Abstracts
a significant functional role in determining susceptibility to insulin secretion and type 2 diabetes (T2D) and its different complications. To assess whether the E23K variant has any impact on occurrence of neuropathy in our Iranian population, we undertook an extensive case– control association study using a total of 204 unrelated Iranian-Guilan T2D patients with neuropathy and 235 T2D without neuropathy as the control subjects with the similar diabetes duration. E23K genotyping was performed through a very high throughput molecular TaqMan assay with the accuracy rate above 99% using ABI7300 system. No significant difference was observed in allele frequency (P= 0.539) between T2D with neuropathy and without neuropathy. Also, by genotype frequency comparison, no difference were observed between frequency of various genotypes regarding to diverse genotype models( recessive , dominant and co-dominant model) of KCNJ11 E23K variant in our Iranian diabetic patients with and without nephropathy. Our results report for the first time an association study between E23K variant and progression of neuropathy among Iranian T2D patients. Confirmation by a larger study is indicated. Keywords: Neuropathy, E23K, Type 2 diabetes, Association study doi:10.1016/j.clinbiochem.2011.08.753
E.poster – [A-10-965-2] The absence of gender specific of KCNJ11 E23K variant with risk of type 2 diabetes in a population in Iran, Guilan Ghasemi Malaekea, Keshavarz Parvanehb, Habibi Raziehb, Hedayati emami Mohammad Hassanb, Kazemnejad Ehsanb a Tehran, Iran b Rasht, Guilan E-mail addresses:
[email protected] (K. Parvaneh),
[email protected] (K. Ehsan) Introduction: The KCNJ11gene encodes for the Kir6.2 subunit of the inward rectifier K+ channel family, and glucose and insulin metabolism. Data from a few studies was basically confirmed that the KCNJ11 E23K polymorphism is more frequent in type 2 diabetic (T2D) women compared with T2D men. Our objective was to investigate the frequency of KCNJ11 E23K variant among diabetic women and diabetic men and its association with progression of T2D in women in a population in Guilan. We used 282 T2D women and 168 diabetic men in our study. E23K genotyping was accomplished by using Allelic discrimination real time PCR assay using the ABI7300 system. The diabetic women and men had similar frequency of the allele K (39.3% vs. 35.2%,). There was no significant difference in allele K frequency between diabetic women and men(OR = 1.2, p = 0.273). Also, by genotype frequency comparison, no difference were observed between frequency of E23K genotypes in diabetic women and men (P= 0.165). Although, diabetic women who carriers the K allele (KK + EK) showed slightly higher mean FBS than allele K carriers diabetic male there was no significant difference between them regarding to FBS levels(p = 0.118). Conclusion: Even though, the frequency of E23K polymorphism appear to slightly differ between T2D women and men and the K allele carrier women showed slightly higher FBS levels compared with men, the present study lacks any association between the E23K variant with occurrence of T2D in women. However, this finding may be inconclusive due to limitation of sample size. A subsequent study with a larger sample size is recommended. Keywords: E23K, Association study, Gender, Polymorohism, Iranian population doi:10.1016/j.clinbiochem.2011.08.754
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Poster – [A-10-968-2] Heparan sulfate promotes differentiation of bone marrow mesenchymal stem cell-derived hepatocyte-like cells Sahar Khajeha, Abbas S. Lotfib, Afshin Mohsenifarb, Vahid Razbanc, Masoud Soleimanib a Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran b Faculty of Medicine, Tarbiat Modares University, Chamran-AleAhmad Cross, Tehran, Iran c National Institute of Genetic Engineering & Biotechnology, Pajoohesh Town, Tehran- Karaj Highway, Tehran, Iran E-mail addresses:
[email protected] (S. Khajeh),
[email protected] (V. Razban) Introduction: Liver differentiation potential of stem cells such as bone marrow mesenchymal stem cell (BMSC) is a new perspective in curing patients with liver disease. The aim of this study is to investigate the effects of immobilized HS on hepatic differentiation of human BMSCs. Methods: HS was covalently immobilized on pre coated-collagen plates by EDC. BMSCs were seeded on polystyrene, collagen and collagen-HS matrices and differentiated to hepatocyte for three weeks. Immunocytochemistry for albumin (Alb) and alpha-fetoprotein (AFP), RT-PCR for CK18 (hepatic marker, that its expression decreased after day 21 of differentiation) and CK19 (biliary marker), activity for CYP3A4 (present in the largest quantity of all the CYPs in the liver) were evaluated at day 21 of differentiation. Results: Immunocytochemistry revealed higher expressions of both AFP and Alb in cells differentiated on HS matrix compared to collagen surface seeded cells. CYP3A activity was strongly (2.5 folds) higher in HS compared to polystyrene and collagen matrix. After differentiation, RT-PCR detected expression of CK18 only in cells differentiated on polystyrene and collagen. CK19 had no expression on the cell surfaces. Conclusion: Based on previous studies, we expected that HS interacts with HGF and recruit it from solution to solid phase and strengthen signaling pathways involved in hepatic differentiation. Results for CK18, CK19 and higher expression of Alb and AFP in concomitant of significant increase in CYP3A activity in HS matrixdifferentiated cells compared to collagen matrix indicates effective role of HS in improvement of hepatic differentiation. Keywords: Hepatic differentiation, Heparan sulfate, mesenchymal stem cell doi:10.1016/j.clinbiochem.2011.08.755
Poster – [A-10-968-3] Improved hepatic differentiation of mesenchymal stem cell by enhancing cell attachment and HGF receptor expression via a 2-dimensional matrix Sahar Khajeha, Abbas S. Lotfib, Afshin Mohsenifarb, Vahid Razbanc, Masud Soleimanib a Faculty of Medical Sciences, Tarbiat Modares University, Chamran-AleAhmad cross, Tehran, Iran b Faculty of Medicine, Tarbiat Modares University, Chamran-AleAhmad Cross, Tehran, Iran c Pajuhesh Town, Tehran_Karaj Highway, Tehran, Iran E-mail addresses:
[email protected] (S. Khajeh),
[email protected] (V. Razban) Introduction: Tissue engineering provides an alternative approach to liver transplantation in patient with liver diseases. In natural microenvironment of hepatocytes, ECM-linked growth factors have effectively interact with cell receptors. To have more functional
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Abstracts
cells during in vitro differentiation, mimicking the structure and biological function of the liver ECM may result in more effective signaling of HGF and overcome detachment and death of differentiating stem cells. The aim of this study is to seek the influence of a Heparan Sulfate-collagen matrix on hepatic differentiation of bone marrow mesenchymal stem cells. Method: Heparan sulfate (HS) was covalently immobilized on precoated-collagen IV plates by EDC. Cells were cultured on HS-collagen matrix and polystyrene, as control. Cell attachment was evaluated 60 min after plating by washing with PBS followed by trypsinization and counting attached cells using hemacytometer. Flowcytometry for c-Met (HGF receptor) at days 0, 10 and 21 of differentiation and assessing CYP3A4 enzyme activity at day 21 was performed. Results: Cell attachment on HS-collagen matrix was evaluated 1.5 times more than polystyrene plates. Flowcytometry revealed that cells differentiated on HS-collagen matrix expressed c-Met significantly higher than cells differentiated on polystyrene plates (about 2.5 folds). Assessment for CYP3A4 showed near 3 times more activity in cells differentiated on HS-collagen matrix compared to differentiated cells on polystyrene. Conclusion: Based on these results, HS-collagen matrix improves cell attachment and provides a suitable surface for hepatic differentiation by enhancing c-Met expression and activity of CYP3A4 (the predominant enzyme in the liver) via HGF and HS interaction. Keywords: Hepatic differentiation, cell attachment, HGF, c-Met, Heparan Sulfate, Collagen IV doi:10.1016/j.clinbiochem.2011.08.756
Poster – [A-10-1015-2] Evaluation of Mycoplasma arginini abundance in slaughtered camels with pneumonia confirm with nested PCR method for the first time in Iran Izadi Maryama, Navid Mehr Jafarb, Lal Alizadeh Samanec, Bolori Mehrind, Bahmani Somayee a Mashad, Iran b Mashad-AhmadAbad, Iran c Mashad-Ferdosi, Iran d Mashad-Pirozi, Iran e Mashad-Razi, Iran E-mail addresses:
[email protected] (I. Maryam),
[email protected] (L.A. Samane),
[email protected] (B. Mehrin),
[email protected] (B. Somaye) Mycoplasmas belong to members of Mycoplasmatacea family in Mulicutes class that lack of cell wall and have spontaneous reproduction. Mycoplasma arginini has been isolated from animals with pneumonia and pleuropneumonia specially cow, sheep and goat. This microorganism has been reported from camels with pneumonia. The following research considers isolation and identification of M. arginini of camels with pneumonia in Mashhad's abattoir. Sampling from the lung camels with pneumonia was implement cultivated in specific media. 48 h after incubation incubated in 37 °C, samples were filtered. Samples were then streaked on solid media and incubated in 37 °C during 3–5 days in wet chambers in order to see the colonies. After extraction DNA Nested PCR was performed for M. arginini by specific primers. Existence of M. arginini in camels with pneumonia in Saudi Arabia was reported in 2001. We have performed nested PCR for 100 lungs with pneumonia in order to search Mycoplasma. 19 samples were positive for M. arginini.The role of some Mycoplasma spp. in pulmonary diseases for human being and some livestocks is obvious.
Keywords: Camel, Nested PCR, Mycoplasma arginini, Pneumonia doi:10.1016/j.clinbiochem.2011.08.757
Poster – [A-10-1017-1] Comparison of the genetic similarity using two different DNA, bulked- and individuals, among different populations of Onobrychis viciifolia (Fabaceae) at close geographical distances using ISSRs Nosrati Hoshanga, Hosseinpour-Feizi Mohammad-Alia, Latifiyan Farzanehb, Razban-Haghighi Ahmadc a Department of Plant Science, University of Tabriz, Iran b University of Tabriz, Tabriz, Iran c Research Centre for Agriculture and Natural Resources, Tabriz, Iran E-mail addresses:
[email protected] (N. Hoshang),
[email protected] (H.-F. Mohammad-Ali),
[email protected] (L. Farzaneh),
[email protected] (R.-H. Ahmad) Introduction: In this study the patterns of genetic similarity was compared among five different populations of Onobrychis viciifolia from East-Azerbaijan Province, Iran at close geographical distance using ISSR markers. The levels of similarity among the populations were measured in two different ways: using bulked- and individual-DNA. A number of 4 arbitrary ISSR primers were used, and a number of 10 plants from each population were randomly included in the study. For both bulked- and individual-DNA the genetic similarity among the populations was estimated on the basis of Nei's Distances, and UPGMA dendrogram. Then, the resulting data were compared to assess the level of compatibility between two DNA sources. The different genetic patterns, among population were obtained from two different DNA (bulked and individual DNA). Our results suggest that both bulked and individual DNA must be used in study of genetic similarity among populations of a given species at close geographical distance. Keywords: Bulked-DNA, Fabaceae, Genetic similarity, Onobrychis viciifolia, Individual-DNAs, ISSRs doi:10.1016/j.clinbiochem.2011.08.758
Poster – [A-10-1042-2] An investigation on results of the usage of viral vectors in gene therapy Mehrdad Asgharia, Nima Esmaeel Nasabb, Mojtaba Mohaddes Ardebilic, Abdolnasser Rafia a Laboratory Sciences Department, Tabriz University of Medical Sciences, Tabriz, Iran b Medical School, Tabriz University of Medical Sciences, Iran c Biochemistry & Clinical Laboratory Department, Tabriz University of Medical Sciences, Iran E-mail addresses:
[email protected] (M. Asghari),
[email protected] (N. Esmaeel Nasab),
[email protected] (M. Mohaddes Ardebili),
[email protected] (A. Rafi) Introduction: One of the main goals of gene therapy is to add a useful gene to the selected gene in order to compensate the lack of a gene or its inactivity as well as induction of some new features. There are two ways for importing the genes into body, in both of which, gene should be placed on a vector to enter the cell. The key factor of being successful in all of the gene therapy methods is a proper vector which would deliver the gene in the best way. Materials and methods: This study is a reviewing research which investigates the results and reports of the usage of viral vectors in
