54% and 20.95% and in all the cases they did not affect the personal habits and social life of patients. In conclusion we could say that the intervention is not painful and rarely leads to complications. It is well accepted by patients and does not require a long period of rehabilitation. Key words: lipofilling, postoperative pain

SU 13.

TISSUE EXPANSION OF SOFT TISSUE DEFECTS IN RECONSTRUCTIVE

SURGERY V. Anastasova1, I. Traikov1, B. Hadjiev2, E. Zanzov1, K. Janev1, 1Department of Propaedeutics of Surgical Diseases, Division of Plastic, Reconstructive and Aesthetic Surgery, 2Department of General Surgery, Medical University - Plovdiv, Bulgaria Stretching tissues is a physiological phenomenon and is as pregnancy, breast growth and development of tumor formation, the formation of a seroma or hematoma. The last decade has seen


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an increase in the number of patients with postburn and posttraumatic defects and contractures. After submission by Neumann, surgical treatment with tissue expanders has become a routine method for the treatment of large defects where there is not enough surrounding tissue to form the flap. This technique allows the coating of tissue defects with similar color, thickness and elasticity. In combination with other methods of plastic surgery, tissue expansion facilitates the coating of large soft tissue defects of various etiologies with minimal formation of new scars and minimal donor sites. In this study, we present several cases treated at the Clinic for Plastic, Reconstructive and Aesthetic Surgery, Saint George University Hospital on the occasion tissue defect by expander technique. We discuss the advantages and limitations of the tissue stretching in comparison to other methods of reconstruction. Key words: reconstructive surgery, post-burn reconstruction, tissue expansion, soft-tissue defects

SU 14.

S41.0 - S61.0 - WOUNDS OF UPPER LIMB. CHOICE OF SURGERY

Krasimir Yanev Department of Propaedeutics of Surgical Diseases, Medical University - Plovdiv, Clinic of Plastic, Reconstructive and Aesthetic Surgery, St. George University Hospital - Plovdiv, Bulgaria The hand of man, at any age performed his social adaptation. They use it in labor. Therefore, the upper limb is subjected to injuries of various kinds. Severe burns or injuries with no skin and subject fabrics are responsible for distortions in the function and cosmetic defects. Investigation - stiffness and in deep lesions, partial amputations or loss of an entire limb. Important locations are - dorsal side of the hand with fingers, wrist and elbow joint. To avoid severe consequences, prompt treatment is needed, including early surgical intervention and early rehabilitation. Watchful waiting or inadequate surgery usually lead to secondary infection in the wound and other kind of surgery. Deepening the harm coverage of deep-seated anatomical elements, lead to disability. Superficial injuries and burns require early (3-4 day) surgical excision and instantaneous free skin autoplastics. Deep damage (with exposed by the wound bed tendons, bones or nerve vascular bundles), the viability of the hand is greatlt threatened. Only adequate solution for rescuing a cover flap (skin, fascitis cutaneous, muscle or myo cutaneous). The protection allows them to immediately exercise. The operational equipment must be adapted to the depth of the lesion location and opportunity to use the local tissues. Planned and executed, operating machinery should not interfere and hampers subsequent reconstructive interventions. Key words: S41.0 - S61.0

SU 15.

SEPTIC ARTHRITIS OF THE INTERVERTEBRAL JOINTS OF THE LUMBAR

SPINE - CASE REPORT AND LITERATURE REVIEW Margarita Kitova-John1, Ali Azim-Araghi2, Borislav Kitov3, Tanya Kitova4 1South Wight Medical Practice, Isle of Wight, UK; 2Wessex Deanery, Hamshire, UK; 3Department of Neurosurgery, Faculty of Medicine, Medical University - Plovdiv, Bulgaria; 4Department of Anatomy, Histology and Embryology, Medical University - Plovdiv, Bulgaria The septic arthritis of intervertebral joints (SAIJ) is a rare and not well known condition. Up to date there are only a few dozen reported cases in the literature.

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The presented case is of a 48-year-old woman who reported severe lower lumbar pain of one month duration associated with right sided foot drop. The patient had developed the symptoms while abroad and initially consulted a chiropractor in Spain. She had been generally tired and had a mild fever, which was attributed to the severe back pain. The examination revealed a vertebral syndrome - pain on palpation at the level of L4, L5, S1, radiating to the back of the thigh, side of calf, reaching the sole of the foot. There was also sensory loss along L4, L5, S1 and loss of muscular power (3/5) of plantar and dorsal flexion of the right foot. The left foot was unaffected. The routine blood count revealed slightly raised WBC (11.7) and CRP (60). The X-ray performed by the local emergency department was reported as unremarkable. This case prompted us to do a literature review, which highlighted some cases of lower lumbar back pain as a result of facet joints infections. Their diagnosis could be challenging and adequate timely investigations, including imaging, is essential for the prognosis, duration of treatment and recovery. Key words: septic arthritis of intervertebral joints

SU 16.

SIGNIFICANCE OF OPERATIVE TREATMENT FOR DEGENERATIVE

SPINAL STENOSIS Aneta Petkova, Borislav Kalnev, Hristo Jelyazkov, Atanas Davarski, Borislav Kitov Department of Neurosurgery, Medical Faculty, Medical University - Plovdiv, Bulgaria AIM: To show the significance of operative treatment for degenerative diseases of the spine. METHODS: The present study included 77 cases of degenerative lumbar stenosis who underwent surgery at the Clinic of Neurosurgery at St. George University Hospital in Plovdiv in 2014. Eleven cases of degenerative stenosis in the cervical spine were studied, 4 of them - in the thoracic area and 62 - in the lumbar area. Clinical status, imaging, surgical treatment and post-operative results were analyzed. R ESULTS: The main aim of the operative treatment was the decompression of the neural elements, without any further damage and without disruption of the spinal cord. In the cases of cervical pathology the operative techniques of anterior decompression was used - discectomies, osteophytectomy, corporectomy, followed by fusion and anterior stabilization. In the cases of thoracic and lumbar stenosis, we used posterior decompression - interlaminotomy, hemilaminectomy, laminectomy. In 11 of them, having considered the presence of multilevel lumbar stenosis, the wide decompression was combined with posterior transpedicular stabilization. A considerable abatement of the pain syndrome was registered with all patients, as soon as the early post-operative period (from day one to day three). CONCLUSION: When timely conducted and correctly indicated and performed, the operative treatment eliminates pain, allows the restoration of the physical activity and improves the patients’ quality of life. Key words: operative degenerative spinal stenosis decompression


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SU 17.

UPPER URINARY TRACT CONGESTION DURING PREGNANCY – SYNDROME

OF THE OVARIAN VEIN? CAUSES, DIAGNOSIS AND MANAGEMENT Angel Shopov, D. Delkov Department of Urology, Medical University - Plovdiv, Bulgaria AIM: To find the causes for congestion in the upper urinary tract during pregnancy. MATERIAL AND METHODS: Between 2007 and 2011, we diagnosed 66 cases with upper urinary tract congestion during pregnancy. In 61 patients (92%) it was right-sided, in 2 (3%) – left-sided and 3 patients (5%) had the stagnation bilaterally. The clinical symptoms were pain in the relevant waist area, fever and high temperature. The diagnose was confirmed by ultrasound study; in addition to drainage failures in the upper urinary tract, we investigated the ovarian vein split status and the position of uterus. The treatment was drainage of the relevant kidney by implanting ureteral stent, which was removed in the puerperium. CONCLUSION: We think that apart from enlarged uterus, the main cause for the upper urinary tract stagnation during pregnancy, predominantly in the right side, is the ovarian vein syndrome. In these cases the syndrome is temporary. It occurs mainly during the last trimester of pregnancy and disappears after the puerperium. Key words: stagnation, pregnancy, ovarian vein

SU 18.

SURGICAL MANAGEMENT OF HIGH-RISK LESIONS OF THE MAMMARY

GLAND G. Baitchev1, I. Inkov1, I. Ivanov2, N. Kyuchukov1, A. Goncharov1, 1Clinic of Thoracic Surgery, Military Medical Academy, Sofia, Bulgaria, 2Department of Medical Diagnostic Research, Medical University - Pleven, Bulgaria The management of premalignant breast lesions has become a significant clinical problem for the past 20 years, particulary due to their increased detectability. The aim of this study is to analyze the frequency, morphology and management of such cases. The subjects in this prospective study are 105 female patients, who were operated at the Thoracic Surgery Clinic of the Military Medical Academy in Sofia, Bulgaria between 2010 and 2014. Of all diagnosed cases, 69 had ductal intraepithelial neoplasia (DIN) of which 38 were with atypical ductal hyperplasia and 31 - with intraductal carcinoma; 27 had lobular intraepithelial neoplasia (LIN), 16 patients of these had atypical lobular hyperplasia and 11 were with lobular carcinoma in situ. Finally 9 cases were diagnosed with papillary lesions of which 7 were with atypical papilloma and 2 were with papillary carcinoma in situ. The purpose of the clinical examination, excluding cases of papillary lesions presenting with nipple discharge, is limited. Of all cases with DIN, 39.1% were pre-operatively mamographically diagnosed with microcalcifications. The cases with LIN were most often detected following a biopsy. The main dilemma facing surgeons is the balance between the risks of a high-volume resection, leading in some cases to unnecessary cosmetic defects and a minimal excisional biopsy associated with an increased risk of recurrence or a consequent neoplasia. Additional knowledge of predictive and prognostic factors as well as the development of genetic techniques will give an answer to these questions. Key words: premalignant breast lesions

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DIFFERENTIAL DIAGNOSIS BETWEEN SPONTANEUS AND BACTERIAL

SU 19.

PERITONITIS IN PATIENTS WITH LIVER CIRRHOSIS AND ASCITES Nikolay Krastev1, Ventseslav Djurkov1, Penka Akrabova2, Tanka Kurparova2, Georgi Kiprin2, Krasimir Asenov2, 1Eurohospital, Plovdiv, Department of Internal Medicine, Second Department of Internal Medicine, Division of Gastroenterology, Medical University – Plovdiv, 2Department of Internal Medicine, Eurohospital - Plovdiv, Bulgaria Differentiation of spontaneous bacterial peritonitis from secondary (SeBP) in patients with cirrhosis and ascites is important and often decisive for the outcome of the complication. Objective of the study is to identify patients with SBP and to differentiate between cases with nonperforative SeBP. We studied 326 patients with liver cirrhosis and ascites for 10 years, SBS (including culture are more negative neutrophilic ascites) is found in 48 of them (14.7%) and secondary bacterial peritonitis - in 37 (11.3%). Perforative peritonitis was found only in one patient with cirrhosis and ascites. A diagnosis of SBS is made in polymorphonuclear leukocytes in ascites > 250 mm3, while the bacterial cultures of routine microbiological testing ascites were positive in 7 patients (14.6%). These results are similar to those obtained in another Bulgarian study. The diagnosis of SeBP is set according to the criteria of Runyon. The total protein in the ascites was > 10 g/l in all patients, and > 30 g/l in 64.8% (24/37) of these. In ascitic fluid LDH > 240 U/l was also established in all patients, but glucose in ascites < 2.78 (consumed by the bacteria) was observed only in 5.4% (2/37) of patients with SeBP. Many other criteria can support the diagnosis SeBP and the use of Multistix (sets esterase activity of neutrophils) is justified to a greater extent in suspicion for SeBP than SBP, due to the large number of polymorphonuclear leukocytes in the ascites. Key words: cirrhosis, ascites, SBP, SeBP

SU 20.

DIABETIC RETINOPATHY IN PATIENTS REVIEWED IN TEPC FOR EYE

DISEASES – PLEVEN Аnna Angelova1, D. Stateva2 1TEPC - University Hospital “G. Stranski” - Pleven, 2Eye Clinic - University Hospital “G. Stranski” Pleven, Bulgaria INTRODUCTION: Diabetic retinopathy is one of the complications of diabetes mellitus and as such it is a major cause of eye impairments that leads to visual loss (advanced proliferative diabetic retinopathy (PDR) and diabetic macular edema). PURPOSE: To determine the incidence of patients with diabetic retinopathy and the degree of impairment of visual function among the people with diabetes mellitus, reviewed by the Territorial Expert Physicians’ Committee (TEPC) of eye diseases – Pleven. MATERIALS AND METHODS: In the first three months of 2013 all patients with diabetes mellitus that were referred to TEPC were examined. R ESULTS: For this period a total of 1307 patients from 9 regions of North Bulgaria – Pleven, Lovech, Gabrovo, V. Tarnovo, Russe, Targovishte, Vratza, Montana and Vidin - were examined. 473 (36%) of them had diabetes mellitus. According to their visual acuity and the presence of diabetic retinopathy these patients can be distributed in the next groups: with intact visual acuity and without retinopathy – 29 %, with intact visual acuity and non proliferative diabetic retinopathy (NPDR) data – 20 %, with intact visual acuity and presence of PDR – 0.8 %, with decreased visual acuity and NPDR – 24%, with decreased visual acuity and PDR – 16 %, with other eye diseases and reduced visual acuity, but


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without retinopathy – 10.2 %. Laser treatment was administered to 63% of patients with NPDR and 85% of patients with PDR. CONCLUSION: TEPC certification in many cases starts with a laser treatment or diagnosed DR. Key words: diabetic retinopathy, visual acuity, diabetes mellitus

ASSESSMENT OF VITREO-MACULAR INTERFACE IN PATIENTS

SU 21.

WITH DIABETIC MACULAR EDEMA BY USE OF SPECTRAL-DOMAIN OCT Desislava Koleva-Georgieva, Nelly Sivkova Department of Ophthalmology, Medical University - Plovdiv, Bulgaria AIM: To examine and analyze changes in vitreo-macular interface in patients with diabetic macular edema (DME) and assess the types of traction by use of spectral-domain OCT. PATIENTS AND METHODS: The study included 137 patients with diabetes (256 eyes) and 120 healthy subjects. All received visual acuity check-up, stereo-ophthalmoscopy and spectral-domain OCT. The characteristics of vitreo-macular interface were assessed (stages of posterior vitreous detachment – PVD and types of epiretinal membrane – ERM) in diabetic patients without diabetic retinopathy (DR), with DR and no DME and with DME, as well as in healthy subjects. The severity of changes in the underlying retina and visual acuity were assessed among the different types of traction. R ESULTS: The process of PVD in eyes with DME is characterized with greater proportion of incomplete PVD (54.7%), which probably plays role in the pathogenesis of edema. For evaluation of the strength of traction it is significant not only to pay attention to the different types of PVD (no PVD, incomplete PVD with peripapillary and perifoveal adhesion and complete PVD) and ERM (focal and global adherent), but also to analyze the changes in the retinal contour for detecting distortion at points of adhesion. Eyes with apparent distortion were with significantly greater retinal thickness, greater proportion of cystoid edema, impaired integrity of neuroreceptors, and lower visual acuity. CONCLUSION: The detected on OCT macular traction in cases with distortion of the retinal contour probably plays role as an aggravating factor in the pathogenesis of DME. Its diagnosis is important for the proper determination of the therapeutic approach conservative or surgical. Key words: vitreo-macular traction, OCT, macular edema

SU 22.

CLINICAL APPLICATION OF ANTERIOR-SEGMENT OPTICAL COHERENCE

TOMOGRAPHY (AS-OCT) Ralitsa Kermedchieva, Marieta Konareva - Kostianeva, Snezhina Kostianeva, Marin Atanasov Department of Ophthalmology, Medical University - Plovdiv, Bulgaria Optical coherent tomography is modern, noncontact, noninvasive diagnostic method with high resolution ability that enables imaging of eye structures through topographic transverse cuts (optic biopsy of the tissues). Images of anterior segment by OCT are demonstrated for the first time in 1994 by Izatt et al, using wavelength of 830 nanometers. Anterior segment OCT makes approximately 20,000 – 40, 000 scans per second and that decreases the chance of missing lesions. Anatomical structures which may be quantitative and qualitative defined are tear meniscus, anterior chamber, iridocorneal angle and lens. The purpose of the study is through literary database and examinations of our own to present the role of Spectral-Domain AS-OCT (Оptovue, RTV-ue) as high technology innovative method in diagnostics and treatment of different diseases of anterior eye segment.

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CONCLUSION: The diseases of anterior eye segment lead to impaired vision in large percentage of patients. The results obtained by AS-OCT are particularly useful in diagnostics, treatment and follow-up of diseases of the cornea and traumas, surgical interventions in glaucoma and cataract, as well as refractive surgery. Key words: AS-OCT, anterior eye segment, cornea, glaucoma

SU 23.

COMPARATIVE ANALYSIS OF OCT DATA IN PATIENTS WITH OCULAR

HYPERTENSION AND HEALTHY SUBJECTS Aleksandar Gerdzhikov Department of Ophthalmology, Medical University - Plovdiv, Bulgaria AIM: This study aims towards comparison of the data, acquired by examination with optic coherence tomograph (OCT) of patients with ocular hypertension and healthy subjects. MATERIALS AND METHODS: 15 patients (30 eyes) with ocular hypertension (OH) were examined. All patients received a thorough ophthalmological examination, including standartized automated computer perimetry (HFA II; Carl Zeiss) and optical coherence tomography (Optovue RTVue-100). The control groups consists of 30 patients (60 eyes) with no ocular pathology found. R ESULTS: Data acquired from examination of retinal nerve fiber layer (RNFL) and ganglion cell complex (GCC) was compared and analyzed. Key words: ocular hypertension, OCT

SU 24.

NON DRAINAGE SURGERY FOR RHEGMATOGENOUS RETINAL

DETACHMENTS WITH MULTIPLE BREAKS Arsenia Baliyan1, N. Gineva1, T. Hristova2 1Department of Ophthalmology, Medical University - Plovdiv, 2Clinic of Ophthalmology, St. George University Hospital, Plovdiv, Bulgaria INTRODUCTION: Non drainage surgery for rhegmatogenous retinal detachment (RRD) was introduced by Custodis in 1953. In 1963 Linkoff replaced diathermy by cryopexy and the polyvinylchloride intrascleral buckles by episcleral sponge buckles. The subretinal fluid is not drained. PURPOSE: To evaluate the anatomical and functional results of non drainage surgery in patients with RRD with multiple breaks and different stages of PVR (B and less). MATERIAL AND METHODS: Retrospective study of 32 surgeries (31 patients) for RRD with multiple breaks and different stage of PVR (B and less) performed by us over four years (2003-2007). The pre- and postoperative examination includes: BCVA, slit-lamp examination, fundubiomicroscopy, indirect ophthalmoscopy. The surgical procedure is cryopexy and episcleral buckling by different oriented (radial, limbus parallel and combination of both) silicon sponge, gel and band. The subretinal fluid is not drained. R ESULT: 78% of the surgeries were anatomical success with a fairly good functional result. DISCUSSION: Non drainage surgery is a surgery limited to the breaks which minimizes the procedure and hence the complications. Precisely performed, it is successful in most of the cases. Key words: rhegmatogenous retinal detachments


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SU 25.

TREATMENT OF GLAUCOMA

Kristina Yotova, Victoria Monovska, Desislava Stateva Faculty of Medicine, Medical University - Pleven, Bulgaria AIM: To describe the different methods of glaucoma treatment. METHODS: Different literature sources on glaucoma treatment were studied. Data of patients diagnosed with glaucoma in the Eye Clinic, Pleven from 01.2014 to 12.2014 was reviewed. R ESULTS: Our study showed that conservative treatment of glaucoma is long-term and the result of its effectiveness is that surgical treatment is required in only few cases. CONCLUSION: Early diagnosis, proper treatment and regular follow-up are essential for prevention of vision loss in glaucoma patients. Key words: glaucoma, treatment, trabeculectomy

OPTICAL COHERENCE TOMOGRAPHY CHANGES IN RETINAL NERVE FIBER LAYER AND GANGLION CELL COMPLEX IN DIFFERENT STAGES OF SU 26.

OPEN-ANGLE GLAUCOMA Snezhina Kostianeva, Aleksandar Gerdzhikov, Marieta Konareva - Kostianeva, Marin Atanasov Department of Ophthalmology, Medical University - Plovdiv, Bulgaria PURPOSE: To evaluate the quantitative measurements of ganglion cell complex (GCC) and peripapillary retinal nerve fiber layer (RNFL) obtained by optical coherence tomography (OCT) in different stages of open-angle glaucoma. METHODS: Sixty eyes of 34 patients with open-angle glaucoma and 60 eyes of 30 healthy people were examined using RTV-ue 100 (Optovue Inc., Fremont, CA). According to visual field global indices MD and PSD (Humphrey Analyzer) glaucomatous eyes are categorized into two groups: 33 eyes with early glaucoma and 27 eyes with advanced glaucoma. GCC and RNFL (using two scanning protocols of RTV-ue – ONH and RNFL 3.45) are analyzed in all of the eyes included in the study. R ESULTS: GCC average is with highest values in normal eyes, followed by the glaucomatous eyes in early stage and glaucomatous eyes in advanced stage (96.7 ± 5.3 μ versus 81.0 ± 9.4 μ versus 75.0 ± 16.8 μ). Concerning the indices GCC average, GCC superior and GCC inferior in eyes with early and advanced glaucoma statistically significant difference is not found (respectively P = 0.081, P = 1.00, P = 0.335). However, the thickness of the peripapillary nerve fiber layer, measured by the two scanning protocols, is statistically significantly different in early and advanced glaucoma (p < 0.001). Conclusion: Our results show that measurement of the peripapillary RNFL is more reliable OCT indicator for follow-up of glaucoma progression. Key words: open-angle glaucoma, OCT

SU 27.

ORBITAL TUMORS - CLINICAL AND DIFFERENTIAL DIAGNOSTIC

ASPECTS – 31 CASES AND REVIEW OF LITERATURE Atanas Bogoev1, Petar Miryanov1, Marieta Konareva-Kostyaneva2 1Medical Faculty, Medical University - Plovdiv, 2Department of Ophthalmology, Medical University Plovdiv, St. George University Hospital, Plovdiv, Bulgaria INTRODUCTION: Orbital tumors are relatively rare pathology, presenting with various local symptoms in a clinical examination – ptosis, palpebral retraction, edema, inection, exophthalmos ,

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pain, palpable mass, ipsilateral reduction in visual acuity, ophthalmoplegia, visual field defects, diplopia and etc; different findings in CT and MRI, depending on the location in orbit, histological type and biological behavior of the tumor. The diagnostic and therapeutic management of this pathology involves various specialists - general practitioners, ophthalmologists, neurologists, neurosurgeons and otorhinolaryngologists, making the topic interdisciplinary. AIM: The aim of this work is to direct clinical and differential diagnostic thinking in a discussion of symptoms and the information from imaging studies in intraorbital tumors, and to present a comparative analysis of the frequency of ocular symptoms, histology and biological behavior within our own cases and literature data. MATERIAL AND METHODS: 31 cases between 2005 and 2014 with orbital tumors are presented, with different clinical expression, localization in orbit and histological findings. Some of the patients are hospitalized аgain due to relapse. A thorough literature review of scientific topic was performed. CONCLUSION: In the differential diagnosis with similar orbital tumors the symptoms can present with various inflammatory/specific and non-specific/, infiltrative, infectious, parasitic, and endocrinological diseases due to non-specific expression. Knowledge of clinical signs, proper interpretation and good comparison between the findings of the imaging results determines the choice of adequate treatment strategy. Key words: orbital tumors, exophthalmos, diplopia, diagnosis

SU 28.

FOSTER-KENNEDY SYNDROME AND PSEUDO-FOSTER-KENNEDY

SYNDROME Silviya Krumova, Nelly Sivkova, Desislava Koleva-Georgieva, Vesela Mitkova, Velin Stoyanov Department of Ophthalmology, Medical University - Plovdiv, Bulgaria INTRODUCTION: Foster-Kennedy syndrome is a rare neuro-ophthalmological condition characterized by: 1. Optic disc atrophy of the ipsilateral eye, 2. Optic disc oedema of the contralateral eye, 3. Central scotoma, 4. Anosmia. Pseudo-Foster-Kennedy syndrome includes: optic disc oedema of the ipsilateral eye, optic disc atrophy of the contralateral eye without a tumoral process. PURPOSE: Two clinical cases are presented: one - with Foster-Kennedy syndrome and another with Pseudo-Foster-Kennedy syndrome. Methods: The ophthalmological examination includes: fundus biomicroscopy, computerized tomography (CT) with and without contrast, computerized perimetry (CP) with Humphrey Field Analyzer – II series. R ESULTS: First case: female aged 56 years who has visual acuity of the right eye 0.001 and VA of the left eye P.P.L.I. Fundus examination reveals optic disc oedema of the left eye and partial optic disc atrophy (temporal). CT shows bifrontal tumoral process. Second case: male aged 58 years who has normal VA. Fundus examination reveals optic disc oedema of the right eye and partial optic disc atrophy of the left eye. CT does not show tumor process, CP shows altitudinal scotoma found in anterior ischemic optic neuropathy. CONCLUSION: The presented cases are clinical and diagnostic challenge due to the connection between the pathological changes in the ocular fundus and the following bilateral partial lost of vision. Delayed diagnosis of a brain tumor is a life threatening condition. Key words: Foster-Kennedy syndrome, papilledema, altitudinalen scotom


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WEIGHT GAIN SIXTH WEEK AFTER BIRTH AS A PROGNOSTIC FACTOR

SU 29.

FOR DEVELOPMENT OF RETINOPATHY OF PREMATURITY Vasil Мarinov1, N. Sivkova1, Т. Boeva2, М. Krusteva3 1Department of Ophthalmology, Medical University - Plovdiv; 2University Eye Clinic, St. George University Hospital, Plovdiv; 3Department of Obstetrics and Gynecology, Medical University - Plovdiv, Bulgaria INTRODUCTION: Weight gain sixth weeks after birth is considered a significant and independent risk factor for retinopathy of prematurity. AIM: To examine the prognostic value of weight gain sixth weeks after birth for development of ROP. PATIENTS AND METHODS: We screened 132 prematures from 4-th week after birth and calculated absolute and relative weight gain at six weeks after birth. Patients were divided into: group I – no signs of ROP (N = 92) and group II – any signs of ROP (N = 40). Group II was divided into: IIA – spontaneously regressed (N = 24), and IIB – progressed to treatment (N = 16). We investigated 15 maternal and 20 newborn presumable risk factors for development and progression of ROP (Mann-Whitney U, χ2 or Fishers Exact Test). Logistic regression was performed for finding significant and independent risk factors for manifestation and progression of ROP. R ESULTS: Absolute and relative weight gain at 6 weeks after birth are significant risk factors for manifestation of ROP (р = 0.00021 и р = 0.0019, respectively), but not for progression to stages, requiring treatment (p = 0.901 and р = 0.825, respectively). CONCLUSION: Delayed absolute weight gain at 6-th week after birth is significant and independent risk factor for manifestations of any signs of ROP. Key words: retinopathy of prematurity (ROP), risk factors, weight gain

SU 30.

DO HLA-B27 POSITIVE ANTIGEN HAVE INFLUENCE ON MACULAR

THICKNESS AND THE DEVELOPMENT OF MACULAR OEDEMA IN PATIENTS WITH ANTERIOR UVEITIS? Vesela Mitkova-Hristova, Marieta Konareva–Kostianeva, Nina Stoyanova, Snezhina Kostianeva Department of Ophthalmology, Medical University - Plovdiv, University Eye Clinic, St. George University Hospital, Plovdiv, Bulgaria PURPOSE: To determine HLA-B27 positive antigen and its effect on the macular thickness and the development of macular oedema (MO) in patients with anterior uveitis (AU). METHODS: Ninety one patients (91 eyes) with AU participated in the study. Every patient had a complete eye examination and spectral optic coherent tomography (SD-OCT) of macula with RTV-ue 100 (Optovue) using MM6 and EMM5 scanning protocols. Ethiologic investigations and HLA-B27 typing were conducted according to which patients were divided into two groups: HLA-B27 positive and HLA-B27 negative. R ESULTS: From all 91 patients with AU 33 (36.3%) are HLA-B27 positive, as 26 (78,.%) of them are men. It is observed in the HLA-B27 positive group using both scanning protocols that the macula is thicker only in the area of the fovea (Independent-Sample T-Test; p < 0.05). In 29 of the patients with AU MO is proved by SD-OCT, 15 of which has HLA-B27 positive antigen. MO is more common in the HLA-B27 positive group (u = 2.08; p < 0.05). CONCLUSION: Frequency of HLA-B27 positive antigen is high in patients with AU, as men are affected more than women. If HLA-B27 typing is positive, the probability of development of MO is increased. Key words: HLA-B27 positive antigen, anterior uveitis, macular oedema, optic coherence tomography

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EXTRAOCULAR MUSCLES THICKNESS IN PATIENTS IN DIFFERENT

SU 31.

STAGES AND PHASES OF ACTIVITY OF THYROID–ASSOCIATED ORBITOPATHY Nina Stoyanova, Marieta Konareva-Kostianeva, Vesela Mitkova-Hristova, Snezhina Kostianeva, Department of Ophthalmology, Medical University - Plovdiv, University Eye Clinic, St. George University Hospital, Plovdiv, Bulgaria PURPOSE: To determine the thickness of the extraocular muscles using computed tomography (CT) in patients with thyroid - associated orbitopathy (TAO) according to the activity and the severity of the disease. METHODS: Forty five patients with TAO were examined. Every patient had a complete eye examination and CT of orbita (16 slices CT) with measuring the total extraocular muscles thickness (TEMT) of both eyes in millimetres. According to the activity of TAO patients are divided into two groups – with and without activity, and according to the severity – in 5 groups – “O” – only subjective symptoms; “S” – symptoms of the soft tissues; “P” – proptosis; “E” – disturbance in eye movements; “C” – corneal affection. R ESULTS: Activity of TAO is found in 20 patients (44.4%), in which TEMT is 46.47 ± 7.07. Twenty five patients (55.6%) with TEMT 38.62 ± 5.67 are in inactive phase. Statistically significant difference in TEMT according to the activity of TAO (Independent Sample T-Test, p > 0.05) is not found. With “O” are 12 patients (26.7%), with “S” – 5 (11.1%), with “P” – 22 (48.9%), with “E” – 5 (11.1%), with “C” – 1 (2.2%); TEMT for the groups is respectively – “O” – 35.35 ± 4.58, “S” – 41.76 ± 3.87, “P” – 45.0 ± 7.57, “E” – 44.44 ± 4.96 and “C” – 49.5. Statistically significant difference in TEMT between groups “O”–“P” and “O”–“E” is found (ANOVA, LSD, p < 0.05). CONCLUSION: Degree of severity of TAO is what matters for TEMT, but the activity of the disease is not. Key words: thyroid-associated orbitopathy, extraocular muscles, computed tomography

SU 32.

CATARACT SURGERY IN PATIENTS WITH PENETRATING KERATOPLASTY

(PKP) Tatiana Hergeldzhieva, P. Vassileva, I. Shandurkov, Y. Kirilova, N. Surchev Acad. Pashev Specialized Eye Hospital Sofia, Medical University, Sofia, Bulgaria PURPOSE: To follow cataract formation in patients with PKP and to present our approach in the surgical treatment of patients with corneal graft and cataract. METHODS: We have performed a retrospective study of all phakic patients with PKP for the period from September 2003 to December 2013. Triple procedure or consecutive surgeries (PKP followed by cataract extraction) were done depending on the lens status. Phacoemulsification and IOL implantation before suture removal were performed in patients with intumescent cataracts after PKP. In cases with planned surgery, lens extraction was undertaken at least 3 months after suture removal. R ESULTS: For a period of 10 years 200 PKPs (189 patients with a mean age of 57.5 years) were performed at our hospital. The phakic cases with PKP were 106 (53%). Triple procedure was performed in 14 of them (13.2%): “open sky” cataract extraction and IOL implantation - 11 cases, or phacoemulsification and IOL implantation, followed by PKP - in 3. Cataract progression was observed in 44 eyes (41.5%) at various time intervals after PKP. Cataract extraction was done in 40 of them (37.7%). Corneal graft remained transparent in 39 patients with cataract extraction after PKP (97.5%). Visual acuity ≥ 0.3 was achieved in 30% of cases with triple procedure and in 74% of cases with consecutive surgeries. No lens opacities were diagnosed in 48 patients with PKP (45%) at the end of follow up. Folia Medica 2015; 57; Suppl. 1 © 2015 Medical University, Plovdiv, Bulgaria

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CONCLUSIONS: Our experience from the surgical treatment of patients with PKP and cataract shows excellent results in cases with consecutive surgeries. Key words: cataract

POSTERS

Dental Medicine DM 1.

3D ARCHIVE IN ORTHODONTICS

Miroslava Dinkova Department of Orthodontics, Faculty of Dental Medicine, Medical University - Sofia, Bulgaria In orthodontic practice diagnostic plaster models are used. Their long-term archive conservation is an obligatory requirement. They are heavy, fragile, non-durable and require specific conservation location. The AIM of the study is to present the development of 3D digital archive by scanning and archiving of models. MATERIAL AND METHODS: The plaster model is scanned by a 3D scanner. The information is processed, recorded and archived in an appropriate format by software created by us. Thus the file allows indefinite conservation of the information in 3D format. R ESULTS: The advantages of the digital archive are as follows: Possibility to materialize models by CAD/CAM technology at any time through the created file. 3D digital technology allows manipulation of the models. Simulation of the expected treatment outcome is possible. That could be included in the patient’s motivation program before the start of the treatment. Maximum reduction of plaster models application. The need of plaster models conservation is dying out. Kew words: 3D archive, orthodontics, CAD/CAM

DM 2.

A 10-YEAR RETROSPECTIVE STUDY ON ORAL CANCER

Konstantin Tonchev1, Boyan Vladimirov2 1Faculty of Dental Medicine, Medical University - Plovdiv, 2Clinic of Maxillofacial Surgery, St George University Hospital, Plovdiv, Medical University - Plovdiv, Bulgaria Oral cancer is the eighth most common cancer worldwide and presents more serious health problem in countries with higher alcohol consumption and smoking. The aim of the present study was to analyze the clinicopathological data of patients with oral cancer diagnosed and/or treated at a single centre in Bulgaria. The clinical records of patients with oral cancer admitted to the Clinic of Maxillofacial surgery, St. George University Hospital, Plovdiv, Bulgaria, from 2004 to 2013 were used for this retrospective study. Data about the incidence, distribution by age and sex, T-stage and localization as well as information about primary symptoms, delay of the final diagnosis, treatment and mortality were analysed. The study confirmed that oral cancer remains serious problem for clinicians in terms of incidence, delayed diagnosis, suboptimal treatment. Key words: oral cancer, retrospective study

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DM 3.

A CLINICAL CASE OF GINGIVAL RECESSION IN A PATIENT WITH

RETROGNATHIA Stiliana Krusteva, E. Popova, S. Krusteva 1Department of Periodontology and Оral Мucosal Diseases, Faculty of Dental Medicine, Medical University - Plovdiv; 2Department of Periodontology and Оral Мucosal Diseases, Faculty of Dental Medicine, Medical University - Plovdiv; 3Department of Orthodontics, Faculty of Dental Medicine, Medical University - Plovdiv, Bulgaria Gingival recession is a change in the position of margo gingivalis in apical direction uncovering the tooth root. One of the factors for that appearance are malocclusions, including retrognathia of the lower jaw. It’s a change of the occlusion where the lower jaw is back against the base of the skull, where in most cases producing elongation of the frontal teeth (overjet). This type of deformation become from the result of difficult nasal breathing. In the treatment of these recessions requires a combined approach - orthodontic treatment, treatment of nasal breathing difficulties and surgical treatment of recessions. MATERIAL AND METHODS: The patient P.H. -14 years old from Plovdiv, were established gingival recessions of teeth 31, 32, 41, 42 Class 1 Miller and shallow and broad of the classification of Sullivan and Atkinson. After the consultation with an orthodontist it is established a nasal difficult breathing and the diagnosis retrognatiya with huge frontal distance (overjet). Based on this it had to correct the main diagnosed factors: the nasal difficult breathing and the orthodontic deformation. R ESULTS: After correcting the overjet and normalization of nasal breathing,it had to contributed to reducing the volume of gingival recessions. CONCLUSION: In a manifestation of gingival recession it is necessary to find the etiological factor. That is the way that the clinicist could avoid surgery to correct them, as in the clinical case. Key words: gingival recession, retrognathia

DM 4.

A SURVEY OF DENTISTS ON THE APPLICATION OF ENDOCROWNS IN

THEIR DAILY CLINICAL PRACTICE Viktor Hadzhigaev1, Stefan Zlatev1, Neshka Manchorova2, Yavor Kalachev3 1Department of Prosthetic Dentistry, 2Department of Operative Dentistry and Endodontics, 3Department of Prosthetic Dentistry, Faculty of Dental Medicine, Medical University - Plovdiv, Bulgaria INTRODUCTION: The choice of method for rehabilitation of endodontically treated teeth with severely compromised tooth substance is of great importance, for ensuring good and long-lasting clinical results. One of the possibilities is the usage of endocrown. AIM: To survey the opinion of dentists in Bulgaria on the usage of endocrowns in their daily clinical practice. MATERIAL AND METHODS: A group survey was conducted among 116 dentists during a general assembly of the Bulgarian Dental Association. It was anonymous and consisted of 9 questions. Five of these are about the usage of endocrowns as a method of choice in the clinical practice. Four of the questions are alternative with a positive or a negative answer. Two of the questions are about the difficulties which dentists encounter during the application of endocrowns. For statistical data processing the Student-Fisher and chi-square criterion were used. R ESULTS: With respect to gender respondents are equally distributed. The most frequent age group ranges from 31 to 40 years, and the least frequent is below 30 years. 72.41% of the participants have clinical specialty. Three quarters of the respondents have never used endocrowns. Dentists, who have difficulties using endocrowns are 21.57%. More than half of the respondents think that this type of construction will be used


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frequently in the future. CONCLUSION: With the development of dental materials the usage of endocrowns is becoming easier and the clinical indications are increasing. Key words: survey, endocrown

DM 5.

APICALLY EXTRUDED MATERIAL DURING REMOVAL OF GUTTA-PERCHA

FROM THE ROOT CANAL WITH NICKEL TITANIUM INSTRUMENTS WITH DIFFERENT CHARACTERISTICS - COMPARATIVE IN VITRO STUDY Liubomir Vangelov Department of Operative Dentistry and Endodontics, Faculty of Dental Medicine, Medical University – Plovdiv, Bulgaria INTRODUCTION: extrusion of debris and filling material in endodontic re-treatment is a problem with clinical importance because it can cause immediate postoperative flair-up, acute inflammation and edema. In the long term there is a possibility for the healthy periodontium to develop pathology and in case of existing periapical pathology, it is less likely to heal. OBJECTIVE: To compare the amount of material extruded through the apex after removal of guttapercha with nickel - titanium instruments with cutting edges or radial lands. MATERIAL AND METHODS: sixty-three extracted single rooted anterior teeth are prepared to 06/30 with ProFile 0.06 taper (Dentsply Maillefer, Switzerland) and obturated with lateral condensation. For the measurement of extruded debris a modified methodology of Hulsmann and Stotz was used. For the removal of gutta-percha three groupes are formed: Group 1 - Pro Taper Universal; Group 2 - GTX Rotary Files ;Group 3 – K files(control). After cleaning, the weight of the extruded material is weighed on an analytical microbalance „Sartorius PM 210S“ (Germany / GmbH) in grams. For statistical analysis one-way Anova test was used as the level of statistical significance is P < 0.05. R ESULTS: There was no statistically significant difference between the groups (P > 0.05). CONCLUSIONS: The amount of material extruded through the apex does not depend on differences in the design of instruments used. Key words: retreatment, gutta-percha, extrusion, nickel-titanium instruments

DM 6.

APPLICATION OF CAD/CAM TECHNOLOGY IN THE CONSTRUCTION OF

TEMPORARY FPD’S AS A STAGE OF THE PROSTHETIC REHABILITATION OF THE DENTITION Viktor Hadzhigaev1, Stefan Zlatev1, Georgi Todorov1, Desislava Georgieva2, Veselin Petrov2 1Department of Prosthetic Dentistry, 2Faculty of Dental Medicine, Medical University - Plovdiv, Bulgaria INTRODUCTION: An important stage in the construction of FPD is the preparation of temporary construction. They ensure keeping of function, esthetics and the ability to conduct proper oral hygiene. Temporary constructions have a prophylactic effect during the treatment thus preserving the vitality of the prepared teeth. They prevent any movements of the prepared, adjacent and antagonist teeth in the area of the defect. With the utilization of a temporary construction a controlled regeneration of soft tissues is ensured. MATERIALS AND METHODS: The patient V. I. 43 years old contacted us with complaints of difficulties during eating. The clinical examination established two defects of the dentition with missing 26 and 46 teeth. Rehabilitation with two fixed partial dentures was proposed. A preliminary impression was taken and a diagnostic model

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was poured. The model was scanned with 3D laboratory scanner – D800. The virtual models were analyzed in the software of 3Shape dental and two FPDs were modelled. The bridges were constructed in a 5-axis milling machine from PMMA. The FPD’s are cemented with temporary cement (MetaBiomed, Korea). R ESULTS: The temporary bridges have with high precision. The have excellent color stability over a long period of time and good mechanical properties because of the pre-polymerized PMMA. Because of the ability to be polished perfectly, they improve hygiene, soft tissue regeneration and formation. Clinical and laboratory time is reduced. CONCLUSION: The utilization of CAD/CAM technology in the construction of temporary constructions gives excellent results. Key words: provisional restoration, CAD/CAM

DM 7.

APPLICATION OF INFRARED SPECTROSCOPY IN DETERMINING

THE DEGREE OF CONVERSION OF DENTAL COMPOSITES Yordanka Uzunova, Mariana Katsarova, Lyudmil Lukanov Department of Chemistry and Biochemistry, Faculty of Pharmacy, Medical University - Plovdiv, Bulgaria In this study spectroscopy in the near and mid regions of the infrared spectrum was used in order to find the better technique for determining the degree of conversion (DC) of two ormocer dental composites used as direct and indirect restoratives (laboratory-processed composite). Specimens (n = 10) were prepared from both composites using special mold (2x8 mm) and were irradiated for 40 seconds with Bluedent LED Smart device (Bulgaria) having intensity 800 mW/cm2. The specimens made of laboratory-processed composite were additionally polymerized for 90 seconds at 80ºC. The degree of conversion was studied by fourier transform infrared spectroscopy (FTIR), Raman spectroscopy (400-4000 cm-1), and NIR spectroscopy (4000-7000 cm-1). Infrared spectra were recorded before and after polymerization. It was found that the degree of conversion of the composite used for direct restoration determined by FTIR technique (68.5%) was statistically different (p < 0.05) from DC determined with Raman spectroscopy (72%) and NIR spectroscopy (74.1%). FTIR technique was inapplicable for indirect composite. It achieved/displayed higher degree of conversion than direct composite (NIR–75.8%, Raman–74.2%). Although infrared spectroscopy is a widely utilized technique for determining DC, a scientifically based approach is mandatory when choosing the best methodology, so as to obtain the spectra that will enable a reliable evaluation of the degree of conversion. Key words: dental composites, degree of conversion, FTIR spectroscopy, Raman spectroscopy, NIR spectroscopy

DM 8.

COMBINED ORTHODONTIC SURGICAL TREATMENT OF A PATIENT WITH

PATHOGALVANISM Kristina Mihaylova1, Ivan Nachkov2, Donka Shetkova1 1Department of Orthodontics, 2Department of Periodontology, Faculty of Dental Medicine, Medical University - Plovdiv, Bulgaria INTRODUCTION: Pathogalvanism is condition to give an ion exchange between the different metals in the electrolyte environment, such as saliva. The pathogalvanism can be expressed clinically mainly by a change in the oral mucosa. CASE STUDY: A patient 28 years old, sought medical aid at the department of Periodontology and Oral Diseases, with a history of mucositis and glossopyrosis. CLINICAL STATUS: The initial examination found a fixed orthodontic apparatus and


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placed two endosteal implants in the field of 12 and 22 missing teeth. Over the implants we fixed transmucosal gingival formers. Buccal, labial and lingual mucosa were painful and hyperaemic. Paraclinical examinations: The corrosion potentials were measured: total, -1840 mV, at 12 and 22 tooth -200 mV. THERAPEUTIC APPROACH: The fixed device was saved. The steel arcs, placed on stability of the result at the end of orthodontic treatment, were replaced by rectangular NiTi aesthetic arcs. The supramucosal parts of the implants were isolated with orthodontic wax. We preferred a topical medication. R ESULTS: Early and late results were reported. After medical approach to establish sustained remission of the disease. DISCUSSION: In the combined treatment of maxillofacial congenital and acquired defects using different metal elements-brackets, arches, implants, which can be a cause of bimetal and cause disease of the oral mucosa. It should be taken into account when looking for clinical decisions with alternative materials and instrumentations. Key words: pathogavanism

DM 9.

DENTAL FEAR AND ANXIETY IN CHILDREN AND THEIR ASSOCIATION

WITH ORAL HEALTH Maria Shindova, Ani Belcheva Department of Pediatric Dentistry, Faculty of Dental Medicine, Medical University - Plovdiv, Bulgaria AIM: The aim of this study is to assess dental fear and anxiety and analyze their relationship with oral health, dietary habits and oral hygiene in children. MATERIAL AND METHODS: The study was conducted on 92 randomly selected children between 6 and 12 years old, treated at Department of Pediatric Dentistry, Plovdiv, Bulgaria. During the oral examination the dental status was determined by using the WHO criteria to obtain dft and DMFT indexes. A questionnaire was designed to investigate the gender, age, level of dental fear and anxiety, oral hygiene frequency and carbohydrate intake of the participants. R ESULTS: The mean scores dft and DMFT are 4.66 ± 3.73 and 1.20 ± 1.92, respectively. The children with high dental anxiety have significantly higher dft score than those with low or no anxiety levels (p < 0.05). The majority of children with moderate or high dental fear brush their teeth rarely or once a day (58.10%) and consume carbohydrates at least twice daily (74.20%). There is a significant association between dietary and oral hygiene habits and dental fear (p < 0.05). CONCLUSIONS: Dental anxiety causes avoidance of dental treatment, resulting in declines of oral health. Patients with more carious lesions undergo more painful and invasive dental procedures than caries-free children, which leads to the development of dental anxiety. Poor oral hygiene and frequent carbohydrate intake are risk factors for dental fear. They lead to deterioration of oral health, resulting in higher dental fear according to the `vicious cycle of dental fear. Key words: fear, anxiety, caries, hygiene, diet

DM 10.

ORAL PRECANCER AND EARLY CANCER DETECTION IN THE DENTAL

OFFICE – REVIEW OF NEW TECHNOLOGIES Nikolay Nikolov1, Svetlana Bachurska2, Ivan Nachkov3, Georgi Tomov1 1Division of Oral Pathology, Department of Periodontology and Oral Diseases, Faculty of Dental Medicine, Medical University Plovdiv; 2Department of General and Clinical Pathology, Medical University - Plovdiv; 3Department of Periodontology, Faculty of Dental Medicine, Medical University - Plovdiv , Bulgaria BACKGROUND: New technologies have provided an exciting new array of clinical diagnostic tools for localizing or emphasizing abnormal mucosa in the dental office, especially leukoplakia and

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dysplasia. Some of these technologies claim to identify atypical cells prior to biopsy, even before there are clinically visible mucosal changes, hence, can allow a more confident assessment of risk and localization of the most “suspicious” area to biopsy. In essence, molecular-level detection of dysplastic oral mucosal change appears to be moving into the practitioner’s office. METHODS: Extensive literature review, personal experience and discussion with other professionals with clinical experience with these technologies were used in order to provide a critical summary and evaluation of available technologies. CONCLUSIONS: Most technologies are beneficial but must be used with intelligence and must be considered adjunctive tests rather than stand-alone diagnostic tools. Loss of autofluorescence seems to hold the most promise for identifying mucosal dysplasia, but several nondysplastic lesions may also be nonfluorescent and occasional false positive results do occur. Key words: oral cancer, oral diagnosis, leukoplakia, erythroplakia, cytology

DM 11.

STUDY OF THE RELATIONSHIP BETWEEN THE ROTATION OF THE UPPER

FIRST MOLARS AND NON-SKELETAL CLASS II MALOCCLUSION Vladimir Petrunov, Ivanka Gigova Department of Orthodontics, Faculty of Dental Medicine, Medical University - Sofia, Bulgaria Many clinicians consider the position of the upper first molars as key for normal occlusion. It is known that these teeths are rhomboidal in form and wider buccolingually than mesiodistally. The mesiopalatally rotated molars occupy more space. The rotation of the upper first molar impair the normal occlusion and it is one of the etiological causes of non-skeletal ClassII malocclusion. The aim of this study was to establish a correlation between the extent of the non-skeletal Class II malocclusion and the rotation of the upper first molars. We studied a group of 82 untreated patients with mixed and permanent dentition, and a control group with Class I patients. To determine the position of the upper first molars we used the Ricketts’ method of assessment rotation of the above teeth. According to this method, a line is drawn from the distal buccal cusp through the mesial palatal cusp of the molar, that should pass through the opposite canine in the distal third of its crown. In case of mesiopalatal rotation, the line passes distally of the crown of the opposite canine. In all 82 models of untreated patients with Class II malocclusion we found mesiopalatal rotation of the upper first molars. In 6 of them (7%) the rotation was close to the border zone. The mesiopalatal rotation of the upper first molars is a main etiological causes for non-skeletal Class II malocclusion. A proper treatment plan will correct the rotation and the Class II malocclusion and it will increase the arch length and resolve the space deficiency. Key words: class II malocclusion, molar rotation

DM 12.

PHOTODYNAMIC LASER THERAPY – AN ALTERNATIVE THERAPEUTIC

APPROACH IN ORAL LICHEN PLANUS TREATMENT Maria Mutafchieva1, Milena Draganova-Filipova2, Svitlana Bachurska3, Plamen Zagorchev4 1Division of Oral Pathology, Department of Periodontology and Oral Disease, Faculty of Dental Medicine; 2Department of Medical Biology, Medical Faculty; 3Department of General and Clinical Pathology, Medical Faculty; 4Department of Medical Physics and Biophysics, Faculty of Pharmacy, Medical University - Plovdiv, Bulgaria Oral lichen planus (OLP) is a chronic inflammatory disease of unknown etiology and multifacto-


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rial pathogenesis. Corticosteroids are treatment of choice, but they have adverse effects and the results are often insufficient. Photodynamic laser therapy (PDLT) applied in patients with OLP shows good clinical results. The purpose of the current study was to establish the effectiveness of PDLT in OLP patients evaluating morphological changes in biopsy. A 56-year-old woman complaining of pain and burning sensation in the gingiva was clinically and histologically diagnosed with lichen planus- reticular form. The patient underwent continuous treatment (3 times per week for one month) with PDLT, using MB solution (500 μg/ml) and diode laser (810 nm) with following parameters 0.5 W; 30s; 1.2 J/cm2. The progression of the inflammatory process was evaluated by immunohistochemical method for detection of COX2. Its levels of expression before and after treatment were measured by semiquantative scale. The symptoms disappeared at the end of the first week. The keratotic lesions showed a partial response. The histological analysis demonstrated reducing of the hyperplastic processes in the epithelium and decreasing of the lymphocyte infiltrate after treatment. The expression of COX2 in the control specimen (+) was lower in comparison with the initial level (+++). The reported case proves the ability of PDLT to reduce the clinical symptoms of OLP. Morphological and IHC analysis gives evidence about the possible role of the laser energy in regulating the pathogenetic mechanisms of OLP. The results are basis for forthcoming studies, regarding more effective treatment of the disease. Key words: OLP, PDLT, COX2

DM 13.

PROVEN FOCAL-ASSOCIATED DISEASE

Marieta Todorova1, Stoyanka Velinova1, Antoaneta Gjurova2 1Department of Operative Dentistry and Endodontics, Faculty of Dental Medicine, Medical University Plovdiv, 2Department of Dermatology and Venerology, St. George University Hospital, Bulgaria Alopecia is a disorder, associated with hair-loss in areas where normally hairs grow. It can be congenital or acquired, scarring and nonscarring. Alopecia areata is a relatively frequent disease from the group of the acquired conditions. It is manifested as a loss of hairs in one or several well-circumscribed patches in hair-growing regions, with no subjective symptoms. The evolution of the disease is diverse. The infectious factors in the epidemiology of alopecia are often the most accessible to examine and prove, although they are questioned by some authors. We describe a case of a 30-year-old male patient with Alopecia areata in the region of the beard. Routine tests were conducted and revealed active foci in 38 and 48-semi-impacted third molars and potential focus in the area of 16 tooth with diagnose periapical lesion. There was slow reversible recovery of hair growth once the dental cause was removed. Key words: alopecia areata, focal disease

DM 14.

RESTORATION OF ENDODONTICALLY TREATED TEETH WITH

RADICULAR POSTS: A SURVEY AMONG BULGARIAN DENTISTS Ekaterina Karteva1, Teodora Karteva1, Neshka Manchorova1, Donka Keskinova2 1Faculty of Dental Medicine, Medical University - Plovdiv, 2Paisii Hilendarski University, Plovdiv, Bulgaria INTRODUCTION: There is an ongoing discussion among dentists on the use of radicular posts when restoring endodontically treated teeth (EТТ). AIM: The purpose of this survey was to determine the preferred techniques and materials used by dental professionals when restoring EТТ, and their opinion on the advantages and disadvantages they present. MATERIAL AND METHODS: A survey containing 12 multiple choice questions was distributed among Bulgarian dentists both online and during continuing education lectures. A total of 198 questionnaires were returned. The

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questions asked for information on when the clinicians used posts and what type of posts they preferred. They were also asked whether, in their opinion, vertical root fractures were caused by the placement of posts. The data obtained were processed using SPSS software. R ESULTS: Irrespective of their work experience, 96.9% of the surveyed dentists use posts when restoring EТТ. Almost 70% of them use posts routinely when restoring premolars and molars. The material of choice is a prefabricated metal post (70.3%), followed by a cast metal post (46.2%). Only 37.4% use non-metal posts, preferably fiber posts. The majority of dentists (54.4%) believe that vertical root fractures are caused by post placement. Eighty per cent prefer to place a crown after restoration with a post. CONCLUSION: In contrast to the recommendations in the literature, metal posts are still widely used as opposed to fiber ones. The reasons behind this might be a lack of information among dentists on the advantages of the latter and the clinical technique required. Key words: radicular posts, survey, fiber posts

DM 15.

STRESS IMPACT ON DENTAL MEDICINE STUDENTS’ PSYCHOLOGICAL

HEALTH Maria Manolova1, Neshka Manchorova1, Vesela Stefanova1, Donka Keskinova2, Ivan Panajotov1 1Faculty of Public Health, Medical University - Plovdiv; 2Plovdiv University „Paisii Hilendarski“, Bulgaria INTRODUCTION: The stress is the effect of external influences (physical and mental) upon the physical and psychological health of the individual. Dental medicine is associated with high levels of stress. The prime source can be found in the process of dental education. The present-day academic cycle requires numerous skills, including theoretical knowledge, clinical experience and interpersonal communication ability to be gained. The stress leads to mental manifestations: depression, anxiety, which influence over physical health. A negative correlation between stress and academic performance was founded. AIM: To study the impact of stress on psychological health of students in Dental Medicine. MATERIAL AND METHODS: In this research 104 third year students from the FDM in Plovdiv are involved. The questionnaire is anonymous. It consists of 22 questions that evaluate six emotional conditions: anxiety, depressive mood, positive well-being, self-control, general health, vitality. All responses are scaled from 0 to 5 points. The minimal score is 0, and the maximal is 110. The higher result shows a higher level of psychological health. For data processing a descriptive analyses is used. R ESULTS: 21% of the respondents have score in the 0-60 points’ interval, 31% of the students-in 80-110. 48% have a result between 60-80 points. CONCLUSION: Third year students show good psychological health. For the most of them stress effects positively by stimulating their creativity and energy. If everyday requirements reach levels which exceed the compensation opportunities of the organism negative consequences for mental and physical health are reached. Key words: stress, students, dental medicine, psychological health

DM 16.

SUBJECTIVE PAIN EVALUATION DURING INITIAL PERIODONTAL

TREATMENT USING GRACEY CURRETS AND ER:YAG LASER Elena Firkova1, Blagovesta Yaneva1, Emilia Karaslavova2 1Department of Periodontology and Oral Diseases, Faculty of Dental Medicine, Medical University Plovdiv; 2Faculty of Biology, Plovdiv University “Paisii Hilendarski”, Plovdiv, Bulgaria Mechanical root debridement is the corner stone of cause-related periodontal therapy, aimed at removal of subgingival biofilm and calculus, аnd controlling them to levels, allowing regeneraFolia Medica 2015; 57; Suppl. 1 © 2015 Medical University, Plovdiv, Bulgaria

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tive processes in the affected periodontal complex. AIM: To assess the subjective intensities of pain during initial periodontal treatment using Gracey currets and Er:YAG laser in patients with moderately advanced periodontitis. MATERIAL AND METHODS: 22 patients were included in the study. Periodontal treatment was performed following the clinical protocols. Subgingival scaling and root planning of teeth in 1 and 3 quadrants was performed with Gracey currets, the contralateral – with Er:YAG laser. A visual analogue scale was used by patients who were asked to define the intensity of pain experienced during clinical procedures in quadrants immediately after completion of the procedures. Comparison of mean levels was done by parametric Student’s t-test. R ESULTS: Subjective pain evaluation, experienced by patients during laser instrumentation is almost 2 times weaker then the pain, experienced with hand instruments (t = 3,594, р = 0,001). CONCLUSION: The results of this study show that root debridement of periodontally involved root surfaces performed by Er:YAG laser is nearly two times less painful than currets’ instrumentation. This increases the patients comfort during the treatment procedure and his compliance during initial and supportive periodontal therapy. Key words: periodontal treatment, pain, currets, laser

DM 17.

SURGICALLY ASSISTED ORTHODONTIC TREATMENT OF PROGENIA WITH

LATEROGNATHIA (A CLINICAL CASE) Kristina Mihailova, Silvia Krusteva Department of Orthodontics, Faculty of Dental Medicine, Medical Univerity - Plovdiv, Bulgaria Progenia (mandibular prognathism, III class malocclusion), a severe malocclusion, is a sagittal occlusal deviation characterized by medial occlusion of the lower dental arch toward the upper arch. This disorder is often combined with laterognathia. Treatment is difficult and at later age is a subject of common orthodontic-surgical correction. AIM: Preparation of an adequate treatment plan based on placement of comprehensive and accurate diagnosis. MATERIALS AND METHODS: A 14-year-old girl from Assenovgrad was referred to the Department of Orthodontics. After specific clinical and para-clinical studies (orthopantomography and teleradiography frontal and side views) the form and severity of the deformity were determined. The girl was diagnosed with progenia combined with laterognathia and hypodontia in both jaws. The planned orthodontic and surgical treatment resulted in optimal occlusion-articulation ratio with improvement of function and aesthetics. CONCLUSION: The specific orthodontics diagnostic methods made us choose surgically assisted orthodontic treatment but not orthognathic surgery indicated for this serious condition Key words: progenia, laterognathia, cephalometrie

DM 18.

SURVEY OF DENTAL STUDENTS’ PERCEPTIONS ABOUT THE MISSION OF

DENTAL SCHOOLS TO ADVANCE GLOBAL DENTISTRY AND PHILANTHROPY Bogomil Andonov1, Krasimira Yaneva2, Nina Musurlieva3, E. Athena, C. S. Ivanoff 1Department of Operative Dentistry and Endodontics, Faculty of Dental Medicine, Medical University - Plovdiv; 2Faculty of Dental Medicine, Medical University - Sofia; 3Department of Social Medicine and Public Health, Faculty of Public Health, Medical University - Plovdiv, Bulgaria The most visible mission of global dentistry is to educate students to work in a multicultural environment. AIM: To examine dental students’ perceptions about the mission of dental schools to

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advance global dentistry and philanthropy. MATERIALS AND METHODS: A survey among 228 third, fourth, fifth and sixth year dental students was conducted at the Medical University of Plovdiv, Faculty of Dental Medicine as part of an international project called the Global Oral Health Initiative (GOHI). The original questionnaire for all countries participating in the GOHI survey was translated into Bulgarian. A standard Pearson chi-square test was used to compare categorical variables (p < 0.05). SPSS v.14 was used for data analysis. R ESULTS AND DISCUSSION: Based on the total number of respondents, the majority of students (90 %) believe that volunteerism and philanthropy are important attributes of a well-rounded dental practitioner and that their dental school does teach them about these virtues. However, 54% of the respondents felt that their dental school does not encourage them in volunteerism, nor does enough to promote programs to care for the underserved. On the other hand, the multicultural environment at the dental school encourages students to respect and tolerate people of different cultures. The majority also reported readiness to participate in international exchange programs that would enhance their cultural competence training and promote understanding of the factors challenging disparate groups to accessing equal health care. Conclusion: This study indicates that students want their school to provide them with increased educational experiences in global dentistry and community service. Key words: curriculum, cultural competence

DM 19.

TEMPERATURE CHANGES ON THE TITANIUM SURFACE DURING LASER-

ASSISTED THERAPY OF PERIIMPLANTITIS WITH ER:YAG LASER Ivan Nachkov1, Nikolay Nikolov1, Plamen Zagorchev2, Georgi Tomov1 1Division of Oral Pathology, Department of Periodontology and Oral Diseases, Faculty of Dental Medicine, Medical University - Plovdiv, 2Department of Medical Physics and Biophysics, Faculty of Pharmacy, Medical University - Plovdiv, Bulgaria INTRODUCTION: Peri-implantitis is defined as an inflammatory process affecting the tissues around the integrated implant, resulting in loss of function and supporting alveolar bone. As microbial colonization plays a major role as an etiological factor, it is assumed that the removal of bacterial biofilm from the implant surface is a prerequisite for acquiring periimplant infection. There are reports on the use of ER: YAG lasers for decontamination of the implant, without surface modification. AIM: The aim of this study is to evaluate the temperature changes in the endosteal titanium implants and surrounding biological tissues when treated with ER: YAG (2940 nm) laser. MATERIAL AND METHODS: Biological material (pig jaws) with placed submerged implants were used (preheated up to (32 ± 0.5°C). Via thermocouples placed in and around the implants were measured temperature changes during 30-min irradiation with Er: YAG laser at two modes: 400 mJ / 17 Hz / 6.80 W and 50 mJ / 30 Hz / 1.50 W. R ESULTS: After treatment with Er: YAG laser were recorded minimal temperature variations ranging 30.2÷32.5°C . DISCUSSION: Unlike other lasers, Er:YAG laser does not modify thermally the implant interface, which should be taken into account during the clinical use of lasers for uncovering implants and treatment of periimplantitis. Er:YAG lasers may also be suitable for treatment of the implant surface, because their specific wavelength is absorbed poorly by the titanium and, accordingly, the temperature of the implant does not grow significantly. Key words: periimplantitis, ER:YAG laser


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DM 20.

THE NECESSITY OF STUDYING “COMPUTER TECHNOLOGY” FOR DENTAL

STUDENTS Tania Boeva, Kristina Kilova Department of Medical Informatics, Biostatistics and E-Learning, Faculty of Public Health, Medical University - Plovdiv, Bulgaria INTRODUCTION: Using computers in dental practice has already become a common skill worldwide. The too narrow professional specialisation has long become a fact of the past. To get successful in one’s field dentists should be specialists with a wide range of knowledge and skills. AIM: To survey the opinions of dental students and their teachers at the Faculty of Dental Medicine, Medical University - Plovdiv about including the subject of ‘computer technology’ in their schedules, and about the topics they should be trained in and the forms of studying the new subject. MATERIAL AND METHODS. A direct anonymous survey was conducted among students and lecturers at the Faculty of Dental Medicine, Plovdiv. Three different questionnaires were used – for first year students, for second to fifth year students and for their lecturers. The results were analysed statistically using SPSS 17.0. R ESULTS AND DISCUSSION: We found a statistically significant difference between the opinions of junior students and senior students and their lecturers about the need to study computer technology. CONCLUSION: The subject of computer technology should without a doubt be included in the syllabus of students in the major ‘Dental Medicine’. To know better and apply the modern rapidly evolving technologies, it is necessary to constantly update the educational content. Key words: computer technology, education, dental medicine

DM 21.

THE USE OF DENTAL OPERATING MICROSCOPE IN REMOVING

GUTTA-PERCHA FROM THE ROOT CANAL – COMPARATIVE IN VITRO STUDY Liubomir Vangelov Department of Operative Dentistry and Endodontics, Faculty of Dental Medicine, Medical University Plovdiv, Bulgaria INTRODUCTION: : Dental operating microscope (DOM) has established itself as an indispensable tool for most stages of primary and secondary endodontic treatment. Data on the effectiveness of cleaning the gutta-percha from the canal using the DOM are insufficient and contradictory. The aim of this study was to clarify the effectiveness of cleaning the gutta-percha from the root canal using DOM, compared to cleaning without any magnification. MATERIAL AND METHODS: 51 extracted single rooted teeth were selected for the study. They were prepared to 06/30 and obturated with lateral condensation of gutta-percha and sealer AH-26. Gutta-percha was removed with the following instruments: Group 1-H- files (control group); Group 2-Pro Taper Retreatment (PTR); Group 3- PTR+DOM. Residual gutta-percha was registered with cone-beam three-dimensional computed tomography in the coronal, middle and apical part. Three computer slices were processed for each part - transverse, sagittal and frontal. Residual gutta-percha was evaluated using indexes specially developed for the study. One-way ANOVA was used in the study with level of statistical significance at p < 0.05. R ESULTS: In all groups there remains a certain amount of gutta-percha in the canal. The least amount remains after cleaning with DOM, the difference with the other groups is statistically significant (p> 0.05)) CONCLUSION: Using DOM has the advantages of accomplishing a better cleaning of gutta-percha from the root canal. Key words: retreatment, gutta-percha, microscope

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DM 22.

THREE DIMENSIONAL PREPARATION OF THE DENTAL ARCH FOR

IMPLANTOLOGY. A CLINICAL CASE Greta Yordanova1, Martin Mladenov2 Department of Orthodontics, Faculty of Dental Medicine, Medical University - Sofia; 2Private Practice, Sofia, Bulgaria In the cases of multiple tooth agenesis, a good choice of treatment is restoration of the missing teeth with implants. This is possible if the alveolar ridge has enough volume in the three dimensions and lack of overerupted antagonists. AIM of this study is to analyze the clinical management in the interdisciplinary treatment orthodontic-surgery preparation of the upper dental arch for implantology. MATERIALS: We present a clinical case of 24 years old patients. She has tooth agenesis of the upper left lateral incisor and all of the upper premolars. The primary lateral incisors, which has smaller dimensions rather than the permanent is persistent in the frontal segment. The permanent canines are in incorrect position. In the upper right segment there is a persistent primary ankylozed molar and lack of space for two permanent premolars. In upper left segment there are two primary ankylozed molars persistent. In the areas of tooth ankylosis the bone is underdeveloped and the adjacent teeth are inclined to the primary teeth. R ESULTS. We distalized the first molars with the Pendulum appliance. We used the ankylozed teeth as an absolute anchorage. Alignment of the frontal teeth and gaining space for the lateral incisors was done. Extraction of the ankylozed teeth led to bone loss so bone grafting was done by the surgeon. During the healing period the teeth were stabilized in their new position. CONCLUSION: Without orthodontic treatment there wouldn’t be enough stability of the dental arch for appropriate surgery and implantology. Key words: multiple tooth agenesis, implantology, ortodonthics

DM 23.

ULTRASONICS IN ROOT CANAL FILLING PASTES REMOVAL -

A COMPARATIVE IN VITRO STUDY Liubomir Vangelov Department of Operative Dentistry and Endodontics, Faculty of Dental Medicine, Medical University Plovdiv, Bulgaria INTRODUCTION: Filling pastes in the root canal are serious obstacle in endodontic retreatment. In these cases ultrasonic tips may be the only successful method. There is evidence that their use as a final step in combination with other instruments results in better cleaning of the canal. AIM: To determine the effectiveness of the cleaning of root canal filling paste with hand instruments with and without ultrasonics. MATERIALS AND METHODS: Forty-five single rooted teeth were prepared to 30/02 and obturated with paste cortisomol. The teeth are divided into three groups: 1st group - removal of the paste is done with H-files only; 2-nd group - H-files with ultrasonics (Pro Ultra-Dentsply Maillefer); 3 - Group III (control) - not cleaned paste. The teeth are cut longitudinally and photographed with Canon 500D camera and macro lens. The area of the residual paste was measured using software developed for the purpose of study - Area Calculator. For statistical analysis one-way Anova test was used as the level of statistical significance is P < 0.05. R ESULTS: The amount of residual paste in the 2nd group is less than the 1st group. The difference between the groups for coronal and middle third is statistically significant (p < 0.05).


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CONCLUSIONS: The removal of filling paste with hand instruments in combination with ultrasonic tips results in better cleaning of the root canal as compared to using only hand instruments. Key words: endodontic retreatment, pastes, ultrasonic removal

DM 24.

THE IMPACT OF HALITOSIS ON QUALITY OF LIFE

Nina Musurlieva1, Maria Stoykova1, Donka Kirova2 1Department of Social Medicine and Public Health, Faculty of Public Health, Medical University - Plovdiv; 2Department of Oral Surgery, Faculty of Dental Medicine, Medical University - Plovdiv, Bulgaria The prevalence of halitosis in general population is in the range of 22-50%. Halitosis has a multietiologic origin. Although most of the cases are from the oral cavity, sometimes other etiologies can contribute to oral halitosis - gastro-oesophageal reflux; haematological, renal, endocrine diseases. Halitosis is a common social condition that affects a considerable portion of the general population. This oral condition may cause embarrassment, depression and make relationships more difficult. Halitosis may lead to changes in the behavior and social life of those affected by it. Key words: halitosis, quality of life

POSTERS

Science and Youth SY 1.

LUNG RESECTIONS IN PURULENT–DESTRUCTIVE DISEASE IN CHILDHOOD

Biser Ivanov, D. Dachev, S. Lupanov, M. Simeonov, I. Kirev, M. Nenov, P. Stefanova, A. Yonkov, E. Moshekov, D. Mitkovski Department of Pediatric Surgery, St. George University Hospital, Plovdiv, Bulgaria BACKGROUND: Inflammatory lung diseases are a serious problem in pediatric thoracic surgery. The number of patients with acute purulent lung disease become inreasingly greater. MATERIAL AND METHODS: 116 children with purulent-destructive lung disease were operated over a period of 15 years (2000-2014) in the Department of Pediatric Surgery. R ESULTS: Pulmonectomy was performed in one child, bilobectomy - in 2 children, 3 children had lobectomy, 2 - segmentectomy, and 7 children undergone atypical lung resection. Postoperatively, the children developed no complications. CONCLUSION: Purulent-destructive lung diseases in childhood is characterized by dynamic development and many complications which frequently require surgical treatment. Key words: lung, resection, children

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SY 2.

METHODS FOR DIAGNOSIS AND SURVEILLANCE OF PATIENTS WITH

GLUTEN-SENSITIVE ENTEROPATHY – A CLINICAL CASE Milena Kerezova1, Stanislav Churchev2 1Medical University - Sofia, 2Clinical Center of Gastroenterology, Department of Gastroenterology, Queen Joanna University Hospital, Sofia, Bulgaria INTRODUCTION: Gluten-sensitive enteropathy is a T-cell mediated disease, occurring in genetically susceptible individuals, that is induced by the ingestion of one of several proteins found in wheat (gliadin), barley (hordein) and rye (secalinin). Considered to be a relatively uncommon disease in the past, recent studies show that celiac disease affects between 1% and 2.5% of the population. Symptoms classically include episodic diarrhea, abdominal pain and distention and weight loss, but other signs of malabsorption may be present. Оnly one half of all patients with gluten-sensitive enteropathy have symptoms referable to the gastrointestinal tract. Тhe diagnosis is based on the combination of positive serological markers (anti-tTG, anti-DGP, anti-endomisial antibodies), intestinal villous atrophy confirmed by histologic examination of small-bowel biopsy specimens and clinical and serological answer to gluten-free diet. CASE REPORT: A 41-year-old woman with accompanying autoimmune pathology was admitted to hospital on the occasion of malabsorption. A diagnosis of gluten-sensitive enteropathy was made on the basis of clinical, laboratory, imaging techniques data and histological findings. Pathomorphological examination of the mucosa showed subtotal villous atrophy, and serological results confirmed - anti-tTG > 200 U/ml and anti – DGP antibodies >100 U/ml. Significant distortions of coagulation parameters, with no bleeding despite the reduced absorption of Vitamin K, which responded well to intravenous application of Vitamin K (Konakion). A favorable therapeutic result and decrease in serum antibodies was observed after strict gluten-free diet was introduced. CONCLUSION: Celiac disease should be suspected and excluded in every presenting patient presenting with malabsorption. Key words: gluten-sensitive enteropathy, intestinal villous atrophy

SY 3.

SENSITIZATION TO DRUGS, USED IN ANESTHESIOLOGY PRACTICE

Plamena Novakova1, Vanya Asparuhova2, Mariya Atlieva2, Valeri Goergiev1, 1Medical Faculty, Medical University - Plovdiv, 2Faculty of Public Health, Medical University - Plovdiv, Bulgaria INTRODUCTION: Both underdiagnosis and overdiagnosis of sensitization to drugs are problems in modern allergological practice. The AIM of the present study was to investigate the sensitization to drugs used in anesthesiology practice. PATIENTS AND METHODS: In the present prospective study 356 patients [men 109 (30.62%); mean age 51.98 years (SD 20.19)], tested in the Allergology Section of the Internal Consulting Department, St. Georgi University Hospital - Plovdiv (August 2014 – January 2015), reported having sensitization to anesthesiology drugs from the previous skin testing. A detailed clinical history was taken and dermal prick-tests with anesthetic drugs were performed as recommended by International consensus on drug allergy of European Academy of Allergy and Clinical immunology (EAACI). R ESULTS: Sensitisation was found in 107 (30.06%) of patients. Monosenzitised patients were 42 (39.25%) and the polysenzitised patients were 65 (60.75%). The following sensitizations were found: thiopental – 2; fentanyl – 10; chyrocain – 7; atropine sulfate – 15; ketamine – 14; suxamethonium – 12; pancuronium - 2; pipecuronium – 12; diazepam – 4; galantamine – 59; lidocaine – 7; bupivacain – 5; propofol – 5; midazolam – 2; pethidine – 59. Folia Medica 2015; 57; Suppl. 1 © 2015 Medical University, Plovdiv, Bulgaria

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CONCLUSION: The study established overdiagnosis in assessment of sensitization to anesthetic drugs. The main reason for that, on our opinion, was the incorrect skin-testing, which should be performed by a prick-test as recommended by EAACI and carried out in specialist centers by highly skilled stuff. Key words: sensitization, anesthetics, prick-test

SY 4.

PROCOAGULANT MICROPARTICLES IN PATIENTS WITH SOLID TUMORS –

CLINICAL AND PROGNOSTIC CORRELATIONS Elina Beleva1, T. Deneva2, E. Karaslavova3, S. Stoencheva2, V. Popov4, M. Valcheva1, J. Grudeva-Popova1 1Division of Medical Oncology, Department of Clinical Oncology, Medical Faculty, 2Department of Clinical Laboratory, Faculty of Pharmacy, St George University Hospital St. George, Plovdiv; 3 Paisii Hilendarski Plovdiv University, 4Medical University - Plovdiv, Bulgaria INTRODUCTION: Microparticles (MPs) are highly procoagulant submicron vesicles released from activated cells that serve as a source of functional tissue factor (TF) in circulation. Elevated levels of tumor derived TF-bearing MPs have been found in cancer patients and they have been correlated to tumor burden. AIM: to determine changes in MP-TF procoagulant activity during chemotherapy and find correlation with the clinical course of disease. MATERIAL AND METHODS: Single institution observational study including 128 cancer patients who underwent chemotherapy in 2013/2014 for breast (44), lung (28), colon (37), and ovarian (19) cancer. Patients were subsequently followed up: before chemotherapy initiation, after three cycles, after six cycles and three months post-therapy cessation. A control group of 30 healthy volunteers was studied as well. Serum levels of TF-antigen were determined by ELISA and MP-TF procoagulant activity was measured by a combined immune-chromogenic method (zymuphen MP-TF). R ESULTS: MP-TF procoagulant activity was twice higher in cancer patients compared to controls (0.9421 vs. 0.4300 pg/ml, p < 0.0001). Sequential testing of MP-TF testing in patients with lung, colorectal and ovarian cancer revealed decrease in their procoagulant activity during treatment followed by a significant rise in activity after therapy cessation (0.9421 vs. 1.4103 pg/ml, p < 0.0001). No significant dynamics in MP-TF was observed in patients with breast cancer (0.9814 vs. 0.8733 pg/ml, p = 0.785). CONCLUSION: MP-TF procoagulant activity could serve as a surrogate marker for biologic activity and therapeutic efficacy. Key words: procoagulant microparticles

SY 5.

DUODENAL ATRESIA COMBINED WITH OTHER GASTRO-INTESTINAL

ANOMALIES S. Lupanov, I. Kirev, B. Ivanov, M. Simeonov, D. Dachev, E. Moshekov, P. Stefanova, Department of Pediatric Surgery, St. George University Hospital, Plovdiv, Medical University - Plovdiv, Bulgaria BACKGROUND: Duodenal atresia is a relatively rare anomaly with an incidence of approximately 1:5000 live births. In 1-2% of cases with duodenal atresia, a combination with other gastro-intes-

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tinal anomalies can be present. This combined anomaly leads to many problems in the diagnostic and treatment process. MATERIAL AND METHODS: Over a period of 20 years between 1994 and 2014 in the Clinic of Pediatric Surgery at St. George University Hospital in Plovdiv, 50 children were operated, with different forms of duodenal atresia. In three cases other gastrointestinal anomalies were found. R ESULTS: The three reported children were operated successfully, continuity of the gastro-intestinal tract and per oral nutrition were restored. Mortality rate is too high, due to serious prematurity and immaturity, aggravating the severe postoperative sepsis. DISCUSSION: The reported gastrointestinal anomalies in our patients are most probably due to abnormalities in development. The high mortality in the early postoperative period is a result of various factors. There are only a few similar cases in literature. The difficulties in diagnostic methods, surgical approach, and postoperative intensive care are discussed. The authors share their experience in treating these severe combined gastrointestinal anomalies. Key words: duodenal atresia, children, combined anomalies

SY 6.

SEXTING AND INTERNET ADDICTION

Zlatoslav Arabadzhiev1, Vartuhi Kadiyan2, Andreas Hadzigiannis3 1Department of Psychiatry and Medical Psychology, Medical University - Plovdiv, 2Psychological Consultant in London, UK, 3Faculty of Medicine, Medical University - Plovdiv, Bulgaria In modern society the expression of the ‘sexual’ in the context of interpersonal relationships is more accessible and commercial than ever. The Internet is ubiquitous throughout the world and with it the access to pornographic web sites. It was found that sexual content in the media increases and accelerates the sexual activity of adolescents, along with increased sexually-tolerated attitudes and stereotypical gender perceptions. Adolescents who visit pornographic web sites often have wider and risky sexual encounters with more sexual partners. There is a positive correlation between the content of porn media and off-line sexual tendencies and behavior. We all have witnessed shocking stories with circulated pictures and videos of young people in social media, which have been detrimental to them and have caused them to drop out of school and even commit suicide. Nevertheless, in modern society sexting has become part of a normal sexual maturation of young people. Sexting is currently seen as the electronic version of the ‘if you show me yours, I’ll show you mine’. Key words: cyber psychology

SY 7.

APPROPRIATE SELF-TREATMENT AS AN IMPORTANT ASPECT OF SWITCH-

ING PRESCRIPTION DRUGS TO OVER-THE-COUNTER DRUGS Valentina Belcheva1, Evgeni Grigorov2, Tihomira Zlatanova3, 1Faculty of Public Health, Medical University -Sofia, 2Faculty of Pharmacy, Medical University - Varna, 3Faculty of Public Health, Medical University - Sofia, Bulgaria Self-care refers to unorganized health activities and health related decision made by individuals, families, friends, colleagues at work, etc.; it includes self-medication, self-treatment, social support in illness, first aid in a ‘natural setting’, i.e., the normal social context of peoples’ everyday lives. Self-care is definitely the primary health resource in health care system. This type of care is definitely one of the main resources for health in the healthcare system. For their effective implementation a vital role play the drugs obtained without a prescription. They are the basis for proper healing of patients. Folia Medica 2015; 57; Suppl. 1 © 2015 Medical University, Plovdiv, Bulgaria

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Globally, there is an increase in the number of prescription drugs that are in the process of changing their status of prescription and switch to over-the-counter status (OTC). These changes, called switches, are motivated mainly by three factors: the willingness of pharmaceutical companies to extend the validity of their trademarks; attempts by funding health institutions to contain drug costs; the role of self-treatment by patients. Benefits of self-treatment are the foundation of the pyramid of health and allow the system to focus its scarce resources to the diagnosis and treatment of serious, socially significant diseases, new research and innovative services. OTC drugs are an indispensable element of self-care. CONCLUSION: OTC switches can be an important policy tool for improving public health in drug classes where a significant proportion of the population is untreated and where consumers can effectively manage treatment with limited physician supervision. Key words: self-medication, medicines, switches

SY 8.

A FETOPATHOLOGICAL AND CLINICAL STUDY OF DANDY-WALKER

SYNDROME AND A LITERATURE REVIEW T. Kitova1, B. Kitov2, H. Zhelyazkov2, D. Chelli3, A. Masmoudi4, S. Gaigi4, A. Tenev5, 1Department of Anatomy, Histology and Embryology, 2Clinic of Neurosurgery, Medical University - Plovdiv, Bulgaria, 3Department of Obstetrics and Gynecology, Maternity and Neonatology Center, Tunis, Tunisia; 4Embryo-foetopathology Unit, Maternity and Neonatology Center, Tunis, Tunisia, 5Faculty of Medicine, Medical University - Plovdiv, Bulgaria Dandy-Walker Syndrome is a rare abnormality of the central nervous system pertaining to the group of cystoid malformations. The frequency of occurrence of the syndrome ranges from 1/25000 to 1/35000 live births. The AIM of this study was to investigate Dandy-Walker Syndrome in two cases - one by autopsy after abortion due to medical indications during the 25th gestational week at the Clinic of Embryo and Fetopathology, Center for Maternity and Neonatology, Tunis, Tunisia, and another case of a one-year-old infant diagnosed with an acute internal hydrocephalus at the Clinic of Neurosurgery, Medical University - Plovdiv, Bulgaria, and to compare the results with those in literature. R ESULTS: Both cases are isolated Dandy-Walker Syndrome without any associated abnormalities of the central nervous system and without incompatible malformations of other organs and systems. After the placement of a ventriculoperitoneal shunt in the second case the childs condition was greatly improved. CONCLUSION: The question arises whether the prenatal diagnosis of an isolated Dandy-Walker Syndrome is an indication for pregnancy termination, and who and after what research has the right to make this decision, given the favorable outcome after surgical treatment. Key words: Dandy-Walker syndrome, prenatal diagnosis

SY 9. NEUROENDOCRINE TUMOR OF APPENDIX OR MORBUS CROHN DIFFERENTIAL

DIAGNOSIS AND OPERATIVE APPROACH WITHOUT PREOPERATIVE HISTOLOGY P. Vladova1, S. Iliev1, Iv. Presolski1, K. Nedyalkov1, S. Popovska2 1Department of Coloproctology and Purulent-Septic Surgery, 2Department of Clinical Pathology, University Hospital - Pleven, Bulgaria INTRODUCTION: The neuroendocrine tumors (NET) of the appendix and Morbus Crohn belong to the group of rare diseases according to the widely accepted European definition of a rare disease.

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Usually they are discovered accidentally because of the lack of specific symptoms and because the clinicists are not well acquainted with them. The patients usually come to the hospital with manifestation of intestinal obstruction and they undergo urgent surgery. Both diseases require significantly different surgical approach and subsequent treatment. MATERIALS: We present 3 cases of patients operated urgently or in conditions of urgency with delayed operation with manifestations of intestinal obstruction and palpable tumor formation in the ileocecal area. Different operative interventions were performed. The patients were followed up over a period of 2 years. R ESULTS AND DISCUSSION: Its noteworthy that the case histories and the clinical data suggested malignant neoplasm of appendiceal origin while the intraoperative findings suggested chronic inflamatory process of the terminal ileum with enlarged lymphatic nodules in the pool of arteria ileocolica which is supported by the histological findings. In 2 of the cases the histological diagnosis was Morbus Crohn and in 1 of the cases - NET of the appendix. CONCLUSION: Knowledge of the clinical picture and the macroscopic findings in Morbus Crohn and NET of the appendix and cecum is very important for the choice of appropriate operative tactics in order to avoid subsequent complications and unfavourable outcome of the treatment. The frequency of these diseases increases and, on the other hand, the application of approved regimens and algorithms of treatment is a prerequisite for good results and prognosis. Key words: NET of the appendix, Morbus Crohn, differential diagnosis

SY 10.

INFESTATION WITH GANGLYONELA PULCHRUM – A CASE REPORT

Tsvetomira Dobreva1, Simona Nikolaeva1, Yana Manolova2 1Medical University - Varna, 2Eye Hospital, Varna, Bulgaria Gongylonema pulchrum is the only parasite of the genus Gongylonema capable of infecting humans. Gongylonema pulchrum infections are due to humans acting as accidental hosts for the parasite. The G. pulchrum parasite is a nematode worm of the order Spirurida. It is a relatively thin nematode, and like other worms in its class, it has no circulatory or respiratory system. This parasite is multi-cellular and motile. They have numerous rear mucosal projections, which are believed to assist propulsion through the thin layer of skin on the inside of the human hosts mouth. Rarely, it can be found in the eye. We present a case report of female patient of infestated with Gongylonema pulchrum. She presented with headache and tingling in various parts of the eye tissue after a hit with a metal rod. Later the patient was admitted to the Eye Hospital in Varna. Her main complains were redness and oedema in the left eye. After a consultation with an ophthalmologist an operation was performed and a worm 13 cm long and 3 cm wide was extracted from the left eye. The result of the parasitology analysis showed it to be Gonglyonema pulchrum. During the control physical examination a number ducts and a perforation in the orbit septum were found. After the surgery the patient was referred to a psychiatrist and a neurologist. Key words: ganglyonela pulchrum, infestation, worm

SY 11.

TH1/TH2 CYTOKINE PROFILE IN HIV/HCV CO-INFECTED PATIENTS,

NAIVE TO CART Marina Alexandrova1, Mariya Muhtarova1, Olga Angelova1, N. Yancheva2, Maria Nikolova1 1NRL of Immunology, National Center of Parasitic and Infectious Diseases, Sofia, 2 Professor Ivan Kirov Hospital for Infectious and Parasitic Diseases, Sofia, Bulgaria BACKGROUND: Chronic HIV and HCV infections are characterized by inefficient T cell response and ongoing immune activation, associated with alterations in effector and regulatory cytokines. Folia Medica 2015; 57; Suppl. 1 © 2015 Medical University, Plovdiv, Bulgaria

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Due to shared transmission routes, HCV co-infection affects up to 30% of the HIV+ individuals, leading to unpredictable clinical course and response to combined anti-retroviral therapy (cART). AIM: To establish whether HCV co-infection specifically affects cytokine milieu, and hence T cell response in HIV+ patients . METHODS: Cytokine profiles of cART-naïve HIV/HCV co-infected patients (n=6) were compared to mono-infected (HIV+, n=7; HCV+, n=5), and healthy controls (HC, n=6). IFNγ, TNF, IL-2, IL-4, IL-6, and IL-10 levels were determined after 18h phytohemagglutinin stimulation of whole blood, using a standard flow-cytometry bead array test (CBA Human Th1/Th2 cytokine kit, BDTM). R ESULTS: Although Th1 cytokine response (IFNγ,TNF,IL-2) dominated over Th2 (IL-4), the levels of IFNγ and IL-2 were significantly decreased in all patients’ groups. Importantly, IFNγ/IL-10 ratio describing the efficiency of regulated Th1 effector response was significantly decreased as compared to HC: mean 4.3 (HIV); 2.1 (HIV/HCV); 1.8 (HCV) vs. 9.5 (HC), p < 0.05. The proinflammatory IL-6 cytokine was significantly increased in HIV, and even more so in HIV/HCV co-infection: mean 36866 and 39234 vs. 22453 (HC). Noteworthy, HIV/HCV co-infection was distinguished by a high IL-6/IL-2 index: 1961 vs. 1381 (HIV), 484 (HCV), 112 (HC), p < 0.05, reflecting the insufficient control of chronic activation by regulatory T cells (Treg). CONCLUSION: HCV co-infection contributes to a dysfunctional immune response in chronic HIV, most probably by affecting the levels and differentiation state of Treg. Key words: HIV/HCV co-infection, cytokine profile

SY 12.

PREVALENCE OF CRIMEAN-CONGO HEMORRHAGIC FEVER VIRUS IN

TICKS IN BULGARIA Elitsa Panayotova, Iva Christova National Center of Infectious and Parasitic Diseases, Sofia, Bulgaria INTRODUCTION. Crimean-Congo hemorrhagic fever (CCHF) is an acute severe viral disease transmitted by ticks. It is often associated with hemorrhagic presentations and high mortality rate. Bulgaria is an endemic area for CCHF and in the last 4 years 21 cases have been confirmed. They were registered in 5 districts of the country – Burgas, Yambol, Haskovo, Kardzhali and Blagoevgrad. This study was conducted to determine the rate of CCHF virus infection in ticks, collected from livestock in these 5 endemic districts. Similar studies are very few in Europe. MATERIAL AND METHODS. A total of 2315 ticks were collected. Real time reverse transcriptasepolymerase chain reaction (RT-PCR) was used for the detection of the CCHF virus genome in the collected Hyalomma marginatum ticks. R ESULTS: CCHF virus genome was detected in 7.3% of H. marginatum ticks (17 positive ticks out of 233) collected from livestock in Kardzhali district. The animal infectivity is estimated as 6.3% (5 CCHF virus positive out of 79). CCHF virus genome was also detected in ticks collected from livestock in Burgas district. 8.8% of H. marginatum ticks were found positive (19 out of 217). Animal infectivity here was 6.7% (4 CCHF virus positive out of 60). CONCLUSION: This study elucidates actual rate of CCHF virus infestation of the ticks in endemic regions of Bulgaria. Such rate of infestation facilitates high virus transmission between ticks and their animal reservoirs, enhancing the risk of infection for people in close contact with the infected livestock. Key words: CCHF, tick, Bulgaria, Hyalomma, RT-PCR

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SY 13.

MULTIPLEX PCR METHOD FOR IDENTIFICATION OF CLINICAL CAMPY-

LOBACTER SPP. ISOLATES AND DIFFERENTIATION BETWEEN CAMPYLOBACTER JEJUNI AND CAMPYLOBACTER COLI Maria Pavlova, Elina Dobreva, Katjusha Ivanova, Galina Aseva, Ivan Ivanov, Peter Petrov, Tzvetan Velinov, Todor Kantardjiev National Center of Infectious and Parasitic Diseases, Sofia, Bulgaria Campylobacter causes infections in children, adults and patients with suppressed immunity. C. jejuni and C. coli are frequent agents of campylobacteriosis, which presents clinically like mild to severe gastroenteritis or enterocolitis and may lead to life-threathening complications. OBJECTIVE: Evaluation and optimization of Multiplex PCR method for identification of Campylobacter isolates and differentiation between C. jejuni and C. coli. MATERIALS AND METHODS: 85 samples from diarrheal patients were tested between February 2014 and January 2015. Fecal samples were investigated with conventional microbiological methods. Isolates were identified biochemically using hydrolysis of sodium hyppurate and indoxyl acetate tests. DNA from 20 isolates was extracted with QIAampDNA Blood Mini kit (QIAGEN). The conducted multiplex PCR was based on the genus-specific cadF gene as well as the species-specific hipO for C. jejuni and asp for C. coli respectively. The obtained amplicons were visualized with capillary gel electrophoresis. R ESULTS: Fourty Campylobacter spp. isolates have been obtained from cultures. Results from the biochemical identification were compared to these from the Multiplex PCR analysis. Sixteen out of twenty isolates, which were biochemically identified as C. jejuni, revealed the hipO gene. Three of twenty isolates were biochemically determined as C. coli. Two of them possessed the species-specific asp gene, while the third was genetically determined аs Campylobacter spp., because only the genus-specific cadF was present. One of the twenty isolates, determined biochemically as Campylobacter spp., was confirmed genetically as well. CONCLUSION: The comparative analysis of results has proved that Multiplex PCR method is relevant and reliable for differentiation between C. jejuni and C. coli. Key words: diarrhea, campylobacter, multiplex PCR, gastroenteritis, enterocolitis

SY 14.

STATISTICAL ANALYSIS OF DIFFERENT FACTORS AFFECTING ACUTE

HEPATITIS A D. Petrov1, L. Pekova2 1Department of Social Medicine and Health Management, Medical Faculty, Trakia University, Stara Zagora, 2Clinic of Infectious Diseases, Prof. Stoyan Kirkovitch Hospital, Stara Zagora, Bulgaria Acute hepatitis A (AHA) is an acute infectious disease which is typical for persons in the age range 1-18 years, but lately quite often cases outside this range were observed. The study aims to determine the set of factors by examining the significance on given dependent variable, and study between variables. This will determine the best predictor of the dependent variable. Creation of regression models determines which factors predict the probability of an impact on the dependent variable. Between 2010 and 2012, 1601 patients with AHA were treated in the Clinic of Infectious Diseases, at Prof. Stoyan Kirkovitch Hospital in Stara Zagora. Samples from 214 patients were randomly obtained, 89 (41.6%) women and 125 (58.4%) men. The Kolmogorov-Smirnov test, standard multiple regression and logistic regression analyses were used in the study. Key words: regression models, multiple regression, logistic regression, predictors, infection diseases


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SY 15.


THE ROLE OF MAGNETIC RESONANCE SPECTROSCOPY IN THE

DIAGNOSIS OF HIV-ASSOCIATED NEUROCOGNITIVE DISORDERS Vanya Georgieva1, Mariana Stoycheva1, Borislav Milev2 1Department of Infectious Diseases, Parasitology and Tropical Medicine, Medical University - Plovdiv, Bulgaria; 2 Rusev Diagnostic Laboratory HIV associated neurocognitive disorders are an important issue among HIV+ people, because of their increasing frequency (35-50%) and the fact that they deteriorate the quality of patients’ life. Diagnosing the condition is difficult requiring complex diagnostic approach. Magnetic resonance spectroscopy is a method, which allows clinicians to specify the type and location of the disorders of cerebral metabolism. The method has many advantages over the magnetic resonance tomography - it makes it possible to stage and monitor the patients with HIV-associated neurocognitive disorders. The aim of the present study was to explore the significance of magnetic resonance spectroscopy in diagnosing HIV-associated neurocognitive disorders. MATERIALS AND METHODS: Magnetic resonance tomography and spectroscopy were performed in 8 HIV+ patients with neuropsychological evidence of cognitive deficiency. The HIV infection was confirmed in the national reference laboratory for HIV/AIDS. The patients were monitored in the Clinic of Infectious diseases, Plovdiv. The tomography and spectroscopy were performed in Rusev Diagnostic laboratory. R ESULTS: The magnetic resonance tomography did not find any significant changes: there was no cortical atrophy; the CSF spaces had normal location and capacity. The results from magnetic resonance spectroscopy: 5 patients were found to have significant elevation of the choline/creatinine ratio. The N-acetyl aspartate spectra were depressed in 1 patient, which can be interpreted as a lymphoid transformation. The spectroscopy showed no changes in 2 patients. CONCLUSION: The magnetic resonance spectroscopy provides an opportunity to visualize the discrete changes in the cerebral metabolism in HIV+ patients with neurocognitive disorders. Key words: HIV, magnetic resonance spectroscopy.

SY 16.

USE OF 18F(FDG) PET IN DIAGNOSIS OF CEREBELLAR ATAXIA

A. Kaprelyan, M. Grudkova Department of Neurology, Saint Marina University Hospital - Varna, Bulgaria Cerebellar ataxia is hereditary neurodegenerative disease characterized by malformation of the cerebellar function. It is subcategorized into several types: Friedreich ataxia (with early onset), Holmes, Menzel, Marie, Dejerine-Thomas (with late onset). The diagnosis is usually suggested by the presence of atrophy of the cerebellum and brainstem on PET-CT scan. Main symptoms are: dysdiadochokinesia, dyssynergia, dysmetria, positive Babinski reflex; dysarthria, ataxia of stance and gait, cardiomyopathy; nystagmus, optic nerve atrophy. Cerebellar ataxia is characterized by skeletal deformities and nerve conduction velocity in motor fibers of < 40 m/s in the hands. Over 40% of cerebelar ataxia patients have diabetes mellitus. After 18F-FDG is injected into a patient, a PET scanner can generate 2D or 3D images of the distribution of 18F-FDG within the body. Since its development in 1976, 18F-FDG has had a profound influence on research in the neurosciences. Key words: 18F(FDG) PET

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SY 17.

INFLUENCE OF MATERNAL SMOKING, DIET AND LEVEL OF TRACE

ELEMENTS IN ADOLESCENTS WITH PRIMARY HYPERTENSION Nadia Kolarova-Yaneva1, M. Angelova2, V. Nedkova1, A. Bozhinova2, K. Gospodinov 3, S. Tisheva3, M. Tzonzarova4 1Department of Pediatrics, Medical University - Pleven, Bulgaria, 2Department of Chemistry and Biochemistry & Biophysics and Physics, 3Department of Cardiology, Pulmonology and Endocrinology, Medical University- Pleven, 4National Hospital of Cardiology - Sofia, Bulgaria Primary hypertension (PH) frequency in children is determined by cardiovascular risk factors such as obesity, smoking and inappropriate diet. The success of strategies for its prevention depends on a plurality of perinatal and postnatal risk factors. The AIM of this study was to investigate the influence of potential risk factors like maternal smoking during pregnancy (MSDP) and the diet throughout the first year of the child for PH and the levels of certain trace elements (TE) in children with PH. METHODS: The study was conducted among 75 students aged 10-17 years with hypertension and a control group of 25 normotensive children. MSDP and the diet of the children were determined and studied by conducting a survey. Spectrophotometric methods were used to determine the serum levels of the Zn, Cu, Cr. The data were analyzed statistically. R ESULTS: We found that 53.3% of the mothers of hypertensive children smoked during pregnancy and 77.3% of those children were formula fed. Significantly lower levels of sZn (9.90 ± 1.63 μmol/l) were found in 80 % of the children with hypertension. 73.3% of children with PH showed lower values sCu (10.10 ± 0.41 μmol/l) and in 78.7% of them significantly lower sCr (0.77 ± 0.40 μmol/l) was measured. CONCLUSION: MSDP and formula feeding during the first year of life can probably be considered risk factors for early hypertension manifestation. The status of Zn, Cu, Cr showed a deficit in children with hypertension. Key words: smoking, primary hypertension,trace elements

SY 18.

NON-HODGKIN’S LYMPHOMA OF MUCOSA ASSOCIATED LYMPHATIC

TISSUE OF THE STOMACH. CLINICAL AND THERAPEUTIC DATA OF A 5-YEAR FOLLOW-UP Vasko Graklanov1, Veselina Goranova-Marinova2, Emil Spasov2, Katya Sotirova2, Stefan Goranov2 1Clinic of Hematology, St. George University Hospital, 2Division of Clinical Hematology, Medical University - Plovdiv, Bulgaria BACKGROUND: Non-Hodgkin lympomas of mucosa-assoviated lymphatic tissue (HL-MALT) of the stomach are rare disorders with unique pathogenesis. They are a model for transformation from antigen-dependant lymphocytic proliferation in chronic inflammation to a uncontrolled clonal malignancy. AIM: to analyse the incidence, clinical features, therapeutic options and therapeutic outcome in gastric NHL-MALT. PATIENTS AND METHODS: between 2010 and 2014, 12 patients with gastric NHL-MALT were analyzed [male 4 (3.33%), female 8 (66.7%), mean age 55.4± 6.9 yrs]. The diagnosis in all patients was made after fibrogastroscopy with biopsy. Staging was performed according to the staging system for stomach NHL-MALT. The patients were treated by surgery, polychemotherapy and antibiotics combination for eradication of Helicobacter Pylori. The major disease associated parameters were analyzed, the response rate, the event-free-survival and survival of the patients. Methods of classic statistics were applied (SPSS v 22.0). R ESULTS: Clinical stage I had 2 (16.2%) patients, 9 (75.0%) in stage II, 1 patient - stage IV (8.3%). Six patients (50.0%) were found to have the В symptoms. FGS proved a solitary or


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multiple lesions in 6 (50.0%), polyps in 4 (33.3%) and Tu-mass in 2 (16.7%) patients. HP infection was found in 7 (58.3%). Partial or total gastrectomy was performed in 10 (83.4%). In 7 (58.3%) polychemotherapeutical regimen CVP was applied, in 3 (25.0%) - CHOP and in 2 (16.7%) – Bendamustin. In all patients a complete therapeutic response was achieved. In 2 (16.7%) a relapse was registered at 38 and 44 months. Overall survival at 5 year was 63%. Key words: malt lymphoma helicobacter pylori

ANTIDEPRESSANT-LIKE EFFECT OF ARONIA MELANOCARPA FRUIT

SY 19.

JUICE IN RATS Miroslav Eftimov1, Cvetomira Dobreva2, Deyana Velkova2, Krasimir Kuzmanov3, Stefka ValchevaKuzmanova1 1Department of Preclinical and Clinical Pharmacology, Medical University - Varna, 2Faculty of Medicine, Medical University - Varna, 3Vivarium, Medical University - Varna, Bulgaria The main constituents of Aronia melanocarpa fruit juice (AMFJ) are polyphenolics that in recent years have been investigated for effects on central nervous system functions. The aim of the present studies was to investigate the effect of the subchronic AMFJ administration on the immobility time in the forced swim test (FST) in Wistar rats. AMFJ was administered orally at doses of 5 ml/kg and 10 ml/kg to: i) healthy male rats for 30 days, ii) socially isolated (SI) male rats in the course of 4 weeks after previous isolation for 4 weeks, and iii) alcohol-treated female rats for 14 days. In the experiment on healthy rats, AMFJ caused a dose-dependent decrease in the immobility time which at the dose of 10 ml/kg was significantly lower (p < 0.05) than the control ones. In the second experiment, social isolation caused a tendency to increase immobility time. AMFJ prevented that effect and the immobility time of SI rats treated with the dose of 10 ml/kg was significantly lower (p < 0.05) than that of the SI group. In the third experiment, ethanol at a daily dose of 8 g/kg induced a depressive-like behavior proven by the prolongation of immobility time (p < 0.01 vs controls). The immobility time of the animals treated with the two AMFJ doses did not differ significantly from the control time. In the three experiments, AMFJ caused a decrease in immobility time in the FST which suggested an antidepressant-like effect in healthy and depressed rats. This effect might be due to AMFJ polyphenolic ingredients. Key words: Aronia melanocarpa, depression, rats

SY 20.

DISTRIBUTION OF HIV-1 SUBTYPES AMONG MEN WHO HAVE SEX WITH

MEN IN BULGARIA (PRELIMINARY ANALYSIS) Vanya Dafova1, Reneta Dimitrova2, Anna Gancheva2, Asia Kostadinova2, Danail Beshkov2, Ivaylo Elenkov3, Mariyana Stoycheva4, Ivailo Alexiev2 1 St. Kl. Ohridski University of Sofia; 2National Center of Infectious and Parasitic Diseases, Sofia; 3Hospital for Infectious and Parasitic Diseases, Sofia; 4Department of Infectious Diseases, Medical University - Plovdiv, Bulgaria OBJECTIVES: Since the beginning of HIV-1 epidemic in Bulgaria, for nine years (1986 - 2004) only 30 MSM with HIV-1 were registered in Bulgaria. By 2012, the MSM number with HIV-1 had risen to 277 (17.2%) of all 1606 registered HIV/AIDS cases. The aim of the present study was to analyze the diversity of HIV-1 subtypes among MSM in Bulgaria.

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MATERIAL AND METHODS: In this study 637 HIV-1 pol sequences 146 of which from MSM were obtained with TruGene DNA Sequencing System. HIV-1 sequences were analyzed with Internet based subtyping tools: REGA3 and COMET1.0. Phylogenetic analysis was conducted with Maximum Likelihood method implemented in FastTree2 program. R ESULTS: The most prevalent HIV-1 subtype in MSM was found to be subtype B - 75.2%, followed by URFs - 13.1%, CRF01_AE - 4.8%, CRF02_AG - 2.1%, CRF12_BF - 0.7%, CRF14_BG 0.7%, and G/B recombinant - 0.7%. Phylogenetic analysis reviewed 8 clusters with short evolutionary history and outbreaks within subtype B and 2 other clusters in CRF01_AE and CRF02_ AG composed of sequences isolated from individuals who were both MSM and IDUs. Most of MSM were found in Sofia (58.6%). CONCLUSIONS: The analysis reviewed rapid increase of HIV-1 cases among MSM in Sofia and significant domination of HIV-1 subtype B in this population. Phylogenetic analysis showed multiple clusters demonstrating rapid development of sub-epidemics within MSM and possible spillover of viruses between MSM and IDUs groups. Our analysis demonstrated that molecular epidemiological surveillance among MSM is of great importance to public health and to better control HIV-1 epidemic in this group of population. Key words: HIV-1, homosexsual, epidemic, Bulgaria

SY 21.

UTERINE ARTERY DOPPLER VELOCIMETRY AS EARLY SCREENING TEST

FOR SPONTANEOUS ABORTION Ilian Traianov Department of Obstetrics and Gynecology, Medical University - Plovdiv, Bulgaria INTRODUCION: Only 50-60% conceptions advance beyond 20 weeks of gestation. In this period the main change in the uterine is blood flow. The uterine perfusion increase in the pregnant, and the beginning of this process is at the first trimester. This is detected by Doppler-ultrasound test. AIM: To assess the prognostic value of left and right uterine artery Doppler test as early screening test of threatened abortion outcome. MATERIAL AND METHODS: it is a comparatively prospective study which includes 57 women - 36 patients with threatened abortion and a control group of 21 patients with normal pregnancy. The groups were age and gestation age matched. The parameters that were examined were: index of resistance and radio systole / diastole. R ESULTS: It was found that 26% (72.22) of patients with threatened abortion have significant difference in index of resistance of left and right uterine artery. In control group this difference is only in 7 (0.33%) pregnant women. In all 6 cases of threatened abortion, there is a magnificent dissonance in index of resistance of left and right uterine artery. CONCLUSIONS: It was found, that significant difference in index of resistance of left and right uterine artery is connected with interruption of pregnancy before 20 gestation week and with complications such as uterine bleeding and abdominal pain in the same term. Preliminary conclusions show the need for further research which may determine the diagnostic value. Key words: threatened abortion, Doppler test, index of resistance, left and right uterine artery, first trimester

SY 22.

HEMOPHAGOCYTIC LYMPHOHISTIOCYTOSIS

Kristina Petkova, Hristina Hristozova Faculty of Medicine, Medical University - Varna, Bulgaria The hemophagocytic lymphohistiocytosis is an extremely rare disease and often fatal for histiocytes and lymphocytes, which is spread across all age groups. Clinically, it presents with high Folia Medica 2015; 57; Suppl. 1 © 2015 Medical University, Plovdiv, Bulgaria

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temperature, hepatosplenomegaly, lymphadenomegaly, pancytopenia and rash. In most clinical cases there are changes in the skin in the form of erythrodermia, macula and papula generally. This condition may be found in combination of malignant, genetic and autoimmune diseases. Basic pathogenetic sign in histiocytosis of mononuclear phagocytes, which differ from cells of Langerhans, is aggressive proliferation of activated Ma and Hi. These cells eat other cells - erythrocytes (anemia), leucocytes, platelets. The process attacks bone marrow, spleen, lymph nodes, liver, skin, and brain. NK cells are important. This disease has a familial form and a secondary form of infection. We report a child born on 19.01.2013 of first, normal pregnancy. Four months later the child was pallid, running a temperature of 38°C, Hb 60, Leu 10.10, thrombocytes 91.10. The child was hospitalized in Russe and treated with medakson, dexametazon, methylprednisolon, a single transfusion of erythrocytes and thrombocytes. Later the child was transferred to a Varna hospital for a different treatment. The patient is still without a correct diagnosis for which the family will go to Germany for treatment. Key words: HLH

SY 23.

EFFECT OF SHOCK WAVE THERAPY ON MUSCLE SPASTICITY IN

CHILDREN WITH CEREBRAL PALSY Mariya Gonkova, Elena Ilieva Division of Physical Therapy and Rehabilitation, Medical University - Plovdiv, Bulgaria INTRODUCTION: Extracorporeal shock wave therapy is a method of treatment, which is used in medical practice mainly in the management of muscular-skeletal disorders. AIM: The aim of this study is to evaluate the effect of shock wave therapy on muscle spasticity of plantar flexors in children with cerebral palsy. MATERIAL AND METHODS: Twenty-five children, mean age 4.84 ± 3.11 years, were included in the study. They all had a diagnosis of spastic cerebral palsy. This was an open observational, placebo controlled double-blinded study in which each child served as its own control. Before the active stimulation a placebo session was applied. For the assessment of the results before, after, two weeks and four weeks later, the following methods were used: passive range of motion (pROM) in the ankle joint, Modified Ashworth Scale (MAS) and static and dynamic baropodometric measurement. R ESULTS: There was no significant difference in the results after the placebo session. After the active stimulation of RSWT, significant changes were observed between the results and baseline: a significant increase in pROM: from 33.25 ± 2.20 to 47.00 ± 2.29 (p < 0.001) degrees; significant change in muscle tone proved by changes in MAS: from 2.77 to 2.00 points (p < 0.001). Baropodometric measurement showed a significant increase in the contact plantar surface area of the affected foot and in heel pressure. No side effects were observed after the application. CONCLUSION: RSWT could be used for reducing muscle spasticity in children with cerebral palsy, but further investigation is needed to clarify the most appropriate treatment protocol. Key words: cerebral palsy, extracorporeal shock-wave therapy,spasticity

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SY 24.

TINEA CAPITIS

Anastasiya Chokoeva, Liliya Zisova Department of Dermatology and Venereology, Medical University - Plovdiv, Bulgaria Tinea capitis is the most common fungal infection in childhood, which is significantly less often found in adults. It is caused by dermatophytes, as the etiological structure shows some variations in different latitudes, which is due to the various environmental factors in one hand and the socio-economic status on other. In urban areas of Europe, the dominant etiological agent are anthropophilic dermatophytes, as the infection may occur sporadically or cause small epidemics in kindergartens or other social institutions. In rural areas, among isolated agents of tinea of the scalp prevail zoophylic dermatophytes whose common carriers are agricultural animals and rodents. Microsporum canis remains the most common causitive agent of Tinea capitis in Europe. The Tinea, caused by Microsporum spp. showed no significant differences in the epidemiological structure in different areas. In recent years a significant resistance to certain zoosexuality dermatophyte species to antifungal therapy has been established. We conducted a retrospective study for a 10-year period (2004-2013) on patients aged 0 to 18 years, with proven Tinea capitis, confirmed and identified by direct mycological examination and culture examination of Sabouraud agar, based on data from mycological lab of Dermatology clinic in St. George University Hospital, Plovdiv. We present the results of the study, whose aim was to establish, analyze and compare the etiological structure of fungal infection of the scalp in childhood, over the years in patients of Plovdiv and the region, and to identify the most commonly affected age, therefore to compare the results with a short literature review. Key words: tinea capitis, fungal infection, children

SY 25.

EVALUATION OF ERECTILE DYSFUNCTION USING THE SURVEY METHOD

-IIEF - INTERNATIONAL INDEX OF ERECTILE FUNCTION, IN PATIENTS TREATED WITH OPIOID AGONISTS Sava Petrov1, Maria Orbetzova1, Yanko Iliev2 1Division of Endocrinology, Second Department of Internal Diseases, Medical University - Plovdiv; 2Clinic of Clinical Toxicology, St. George University Hospital, Plovdiv, Medical University - Plovdiv, Bulgaria Methadone is a synthetic opioid. It is used as a means of pain relief or as maintenance therapy in patients with opioid dependence. It was developed in 1937 in Germany. The chronic methadone maintenance therapy started in 1964 as an attempt to deal with the outbreak of heroin use in New York after World War II. Methadone therapy reduces or eliminates the use of heroin, decreases mortality and criminal activities related to heroin use and allows patients to improve their general health and social situation. Methadone represents an acyclic analogue of morphine and heroin, as it binds to the same opioid receptors inducing the same effects. Modern conceptions of the negative effects of opioids on gonadal function define two main mechanisms of lowering testosterone levels and sexual dysfunction. The first mechanism is in disorder with pulse gonadotropin secretion, and the second one is impact on the response of the anterior pituitary to the gonadotropin-releasing hormone. Chronic use of opioids in men - heroin, morphine, methadone - leads to symptoms such as delayed ejaculation, erectile dysfunction and significant reduction of sexual libido.


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International survey on erectile dysfunction (IIEF - international index of erectile dysfunction) is a widely used comprehensive survey that is completed by the patient alone. It was developed for diagnosis and assessment of the degree of erectile dysfunction. The aim of this study is evaluation of erectile dysfunction, using the survey method in patients undergoing methadone maintenance therapy. Key words: erectile dysfunctiom, methadone, opioid

SY 26.

INDICATORS FOR NEURODEVELOPMENT IN 5-YEAR OLD CHILDREN

Ralitsa Yordanova, Ivan Ivanov Department of Pediatrics and Medical Genetics, Medical University - Plovdiv, Bulgaria INTRODUCTION: The elaboration of a Bulgarian pediatric screening method for neurodevelopmental disorders at 5-year old children requires the determination of abnormal results in different test items among the general population. AIM: Report of the proportion of abnormal results in different neurodevelopmental indicators among 5-year old children in Bulgaria. METHODS: Test items from a Finnish method were adapted in Bulgarian and updated with 29 contemporary items. The new test panel contains 89 items grouped in 12 domains. It is applied to 405 5-year old children (male : female = 1 : 1) from 39 randomly chosen settlements in Bulgaria. R ESULTS: 12% of the children had less than 0.8 vision with at least one eye, 37% had difficulties on hopping on one foot; 26% could not stand either on the left or right foot. No children with central motor neuron impairment were found. Abnormal pencil grip was found in 8.8% of the children, abnormal finger-nose pointing - in 4%. Impaired construction of a staircase with 10 cubes and cutting a circle was found in 42%. Difficulties in repeating or forming a sentence was registered in 16%. Difficulties pronouncing more than three consonants were found in 12%; 44% cannot explain differences between objects. Only 2% obtained optimal results for all items. Girls had better scores on vision, standing and hopping on one foot, pencil grip and forming a sentence (p < 0.05). CONCLUSION: A significant variation in the proportion of abnormal results was established - from 0 to 34%. This results will be used to determine the cut-off points and the final set of the different items. Key words: neurodevelopmental screening

SY 27.

A FETUS FROM VARNA WITH A CYSTIC HYGROMA OF THE NECK

Tania Kitova1, R. Minkov 2, D. Milkov3, A. Tenev 3, K. Kilova4 , P. Darmonska1 1Department of Anatomy, Histology and Embryology, Medical University - Plovdiv, Bulgaria, 2 Prof. Dr. D. Stamatov Specialized Hospital of Obstetrics and Gynecology, Varna, Bulgaria, 3Medical University - Plovdiv, Bulgaria, 4Department of Medical Informatics, Biostatistics and E-Learning, Faculty of Public Health, Medical University - Plovdiv, Bulgaria The cystic hygroma of the neck in fetuses is а congenital anomaly, usually detected by ultrasound during prenatal morphological study. It is a rare anomaly that is diagnosed relatively easy by an examination of the nuchal fold. The frequency of the cystic hygroma in the world is approximately 1/6000 live births, but there is no data for diagnosed cases in Bulgaria. The PURPOSE of this work is to present a case of cystic hygroma of the neck in a fetus from Varna, Bulgaria. The METHOD used is an autopsy. The fetus is female weighing 250 g, the result of a spontaneous abortion during the 19th gestational week.

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R ESULTS: The autopsy found a cystic bilocular hygroma of the neck and a unilateral ptosis of the kidney. CONCLUSION: The cystic hygroma of the neck can be isolated, but it is often associated with trisomy 21 or with other malformation syndromes, which confirms the importance of morphological examinations of the neck during the prenatal ultrasound. Key words: cystic hygroma, prenatal diagnosis

SY 28.

ANTI-MÜLLERIAN HORMONE - QUALITATIVE DIAGNOSTIC CRITERION

FOR POLYCYSTIC OVARY SYNDROME Nikoleta Parahuleva1 , Blagovest Pehlivanov1, Ekaterina Uchikova1, Maria Orbecova2, Mariana Parahuleva3 1Department of Obstetrics and Gynecology, University Hospital St. George, Medical University - Plovdiv, Bulgaria; 2Department of Endocrinology, University Hospital St. George, Plovdiv, Medical University Plovdiv, Bulgaria; 3Department of Cardiology and Angiology, University Hospital Marburg, Germany INTRODUCTION: Polycystic ovary syndrome is the most common endocrine disorder in women of reproductive age. Given the important role of AMH in the pathophysiology of PCOS, it is of special interest as a new diagnostic criterion syndrome. OBJECTIVE: To assess the possibilities for setting AMH as a diagnostic criterion for PCOS. MATERIAL AND METHODS: of the prospective comparative study case-controls were 219 women aged 18-35 years. They were divided into two groups: group with PCOS - 120 women selected against the criteria of ASRM / ESHRE 2003; a control group of 99 healthy women. R ESULTS AND DISCUSSION: When comparing the areas under ROC-curves hormonal indicators used as predicates in PCOS diagnosis was validated. Statistically significant diagnostic differences between serum AMH and: FSH, LH, testosterone and estradiol. It was found in - significant diagnostic reliability of the AMX for differentiation of PCOS compared to the other studied hormonal parameters. CONCLUSION These data regarding AMH in PCOS allow us to develop a hypothesis for its use as a new and reliable diagnostic criteria for PCOS. Key words: AMH, MIS, PCOS

SY 29.

APPLICATION OF SMALL VIDEO RECORDERS IN OPEN SURGICAL

PRACTICE Dimitar Penchev1, Dolina Gencheva2, Radosvet Gornev3, Miroslav Zashev3, Lilyana Vladova1 1Medical University of Sofia, 2Medical University - Plovdiv, 3Department of General Surgery, Faculty of Medicine, Sofia University, Bulgaria INTRODUCTION: New biomedical technologies are introduced into the modern surgical practice at a very fast rate. The use of small video recorders with high quality of video and pictures in open surgical practice has many applications. The purpose of this article is to present said applications, as well as the advantages of small video recorders in the surgical and medical practice. MATERIALS AND METHODS: For the purpose of this study we used a summary of literature analysis of the method and its application, as well as its necessity and advantages. We recorded 6 open surgeries and 10 outpatient procedures with a small video recorder in the Clinic of General and Operative Surgery in the Lozenets University Hospital. R ESULTS: The video and pictures of two of operational procedures were used for scientific pur-


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poses. Three records were used in the preparation of demonstration videos for the students of the medical faculty at the University of Sofia. One video was used for assessment and control of the quality of the operational procedure following a complication. A surgical procedure was consulted with a pathologist for choosing the optimal spot for express histological examination. CONCLUSIONS: The use of small video recorders (SVR) in open surgical practice facilitates and improves the quality of the operating procedure. It reduces the difficulties of pathological diagnosis and of sample taking. SVRs are an irreplaceable utility in the preparation of research and teaching materials. Key words: GoPro, surgery, recording devices, camera

SY 30.

BURNS FROM ELECTRIC CURRENT - BEHAVIOR AND PROBLEMS

Elena Krasteva, Krasimir Yanev, Yulia Devedjieva Department of Propaedeutics of Surgical Diseases, Division of Plastic, Reconstructive and Aesthetic Surgery, St. George University Hospital, Plovdiv, Bulgaria Nowadays life without electric current is impossible. Hence the high percentage of household traumatism. Considerable part of the injuries results from theft of cables and equipment of power stations which brings additional losses to communities. Electric current with a frequency of 50 Hz exerts a cytolytic effect on the human body. The injuries in the entry and exit site are just ‘the tip of the iceberg’. Quite often under visibly healthy skin devitalized muscles, clotted vessels, necrotic nerves are found. In overlooking the local status and inactivity of the surgical teams the complications lead to necrosis of parts or whole limbs in large percentage of cases, often necessitating later amputations and eventually resulting in disability. In this study, the authors aim to demonstrate modern approaches or just behavior, entered as a routine practice in admission and treatment, as well as in complications of injuries from electric current. Key words: electric burns, electric current, behavior

SY 31.

AUTOIMMUNE PHENOMENA IN PATIENTS WITH SOLID TUMORS

Iv. Nenova1, M. Valcheva2, E. Beleva1, D. Tumbeva2, M. Janeva1, J. Grudeva-Popova1 1Department of Clinical Oncology, Medical Faculty, Medical University - Plovdiv, 2St. George University Hospital, Plovdiv, Bulgaria INTRODUCTION: Solid tumors and malignant hematological diseases are often associated with autoimmune disorders. Autoimmune phenomena are more frequent and well studied in lymphomas where there is a disbalance in the immune system. They are included in the group of paraneoplastic syndromes and are rarer in the case of solid tumors. Their pathogenesis is unknown. The PURPOSE of our study was to analyze the connection between autoimmune disorders and tumor status and localization as well as possibilities for therapeutic effect on them. PATIENTS AND METHODS: 1083 patients with solid tumors diagnosed and treated in St George University Hospital between 2010 and 2014 were investigated. Patients with lymphomas were excluded. R ESULTS: We found higher frequency of autoimmune phenomena in patients with prostate (9.01%) and ovarian carcinomas (7.05%) than in those with other solid tumors. Distribution according to autoimmune disease type showed highest frequency of vasculitis (27.7%) and polyneuritis (16.6%). Autoimmune hemolytic anemia (16.2%). Immune thrombocytopenia, seronegative arthritis, psoriasis, polymyositis have been rarer. Paraneoplastic autoimmune phenomena usually developed metachronic, rarely simultaneously with the tumor. Mainly their clinical manifesta-

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tion was associated with the progression of metastatic malignancy and seemed to respond well to treatment. CONCLUSION: Paraneoplastic autoimmune phenomena are established with higher frequency in cases of prostate and ovarian carcinomas. They appear with progression of malignant tumor and regress with the medical or surgical treatment of the malignancies. Key words: carcinoma, autoimmune phenomena

SY 32.

EXERCISE-RELATED DESATURATION IN PATIENTS WITH COPD –

ANOTHER STEP TOWARDS PHENOTYPING THE DISEASE Roman Kalinov1 , Vladimir Hodzhev1 , Blagoi Marinov2, Stefan Kostianev2 1Clinic of Pulmonology, St. George University Hospital, Plovdiv; 2Department of Pathophysiology, Medical University - Plovdiv, Bulgaria Desaturation during 6MWT is considered as important predictor of mortality in COPD patients. AIM: To investigate relationship among exercise related desaturation (6MWT), symptoms, spirometric indices and number of exacerbations within last year. PATIENTS AND METHODS: Fifty patients (age 64.5 ± 9.2 yrs; BMI 24.6 ± 4.5; smoking pack/years, 39.0 ± 23.3) had lung function assessment and exercise test (6MWT). Desaturation was considered when SpO2 dropped ≥ 4% for at least 3 min. R ESULTS: All patients had pronounced airway obstruction (FEV1% = 40.8 ± 13.8; КСО% = 76.0 ± 27.9; РО2 = 64.4 ± 9.7 mm Hg), moderately decreased functional capacity (6MWT 492 ± 91 m.) and high symptom scores (МRС = 2.3 ± 1.0 ; САТ = 21.4 ± 7.5). They were divided in two groups according to their exercise SpO2 – without desaturation (A; n = 24) and with desaturation (B; n = 26). The two groups differed significantly with regard to FEV1% (47.9 ± 12.4 vs 34.2 ± 11.8; p = 0.001), KCO% (90.8 ± 22.4 vs 63.4 ± 26.1 p < 0.001), PCO2 (39.4 ± 4.5 mm Hg vs 44.3 ± 7.0 mm Hg, p = 0.006), but there was no significant difference in symptoms (MRC and CAT scores), number of exacerbations within the last year and multidimensional indices. The decrease in saturation correlated with basic spirometric and blood gas indices but not with MRC and CAT scores and number of exacerbations during last year. CONCLUSION: Desaturation in patients with COPD is associated with deterioration in lung function but not with symptom severity and number of exacerbations in the last year. Presumably, the patients with exercise (6MWT) related desaturation represent separate phenotype of COPD. Key words: 6MWT, desaturation, COPD

SY 33.

CHANGES IN THYROID FUNCTION TESTS IN WOMEN WITH POSTPARTUM

THYROID DYSFUNCTION Antoaneta Argatska1, Boyan Nonchev1, Maria Orbetzova1, Blagovest Pehlivanov2 1Division of Endocrinology, Faculty of Medicine, Medical University - Plovdiv, 2Department of Obstetrics and Gynecology, Faculty of Medicine, Medical University - Plovdiv, Bulgaria INTRODUCTION: Thyroid dysfunction occurs with increased frequency in the postpartum period and it may present either as transient functional disorder or persistent impairment of thyroid functional status. AIM: To investigate the evolution of thyroid hormonal parameters in women with postpartum thyroid dysfunction. PATIENTS AND METHODS: 51 women with thyroid dysfunction after pregnancy were included in the study (mean age 30.71 years, range 22-38 yrs). 46.4% of them were recognized with hypothyroidism and 60.8% with thyrotoxicosis. Serum levels of TSH, FT4, FT3, TPOAb, TgAb were measured and US examination of the thyroid gland was performed


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at 3 and 9 months after delivery. R ESULTS: Thyroid dysfunction of autoimmune pathogenesis during the early postpartum period was found in 41 (80.4%) of the studied patients. At the follow-up visit 9 months after parturition functional abnormalities persisted in 27 (52.9%) of the patients. In the hypothyroid group spontaneous recovery of thyroid function was seen in 70% and 30% of patients remained with permanent hypothyroidism. Transition from thyrotoxicosis to hypothyroidism was observed in 31.4% of the women. Persistent thyrotoxic syndrome was reported in 16.1% of thyrotoxic patients due to onset or relapse of Graves disease during the postpartum period. In the patients with prolonged thyroid dysfunction immunological and morphological alterations were more pronounced than in those who restored euthyroid state and therefore needed continuous treatment. CONCLUSION: Postpartum thyroid dysfunction can undergo dynamic changes. Thus, a comprehensive clinical assessment and individualized approach are required to determine the need for long-term treatment and follow-up. Key words: thyroid dysfunction, postpartum period

SY 34.

DIABETES MELLITUS TYPE 2 AND PHOTOSENSITIVITY

Raditsa Sokolova1, Rumyana Yankova1, Marianna Murdjeva2, Maria Orbetzova3, Tsvetana Abadzhieva1, Tatyana Popova2, Maria Pavlova3 1Department of Dermatology and Venereology, 2Department of Microbiology and Immunology, 3Second Department of Internal Diseases, Medical University - Plovdiv, Bulgaria Type 2 diabetes mellitus (DM2) is a metabolic disorder characterized by hyperglycemia, insulin resistance and relative lack of insulin. Medications used for the treatment of DM2 and its complications belong to different groups, have different chemical structures, and some of them are capable to induce cross-reactivity, phototoxic and photoallergic reactions. The aim of the present study was to determine the presence of photoallergic and phototoxic reactions in patients with DM2 on oral antidiabetic therapy, with or without clinically manifested аllergic skin diseases. A total of 97 patients (76 women and 21 men) aged between 25 and 80 years was included in the study. DM2 patients on oral antidiabetic therapy were 53, and those without DM2 - 44. Аll patients were photopatch tested with European Photopatch Baseline Series EP-1000. Twenty four hours after application of the test materials on the back of the patients, one of the lines of pair allergens (haptens) was irradiated with UVA (320-400 nm) at a dose of 5 J/cm2. Reading was done at 24, 48 and 72 hours and 7 days after irradiation. Phototoxic reactions were determined in 16.9% of the patients with DM2 on oral antidiabetic therapy compared to the control group of patients where phototoxic reactions were found in 2.2%. Photoallergic reactions in patients with DM2 were not registered. The oral administration of antidiabetic drugs and medicaments for the treatment of concomitant diseases, increasing the possibility of developing phototoxic, but not photoallergic reactions. Key words: diabetes mellitus type 2, photosensitivity

SY 35. SUBCUTANEOUS

ADIPOSE TISSUE OF CHILDREN OF ROMA ETHNIC ORIGIN

Petya Darmonska, Denis Milkov Department of Anatomy, Histology and Embryology, Medical University - Plovdiv, Bulgaria The topical distribution and thickness of subcutaneous fat tissue is of multietiologic nature. The height, weight and thickness of the subcutaneous adipose are most frequently measured. These indicators correlate with ethnicity. The aim of this study was to characterize the topical distribution and thickness of subcutaneous adipose tissue and its correlation with height and weight of Roma children. Our study included 41 Roma children (12 female and 29 male)

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aged 9 years. Height, weight and thickness of skin folds of nine typical places were measured. The data was processed using SPSS 19.0. Our results show that the average height and weight of boys was 133.58 cm and 33.1 kg, respectively. The thickness of the subcutaneous fat tissue in boys was greatest on the thighs (15.91 cm). The average height of girls was 136.66 cm and the average weight - 34.95 kg. The subcutaneous tissue in girls was greatest on the thighs (21.11 cm) and abdominal region (16.13 cm). No significant differences in height and weight were found. A significant difference was found in relation to the topical distribution of subcutaneous adipose tissue. The thickness in the region of the tenth rib, suprailiac region, abdomen and lower limbs in females is significantly greater than in males. In comparison with data of children of Bulgarian origin, Roma boys were significantly shorter in height and had a more developed subcutaneous fat tissue. This data shows that while subordinating on the same mechanisms, the physical development of children is also predetermined by their ethnicity Key words: subcutaneous fat tissue, Roma children

SY 36.

ANTINOCICEPTIVE EFFECT OF ELETRIPTAN AND RIZATRIPTAN IN RATS

K. Saracheva, D. Getovа, Department of Pharmacology and Clinical Pharmacology, Medical Faculty, Medical University - Plovdiv, Bulgaria AIM: Triptans are specific and selective agonists of 5HT 1B/1D receptors, used in the treatment of migraine. The aim of the research is potential antinociceptive effects of eletriptan and rizatriptan in rats. METHODS: Male Wistar rats ( 8 groups, n=8) treated with: controls; metamizol; eletriptan 0.1mg/ kg s.c; eletriptan 0.5 mg/kg s.c; rizatriptan 0.4mg/kg s.c; rizatriptan 0.8mg/kg s.c; dihydroergotamine 0.05 mg/kg s.c; dihydroergotamine 0.1mg/kg s.c. Three nociceptive tests were used. Criterion for hot-plate and tail flick test was the latency of reaction in seconds, for analgesimeter - the paw pressure reaction. The statistic evaluation was done by ANOVA. R ESULTS: In analgesimeter test the rats treated with eletriptan in high dose increased significantly the paw reaction compared with ‘zero time’ of the group and with metamizol. The group treated with rizatriptan in high dose increased significantly the paw reaction compared with the control group and the metamizol group at 60 minutes. In hot-plate test the rats treated with eletriptan in high dose increased significantly the latency reaction at 60 and 120 minutes compared with the control group and with metamizol. In tail flick test triptans increased significantly the latency reaction at 60, 120, and 180 minutes compared to the control group of the same hour. CONCLUSION: The study suggests that triptans possess some antinociceptive effect on the test used. Key words: triptans, nociception, rat

SY 37.

BURNOUT SYNDROME AMONG MEDICAL STUDENTS – A PILOT

CROSS-SECTIONAL STUDY Stanislava Harizanova1, Chris Abousleiman2 1Department of Hygiene and Ecomedicine, Faculty of Public Health, Medical University - Plovdiv; 2Faculty of Medicine, Medical University - Plovdiv, Bulgaria BACKGROUND: Medical students are continuously exposed to psychosocial stressors throughout training that can lead to burnout. Until now, the prevalence of this syndrome is unknown among Folia Medica 2015; 57; Suppl. 1 © 2015 Medical University, Plovdiv, Bulgaria

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medical students in Bulgaria. The AIM of this study was to determine the prevalence and level of burnout among medical students at Medical University - Plovdiv. MATERIAL AND METHODS: A cross-sectional survey was carried out in a sample of 43 medical students using the Maslach Burnout Inventory. Participation was voluntary and anonymous. The average age of respondents was 24.21 ± 0.47 years, males represented 55.81% of the sample, the majority of students were single (88.37%) and 62.79% were in clinical years of medical training. Statistical analysis was performed with SPSS software. R ESULTS: Prevalence of burnout was 16.28%, and 41.86% were at high risk of burnout. Almost one-third of the total sample (32.56%) had a high level of exhaustion, 27.91% had high level of depersonalization and 81.40% had low level of personal accomplishment. Male students reported significantly higher level of emotional exhaustion than female students (χ2 = 8.281, p = 0.016). Given the training year, students in clinical years of training showed a significantly lower score on personal accomplishment compared to students in pre-clinical years of their training (t = 3.03, p = 0.005). CONCLUSION: It is important for medical students to study Balint approach before they become physicians. Individual or group based interventions (workshops) seem to be effective, particularly in reducing exhaustion in the short run. Preventive measures must be adopted and longitudinal studies should be conducted. Key words: burnout, gender, medical students

SY 38.

C-REACTIVE PROTEIN AS A MARKER FOR LOW-GRADE INFLAMMATION

IN HYPERTENSIVE PATIENTS WITH AND WITHOUT TYPE 2 DIABETES MELLITUS K. Kostov1, A. Dimitrova1, S. Tisheva2, A. Ruseva3, A. Blazhev4, M. Atanasova 4, K. Gospodinov2, A. Grigoryan1 1Department of Pathophysiology, 2Department of Cardiology and Rheumatology, 3Department of Clinical Laboratory, 4Department of Biology, Medical University - Pleven, Bulgaria INTRODUCTION: C-reactive protein (CRP) is an acute phase protein, which is synthesized by hepatocytes, under the effect of interleukin-6 in inflammation. The CRP levels may be increased in patients with arterial hypertension (AH) and type 2 diabetes mellitus (T2DM) as a consequence of low-grade inflammation and may be predictors of cardiovascular events. PURPOSE: The aim of the study was to determine whether or not there are significant differences in the serum concentrations of CRP in hypertensive patients with and without T2DM in comparison to normotensive individuals. METHODS: Three groups were formed: Group I - control group (KG) (n = 20, male - 8, female 12; SBP = 124.0 ± 3.7; DBP = 82.2 ± 4.1 mm Hg; mean age 47.92 ± 11.3 yrs); Group II - arterial hypertension group (AHG) (n = 60, male - 24, female - 36; SBP = 155.4 ± 4.8; DBP = 87.15 ± 2.6 mm Hg; mean age 65.3 ± 11.5 yrs); Group III - arterial hypertension with T2DM (AHT2DM) (n = 50, male - 21, female - 29; SBP = 146.83 ± 11.9; DBP = 82.55 ± 9.7 mm Hg; mean age 63.5 ± 11.0 yrs). CRP was determined with COBAS INTEGRA 400 analyzer, by immunoturbidimetric method (Roche Diagnostics kit). Statgraphics centurion XVI was used for statistical analyses. R ESULTS: It was found that the serum concentrations of CRP in AHG (7.51 ± 9.6 mg/l) and AHT2DM (8.93 ± 7.8 mg/l) were higher than those of KG (1.06 ± 0.87 mg/l), and there are statistically significant differences, respectively between AHG and KG (р = 0.0020) and between AHT2DM and KG (р < 0.0001). There was no statistically significant differences between AHG compared to AHT2DM (p = 0.3998). CONCLUSION: The serum CRP concentrations were significantly elevated in AH groups with and without T2DM, confirming pathogenetic link of the disease with low-grade systemic and vascular inflammation. Key words: C-reactive protein, arterial hypertension, low-grade inflammation, type 2 diabetes mellitus

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EFFECT OF FERULIC ACID ADMINISTERED SUBCHRONICALLY ON

SY 39.

ANXIETY IN RATS Antoaneta Georgieva1, Stiliana Belcheva2, Roman Tashev3, Stefka Valcheva-Kuzmanova4 1Medical University - Varna, 2Department of Behavior Neurobiology, Institute of Neurobiology, Bulgarian Academy of Sciences, Sofia; 3Faculty of Pre-School and Primary School Education, Sofia University, Sofia, 4Department of Behavior Neurobiology, Institute of Neurobiology, Bulgarian Academy of Sciences, Sofia, Bulgaria Ferulic acid (FA), called also hydroxycinnamic acid, is a polyphenol compound, found in many foods such as coffee, apples, citrus fruits, tomatoes, rice and wheat. It possesses antioxidant activity but its other activities are not well studied as a whole. The aim of the present study was to investigate the effect of FA on anxiety in rats. The experiment was performed on 80 male Wistar rats divided in 8 groups (n = 10). FA was administered daily orally for periods of 7, 14, 21 and 30 days at a dose of 20 mg/kg prepared as a 10 ml/kg solution. For each testing period, there were two groups – FA and Control (treated with saline). At the end of each experimental period, the state of anxiety was evaluated using the elevated plus-maze test. The results for all testing periods showed that FA did not significantly increase the number of entries into the open arms, the time spent there and the ratio of open/total arm entries as compared to the respective controls. These results demonstrated lack of anxiolytic/anxiogenic activity of FA. FA did not affect the locomotor activity assessed by the total number of arm entries. This suggested that the lack of effect on the indices of anxiety was not due to diminished locomotor activity of rats. In conclusion, the findings from the present study suggest lack of anxiolytic/anxiogenic effect of FA in rats. Key words: ferulic acid, anxiety, elevated plus-maze, rats

SY 40.

EXPERIMENTAL RESEARCH AS EVIDENCE OF DISONTOGENETIC ORIGIN

OF SCHIZOPHRENIA Lilia Grozlekova, Yvetta Koeva Department of Anatomy, Histology and Embryology, Medical University - Plovdiv, Bulgaria Schizophrenia is the most common worldwide psychiatric disease characterized by disorder of thought processes and emotional reactions. It is a complex psychiatric disorder with various clinical presentations and with variations in the clinical manifestation, which is characterized predominantly by psychological disintegration, both intra- and interpersonal interaction. According to the neuroontogenetic theory underlying the emergence of schizophrenia the participate events occur during the intrauterine period, causing disorders in brain structure and function, leading to the appearance of the typical symptoms of the disease decades later. Nowadays, there are many animal models of schizophrenia, which, to a lesser or greater extent, reflect the key symptoms observed in patients suffering from this disease. Key factors determining the pathophysiological expression of schizophrenia with typical symptoms of the disease are neuronal structural abnormalities in some specific brain areas. One hypothesis is that psychotic disorders are generally secondary to malfunctioning of neurotrophic factors. Improper neurotrophic expression during brain development can lead to structural disorganization, leading to sub-optimal structuring of neuronal networks. Application of experimental rodent models show symptoms similar as positive symptoms and negative symptoms of schizophrenia. In the present study we review three experimental models in experimental animals: (1) models created based on pharmacological in-


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terventions, (2) genetic models (3) models using neonatal lesion (NVHL - Neonatal Ventral Hippocampal Lesion) Key words: schizophrenia, developmental model of animal schizophrenia, neuroontogenetic theory

ZN/AG AND ZN/AU WITH 2,6-DIFORMYL-P-CRESOL OR SALEN COMPLEXES

SY 41.

SIGNIFICANTLY REDUCE VIABILITY AND PROLIFERATION OF HUMAN BREAST CANCER CELLS Radostina Alexandrova1, Abedulkadir Abudalleh1, Tanya Zhivkova1, Desislav Dinev1,2, Nikola Simeonov1,3, Ivan Gavrilov4, Gabriela Marinescu5, Daniela-Cristina Culita5 1Institute of Experimental Morphology, Pathology and Anthropology with Museum, Bulgarian Academy of Sciences; Sofia, Bulgaria; 2Faculty of Biology, Sofia University, Bulgaria; 3Faculty of Medicine, Sofia University, Bulgaria; 4National Oncological Hospital, Sofia, Bulgaria; 5 Ilie Murgulescu Institute of Physical Chemistry, Romanian Academy, Bucharest, Romania Globally, breast cancer is the most frequently diagnosed cancer in women, with 1.67 million new cases only in 2012. There are 458 000 deaths per year from breast cancer worldwide making it the most common cause of female death in both the developed and developing world. Breast cancer is a highly complicated disease, and although enormous progress has been made over the last decades, many challenges lie ahead. One of them is the need to search for new antitumour agents with a high efficiency and good biological tolerance. The aim of this study was to evaluate the influence of newly synthesized Zn/Ag and Zn/Au complexes with 2,6-diformyl-p-cresol (Diald) or Salen in cultured human breast cancer cells - permanent cell line MCF-7 and primary culture from invasive ductal carcinoma. The investigations were performed in short-term (24-72 h, with monolayer cultures) and long-term (18 days, 3D colonies) experiments using methods with different cellular/molecular targets and mechanisms of action: thiazolyl blue tetrazolium bromide (MTT) test, neutral red uptake cytotoxicity assay, double staining with acridine orange and propidium iodide, colony-forming method. The results obtained reveal that administered at concentrations of 0.1 - 100 μg/mL these compounds significantly decreased viability and proliferation of the treated cells in a time- and concentration-dependent manner. Acknowledgements: Supported by Grant DFNI - Б02/30 from 12.12.2014, Bulgaria, and a bilateral project between Bulgarian Academy of Sciences and Romanian Academy. Key words: cancer, cell cultures, antitumor activity

SY 42.

GENOME-WIDE EXPRESSION PROFILING IN PERIPHERAL BLOOD OF

CHILDREN WITH AUTISTIC SPECTRUM DISORDERS Hristo Ivanov1, Vili Stoyanova1,2, Nikolay Popov3, Tatyana Kichukova4, Ivan Ivanov2, Tichomir Vychev1,4 1Department of Pediatrics and Medical Genetics, Medical University - Plovdiv, 2Clinic of Pediatrics and Genetical Diseases, St. George University Hospital, Plovdiv; 3Psychiatric Ward for Active Treatment of Men, State Phsychiatry Hospital Pazardzhik; 4Department of Plant Physiology and Molecular Biology, Paisii Hilendarski Plovdiv University, Bulgaria INTRODUCTION: Autism Spectrum Disorders (ASD) is characterized by severe impairment in the areas of development, social interaction and communication and / or stereotyped behavior. ASD includes a wide range of clinical characteristics with a prevalence estimated at about 1 in 88 chil-

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dren. Due to the high social significance of ASD and the arising frequency, clinical practice has long sought to shed light on the etiology and pathogenesis of this disorder. The advent of microarray technologies allows the design and implementation of genome-wide expression profiling. Global expression profiling provides the opportunity to evaluate possible transcriptomic deviations in ASD at both genes and gene-network levels. MATERIALS AND METHODS: The study included 30 children with ASD aged 3 to 11 years and 30 healthy children matched in sex and age with the group with ASD. The diagnosis of ASD was made by using ADI-R, CARS or GARS. The expression profile was studied. A genome-wide sequencing of copy DNA molecules was conducted to obtain information for quantitative expression of all genes subject to this study. R ESULTS: Based on the conducted expression analysis 23 differentially expressed genes with statistical significant change (p < 0.01 and FDR < 0.001) were identified. DISCUSSION: If the results of this work are confirmed in future studies and the identified changes in the study group are individually validated by us or by others in other independent cohorts, we can assume that the differentially expressed genes may help clarify the etiology and pathogenesis of this disorder. Key words: ASD, genome-wide expression analysis

SY 43.

METAL COMPLEXES DECREASE VIABILITY AND PROLIFERATION OF

CULTURED HUMAN NON-SMALL CELL LUNG CANCER CELLS Tanya Zhivkova1, Ivayla Pantcheva2, Elena-Maria Mosoarca3, Ramona Tudose3, Otilia Costisor3, Katalin Nemet4, Mariana Mitewa2, Radostina Alexandrova1 1Institute of Experimental Morphology, Pathology and Anthropology with Museum, Bulgarian Academy of Sciences, 2Faculty of Chemistry and Pharmacy, St. Kliment Ohridski Sofia University, 3Institute of Chemistry, Rоmanian Academy, Timisoara, Romania, 4Institute of Haematolohy and Immunology, National Medical Center, Budapest, Hungary Lung cancer has been the most common cancer globally for several decades and it is the leading malignant cause of death. According to the WHO, non-small cell lung cancer (NSCLC) represents 85% to 90% of all cases of lung cancer. The aim of our study was to evaluate the effect of newly synthesized metal complexes on viability and proliferation of cultured NSCLC cells. Two groups of compounds were investigated: i) monensic acid (MonH) and its biometal (II) complexes [M(Mon)2(H2O)2] (M = Mg, Ca, Mn, Co,)]; ii) mixed ligand Cu (II) complex Cu2(BAMP) (dipy)Cl4 (BAMP = N,N’-bis(4-antipyrylmethyl)-piperazine; dipy = 2,2-dipyridyl. The following permanent human NSCLC cell lines were used as model systems in our study: A549; NCI-H1650 and its clones C11 и G7, which are resistant to the kinase inhibitor gefitinib. The experiments were carried out by MTT test, neutral red uptake cytotoxicity assay, trypan blue dye exclusion technique, Bradford protein assay and colony-forming method. The results obtained reveal that applied at concentrations of 0.5 to 50 μg/ml for 24, 48 and 72 h the compounds investigated decreased significantly viability and proliferation of the treated cells in a time- and concentration-dependent manner. Metal complexes are found to be stronger cytostatic and cytotoxic agents as compared to their ligands. Acknowledgements: Supported by Grant DFNI - Б02/30 from 12.12.2014, Bulgaria, and a bilateral project between Bulgarian Academy of Sciences and Romanian Academy. Key words: NSCLS, drug resistance, antitumor activity


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SY 44.

METAL COMPLEXES OF BILE ACIDS AS POTENTIAL ANTITUMOR AGENTS

AGAINST COLON CANCER Lora Dyakova1, Dana Culita2, Gabriela Marinescu2, Luminita Patron2, Reni Kalfin1, Galina Kurteva3, Radostina Alexandrova4 1Institute of Neurobiology, Bulgarian Academy of Sciences, Sofia, Bulgaria, 2 Ilie Murgulescu Institute of Physical Chemistry, Romanian Academy, Bucharest, Romania, 3Specialized Hospital for Oncological Treatment, Sofia, Bulgaria, 4Institute of Experimental Morphology, Pathology and Anthropology with Museum, Bulgarian Academy of Sciences; Sofia, Bulgaria INTRODUCTION: As of 2012 colon cancer is the second most common cancer in women and the third most common in men with it being the fourth most common cause of cancer death after lung, stomach, and liver cancers. Physiological role of bile acids is well known. There are data about carcinogenicity of some of these acids. At the same time many investigations have shown antitumor properties of various bile acids and their derivatives. AIM: The aim of our study was to evaluate the influence of Cu(II), Zn(II) and Ni(II) complexes of ursodeoxycholic, deoxycholic and chiodeoxycholic acids on viability and proliferation of cultured human colon cancer cells. MATERIALS AND METHODS: The permanent human cell line HT29 was used as a model system in our investigations. The compounds tested were applied at concentrations of 10-200 μg/ml for 24-72 h and their effect on cell viability and proliferation was evaluated by MTT test, neutral red uptake cytotoxicity assay, trypan blue dye exclusion technique, double staining with acridine orange and propidium iodide, colony-forming method. R ESULTS: Our results show that the compounds investigated decrease viability and proliferation of the treated cells in a time- and concentration-dependant manner. Metal complexes express more pronounced cytotoxic/cytostatic activity compared to the corresponding ligands themselves. CONCLUSION: The examined metal complexes exhibit promising cytotoxic/antiproliferative properties against HT29 colon cancer cells and deserve further studies to clarify better their antitumor potential. Acknowledgement: Supported by Grant DFNI - Б02/30 from 12.12.2014, Bulgaria, and a bilateral project between Bulgarian Academy of Sciences and Romanian Academy. Key words: bile acids,cell lines,cytotoxicity

SY 45.

STUDY ON IMMUNOLOGICAL REACTIVITY IN PEOPLE PRACTICING YOGA

Mariya Ivanovska1,3, Petya Gardjeva1, Tatyana Popova1, M. Atanasova1, M. Semerdzhieva2, Marianna Murdjeva1,3 1Department of Microbiology and Immunology, Faculty of Pharmacy, Medical University - Plovdiv, 2Department of Healthcare Management, Faculty of Public Health, Medical University - Plovdiv, 3Technological Centre of Emergency Medicine - Plovdiv, Bulgaria INTRODUCTION: Stress is associated with abnormalities in the immune system. Practicing yoga, a system of physical and mental practices, is a way to achieve meditation, positive thinking, relaxation and thus to overcome stress. OBJECTIVE: To investigate the immunological reactivity in people practicing yoga (yogins) more than 6 months. MATERIAL AND METHODS: The study included 31 yogins (29 women and 2 men) aged between 18 and 65 years. This group was practicing yoga with an instructor 2 times a week for one hour. An informed consent and a questionnaire assessing the scale of stress were filled prior to blood

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collection. The absolute number and percentage of T, B and NK-cells as cell immunity parameters were tested by flowcytometry. Serum concentrations of humoral immune factors IgG, IgM and IgA were detected by nephelometry. R ESULTS: Lymphocyte populations and serum immunoglobulins were found in reference range in significant part of yogins (> 70%). In 51.6% of them an increase in absolute number of lymphocytes was observed while in 19.4% of the group – higher number of total immunocompetent CD3+ T cells was detected. CD4+ T-helper number was elevated in 16.1%, CD8+ suppressor/cytotoxic T lymphocytes and NK cells – in 9.7%. The decrease in absolute number and percentage of lymphocyte populations and subpopulations as well as in serum IgG, IgM and IgA levels was determined only in few of the tested yogins. IgA was increased in 22.6%. CONCLUSION: Practicing yoga maintains basic immune parameters in levels that provide adequate and protective immune response. Key words: yoga, stress, immunе response

SY 46.

EXPRESSIONAL CHANGES OF ENDOTELIN-1, METALLOPROTEINASES-2

AND -9 IN ABDOMINAL AORTA IN PATIENTS WITH ATHEROSCLEROSIS A. Grigoryan1, K. Kostov1, A. Dimitrova1, T.Betova2, M. Apostolova3 1Department of Physiology and Pathophysiology”; 2Department of General and Clinical Pathology; 3Institute of Molecular Biology, Lab Med & Biol Res, Sofia, Bulgaria INTRODUCTION: Endothelin-1 (ET-1) is a vasoactive peptide that is released by endothelial and vascular smooth muscle cells (VSMC). Matrix metalloproteinases -2 and -9 (MMPs-2 and -9) degrade collagen type IV. ET-1, MMP-2 and MMP-9 are directly related to pathogenesis of atherosclerosis since participate in endothelial dysfunction, inflammatory response and oxidative stress on the vessel wall. AIM: To determine the extent of the immunohistochemical expression of ET-1, MMP2 and MMP-9 in the vascular wall of the abdominal aorta in patients with and without atherosclerotic changes. MATERIALS AND METHODS: For a one-year period we studied segments (26) of human abdominal aortas of patients with various diseases, autopsied at the department of Clinical Pathology at the University Hospital in Pleven, 5 of which were designated as controls (free of atherosclerosis). Routine histological methods (hematoxylin/eosin, Van Gieson) and immunohistochemistry were used to determine the expression of ET-1, MMPs-2 and 9. For immunohistochemical detection we used polyclonal antibodies (class IgG, Santa Cruz, USA). The materials were divided into two groups: Group 1 with complicated atheromatous plaque. 2. Control-group. R ESULTS: An immunohistochemical study showed that ET-1, MMPs-2 and -9 are expressed mainly in the endothelium and VSMC of the vascular wall. The expression of ET-1, MMPs-2 and -9 was increased significantly in group 1 in comparison with group 2 (p < 0.05). CONCLUSION: The increased expression of ET-1, MMPs-2 and -9 in group 1 supports the hypothesis that they are leading factors in the destabilization of the atheromatous plaque. Key words: atherosclerosis, ET-1, MMP-2, MMP-9, immunohistochemistry


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SY 48.

ADIPOCYTOKINES AND ASYMMETRIC DIMETHYLARGININE IN WOMEN

WITH POLYCYSTIC OVARY SYNDROME Daniela Koleva1, Maria Orbetzova1, Tanya Deneva2, Iliana Atanassova3 1Clinic of Endocrinology and Metabolic Diseases, St. George University Hospital, Medical University - Plovdiv, 2Department of Clinical Laboratory, St. George University Hospital, Medical University - Plovdiv, 3Clinical Centre of Endocrinology, Acad. Ivan Penchev University Hospital of Endocrinology, Sofia, Bulgaria АIM: to determine the levels of leptin, resistin and interleukin-6 (IL-6) and plasma asymmetric dimethylarginine (ADMA) and its relationship with clinical and metabolic parameters in women with polycystic ovary syndrome (PCOS). MATERIALS AND METHODS: The study included 22 women with PCOS (mean age, 26.64 ± 3.96 yrs; mean BMI, 30.56 ± 4.34 kg/m²) and 17 clinically healthy women (mean age, 29.35 ± 5.30 yrs; mean BMI, 32.20 ± 7.39 kg/m²) as a control group. The following measurements and laboratory tests were conducted: weight, height, waist circumference, fasting plasma glucose, immunoreactive fasting insulin (IRI), total testosterone, leptin, resistin, IL-6, ADMA, total cholesterol, HDL-cholesterol, triglycerides. Body mass index (BMI), waist-hip-ratio (WHR) and homeostasis model assessment insulin resistance index (HOMA-IR) were calculated. R ESULTS: Significantly higher plasma levels of ADMA (0.80 ± 0.31, respectively 0.55 ± 0.28 μmol/l, P < 0.05) and resistin (5.65 ± 1.99, respectively 4.36 ± 1.80 ng/ml) were determined in women with PCOS as compared to those in the controls. Comparable leptin levels were found in women with PCOS and healthy women. IL-6 tended to have lower levels in the controls (P > 0.05). No correlation between ADMA and the other parameters in women with PCOS was found. Leptin showed significantly positive correlation with weight and negative correlation with IL-6. IL-6 was positively correlated with WHR and negatively correlated with HDL-cholesterol. A significant reverse correlation between resistin and testosterone was determined. CONCLUSION: Higher levels of ADMA (a marker of endothelial dysfunction) in the women with PCOS proved increased cardiovascular risk in these patients. Higher resistin levels showed a possible pathophysiological role of resistin in the development of insulin resistance, prediabetes and DM type 2 in PCOS. Key words: leptin, resistin, IL-6, ADMA,PCOS

SY 49.

ANALYSIS OF THE ELECTRONIC PRESCRIPTION IN EUROPE AND ITS

FUTURE IMPLEMENTATION IN BULGARIA Svetlana Galeva1, Evgeni Grigorov2 1National Center of Public Health and Analyses (NCPHA), Sofia, 2Prof. Dr. Paraskev Stoyanov Medical University, Varna, Bulgaria The global tendency of computerization of the health system gradually sets in our country. The implementation of the electronic prescription in Bulgaria is of great importance based on the emphasis on information systems and registers, which the EU set in the programming period 2014 - 2020. Some activities and services are electronized and separate registries and databases have been built so far. However, there is still no complete software solution and integrated information system for the electronic management of the pharmaceutical sector. The electronic prescription is the indispensable integrative component to ensure the transmission of information and combine the selective databases formed in different parts of the health system. Thus, the electronic prescription is not only a digital version of the standard paper recipe, but also a key element for the development of e-health in Bulgaria. For the computerization of the health system it is essential to analyze the experience of countries which already partially or fully use electronic prescription. This study focuses on the models applied in the United Kingdom, the Netherlands, Sweden, Nor-

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way, Denmark, Finland, Estonia, etc. It emphasizes the similarities, differences, advantages and possible obstacles during the implementation of international practical experience in our country. Here we analyze the extent to which Bulgaria is ready to implement the electronic prescription and define the main steps needed to be made. Key words: electronic prescription, electronic registers, e-health

SY 50.

COMPARISON STUDY ON THE INFLUENCE OF RETIGABINE, LAMOTRIGINE

AND VALPROATE SODIUM ON THE COGNITIVE FUNCTION IN PENTYLENETETRAZOLE - KINDLED RATS E. Apostolova, V. Kokova, L. Peychev Deparment of Pharmacology and Drug Toxicology, Medical University - Plovdiv, Bulgaria Retigabine is a new anticonvulsant drug with broad spectrum of activity. Its effect on the memory impairment caused by pentylenetetrazole (PTZ) - kindling in rats is still not studied enough. AIM: The aim of our study is to compare the effect of retigabine (RTG), lamotrigine (LTR) and valproate sodium (VS) on cognitive deficits in PTZ-kindled rats. MATERIALS AND METHODS. Thirty two rats were divided in 4 groups (n = 8) and treated orally respectively with Aqua destillata, RTG in a dose of 30 mg/kg bw, LTR in a dose of 5 mg/kg bw and VS in a dose of 100 mg/kg bw. Kindling was induced using subcutaneous injections of 35 mg/kg bw PTZ every other day. Animals were observed for 30 minutes and the seizure score was determined using a 6-stage scale. Rats, experiencing stage 4 or higher on two consecutive injections, were considered kindled. After reaching this criterion, the influence on cognition was evaluated using automatic reflex conditioner (Shuttle box). R ESULTS: VS treated rats showed increase in the number of active avoidances compared to the control group on days 2 and 3 of training. This tendency remained until the end of testing. Tendency of increasing was evaluated after administration of RTG in comparison to controls during the training session. No significant difference was found in the retention test. LTR has no influence on the number of active avoidances compared to the control group. CONCLUSION: Our results showed that RTG and VS has a potential neuroprotective effect in kindled rats. LTR does not influence the cognition in this case. Key words: retigabine, lamotrigine, valproate sodium, cognitive function, PTZ-kindling

SY 51.

HOSPITAL PHYSICIANS’ QUALIFICATION IMPROVEMENT AND JOB

SATISFACTION Petar Atanasov, Yuliyana Marinova, Boryana Parashkevova Department of Social Medicine and Health Management, Faculty of Medicine, Trakia University, Stara Zagora, Bulgaria Job satisfaction is an important precondition for qualitative and effective work activity. Good qualification is one of the main social factors for medical workers’ satisfaction with the profession. The aim of this investigation was to analyze the role of the continuous physicians’ qualification improvement in some hospitals in Bulgaria for their job satisfaction. In 2013, an inquiry examination among 450 physicians, 235 males and 215 females, at a mean age of 45.6 ± 10.6 years in hospitals in the towns of Stara Zagora, Plovdiv, Sliven, and Yambol was carried out. The results were statistically processed by means of descriptive, variance (ANOVA) and correlation analyses (Pearson’s coefficient). A total of 61.40% of the physicians had acquired a clinical specialty but 24.80% were postgraduate students. Some 66.10% of the respondents had one speciality, 29.10% Folia Medica 2015; 57; Suppl. 1 © 2015 Medical University, Plovdiv, Bulgaria

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of them had two specialities while 4.80% of them had more specialities. Qualification improvement influenced favourably upon work and physician’s job satisfaction of the respondents. A higher general job satisfaction (60.2 ± 22.7) and work satisfaction (63.8 ± 23.7) was achieved. The physicians sharing that, in their hospital, there was current qualification improvement, were more satisfied than those expressing the opposite opinion. In case of absence of such an improvement, job satisfaction (52.3 ± 20.9) and work satisfaction (51.2 ± 25.8) were statistically significantly lower (р < 0,001). Implementation of a variety of current measures directed to personnel’s qualification improvement and, particularly, to physicians’ one in the hospitals in Bulgaria is necessary. This would promote further the enhancement of physicians’ job satisfaction. Key words: job satisfaction, qualification, hospital physicians

SY 52.

HOSPITAL PHYSICIANS’ RESEARCH ACTIVITY AND JOB SATISFACTION

Petar Atanasov, Boryana Parashkevova, Yuliyana Marinova Department of Social Medicine and Health Management, Faculty of Medicine, Trakia University of Stara Zagora, Bulgaria Several contemporary investigations reveal the importance of the individual and collective research for the enhancement of medical workers’ job satisfaction. The objective of the present study was to outline the role of research activity of the physicians in some Bulgarian hospitals for their job satisfaction. A sociological investigation among 450 physicians, 235 males and 215 females, at a mean age of 45.6 ± 10.6 years in hospitals in the towns of Stara Zagora, Plovdiv, Sliven, and Yambol was carried out. The results were statistically processed using analysis of variance (ANOVA) and correlation analysis.They demonstrated that the enhancement of the level of practical clinical work, professional qualification and awareness about world science novelties were considered by the responding physicians as the primary contributions of research towards their job satisfaction. A statistically significant correlation between research accomplishment by the physicians and adequate job satisfaction achieved by them was found (р < 0.05). This satisfaction presented with higher mean values among the physicians in the hospitals where research was done (70.7 ± 19.5) than among those working in the other hospitals (56.4 ± 25.6). However, only 22.60% of the responding physicians conducted research directly, although approximately one half of all the physicians were acquainted with its advantages. The physicians involved in research were more commonly satisfied with their work (in 62.3%) in comparison with their colleagues (in 55.2% of the cases). There was a necessity for creation of optimal conditions for effective research within the large hospitals in Bulgaria. This would contribute to the enhancement of medical workers’ job satisfaction Key words: job satisfaction, research activity

SY 53.

IMMUNOFLUORESCENT INVESTIGATION OF D1 CLASS DOPAMINERGIC

RECEPTORS IN THE RETINA OF COLD-BLOODED VERTEBRATES Desislava Zhekova, Liliya Vitanova Department of Physiology, Faculty of Medicine, Medical University - Sofia, Bulgaria Dopamine which is low-molecular weight neurotransmitter/neuromodulator and the corresponding dopaminergic receptors are widely distributed in the central nervous system (CNS). Since the

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retina is part of the CNS due to its embryonic origin, it becomes a crucial site for investigation of brain disorders, some of which are due to disturbances of dopaminergic neurotransmission. The AIM of the present investigation was to study, using the indirect immunocytochemical method, the distribution of D1 class dopaminergic receptors in the retinas of frog and turtle – major representatives of amphibians and reptiles respectively. These species are not studied enough as compared to mammalia, although the lower vertebrate retinas are thought to posses neuronal schemes typical for mammalian brain. We used cryostat retinal sections that were stained with primary antibodies directed against dopamine D1 receptor (D1a) and dopamine D5 receptor (D1b). The secondary antibody was labeled with fluorochrome. The RESULTS showed that both D1a receptor and dopamine D1b receptor immunoreactivities were well expressed in almost all retinal layers in frog and turtle retinas. Both neuronal and glial type of staining was observed. The two plexiform layers – the inner and outer ones, the places where the retinal neurons make their synaptic contacts, were visualized. Elements of the glial Müller cells were also stained. Our CONCLUSION is that in the retinas of investigated species the D1 class dopaminergic receptors take place both in the neuronal information processing and the neuro-glial interactions. Key words: retina, D1 class dopaminergic receptors,immunofluorescence

SY 54.

IMPLEMENTATION OF THE PROGRAM FOR PREVENTION OF COMPUL-

SORY HEALTH INSURED PEOPLE OVER 18 YEARS OF AGE BY GENERAL PRACTITIONERS AND DISPENSARY OBSERVATION OF PATIENTS WITH CHRONIC DISEASES Penka Kostadinova1, Angelika Velkova2, Petkana Hristova3, Joana Simeonova4, Sergey Kostadinov5, Irena Stoilova6, G. Naydenova7, Stefan Kostadinov8 1Regional Health Insurance Fund - Pleven, 2Faculty of Public Health, Medical University - Sofia, 3Department of Public Health, Faculty of Public Health, Medical University - Pleven, 4Department of General Medicine, Forensic Medicine and Deontology, Faculty of Public Health, Medical University - Pleven, 5Department of Health and Occupational Diseases, Faculty of Public Health, Medical University - Pleven, Bulgaria, 6General practitioner, Diagnostic Consultation Center 2, Pleven; 7Faculty of Medicine, Medical University - Pleven; 8Faculty of Medicine, Medical University - Pleven, Bulgaria INTRODUCTION: The main activities of general practitioners (GPs) in the execution of contracts with the Health Insurance Fund are prevention and outpatient monitoring of compulsory insured people over 18 years. Activities included as mandatory in the outpatient monitoring largely overlap with those of prophylactic examination. Objective: The aim of this study is to establish the necessity for a common prophylactic medical examination of the monitored adults with respect to chronic diseases. MATERIALS AND METHODS: Official data from the National Health Insurance Fund and current legislation have been used. Results: Between 2006 and 2012, the annual medical check-ups in Bulgaria cover 35.8% to 48.3% of the compulsory insured subjects aged over 18 years. In Pleven region the share of covered preventive examinations varies from 33% to 45%. Of these, only 6% have attended regularly (every year) prophylactic examination, 65% had done it only a few years, and 29% never went to check-ups. From the ones that have regularly attended a prophylactic medical examination 70% are dispensary, while from the compulsory insured people that never went to a prophylactic examination only 11% of men and 15% of women are dispensary. CONCLUSION: The high range of dispensary people with general health check is ex-


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plained by the fact that they actively visit GPs in relation to chronic diseases for which they were monitored. Duplication of effort and research in both programs lead to an unwarranted expenditure of resources. It is necessary to revise the legislation. Key words: prophylactic examination, dispensary, GPs, CHIP

MICRONEEDLES – A LOOK INTO THE FUTURE OF VACCINES

SY 55.

Atanas Bogoev1, Ani Kevorkyan2 1Medical Faculty, Medical University - Plovdiv, 2Department of Epidemiology and Disaster Medicine, Faculty of Public Health, Medical University - Plovdiv, Bulgaria INTRODUCTION: Globalization, intensive migration, the increasing world population, the emergence of new pathogens, as well as re-emerging diseases, are some of the prerequisites for the widespread and sometimes hard to control infectious diseases. These serious threats to health are challenges for the epidemiological supervisory and control systems worldwide. The scientific community aims at developing new technologies for the production of vaccines that are not only effective and safe, but also easy to apply, easy to store and affordable means of disease prevention. AIM: The aim of this review is to present the innovative experimental technology based on different types of microneedles for dermal and transdermal administration of some vaccines and to measure the advantages and disadvantages compared to conventional methods. MATERIALS AND METHODS: A thorough review of available literature published between 2000 2014, and referenced in the database PubMed. R ESULTS: In recent years, many research centers in the world have been working on using the advantages of the microneedle delivery systems in introduction of therapeutic and antigenic proteins. Microneedles are subject of intense experimental development in releasing antigens included in the composition of vaccines against diseases prone to pandemic spread like flu, measles and others. CONCLUSION: Microneedles can provide a safe and effective method for the delivery of vaccines at lower doses, allowing production at low cost, simple distribution and use. Key words: microneedles, vaccines, innovation, new technologies

SY 56.

THE CLINICAL SIGNIFICANCE OF SERUM LEVELS OF VASCULAR

ENDOTHELIAL GROWTH FACTOR (VEGF) IN PATIENTS WITH COLORECTAL CARCINOMA Alen Petrov1, Georgi Minkov1, Krasimira Halacheva2, Cvetan Popharitov1, Yovcho Yovchev1 1Department of Surgery, University Hospital, Stara Zagora, 2Department of Immunology, Medical Faculty of Trakia University, Stara Zagora, Bulgaria AIM: Some biological factors play a major role in the initial stimulation of malignant growth, metastasing and subsequent angiogenesis. However, their prognostic value and clinical significance have not been validated yet. The aim of our study was to analyze the clinical significance of preoperative serum levels of vascular endothelial growth factor (VEGF) in patients with colorectal cancer (CRC). MATERIALS AND METHODS: For the period from May 2011 to May 2013, we analyzed VEGF’s preoperative, serum levels in 64 patients with sporadic CRC who subsequently underwent surgery in the Surgical Department of the University Hospital, Stara Zagora. We compared this results with 40 healthy controls.

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R ESULTS: The levels of VEGF in patients with CRC were significantly higher compared to the levels in the healthy controls (p = 0.014). We found significantly high VEGF’s levels in patients with advanced clinical stage (p = 0.017) and with low grade tumors (p = 0.041). CONCLUSION: Preoperative, serum levels of VEGF in patients with CRC are beneficial, positive, noninvasive, prognostic indicators which could be associated with clinical stage, grade, effect of systemic therapy and outcome. Key words: colorectal cancer, serum VEGF, prognosis

SY 57.

RISK CLUSTER ASSOCIATED WITH ACADEMIC ENVIRONMENT AND

GENDER IN ADOLESCENCE Galya Chamova1, Georgi Sarov2 1Department of Social Medicine and Healthcare Management, Medical Faculty, Trakia University, Stara Zagora, Bulgaria, 2Medical Sociology, Medical Faculty, Trakia University, Stara Zagora, Bulgaria INTRODUCTION: Recently scientific evidence has shown that risk behaviors tend to form risk clusters but the determinants of risk clustering are still unknown. OBJECTIVE: To analyze the risk clustering determination of a combination of academic environment and gender. MATERIALS AND METHODS: We conducted a survey using a self-completed questionnaire. From 903 students, aged 15-19 yrs, 499 (55.3%) were students from school demanding high level of academic performance (school2), with equal gender distribution: 248 (49.7%) males (M2) and 251 (50.3%) females (F2). The students from schools without such demands in application procedure (school1) were 404 (44.7%) - 204 (50.5%) of them are males (M1) and 200 (49.5%) - females (F1). These four groups were compared statistically (M1 vs. F1 and M2 vs. F2) concerning smoking, drinking, drugs use, physical activity and sexuality. R ESULTS: M1 and M2 were more likely than F1 and F2 respectively, to drink alcohol regularly (OR = 2.08; OR = 2.95), to play sports (OR = 5.47; OR = 3.83) and have sex with more than 5 partners (OR = 2.65; OR = 3.81). F1 were more likely than M1 to smoke (OR = 1.63), to be heavy smokers (OR = 2.34), and to make sex with a person they love (OR = 1.89), and there were no differences between F2 and M2 in these indices. Conclusions: Academic environment does not impact male’s risk clustering, while low level of academic demands makes girls more vulnerable to smoking in association with increased ability to experience emotionally satisfying sex. Probably low academic engagement makes girls more emotional and sexual and smoking plays some instrumental role in reaching intimate success. Key words: academic environment, gender, adolescents, risk clustering

SY 59.

CYBER PSYCHOLOGY - A DEVIATION OF THE SELF OR AN EXTENSION

OF PERSONALITY IN CYBERSPACE Andreas Hadzigiannis1, Zlatoslav Arabadzhiev1, Vartuhi Kadiyan2 1Department of Psychiatry and Medical Psychology, Medical University - Plovdiv, Bulgaria; 2Psychological Cosultant, London, UK The huge advance science and technology made over the last century has opened many new prospects for humanity. Since the advent of computers, advanced mobile technology, software products and different applications to access the Internet, communication between people is constantly changing. Constantly changing and evolving is also their idea of one another. People are beginning to seek social contacts mainly in cyberspace. At present, cyberspace is a new and


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major source of energy, experience, self-confidence, social status, money and so on. Today’s children prefer to spend time online rather than interact and play with their peers. Modern communication today has been transformed - it takes place between a person, a computer and another person. The development of technology in the modern world, blurring of the boundaries in cyberspace, the ever increasing flow of information and the increasing market competition for education and work, inevitably lead to a high demand for computer literacy. The distance between people, the lack of confidence and love, have undoubtedly led to adaptation of society to ‘the road of least resistance’, namely a virtual social life where anyone can be who they want to be. The lack of a direct visual contact has opened the doors of social impotence and incompetence and everyone is able to relieve the stress of their everyday life and their own limitations. Key words: cyber psychology

SY 60.

PROVIDING THE CHRONICALLY ILL WITH MEDICATIONS

Rumyana Stoyanova1, Gergana Foreva1, Marieta Todorova2, Todor Stoev1, Radost Asenova1, Rositsa Dimova1 1Department of Public Health Management, Health Economics and General Medicine, 2Department of Health Care, Faculty of Public Health, Medical University - Plovdiv, Bulgaria ‘Drug regulation’ is the internationally accepted term for the set of activities designed to provide the public with qualitative, efficient and safe medications. In Bulgaria, a number of laws and regulations are in place. The primary tool for providing the chronically ill with medications is the so called positive medication list. The AIM of this study was to describe the positive and negative aspects of the current drug regulation model. MATERIALS AND METHODS: A form was compiled with information for conducting the focus groups and included a written informed consent. The selection of participants was performed by providers and users of medical care. The audio recordings were transcribed, and three step analyses were performed. R ESULTS: The focus group discussions included two main topics - the policies for medication reimbursement for the chronically ill and the expense of medications. As a whole, the system was determined to be extremely bureaucratic. One of the most important limitations of the current model is the lack of individualization for treating particular patients with a chronic disease. CONCLUSIONS: The general attitude of the participants was that there are multiple disadvantages of the current policy for providing medications to the chronically ill and that change is necessary. Key words: drug policy, reimbursement, chronically ill

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11th National Conference of Medical Biology (15-17 May 2015)

The Conference is held under the auspices of the Rector of Medical University - Plovdiv

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ORGANIZING COMMITTEE of the 11th National Conference of Medical Biology (15-17 May 2015)

Chairperson: Professor Victoria Sarafian, MD, PhD, DMSc Members: Assoc. Prof. Marian Draganov, PhD Milena Filipova, PhD Maria Kazakova, PhD Nikolay Mehterov, PhD Yana Feodorova, PhD Assen Milchev, MD Vesselin Alexandrov Vassil Nizamov

Conference Secretariat: Slavka Rainova, Stella Chobanova, Boryana Deleva, Pavlina Slavova, Krassimira Atanassova

The Organising Committee gratefully acknowledges the support of Medical University - Plovdiv, ELTA 90M Ltd., Medical Technics Engineering Ltd, FOT Ltd, SGP BioDynamics Ltd, Biosystems Ltd.

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WELCOME ADDRESS

Dear Friends and Colleagues, On behalf of the Organizing Committee I am delighted to invite you to the 11th National Conference of Medical Biology. It is organized jointly by the Medical University of Plovdiv and the Bulgarian Society of Medical Biology and is taking place in the Auditorium Complex of Plovdiv Medical University from 15 to 17 of May 2015. The Conference is included in the series of events on the occasion of the 70th anniversary of the establishment of Plovdiv Medical University and is a satellite forum to the Science for Health Anniversary Conference. This traditional assembly provides a fertile meeting ground for representatives of the Departments of Biology at the Medical Universities in Bulgaria, for students and researchers working in the field of biology and medicine. I am deeply convinced that the Conference will feature high quality presentations, fruitful discussions and a lot of friendly meetings. I am extending to you our warm welcome to Plovdiv!

Professor Victoria Sarafian, MD, PhD, DMSc On behalf of the Organizing Committee

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PLENARY LECTURES CHROMATIN ORGANIZATION DURING TERMINAL DIFFERENTIATION OF MOUSE PHOTORECEPTOR CELLS - DYNAMICS AND FORMATION OF INVERTED NUCLEAR ARCHITECTURE Yana Feodorova1, Irina Solovei2, Heinrich Leonhardt2 1Department

of Medical Biology, Medical University - Plovdiv, Bulgaria, 2Department of Human Biology and Bioimaging, University of Munich, Germany Key words: nuclear architecture, chromatin folding, XCI, MECP2

Spatial segregation of functionally distinct chromatin domains is a fundamental characteristic of the eukaryotic nucleus. In interphase nuclei heteroand euchromatin are localized in discrete zones. Heterochromatin is mainly found at the nuclear and nucleolar peripheries whereas euchromatin is confined to the nuclear interior. This spatial distribution is characteristic of most eukaryotic cells which have so called conventional nuclear architecture. Rods in the retina of nocturnal mammals represent a unique exception to this rule. Chromatin in the differentiated spherical rod nuclei possesses a radial symmetrical organization with three distinct concentric zones. A single chromocenter, built from subcentromeric major satellite repeat, occupies the core of the nucleus. The chromocenter is surrounded by a thick layer of LINE-enriched heterochromatin.

The thin euchromatic shell enriched in SINE-repeats is associated with the nuclear periphery. This type of nuclear architecture is referred to as “inverted”. It is established from the conventional one during postnatal development by alterations in large-scale chromatin folding. The aim of the current project was to study the formation of the inverted nuclear architecture during terminal differentiation of mouse rods. For this purpose we tracked the changes in folding of selected small chromosome regions. Furthermore, we studied the spatial distribution of the X chromosome in rod nuclei with an emphasis on X chromosome inactivation. In the last part of the work we considered the role of the protein MECP2 as a potential factor for the formation of the inverted pattern of nuclear architecture and, in particular, for chromocenter fusion.

ORGANIZATION OF INTERMEDIATE FILAMENTS (CYTOKERATINS, VIMENTIN AND LAMINS) AND THEIR CHANGES DURING OOCYTE MATURATION IN THE MOUSE Venera Nikolova Department of Biology, Medical Faculty, Medical University of Sofia, Bulgaria Key words: cytokeratins, vimentin, lamins, oocyte A set of proteins of cytoplasmic and nuclear intermediate filaments (cytokeratins, vimentin, lamin B and lamin A/C) were localized by immunofluorescence and confocal microscopy in mouse oocytes at different maturation stages in order to obtain data about their still unclear intracellular localization during mammalian oogenesis. Oocytes and Folia Medica 2015; 57: Suppl. 1 © 2015 Medical University, Plovdiv, Bulgaria

cumulus-oocyte complexes from prophase I (germinal vesicle, GV) to metaphase II were subjected to immunofluorescence and laser scanning confocal microscopy. Cytokeratins and vimentin showed full colocalization. At GV stage, they were found in the cortical cytoplasm and in perinuclear position, and after the karyosphere was formed, they accessed the

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XI National Conference on Medical Biology

Plenary Lectures

inside of the nucleus and associated with karyosphere heterochromatin. In metaphase oocytes, the cortical reaction for these proteins was diminished, except for the cap, and binding was observed also in the region of metaphase chromosomes. Lamins at GV stage were associated with the nuclear envelope. After karyosphere formation, as the cell was preparing for GV breakdown, they could be found also inside the nucleus. At GV breakdown stage, lamin B localized inside the karyosphere while lamin

A/C was observed in association with karyosphere heterochromatin. During metaphase I and metaphase II, lamin B was found in the meiotic spindle and lamin A/C was detected in a bright diffuse halo around the spindle and, less intensively, throughout the cytoplasm. In conclusion, the studied intermediate filament proteins are present in mouse oocytes and undergo reorganization during oocyte maturation which is correlated with chromatin rearrangements at different stages of meiosis.

IMMUNOBIOLOGICAL EFFECTS OF BULGARIAN PROPOLIS IN VITRO Milena Draganova-Filipova1, Lyudmil Peychev2, Victoria Sarafian1 1Department

of Medical Biology, 2Department of Pharmacology and Toxicology, Faculty of Pharmacy, Medical University - Plovdiv, Bulgaria Key words: propolis, cell lines, immunomodulation

Propolis is a natural product resulting from the life activity of honey bees. It possesses many biological effects including antitumoral, immunomodulatory, anti-inflammatory, antioxidant, antimicrobial effect, among others. The comparison between the effect of the product as a whole and that of its compounds explains the way it functions and participates in the control and regulation of fundamental biological processes. The complex immunobiological characteristics of Bulgarian propolis are still scantily studied. Research in this field can contribute to justifying its various applications. In this study, specificity of the effect that ethanol extract of Bulgarian propolis and chemically synthesised propolis compound CAPE (caffeic acid phenethyl ester) have upon normal and tumor cell lines is determined. While the vitality of normal cells was not influenced, tumor cell

decreased their proliferation. Both products influence critical molecules in cell cycle regulation and induce apoptosis. Cell death is proved by four high selective molecular methods (Comet, TUNEL, PS – analysis and Anexin V). Our attention is focused on the effects of the natural product and its main compound (CAPE) on the immune system. In low concentrations propolis and CAPE influenced both humoral and cellular immunity. This is proved by their effects on the vitality of lymphocyte subpopulations and cytokine secretion in vitro. The changes in the concentrations of IL-1β, TNF-α, IL-6 and IL-10 evidence the anti-inflammatory and immunomodulatory effects of the products. Our in vitro results can serve as a basis for future research on propolis application in clinical practice.

POSITIONAL CLONING OF А GENE INVOLVED IN OXIDATIVE STRESS TOLERANCE IN ARABIDOPSIS THALIANA Nikolay Mehterov1, V. Toneva2, T. Gechev2 1Department

of Medical Biology, Faculty of Medicine, Medical University - Plovdiv, Bulgaria; 2University of Plovdiv, Department of Plant Physiology and Molecular Biology, Plovdiv, Bulgaria Key words: oxidative stress, programmed cell death, transcription assay Annotation: The Arabidopsis thaliana atr7 mutant is tolerant to oxidative stress induced by paraquat

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(PQ) or the catalase inhibitor aminotriazole (AT), while its original background loh2 and wild-type



plants are sensitive. Both, AT and PQ, which stimulate the intracellular formation of H2O2 or superoxide anions, respectively, trigger cell death in loh2 but do not lead to visible damage in atr7. The gene responsive for atr7 phenotype has been mapped and the gene product is a highly conservative protein. The transfection of both Arabidopsis mesophyll protoplast and entire plants with chimeric 35S:ATR7/ GFP construct reveal subcellular localization of ATR7. Additionally it has been shown that ATR7 is strongly induce by oxidative stress and act as a positive regulator of the process of programmed cell death. Finally, to study gene expression during oxidative stress and ROS-induced programmed cell death, two platforms for multi-parallel quantitative

real-time PCR (qRT-PCR) analysis of 217 antioxidant and 180 ROS marker genes were employed. The analyses revealed ROS-induced expression of many ROS-responsive genes mainly in loh2, confirming that an oxidative burst plays a role in the activation of the cell death in this mutant. Some of the genes were specifically regulated by either AT or PQ, serving as markers for particular types of ROS. Many of these genes were upregulated in atr7 compared to loh2 under non-stress conditions at the first time point, indicating that higher basal levels of ROS and higher antioxidant capacity in atr7 are responsible for the enhanced tolerance to oxidative stress and suggesting a possible tolerance against multiple stresses.

UPREGULATION OF T-HELPER-17 RELATED CYTOKINE EXPRESSION AND INDUCIBLE NITRIC OXIDE SYNTHASE IN COLORECTAL CANCER IN ASSOCIATION WITH HISTONE DEACETYLASE-3 Spaska Stanilova1, Noyko Stanilov1, Geo Cirovski2, Totina Cirovska1, Lyuba Miteva1 1Medical

Faculty, Trakia University, 2Trakia Hospital, Medical Faculty, Trakia University, Stara Zagora,

Bulgaria Key words: Th17, cytokine, qPCR, CRC The aim of the present study was to investigate the gene expression of cytokines which are relevant to the differentiation and function of Th17 cells (TGF-β, IL-6 and IL-17) as well as inducible NO synthase (iNOS) and histone deacetylase-3 (HDAC3) in colorectal cancer (CRC) tissues. Total RNA was isolated from tumor and normal mucosa samples from 16 CRC patients. After cDNA synthesis quantitative real-time polymerase chain reaction (qRTPCR) was performed on a 7500 Real-Time PCR System with validated PCR primers and TaqMan MGB probes for TGF-b, IL-6, IL-17, iNOS and HDAC3 mRNA. Relative quantitative evaluation of mRNAs was performed by the comparative ΔΔCt method. Results are presented as n-fold mean difference relative to calibrator (RQ = 2-ΔΔCt). We found significantly increased mRNA in tumor tissue compared to normal mucosa with RQ around


10 times that of TGF-β, IL-17, iNOS and HDAC3 (RQ = 10.01; p = 0.034; RQ = 11.3; p = 0.047; RQ = 9.7; p = 0.028; RQ = 9.7; p = 0.013). The highest increased mRNA in tumor tissue compared to normal mucosa was detected for IL-6 (RQ = 107.3; p = 0.00021). A significantly positive correlation of IL-17 expression with iNOS in colorectal cancer specimens was established (r = 0.6757; p = 0.046). In addition, iNOS gene expression was also strongly correlated with HDAC3 expression (r = 0.8290; p = 0.006). Another positive correlation was found between expression of IL-6 and HDAC3 genes (r = 0.7501; p = 0.02). Conclusion: The results demonstrated gene expression profile which favors Th17 differentiation. Based on these results we suppose that the established Th17 type inflammation can contribute to tumor promotion, including enhancing epigenetic alteration.

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Plenary Lectures


DOWN-REGULATION OF RANKL EXPRESSION ON NEUTROPHILS BY PROTEIN KINASE C INHIBITOR Viktoriya Milanova, Petya Dimitrova Department of Immunology, Institute of Microbiology, Bulgarian Academy of Sciences, Laboratory of Experimental Immunotherapy, Department of Immunology, Sofia, Bulgaria Key words: neutrophils, PKC, RANKL, IFN-γ, MMP9

Neutrophils are activated at early stages of innate immune response. Our previous investigations have shown that neutrophils express RANKL and produce IL-17 and IFN-γ contributing to activation of osteoclasts, joint degeneration and destruction in arthritis. Activation and migration of neutrophils can be regulated by protein kinase C (PKC). Herein we studied the role of PKC for RANKL expression on neutrophils. Mature bone marrow neutrophils were stimulated in vitro by TLR agonists in the presence or the absence of PKC inhibitor (inhPKC). The expression of CD11b, RANKL and IFN-γ on neutrophils was analyzed by flow cytometry. The production of MMP-9 by neutrophils was evaluated by zymography. The level of secreted OPG and TNF-α was determined by ELISA. The effect of inhPKC in vivo was investigated in a model of

collagenase-induced paw inflammation and haemorrhage. inhPKC down-regulated in a dose-dependent manner RANKL and CD11b expression. It failed to affect IFN-γ production at low doses and even increased the intracellular depot of the cytokine at higher concentrations. However, inhPKC reduced the production of MMP-9 and TNF-α and enhanced OPG secretion at low concentration. In vivo the pretreatment of mice with inhPKC for 30 minutes decreased paw and ankle swelling induced by collagenase injection and reduced the pain-related sensitivity score. Bone marrow neutrophils from drug-treated mice showed lower expression of CD11b and failed to express RANKL after restimulation by TLR agonists in vitro. Our data show the effect of inhPKC on RANKL expression on neutrophils and its potential for an inhibition of tissue destruction.

IMMUNOBIOLOGICAL STUDIES OF YKL-40 IN INFLAMMATORY JOINT AND TUMOR PROCESSES Maria Kazakova1, Anastas Batalov2, Victoria Sarafian1 1Department

of Medical Biology, Medical University - Plovdiv, 2Clinic of Rheumatology, Kaspela University Hospital, Plovdiv, Bulgaria Key words: YKL-40, biomarker, arthritis, glioma YKL-40 is a recently discovered protein with still unclear biological functions. It is an extracellular matrix glycoprotein with conservative amino acid sequence similar to that of chitinase proteins but without enzymatic activity. This new biomarker has not been studied yet in Bulgaria. The aim of the study is to find a possible correlation between YKL-40 and the clinical course, imaging diagnosis, conventional clinical laboratory tests and anti- and

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pro-inflammatory cytokines in inflammatory and degenerative joint diseases and tumor processes. Our investigation is the first that demonstrates a significantly elevated serum YKL-40 level in Bulgarian patients with active rheumatoid arthritis compared to healthy controls. We prove that the high YKL-40 concentration in synovial fluid and serum correlates with sonographic parameters, proinflammatory cytokines and conventional clinical



laboratory parameters such as ESR and CRP. We believe that YKL-40 might have a high significance in the prediction of joint inflammation and play a dominant role in the pathogenesis and disease activity in inflammatory joint diseases. The high levels of the glycoprotein in carcinomas of the mammary gland correlate with HER/2 which could indicate that YKL-40 is a marker for the malignant potential

of breast tumors. We found a correlation between the mRNA and protein expression of YKL-40 in astroglial tumors of the central nervous system, which suggests a role of YKL-40 as a potential indicator of the invasiveness and malignancy of the neoplastic process. Our results provide help in expanding the diagnostic instrumentarium and improve the therapeutic approach in inflammatory joint and tumor diseases.

COMPLEX IMMUNOLOGICAL APPROACH FOR PATIENTS WITH INFLAMMATORY BOWEL DISEASE Tsvetelina Velikova1, Zoya Spassova2, Raina Shentova3, Ekaterina Ivanova-Todorova1, Dobroslav Kyurkchiev1, Spaska Stanilova4, Iskra Altankova5 1Department

of Clinical Laboratory and Clinical Immunology, 2Clinic of Gastroenterology, St. Ivan Rilski University Hospital, Medical University, Sofia, Bulgaria, 3University Children’s Hospital, Sofia, 4Department of Molecular Biology, Immunology and Medical Genetics, Medical Faculty, Trakia University, Stara Zagora, Bulgaria, 5Lozenets University Hospital, Sofia University, Bulgaria Key words: inflammatory bowel disease, fecal calprotectin, fecal lactoferrin, ASCA, cytokines, qRT-PCR

Identification and evaluation of intestinal inflammation in patients with inflammatory bowel disease (IBD) are essential for the diagnosis and management of these patients. The AIM of the study was to investigate some immunological markers locally and in the serum. METHODS: We studied 141 patients with IBD and 25 healthy persons. Fecal calprotectin was assessed in stool samples by one step-card test, point-of-care Quantum Blue and ELISA, and lactoferrin - by one step-card test. We assessed also mRNA relative quantities of IL-10, TGFβ, IL-6, IL-23, IL-17A and FoxP3 in inflamed and adjacent normal colonic mucosa samples by performing qRT-PCR assay, and protein tissue and their serum levels by ELISA. Antibodies against Sacharomyces cerevisiae (ASCA) were assessed by ELISA. RESULTS: The results confirmed the higher diagnostic specificity and sensitivity of fecal calpro-


tectin in inflammation assessment in IBD patients (100% and 72.4%, respectively), and much lower of fecal lactoferrin (100% and 48%, respectively). The activated genes in inflamed tissue were predominately genes for FoxP3, TGFβ and IL-10 in comparison to adjacent normal tissue in IBD, and compared to normal mucosa from non-IBD patients (p < 0.05). We found significantly higher protein levels of IL-6, IL-17A и IL-10 in inflamed mucosa in comparison with serum levels in IBD patients. We detect significant differences between higher IL-23 levels in serum of IBD patience compared to non-IBD persons. ASCA performed satisfactory in diagnosis of IBD patients. CONCLUSION. Complex immunological markers of the local intestinal inflammation and systemic involvement could be employed with benefits in assessment of IBD patients.

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POSTERS T-CELL DEVELOPMENT IN THYMIC MULTICELLULAR COMPLEXES: IT ALWAYS TAKES TWO TO TANGO Tsvetanka Marinova1, Lubomir Spassov2 1Department

of Biology, Medical Genetics and Microbiology, 2Department of Surgery, St. Kliment Ohridski Sofia University, Sofia, Bulgaria Key words: T- cell development, thymic complexes

Thymic non-lymphoid (stromal) cells form a specific microenvironment for lymphocytes, which ensures the intra-thymic inductive humoral and selective cellular signals of T-lymphocytes proliferation and differentiation. The T-cell development in the thymus is said to ”take two to tango” (T cells and thymic stromal cells). However, the mechanism of interaction between complex-forming stromal cells and thymocytes remains to be clarified. The present study was focused on the detection and analysis of thymic multicellular lympho-stromal complexes as possible sites of positive and negative T-cell selection in teleost fishes, reptilia and some mammals using light, transmission and scanning electron microscopy. Structural and immunocytochemical peculiarities (expression of adhesion molecules, MHC antigens and growth factors) of lympho-epithelial complexes

(LEC), rosettes, thymic nurse cells-like complexes (TNC): classical epithelial cell-derived TNC and macrophage-derived TNC were detected in normal and pathological mammalian thymus. Predominant involvement of subcapsular LEC in T-cell development during thymus leukemogenesis was demonstrated. Thymic multicellular complexes were not included as an essential part of T-cell ontogenesis in fishes. Myo-epithelial complexes were typical for reptilian thymus. The data provide evidence for significant participation of complex-forming stromal cells in T-cell repertoire generation. It is highly probable that cortical epithelial cell-derived TNC and macrophage-derived TNC are involved in both positive and negative T-cell selection in mammalian thymus, including in human thymus.

CORRELATION BETWEEN ANTI-HELICOBACTER PYLORI ANTIBODIES AND ANTISPERM ANTIBODIES IN DIFFERENT GROUPS OF PATIENTS D. Dimitrova1, R. Hadjiolova2, Iv. Dimova3, P. Andreeva-Gateva4, N. Trifonova1, N. Petkova5, Z. Krastev5 1Department

of Biology, 2Department of Pathophysiology, 3Department of Medical Genetics, 4Department of Pharmacology, Medical Faculty, Medical University, Sofia, Bulgaria, 5Clinic of Gastroenterology, St. Ivan Rilsky University Hospital, Sofia, Bulgaria Key words: anti-H. pylori antibodies, H. pylori infection, immune infertility, anti-sperm antibodies, sperm antigens

Objective: To find a correlation between serum anti-Helicobacter pylori antibodies (anti-Hp Abs) and anti-sperm antibodies (ASA) in patients with gastroduodenal diseases caused by Helicobacter pylori (HpP) and infertile patients (InfP). Study design: HpP, InfP and control group of blood donors

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(BD) were selected. Serum ASA were studied by agglutination tests, sperm immobilization test (SIT), and ELISA. Serum IgG anti-Hp Abs were detected by ELISA. Results: A significantly higher ASA prevalence was found in InfP as compared with BD and HpP, as well as in HpP compared with BD. We Folia Medica 2015; 57; Suppl. 1 © 2015 Medical University, Plovdiv, Bulgaria


found a positive linear correlation between anti-Hp Abs and ASA for HpP by SIT. The absorption with human spermatozoa reduced completely the sperm immobilization values of all positive in SIT sera. Conclusion: This study presents data about the

prevalence of ASA in InfP and HpP as compared to BD, and the correlation between ASA and anti-Hp Abs found only in HpP, assuming that H. pylori infection may play a role in ASA induction.

WOMEN WHO HELPED US TO MAKE THE FIRST STEPS INTO MOLECULAR BIOLOGY Stefka Delimitreva Department of Biology, Medical Faculty, Medical University of Sofia, Bulgaria Key words: women, molecular biology There are not many names of women in the lists of the most prestigious science awards though women were indeed involved in some of the key discoveries in molecular biology. Here are only a few of these: • eukaryotic cells possess nuclei; • the origin of mitochondria and chloroplasts was explained by the endosymbiotic theory; • during meiosis, homologous chromosomes are segregated independently; • DNA molecules are double helix; • DNA caries the genetic information; • the lagging chain of DNA is replicated in fragments; • repeated

DNA motifs disturb the replication; • some viruses can integrate into cell DNA and this can cause cancer; • the specificity of antibodies depends on their amino acid sequence. These are just some of the first topics studied in the first years in many biological and medical faculties, and even in the high schools. The presentation contains biographic data about women involved in the above mentioned discoveries: Eleanor Carothers, Rosalind Franklin, Martha Chase, Tsuneko Okazaki, Yvonne Barr, Lynn Margulis, Gillian Bates, Marian Koshland.

CORRELATIONS OF PHENOTYPE VARIANTS OF SPINDLES, CHROMOSOMES AND ACTIN CAPS IN METAPHASE OOCYTES FROM TWO MOUSE STRAINS Irina Chakarova, Ralitsa Zhivkova, Valentina Hadzhinesheva, Tenyo Tenev, Stefka Delimitreva, Venera Nikolova, Maya Markova Department of Biology, Medical Faculty, Medical University of Sofia, Bulgaria Key words: meiotic spindle, actin, chromatin, oocyte The aim of our study was to analyze correlations of chromatin, tubulin and actin morphology in mouse oocytes. We investigated meiotic spindles, actin caps, chromosomal condensation and alignment in oocytes from two inbred mouse strains, BALB/c and C57/Black. Our study included 931 oocytes, 494 of which were mature (metaphase II) and 437 in metaphase I. To find any correlations between chromatin, tubulin and actin structures, we sought phenotype combinations which were stable and relatively frequent. We used the following methods: ovarian stimulation of the mice, obtaining mature oocytes by stimulated ovulation, dissection of ovarian Folia Medica 2015; 57: Suppl. 1 © 2015 Medical University, Plovdiv, Bulgaria

follicles to obtain immature oocytes, in vitro maturation (IVM), fixation of oocytes, specific labeling and visualization of microtubules, fibrillar actin and chromatin, fluorescent microscopy, measurement of cytoskeletal structures. Chi-square test and Fisher’s test were used to determine the significant differences between distinct phenotypic classes. Based on the obtained data, we calculated the frequencies of different combinations of the following categories: spindle/actin cap, spindle/chromosome condensation, and spindle/chromosome alignment for immature oocytes in metaphase I and mature oocytes in metaphase II. We found the following

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correlations between phenotypic variants: normal spindles with well organized tubulin fibers and without significant deviations from the mean size had normal actin cap and good chromosome align-

ment. Disorganized and destroyed spindles showed correlation with missing or non-homogenous actin cap and poor chromosome alignment.

A STRUCTURE CORRESPONDING TO BALBIANI BODY IN OOCYTES OF PATIENTS WITH OVARIAN POLYCYSTOSIS Valentina Hadzhinesheva1, Venera Nikolova1, Irina Chakarova1, Mariana Panevska2, Ralitsa Zhivkova1, Tenyo Tenev1, Stefka Delimitreva1, Maya Markova1 1Department

of Biology, Faculty of Medicine, Medical University of Sofia, Bulgaria, 2Maychin Dom University Hospital of Obstetrics and Gynecology, Medical University of Sofia, Bulgaria Key words: Balbiani body, oocyte, ovarian polycystosis In mammalian oocytes Balbiani body is an evolutionary relic. It is described as perinuclear aggregate of membrane organelles related to the polarity of non-mammalian oocytes. Some authors report its presence in mouse and human primordial follicles. Despite the importance of cytoskeletal proteins for ovarian function and pathology, very few studies have addressed the presence and distribution of these proteins in polycystic ovaries. We investigated sections of human polycystic ovarian tissue for vimentin and a set of cytokeratins by epifluorescence. Intermediate filaments were revealed using specific antibodies: mouse monoclonal anti-cytokeratin pan Ab I and FITC-conjugated anti-mouse Ab II(SigmaAldrich) and rabbit anti-vimentin Ab I and TRITCconjugated anti-rabbit Ab II (Sigma-Aldrich).

Chromatin was counterstained by Hoechst 33258. The studied proteins showed strong colocalization. Positive reaction was detected in two main ovarian compartments: with weak intensity in follicular cells and very strong in perinuclear position in oocytes of primordial follicles. Epifluorescent study of the oocytes from primordial follicles allowed us to identify the immunopositive structure in oocytes as Balbiani body, a transient organelle responsible for establishing oocyte polarity and ooplasm gradients in non-mammalian vertebrates. Our results suggest functional interaction of different types of cytoplasmic intermediate filament proteins in polycystic ovaries and a possible importance of the Balbiani body for human oogenesis in norm and pathology.

DOWN-REGULATION OF AUTOREACTIVE CELLS BY ANTIBODY THERAPY IN MRL/LPR MURINE MODEL OF SYSTEMIC LUPUS ERYTHEMATOSUS Silviya Bradyanova1, Petroslav Chipinski1, Nikolina Mihaylova1, Stela Chausheva1, Ekaterina Todorova2, Dobroslav Kyurkchiev2, Fulvio D’acquisto3, Andrey Tchorbanov1 1Institute

of Microbiology, Department of Immunology, Bulgarian Academy of Sciences, Sofia, 2Laboratory of Clinical Immunology, St. Ivan Rilski University Hospital, Sofia, 3Centre for Biochemical Pharmacology, William Harvey Research Institute, London, UK Key words: autoimmunity, MRL/lpr mice, annexin A1 antibody Objectives: Systemic lupus erythematosus (SLE) is the prototype systemic autoimmune disease. Annexin A1 (AnxA1) was originally identified as

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a phospholipase A2 (PLA2)-inhibitory protein and second messenger of glucocorticoid pharmacological effects. The purpose of our study is to check Folia Medica 2015; 57; Suppl. 1 © 2015 Medical University, Plovdiv, Bulgaria


if it is possible to down-regulate the activity of autoreactive T and B cells from lupus-prone mice by treating them with a neutralizing monoclonal antibody against AnxA1. Material and methods: Splenocytes from MRL/ lpr mice were incubated with plate-bounded antiCD3 and anti-CD28 antibodies and then incubated in ranging concentrations of anti-Anx-A1 antibody solution. The effect of the antibody on CD25 and CD69 T cell activation markers was tested in vitro. Subsequently we have used two groups of MRL/ lpr mice: 7-week-old mice that are still disease free and a second group of animals (16 weeks old) with advanced manifestations. We injected the mice with the studied antibody. Control groups

were injected with PBS only. Results: The results show that the AnxA1 is expressed by both the B and T cells of the autoimmune mice. We have found a dose-dependent decrease in the expression of the activation markers CD25 and CD69 on splenocytes from MRL/ lpr mice incubated with anti-AnxA1 antibody and retains the levels of anti-dsDNA antibodies, secretion of IL-10 and prevents the appearance of skin lesions in the lupus mice. Conclusion: The generated anti-Annexin A1 antibody could be used to down-regulate the activity of the autoimmune cells in MRL/lpr mice mostly in the animals with the initial manifestations of the disease.

SELECTIVE ELIMINATION OF ALLERGEN-SPECIFIC B LYMPHOCYTES WITH CHIMERIC PROTEIN-ENGINEERED MOLECULES Nikola Kerekov1, Antoaneta Michova2, Georgi Nikolov3, Maria Muhtarova3, Bogdan Petrunov3, Maria Nikolova3, Andrey Tchorbanov1 1Institute

of Microbiology, Bulgarian Academy of Sciences, Sofia, 2National Specialized Hospital for Active Treatment of Haematological Diseases, Sofia, 3National Center of Infectious and Parasitic Diseases, Sofia, Bulgaria Key words: DerP1, allergy, chimeric molecules DerP1 is one of the major allergenic molecules of Dermatophagoides pteronyssinus which causes the widespread house dust allergy. The pathological DerP1-specific B cells produce allergen-specific IgE antibodies that mediate most of the hypersensitivity allergic reactions. The selective elimination of allergen-specific B cells by artificial protein molecules which inhibit Dpt-recognizing IgE antibodies production is a legitimate therapeutic goal for allergy. Co-crosslinking of the immunoglobulin receptor and the inhibitory B cell receptor CR1 (complement receptor 1) on the DerP1-specific B cells by this molecule have the potential to deliver suppressive signals, selectively silencing these B-cells only. A synthetic peptide, DerP1 p52-71, containing a B and T cell epitopes and anti-CR1 monoclonal antibody were used for the construction of DerP1 chimera. We analysed the


effects of the chimeric molecule on PBMC from allergy patients. Suppression of DerP1-specific IgE antibody production was measured by ELISpot. Apoptosis assay was performed by FACS using AnnexinV-FITC/PI staining. We observed significant inhibition of allergenspecific proliferation and reduction of specific IgE antibodies after treatment of PBMC from allergic patients with DerP1-peptide chimera. Culturing of these PBMC in the presence of the same chimera increased the percentage of apoptotic (Annexin Vpositive) B lymphocytes, but not of T lymphocytes. The constructed protein-engineered chimeric molecule binds DerP1 specific B lymphocytes via their BCR and suppresses selectively the production of anti-DerP1 IgE antibodies by co-crosslinking of the BCR and CR1. This way we could alter the allergic immune response towards a mild outcome.

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IN VITRO SUPPRESSION OF CYTOKINE PRODUCTION BY AN INHIBITOR OF HISTONE DEACETYLASES Zlatka Dobreva, Spaska Stanilova Department of Molecular Biology, Immunology and Medical Genetics, Medical Faculty, Trakia University, Stara Zagora, Bulgaria Key words: HDAC, IL-10, IL-12p40, IL-23, SAHA

Histone deacetylases (HDACs) are enzymes that have the ability to remove acetyl groups from specific lysine residues on histone proteins and thus regulate chromatin structure and gene expression. Small molecule inhibitors of histone deacetylases (HDACs) are a new class of drugs used in clinical trials for the treatment of various solid or hematological malignancies. The role of HDACs in the regulation of cytokine synthesis from activated immune cells, however, is not exactly elucidated. Our study was designed to investigate the effect of the HDACs inhibitor suberoylanilide hydroxamic acid (SAHA) on proinflammatory (IL-12p40 and IL-23) and anti-inflammatory (IL-10) cytokine production from stimulated human peripheral blood mononuclear

cells (PBMC). PBMC were isolated from 12 healthy donors. They were stimulated with LPS and C3bgp with or without SAHA. Cytokine production was determined in culture supernatants at 6 and 24 h, by ELISA. IL-12p40, IL-23 and IL-10 synthesis was increased in stimulated with LPS and C3bgp PBMC at 6 and more visible at 24 h. Early at 6 h, we detected significantly decreased IL-12p40, IL-23 and IL-10 production in stimulated PBMC treated with SAHA. This tendency was retained at 24 h when SAHA realized maximum suppressive effect on the stimulated cytokine synthesis. In conclusion, our results demonstrated that inhibition of HDACs leads to suppression of IL-12p40, IL-23 and IL-10 cytokine production regardless of stimuli used.

TRANSFECTION OF SERUM-FREE CELL LINE McCoy-PLOVDIV WITH pMONO-neo-GFP Vesselin Alexandrov1, Samir Naimov2, Victoria Sarafian1, Marian Draganov1 1Department

of Medical Biology, Medical Faculty, Medical University - Plovdiv, 2Paisii Hilendarski Plovdiv University, Bulgaria Key words: McCoy-Plovdiv, pMONO-neo-GFP Production of relatively high quantities of properly folded and post-translationally modified proteins is essential for biomedical research. Due to some protein purification and downstream processing difficulties, expression of human proteins in homologous systems is not a common practice. In this respect the serum free McCoy-Plovdiv cell line offers a number of advantages. However, the data regarding the transfection efficiency of McCoy line and its derivatives is limited. In this study we have tested three different transfection reagents (TurboFect, Lipofectamine 2000 and Alphafectin)

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on McCoy-Plovdiv cell line. The effects of DNA/ Transfection reagent ratio, post transfection culturing time, and the type of culturing media were assessed. The transfection efficiency was estimated by monitoring of fluorescence intensity of the LGFP reporter, delivered by pMONO-neo-GFP plasmid. As a result an optimized standard operating procedure for transfection of serum free McCoy-Plovdiv cell line is developed and reported. The study is supported by grant ДР-04/2013 from Medical University - Plovdiv.



DIFFERENTIAL ROLE OF PROMOTER POLYMORPHISM OF IL12B GENE IN THE DEVELOPMENT OF CONGENITAL ANOMALIES OF THE KIDNEY AND URINARY TRACT Lyuba Miteva1, Emilya Kostadinova2, Spaska Stanilova1 1Department 2Department

of Molecular Biology, Immunology and Medical Genetics, Medical Faculty, Trakia University, of Pediatrics, Medical Faculty, Trakia University, Stara Zagora, Bulgaria

Key words: IL-12B, promoter polymorphism, rs17860508, CAKUT, Bulgaria The aim of the study was to evaluate whether promoter polymorphism in IL-12B (IL12Bpro) gene is associated with the development of congenital anomalies of the kidney and urinary tract (CAKUT) among Bulgarian children. Genotyping of IL-12Bpro polymorphism, resulting from microinsertion combined with AA/GC transition (rs17860508), was performed by allele-specific PCR among 134 patients and 169 healthy children. The genotype and allelic frequencies of IL-12Bpro polymorphism in total group of CAKUT patients were similar to that in controls (chi2 = 0.918; p = 0.632 and chi2 = 0.062; p = 0.804). However, we observed a significantly different distribution when patients were subdivided according to the kind of anomalies. Elevated frequency of homozygous genotype with GC transition was observed in patients with renal hypo-dysplasia (50%) and obstructive uropathies (41%) compared to controls (29%). The frequency of heterozygous

genotype was reduced in patients with renal hypodysplasia (25%) and obstructive uropathies (32%) compared to controls (48.5%). In contrast, heterozygous genotype was significantly overrepresented in patients with double or horseshoe kidney (74%) compared to controls (48.5%) and compared to the rest subgroups of CAKUT patients (37%). Among patients with vesicoureteral reflux, the genotype distribution of investigated polymorphism was close to that in controls. We could assume that different anomalies from CAKUT spectrum are associated with specific genetic background. Here, we present a differential role of IL-12Bpro polymorphism for the development of CAKUT malformations among Bulgarian children. The heterozygous genotype could be accepted as an additional risk factor for the development of double or horseshoe kidney and as a protective for renal hypo-dysplasia and obstructive uropathies.

INVESTIGATION OF MATRIX METALLOPROTEINASES-2 AND -9 IN HYPERTENSIVE PATIENTS WITH TYPE 2 DIABETES A. Blazhev1, K. Kostov2, M. Atanasova1, A. Dimitrova2, S. Tisheva3, A. Nikolov4 1Department 4Department

of Biology, 2Department of Pathophysiology, 3Department of Cardiology and Rheumatology, of Propaedeutics of Internal Diseases, Medical University, Pleven, Bulgaria

Key words: matrix metalloproteinases-2 and -9, arterial hypertension, type 2 diabetes mellitus

Introduction: Matrix metalloproteinases (MMPs) are a family of zinc-dependent endopeptidases which participate in the remodeling of extracellular matrix. Experimental research has shown that MMP-2 and MMP-9 may be related to increased synthesis of collagen and vascular changes in hypertensive patients with type 2 diabetes mellitus (T2DM). Aim: To compare serum concentrations of MMP-2 and


MMP-9 in patients with arterial hypertension and T2DM with those of a control group of healthy individuals. Methods: Two groups were formed: Group I - control group (CG) (n = 20, male 8, female 12; SBP = 124.0 ± 3.7; DBP = 82.2 ± 4.1 mmHg; mean age 47.92 ± 11.3 years); Group II - arterial hypertension with T2DM (AHT2DM) (n = 50, male 21, female 29; SBP = 146.83 ± 11.9;

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DBP = 82.55 ± 9.7 mmHg; mean age 63.5 ± 11.0 years). MMP-2 and MMP-9 were determined by ELISA (R&D Systems). Statgraphics Centurion XVI was used for statistical analyses. Results: We found statistically significant differences in MMP-2 between AHT2DM (36.5387 ± 11.37 ng/ml) and CG (29.8085 ± 7.58 ng/ml) (p = 0.0187), and in

MMP-9 between AHT2DM (39.1386 ± 20.25 ng/ ml) and CG (22.6847 ± 8.04 ng/ml) (p = 0.0013). Conclusion: The serum concentrations of MMP-2 and MMP-9 were increased significantly compared to those of the controls, which demonstrates their important biological role for vascular changes in hypertensive patients with T2DM.

PENDING DROP METHOD: A FAST INFORMATIVE ASSAY FOR IN-VITRO CHARACTERIZATION OF CLINICAL SAMPLES FOR DIAGNOSIS OF PULMONARY DISFUNCTIONS Albena Jordanova1, Asya Tsanova2, Maya Bangyozova3, Eleonora Lontova4, Emilia Christova5, Zdravko Lalchev3 1Faculty

of Medicine, 2Faculty of Medicine, 3Faculty of Biology, 4Faculty of Chemistry and Pharmacy, St. Kliment Ohridski University, Sofia, Bulgaria, 5Faculty of Public Health, Medical University, Sofia, Bulgaria Key words: gastric aspirates, tracheal aspirates, lung maturity, surface tension, prematurely born infants

Gastric aspirates (GA) and tracheal aspirates (TA) taken from prematurely born and full term infants were analyzed. Searching for a fast informative assay to help in diagnosing alveolar immaturity we determined the equilibrium and dynamic surface characteristics of clinical samples using the stateof-the-art pendant drop method. Tensiometer KSV CAM 101 (KSV Instruments Ltd., Finland) was used in the study. Surface tension was determined using the axisymmetric drop shape analysis. By a camera connected to the apparatus series of pictures (from 60 to 300 pictures per minute) were taken and an equilibrium value of the surface tension (γeq, mN/m) was determined. In addition, dynamic surface parameters were detected: maximal value of the surface tension at 100% drop surface (γmax, mN/m) registered as a result of surface readsorption

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of film molecules at the final phase of decompression; and minimal value of surface tension at 20% drop surface (γmin, mN/m) registered at the final phase of surface compression when surface active components are packed with maximal density. The results showed that the most informative parameter to diagnose pulmonary disorders was the γmin value; while lower minimal surface pressure was recorded in GA and TA from healthy full term infants, significantly higher values of this parameter was measured in prematurely born children. Our results could be useful for the clinical practice for fast surfactant immaturity diagnostics by the pendant drop method for prematurely born children with regard to lifesaving therapy by application of exogenous surfactant preparations.



EXOGENOUS MELATONIN AND SOME SPERM PARAMETERS IN RAMS S. Manchev1, P. Taushanova1, R. Stefanov1, G. Anev2, P. Tzvetkova1 1Institute

of Biology and Immunology of Reproduction, Bulgarian Academy of Sciences, Sofia, 2Experimental Agriculture Centre, Targovishte, Bulgaria Key words: ram, melatonin, reproduction, sperm Introduction: The main hormone of the pineal gland, melatonin, has been studied mainly as a participant in the reproductive process at central levels. Studies indicate that the seasonal reproductive cycle in animals is most likely under melatonin regulation. In humans, however, this signal might not have the same significance due to the social environment and the changed conditions of lifestyle. The precise hormone effect on male reproductive function has not been clarified. The direct effect of melatonin on sperm has been studied even less. The aim of the study was to determine whether there is any difference between the effect of melatonin ingested by food and melatonin injected exogenously on the qualitative characteristics of ram spermatozoa. Material and methods: Nine rams

were used in the study divided into 3 groups of 3 animals: controls, a group with melatonin implants and a group of melatonin ingested by food. Ejaculate was obtained three times: on days 30, 45 and 60 after the beginning of the study. Initial assessment of the sperm was carried out within 10 minutes after sample collection. Results: The percentage of motile sperm measured in the control group declined progressively over the studied period while that of the treated animals increased. There was no difference between groups with different addition of exogenous melatonin. Conclusion: Treatment with exogenous melatonin, as well as through food, or in the form of an implant, probably improves the fertility of rams by increasing fertilization important characteristics of their semen.

ANTISPERM ANTIBODIES AND INFERTILITY IN PATIENTS WITH OBESITY V. Lazarov1, D. Dimitrova1, A. Gateva2, Z. Kamenov2, Il. Vatev1 1Department

of Biology, Medical Faculty, 2Clinic of Endocrinology, University Hospital Alexandrovska, Medical University - Sofia, Bulgaria Key words: anti-sperm antibodies; infertility; PCOS; obesity Aim: To study anti-sperm antibodies (ASA) prevalence in women with obesity, to compare it with controls and to assess the correlation between ASA and infertility and some cardiometabolic risk factors. Patients and methods: The study included 72 premenopausal women, divided in 3 groups: obese patients with polycystic ovary syndrome (PCOS), obese patients without PCOS and healthy controls. ELISA was used to detect ASA and the ELISA index/cut-off ratio was used to analyse the results. Results: Patients with obesity with and without PCOS had significantly higher concentration of ASA than controls (0.91 ± 0.2 and 0.89 ± 0.1 vs.


0.56 ± 0.3, p < 0,001). The incidence of the ASA positive sera in the 3 groups was 21.2 %; 30% and 5.9 %, respectively. No correlation was found between ASA and the indices of visceral obesity, insulin resistance, lipid profile and arterial blood pressure. Infertility was present in 5 obese PCOS women; none of them had positive ASA in ELISA. Conclusion: Obese patients have significantly higher ASA compared to C. This is not linked to the insulin resistance and unfavourable cardiometabolic profile of the patients. The role of ASA in PCOS-related infertility needs more investigation in larger studies with different methods for ASA.

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TUMOR TARGETING IN VIVO USING QUANTUM DOT-LABELED POLYMERSOMES Biliana Nikolova1, Severina Atanasova1, Iana Tsoneva1, Ichio Aoki2, Zhivko Zhelev3, Rumiana Bakalova4 1Institute

of Biophysics and Biomedical Engineering, Bulgarian Academy of Sciences, Sofia, Bulgaria, Imaging Center, National Institute of Radiological Sciences, 4-9-1 Anagawa, Inage-ku, Chiba 263-8555, Japan, 3Institute of Biophysics and Biomedical Engineering, Bulgarian Academy of Sciences, Sofia, Bulgaria; Medical Faculty, Trakia University, Stara Zagora, Bulgaria, 4Molecular Imaging Center, National Institute of Radiological Sciences, 4-9-1 Anagawa, Inage-ku, Chiba 263-8555, Japan; Medical Faculty, Sofia University, Sofia, Bulgaria 2Molecular

Key words: cancer, drug delivery systems, quantum dots, in vivo imaging Cancer is one of the leading causes of mortality and the main goal of biological and medical sciences is to increase the effectiveness of early detection, diagnostic and treatment. The present study was designed to investigate whether polyion complex hollow vesicles (polymersomes), based on chemically-modified chitosan, were appropriate for passive tumor targeting in the context of their application as drug carriers. The experiments were performed on colon-cancer grafted mice. Mice were injected intravenously with nanoparticles: (i) QD705labeled polymersomes (average size ~120 nm), or (ii) native non-tagged QD705. Optical imaging was carried out on Maestro EX 2.10 In Vivo Imaging System (excitation filter 435-480 nm; emission filter 700 nm, longpass). In the case of QD705, the fluorescence appeared in the tumor area within 1 min after injection and disappeared completely within

60 min. A strong fluorescence signal was detected in the liver on the 30th minute. Fluorescence of QD705-labeled polymersomes appeared in the tumor area immediately after injection with excellent visualization of blood vessels in the whole body. A strong fluorescent signal was detected in the tumor area within 16 hours. This indicated that QD705labeled polymersomes were delivered predominantly into the tumor due to their long circulation in the bloodstream and enhanced permeability and retention effect. A weak fluorescent signal was found in the liver area. Our data suggest that size-controlled long-circulating polymersomes are very promising carriers for drug delivery in solid tumors including delivery of small nanoparticles and contrast substances, forming an useful matrix for development of nano-formulations with theranostic capabilities.

RELATIONSHIP BETWEEN SERUM AND TISSUE EXPRESSION OF YKL-40 IN HIGH-GRADE GLIOMA Maria Kazakova1, Dessislava Staneva2, Ilian Koev3, Dmitrii Staikov4, Nonka Mateva5, Pavel Timonov4, George Miloshev2, Victoria Sarafian1 1Department

of Medical Biology, Medical University-Plovdiv, Bulgaria; 2Laboratory of Yeast Molecular Genetics, Acad. Roumen Tsanev Institute of Molecular Biology, Bulgarian Academy of Sciences; 3Department of Neurosurgery, Medical University-Plovdiv; 4Division of Forensic Medicine, Medical University - Plovdiv, 5Department of Medical Informatics, Biostatistics and E-Learning, Medical University - Plovdiv, Bulgaria Key words: YKL-40, biomarker, high-grade glioma Malignant gliomas are the most common type of primary malignant brain tumors, characterised by extreme proliferation and aggressive invasion. De-

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spite advances in diagnostic methods and surgical techniques, the survival rate remains unchanged. The YKL-40 gene is one of the most over-expressed



in high-grade gliomas. The high serum levels of the glycoprotein correlate to poor prognosis and malignancy of tumor processes. We aimed to compare YKL-40 mRNA level and protein expression in the tumor site and in the serum of high-grade glioma patients. Expression of YKL40 in 36 patients with glial tumors (astrocytoma III grade and glioblastoma) and 33 age-matched healthy people was measured by gene analysis, immunohistochemistry and ELISA. Significantly increased YKL-40 serum levels in high-grade glioma patients compared to the control group were found. YKL-40 staining in situ was more abundant in glioblastoma tissue than in anaplastic astrocytoma with the lowest level in

normal brain tissue. Gene analysis determined that in general YKL-40 mRNA in glioma patients was over-expressed versus a normal brain. A positive correlation between YKL-40 transcript and protein levels was revealed. In conclusion, these findings support the role of YKL-40 as a player in the pathogenesis of glial tumors and might contribute to glioblastomas’ specific biological characteristics which distinguish them from grade III gliomas. The study is supported by grant DP – 02/2013 from Medical University– Plovdiv and partially by grant DUNK-01-2/2009 from the Ministry of Education and Science.

PROINFLAMMATORY CYTOKINE GENE (II12B AND TNF-ALPHA) POLYMORPHISMS IN BULGARIAN PATIENTS AFFECTED BY CHRONIC INFLAMMATORY ARTHRITIDES Irena Manolova1, Mariana Goycheva2, Rumen Stoilov2, Rasho Rashkov2, Spaska Stanilova3 1Department

of Health Care, Medical Faculty, Trakia University, Stara Zagora, 2Clinic of Rheumatology, University Hospital, Medical University, Sofia, 3Department of Molecular Biology, Immunology and Medical Genetics, Medical Faculty, Trakia University, Stara Zagora, Bulgaria Key words: IL-12, TNF-a, rheumatoid arthritis, ankylosing spondylitis, SNP In the present study, we analyzed the association between two polymorphic loci in the promoter region of IL-12B (IL12Bpro; rs17860508) and TNF-α (-308 A/G, rs1800629) genes and chronic inflammatory arthritides in the Bulgarian population. A case–control study was carried out on 58 patients with ankylosing spondylitis (AS), 138 rheumatoid arthritis (RA) patients and 177 healthy subjects. The allele and genotype frequencies of rs17860508 and rs17860508 were determined using allele specific PCR and restriction fragment length RLFP-PCR assays, respectively. No association between the rs1800629 polymorphisms and RA risk in the Bulgarian population was established. However, we found significant differences in the genotype (p = 0.042) and allele (p = 0.021) frequencies of TNF-α -308A/G polymorphism between AS patients and controls.


There was an increase of the TNFα-308G allele (95.7% vs 87.6%; OR = 3.151) and decrease of the TNFα-308A allele in AS patients versus healthy controls (4.3% vs 12.4%; OR = 0.317). Regarding the allele and genotype distribution of rs17860508 polymorphism of the IL12B in AS and RA cases compared to controls no significant difference was found, although in logistic regression analysis the presence of IL-12Bpro allele 2 in the genotype (1.2 + 2.2 vs. 1.1) was associated with an increasing risk of developing RA (OR = 1.44) and AS (OR = 1.89). Our data support the hypothesis that genetic variation in pro-inflammatory cytokine genes affecting inducible cytokine production may contribute to susceptibility and developing chronic inflammatory arthritides in the Bulgarian population.

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PERIPHERAL BLOOD MONONUCLEAR CELLS ISOLATED FROM HEALTHY DONORS SHOW DECREASED ACTIVATION AFTER CULTURING WITH GLIOBLASTOMA MULTIFORME CELL CULTURES MEDIA Kalina Tumangelova-Yuzeir1, Ekaterina Ivanova-Todorova1, Тsvetelina Velikova1, Emanuil Naydenov2, Dobroslav Kyurkchiev1 1Medico-diagnostic

Laboratory of Clinical Laboratory and Immunology, St. Ivan Rilski University Hospital, Department of Clinical Laboratory and Clinical Immunology, 2Clinic of Neurosurgery, St. Ivan Rilski University Hospital, Department of Neurosurgery, Medical University, Sofia, Bulgaria Key words: GBM, T-lymphocytes, Nestin, GFAP, Sox-2 Glioblastoma multiforme (GBM) is one of the most common primary brain tumours characterized by uncontrolled cell proliferation, increased apoptosis, invasiveness and neoangiogenesis, which makes it very difficult to treat. The most supported theory for the origin of GBM is the cancer stem cells theory which holds that the neural stem cells (NSCs) or progenitors can transform in cancer stem cells (CSCs). CSCs share certain biological features with NSCs such as proliferation, self-renewal and differentiation in neural cells. There are suggestions that CSCs organize the tumour mass and are responsible for its resistance to conventional therapy. CSCs are in the focus of scientific investigations aiming to explain the processes occurring in the GBM microenvironment. We isolated and cultured cells of 14 patients with histologically proven GBM. The resulting cell

cultures were investigated for intra- and extracellular markers typical for NSCs and CSCs - Nestin, CD133, Sox-2, CD44 and GFAP. All cell cultures showed high expression for the referred markers. Peripheral blood mononuclear cells (PBMCs) were isolated from 9 healthy donors and were cultivated with 9 GBMs supernatants for 60 hours. After that we studied their activation ability after stimulation with phytohemagglutinin (PHA) – mitogen used for T-lymphocyte activation. The PBMCs cultured with GBMs media showed decreased activation index as opposed to the control cells. The results suggest that there are factors in the GBM media inhibiting the T-lymphocytes activation and that these factors may be related to the GBM escape from the immune system control.

IDENTIFYING MOLECULAR SIGNATURES IN COLORECTAL CANCER Yana Feodorova1, Desislava Tashkova2, Anton Todorov3, Nikolay Mehterov1, Gancho Kostov4, Rossen Dimov5, Victoria Sarafian1 1Department of Medical Biology, 2Department of General and Clinical Pathology, 3Department of Propedeutics

of Surgical Diseases, Medical University - Plovdiv, 4Clinic of Surgery, Kaspela University Hospital, Plovdiv, 5Department of Special Surgery, Medical University - Plovdiv, Bulgaria Key words: colorectal cancer, gene expression, YKL-40, SETD2 Colorectal cancer (CRC) is one of the most frequent solid tumors worldwide. It has become clear that the multitude of genetic and epigenetic alterations, which gradually accumulate during CRC progression, lead to global impairment of cell signalling. Treatment options are becoming increasingly dependent on the molecular makeup of the tumor and the efficacy has been significantly increased with the introduction

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of targeted substances. Several recent studies aim at identifying intrinsic colorectal cancer subtypes based on molecular signatures but further validation is needed before the results can be translated into the clinic. The aim of our study was to learn more about the molecular mechanisms governing CRC progression. To this end, we determined the expression levels of 14 genes in 24 patients with sporadic Folia Medica 2015; 57; Suppl. 1 © 2015 Medical University, Plovdiv, Bulgaria


CRC by RT-qPCR analysis. We detected different transcription levels between normal and tumor tissue. Interestingly, EGFR and KRAS were downregulated in malignant tissue. The only gene which showed significantly elevated mRNA levels in all patients was YKL-40 whose product has putative functions in angiogenesis and tissue remodeling. In addition, a correlation was found between increased YKL-40 and decreased SETD2 levels. SETD2 is a histone

methyl-transferase which catalyzes the formation of H3K36me3 – a histone modification associated with actively transcribed regions. Our preliminary results suggest that YKL-40 and SETD2 represent a common molecular signature in the cases studied. In addition, altered histone modification patterns could be related to CRC development. The study is funded by grant HO – 02/2013 from Medical University-Plovdiv.

BUILT-IN ADJUVANTICITY OF GENETICALLY AND PROTEIN ENGINEERED CHIMERIC MOLECULES FOR TARGETING OF INFLUENZA A PEPTIDE EPITOPES Iva Ivanova1, Nikola Kerekov1, Desislava Hlebarska1, Nikolina Mihaylova1, Maria Nikolova2, Jozsef Prechl3, Andrey Tchorbanov1 1Laboratory

of Experimental Immunology, Institute of Microbiology, Bulgarian Academy of Sciences, Sofia, Bulgaria, 2National Reference Laboratory of Immunology, National Center of Infectious and Parasitic Diseases, Sofia, Bulgaria, 3Immunology Research Group, Hungarian Academy of Sciences, Hungary Key words: influenza virus, DNA vaccines, gene-engineering

Objectives: We are exploring as a new genetic vaccine, a DNA chimeric molecule encoding a T- and B-cell epitope-containing influenza A virus hemagglutinin peptide joined to sequences encoding a single-chain variable fragment antibody fragment specific for the costimulatory B cell complement receptors 1 and 2 (CR1/2). This recombinant DNA molecule was inserted into an eukaryotic expression vector and used as a naked DNA vaccine in wild type and CR1/2 knockout mice. The DNA vaccination was followed by prime-boosting with the protein-engineered replica of the DNA construct, thus delivering an activation intracellular signal. Methods: Gene and protein engineering; RTPCR, DNA immunization; Cytotoxic assay; FACS,


Sygnal transduction, ELISA and ELISpot assays. Results: The intramuscular administration of the DNA construct resulted in the in vivo expression of an immunogenic chimeric protein, which cross-links cell surface receptors on influenza-specific B cells. Immunization with an expression vector containing the described construct and boosting with the protein chimera induced a strong anti-influenza cytotoxic response, modulation of cytokine profile and a weak antibody response in Balb/c mice. The same immunization scheme did not result in generation of influenza-specific response in mice lacking the target receptor, underlining the molecular adjuvant effect of receptor targeting.

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NEUTRALIZATION OF AUTOREACTIVE T CELLS BY MONOCLONAL ANTIBODY IN PRISTANE-INDUCED MOUSE MODEL OF SYSTEMIC LUPUS ERYTHEMATOSUS Petroslav Chipinski1, Nikolina Mihaylova1, Silvyia Bradyanova1, Stela Chausheva1, Fulvio D’acquisto2, Andrey Tchorbanov1 1Department

of Immunology, Institute of Microbiology, Bulgarian Academy of Sciences, Sofia, Bulgaria;

2Centre for Biochemical Pharmacology, William Harvey Research Institute, Charterhouse Square, London, UK

Key words: autoimmunity, annexin A1 antibody, pristane-induced lupus

Objectives: The protein Annexin A1 (ANXA1) is thought to be a second messenger in some of glucocorticoids’ signal pathways. Researches proved its role as one of the T cell activators. We hypothesized that it could be possible to downregulate the activity of autoreactive T cells from Balb/c mice with pristane-induced lupus using monoclonal antibody against the ANXA1 protein. Methods: Lupus-like symptoms were induced through intraperitoneal injection of pristane in Balb/c mice. Flow cytometry was used for the detection of ANXA1 expression on T and B cells and measuring the effect of anti-ANXA1 antibody on CD25 and CD69 activation markers. We also performed an apoptosis assay. The levels of IL4, IL-10, IFN-γ, anti-double stranded (ds) DNA antibodies in the sera and the number of dsDNA

producing plasma cells were quantified by ELISA and ELISpot. Results: Our model was verified as useful for studying lupus syndrome. We detected decrease of the levels of the cytokines concerned. Furthermore, the number of anti-dsDNA producing B cells was significantly lower in treated groups compared to controls. In vitro data showed that anti-ANXA1 antibody is effective in respect to reducing the activation of the T cells. Anti-ANXA1 antibody also induces the early and late apoptosis of B and T cells. Conclusions: Annexin A1 plays a role in T cell activation and blocking of its activity by specific antibody could be used to reduce the activation of T cells, hence the manifestations of lupus in pristane-induced mouse model.

NOBEL PRIZES AWARDED TO WOMEN IN NATURAL SCIENCES Ralitsa Zhivkova Department of Biology, Medical Faculty, Medical University - Sofia, Bulgaria Key words: Nobel Prize, women Seventeen Nobel prizes in natural sciences have been awarded to women: two in Physics (Nuclear Physics), four in Chemistry (two in Nuclear Chemistry and two in Biochemistry and Structural Chemistry) and eleven in Physiology and Medicine. Only one woman, Marie Curie, has been honoured Noble prize twice, with the 1903 Nobel Prize in Physics and the 1911 Nobel Prize in Chemistry. Particularly notable is the presence of women among the laureates for the last 30 years – time of rapid development of the molecular biology branch in medical sciences. Nobel awarded research of scientific groups

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including women has been related to: discovery of mobile genetic elements (1983, Barbara McClintock, individually); isolation of growth factors (1986, Rita Levi-Montalcini); important principles of drug treatment (1988, Gertrude Elion); genetic control of early embryogenesis (1995, Christiane Nüsslein-Volhard); odorant receptors and the function of olfactory system (2004, Linda Buck); discovery of the human immunodeficiency virus (2008, Françoise Barrè-Sinoussi); investigations on the structure and function of the ribosomes (2009, Ada Yonath); telomere function and discovery of



the enzyme telomerase (2009, Elizabeth Blackburn and Carol Greider), and the present year’s Nobel prize in Physiology and Medicine awarded for the discovery of cells that constitute a positioning sys-

tem of the brain to John O’Keefe and the Mosers - May-Britt and Edvard. Thus, women have their own place in modern science, among the brightest minds that have won Nobel prizes.

A RESEARCH ON THE BRAF MUTATION STATUS OF PATIENTS WITH ADVANCED AND/OR METASTATIC MELANOMA Yoana Stoyanova1, Violeta Dimitrova1, Milka Georgieva2 1Prof.

Dr. Paraskev Stoyanov Medical University, Varna, 2Department of Biology, Prof. Dr. Paraskev Stoyanov Medical University, Varna, Bulgaria Key words: melanoma, BRAF mutations, targeted therapy Melanoma is one of the most aggressive forms of skin cancer. Management of melanoma is evolving rapidly due to an improved understanding of the molecular heterogeneity of this disease and the development of effective, personalized, targeted therapy strategies. Rational molecular testing should be based on an understanding of the events that are

likely to occur in a given tumor and the clinical implications of test results. This presentation briefly summarizes the molecular profiling of melanoma, the molecular effects of activating BRAF mutations and the modern current molecular diagnostic method for targeted treatment of melanoma.

SUPPRESSION OF AUTOREACTIVE B AND T CELLS BY PROTEIN CHIMERIC MOLECULES IN A HUMANIZED MODEL OF AUTOIMMUNE DIABETES MELLITUS Iliyan Manoylov1, Nelly Delcheva1, Iliana Atanassova2, Andrey Tchorbanov1 1Stephan

Angeloff Institute of Microbiology, Bulgarian Academy of Sciences, Sofia, Bulgaria, 2Department of Endocrinology, Acad. I. Penchev Hospital for Endocrinology and Gerontology, Sofia, Bulgaria Key words: autoimmune diabetes mellitus, chimeric molecules, B-cell suppression

Autoimmune diabetes mellitus (ADM) is an organspecific chronic disease characterized by T and B cell autoreactivity leading to the progressive destruction of pancreatic beta cells, followed by hyperglycemia and the generation of autoantibodies against self antigens. GAD65 is a membrane bound enzyme that catalyses the convertion of the glutamic acid to gamma aminobutyric acid. Auto-reactive B cells secrete antibodies recognizing GAD65 as a major auto-antigen in ADM, but they are also important auto-antigen-presenting cells.The elimination of B and/or T cells involved in the pathological immune response is a reasonable approach for effective therapy of diabetes mellitus. It has been recently shown that complement


receptor type 1 (CR1) on human B and T-lymphocytes has suppressive activity. The co-crosslinking of this receptor with BCR inhibits B cell activation and proliferation and it is an attractive new target for negative signal delivery. We hypothesize that it may be possible to suppress selectively GAD65 – specific B cells from ADM patients by treatment with chimeric molecule, containing a monoclonal antibody against the inhibitory B-cell receptor CR1 coupled to peptides carrying-out GAD65 B/T epitopes. This engineered molecule is expected to be bound selectively by the immunoglobulin receptors of B-cells with antiGAD65 specificity and to deliver selectively to them a strong suppressive signal.

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Posters


We observed significant inhibition of cells proliferation and reduction of specific GAD65 IgG antibodies after treatment of PBMCs from ADM patients with GAD65-peptide chimeras. Cultur-

ing of these PBMCs in the presence of the same chimeras increased the percentage of apoptotic B lymphocytes.

p53 AND Ki67 EXPRESSION IN ORAL LICHEN PLANUS PATIENTS BEFORE AND AFTER DIODE LASER (810 NM) THERAPY Milena N. Draganova-Filipova1, Maria Mutafchieva2, Svitlana Bachurska3, Plamen Zagorchev4 1Department

of Medical Biology, Medical Faculty, Medical University - Plovdiv, 2Department of Periodontology and Oral Diseases, Dental Faculty, Medical University - Plovdiv, 3Department of General and Clinical Pathology, Faculty of Medicine, Medical University - Plovdiv, 4Department of Medical Physics and Biophysics, Faculty of Pharmacy, Medical University - Plovdiv, Bulgaria Key words: oral lichen planus, PDLT, Ki67, p53 Oral lichen planus (OLP) is a chronic inflammatory disease of unknown etiology and autoimmune pathogenesis. The autoreactive T-Lymphocytes distract the basal epithelial cells but the antigens are unknown. Corticosteroid therapy is often insufficient, which requires application of alternative treatment methods. Recently, photodynamic laser therapy (PDLT) has been used for treatment of inflammatory diseases because of its analgesic, anti-inflammatory and regenerative effects. The aim of the study is to establish the local effect of PDLT on the proliferation activity of keratinocytes. Ten patients (7 women and 3 men) underwent PDLT with methylene blue and diode laser (810 nm; 0.50 W; 30s; 1.2 J/cm2) 3 times weekly for a month. The control examination was 10 days after the last treatment. The proliferative activity

of keratinocytes was studied by immunohistochemical detection of Ki67 and p53 in biopsy of oral mucosa before and after therapy. The expression of p53 in basal cells was more intensive before therapy while the number of Ki67positive cells in the patients´ biopsies remained the same after PDLT. The observed changes in the expression of pro-apoptotic marker (p53) and maintained levels of the proliferation marker (Ki67) were in correlation with clinical signs such as lesion size reduction and diminished pain after PDLT. Our results demonstrate a new approach for the treatment of OLP, providing a good clinical response, reduction of lesions size and long-lasting remission. The investigation is supported by grant HO03/2014 from Medical University-Plovdiv.

A BIOINFORMATICAL APPROACH TO THE IDENTIFICATION OF miRNAs AND THEIR POTENTIAL TARGETS IN ORAL SQUAMOUS CELL CARCINOMA Nikolay Mehterov1, Boyan Vladimirov2, Victoria Sarafian1 1Department

of Medical Biology, Faculty of Medicine, Medical University - Plovdiv, 2Department of Oral and Maxilofacial Surgery, Faculty of Dental Medicine, Medical University - Plovdiv, Key words: oral squamosus cell carcinoma, microRNA, messengerRNA, bioinformatics

MicroRNAs represent a class of small (18-24nt in length) RNAs that mediate gene expression at the posttranscriptional level. Recent studies propose that

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30 to 80% of all human genes are predicted to act as targets of miRNA. As a result, dysregulation of miRNAs is potentially dangerous and may lead to



various malignant processes including oral carcinogenesis. The aim of the presented analysis was to identify new miRNAs which are directly linked to disease-associated target transcripts in OSCC. Methods: On the basis of both published microarray experiments and a literature survey of OSCC, a dozen differentially regulated mRNA transcripts were selected. Identification of potential miRNA targets in their 3’UTRs was carried out by means of web tools (miRTarBase; microRNA. org; targetscan.org) designed for identification of miRNA:mRNA interactions dependant on base-pair complementarity and target prediction score. Results: The bioinformatical approach used identified 11 miRNAs with a possible role in OSCC by locating miRNA binding sites in 3’UTRs

of transcripts such as NOTCH1, PIK3CA, HRAS, PTEN, MMP9, SMAD4, MYC, CCND, MAP4K3, EP300, CREBBP, KDM6B, PROX1 and EphA2. One of them, miR-21, has already proved to play an essential role in various carcinomas, including OSCC. Subsequent validation of the bioinformatical results is planned and will involve RT-qPCR in tumor tissues, blood and saliva from patients with OSCC versus controls. Conclusions: We propose that selected miRNAs (as well as their likely target mRNAs) might show aberrant expression in OSCC versus normal samples, making them ideal novel tools with diagnostic, prognostic and therapeutic value. The study is supported by grant HO-18/2014 by Medical University - Plovdiv

FINGERPRINT MORPHOLOGY IN TWINS Lyudmila F. Belenska1, Ivan I. Maslarski2 1Department

of Biology, Medical Genetics and Microbiology, Medical Faculty, 2Department of Human Anatomy, Histology and Pathology, Faculty of Medicine, St. Kliment Ohridsky University, Sofia, Bulgaria Key words: fingerprint analysis, twins, predisposition

Objectives: Twin concordance studies are suitable for investigation of morbidity risk of disease development determined by interaction of genes with environmental factors. Such study is the fingerprint analysis using qualitative indicators together with certain genetic markers that define the specific papillary drawings. Although there is a link between quality indicators and the predisposition to various diseases or different behavior, many issues are not well elucidated. Methods: Data was collected from both palms of 21 pairs of monozygotic (MZ) and 22 pairs of dizygotic (DZ) twins. Fingerprints and papillary images were obtained by the standard method. Qualitative parameters were observed and distribution analyses combined by diagnostic statistical analysis were used.


Results: The frequency of ulnar loops on fingers in MTs and DTs appeared to be the highest on the 5th finger (87.5% and 69.04%, respectively) and lowest on the 4th one in MTs (25%) and on the 2nd finger in DTs (35.81%). Radial loops in MTs and DTs had the highest percentage frequency on the 2nd finger (12.5% and 5.95%, respectively), while they were completely absent on the 1st and 3rd fingers in MTs and on the 3rd, 4th and 5th fingers DTs. The frequency of ulnar loops in MTs was almost identical on both hands with a slight prevalence on the left hand (64.55%). In DTs similar prevalence appeared on the right hand (51%, to 50.10%) Conclusion: Present data extend our understanding of the relation between quality indicators such as fingerprints and predisposition to various diseases or behavior.

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SYSTEMIC INFLAMMATORY RESPONSE IN PREECLAMPSIA N. Trifonova1, S. Stoeff2, Sv. Jovtchev2, B. Bechev2, I. Buteva1, I. Dikov3, M. Vretenarska4, A. Mladenova5 1Department

of Biology, 2Department of Medical Physics and Biophysics, 3Department of Obstetrics and of Nephrology, 5Department of Clinical Laboratory and Immunology, Medical Gynecology, Faculty, Medical University, Sofia, Bulgaria 4Department

Key words: preeclampsia, ESR Introduction: ESR (previously an indirect index of fibrinogen level, FBGLVL) and CRP (recent convincing marker for systemic inflammatory response) have been approved as objective lab inflammation indices. There are certain drawbacks characteristic of ESR (age and Hct dependency) and CRP (a costly assay). The plasma viscosity (PV) measurements are usually strongly degraded. For that reason ICSH recommends the Zeta sedimentation ratio (ZSR) like a refined ESR tool insensitive of age and Hct (within 0.25-0.47). The objective is to elaborate the clinical relevance of the ZSR on a sample of women with moderate stage of preeclampsia. Materials and methods: The study included healthy women (HS, n = 53), women with symptomatic (SH n = 26) and essential hypertension (EH, n = 21) and pregnant women-healthy (HP, n = 55) and with preeclampsia (PP, n = 40); Blood

collection and measurements of ESR, ZSR, PV and FBGLVL; Comparative statistics (box plot images BPI) and ROC analysis by SPSS as well as Two Graphs ROC (TG ROC) analysis by Excel. Results: BPI showed a statistically significant (SS) increase in ESR, ZSR and FBGLVL (PP > HP > SH > EH > HS) but BPI of PV increases SS only in EH. The area under curve (AUC) of ROC is larger for ZSR (PP > SH >EH), while for PV AUC is smaller and with degraded shape (in PP and SH). The ZSR’s TG ROC reading for the cut off value of PP shows coincidence with the 95th percentile. All other TG ROC readings indicate narrowing of the reference range though. Conclusion: In preeclampsia the ZSR ROC analysis refer to liable systemic inflammatory response corresponding with highest FBGLVL.

PREGNANCY AS A RISK FACTOR FOR TRIGGERING AUTOIMMUNITY. ANALYSIS OF ANTI-C1q ANTIBODIES IN HEALTHY PREGNANT WOMEN’S SERA Vishnya Stoyanova1, Magdalena Tchorbadjieva2, Svetla Petrova2, Ivanka Tsacheva2 1Department 2Department

of Chemistry, Biochemistry, Physiology, and Pathophysiology, Faculty of Medicine, of Biochemistry, Faculty of Biology, Sofia University, Bulgaria

Key words: autoimmunity, pregnancy, neo-epitopes, anti-C1q antibodies

Autoimmune diseases affect predominantly females of reproductive age, which indicate the role of sex hormones as a risk factor for triggering autoimmunity. Increased levels of estrogens, estradiol, progesterone and prolactin are detected during pregnancy, and these stimulate the humoral immune response and production of autoantibodies. Women with SLE (systemic lupus erythematosus) and C1q-deficiency have an increased risk of preeclampsia, miscarriages and premature birth. Natural IgM antibodies with low titer are detected in the circulation: they are

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low affinity, polyreactive, nonspecific, nonpathogenic antibodies, produced by В1-lymphocytes. In SLE and LN (lupus nephritis) the pathologic anti-C1q antibodies are of the IgG class and correlate with severe inflammatory processes in kidney. We analyzed the interaction of sera from 31 healthy pregnant women with the native C1q molecule and its fragments (CLR – collagen-like region and gC1q - globular head of C1q) for the presence of IgG and IgM autoantibodies. We used for reference a control group of pooled 56 healthy donors. ELISA assays were Folia Medica 2015; 57; Suppl. 1 © 2015 Medical University, Plovdiv, Bulgaria


carried out at high ionic strength only for detection of C1q - anti-C1q interaction. We detected anti-C1q autoantibodies from IgG isotype in healthy pregnant females. The absence of IgМ anti-C1q rejects the suggestion that anti-C1q autoantibodies originate from the natural IgМ autoantibodies. Only gC1q

displayed autoantigen specificity in the analyzed 31 pregnant women. Our suggestion is that the globular fragment of C1q is immunogenic in the initial stages and exposes specific neo-epitopes, which are important for the triggering of pathological process at a subsequent stage.

THE LIFE AND CONTRIBUTIONS OF PROFESSOR GEORGI MARKOV, MD Maya Markova Department of Biology, Medical Faculty, Medical University of Sofia, Bulgaria Key words: Georgi Markov, molecular biology, Bulgaria Professor Georgi Markov was born in 1924. He studied medicine and then worked as a practicing physician for 5 years. In 1955, he devoted himself to science and worked as a researcher for the next 35 years. He started his research career in the Department of Cytology at Metodiy Popov Institute of Biology with the Bulgarian Academy of Sciences. Later, he became one of the founders of the Institute of Molecular Biology. His research was focused on the genome organization and gene expression regulation in eukaryotes, as well as on its changes associated with the cell cycle, cell


differentiation and carcinogenesis. Apart from his role as one of the pioneers of cell and molecular biology in Bulgaria, Georgi Markov is known also as a teacher and science popularizer. He introduced courses of Cell Biology and Cell Proliferation for biology students and Pathobiochemistry for medical students. His best known work is the book “Secrets of the Cell” which explains basic molecular biology in an understandable and entertaining way for a general audience. In 2014, Prof. Georgi Markov celebrated his 90-year jubilee.

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Back cover: One of the most beautiful reliefs of the healing cults in the Greco-Roman pantheon is the unique “Frieze of the healing family” exhibited in the Archeology Museum in Plovdiv. It was excavated in the foundations of an old ruined Turkish mosque in 1921. The correct identification of all figures was performed by Professor Zapryanov* in 1964 - Department of Social Medicine. The frieze, according to him, used to adorn a Roman valetudinaria - a military hospital - off the walls of the east entrance of the ancient city which was called Trimontium by the Romans in the late III century. It weighs about 3000 kg and is 2.80 m long and 1.08 m high. The figures on it are framed in a wide rim; it bears the personified images of the Moon (on the left) and the Sun (on the right). Presented on the frieze are (from left to right): Jaso and Panacea - Asclepios’ daughters, Telesphor - the fortunate genius of the healing process, Asclepios - the god of healing art, Hygeia - his daughter, Epione - Asclepios’ wife, Machaon and Podaleirios - his sons worshipped as military physicians. All figures, except Panacea, are entirely in full face which is very rare in a general composition picture. The frieze’s sculptor depicted in great detail the figures’ anatomic features, clothes and peculiar attributes. All deities in the composition are on a par with the only association seen between Panacea and Asclepios (Panacea touches a bundle of herbs next to Telesphor’s cowl with her left hand, while pouring the cure all (panacea) in Asclepios’ bowl).

_______________________________ * Folia Medica 1964; 6(3): 152 - 156

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