J. A. Shelton,b E. J. Sarcione.a aLifeCell Dx, e-fertility Diagnostics, Buffalo,. NY; bGynecology & Obstetrics, School of Medicine, University at Buffalo,. Buffalo, NY ...
P-1007 Thursday, October 17, 2013 CONSISTENT PREDICTOR OF PREGNANCY (PREG) AND SPERM FERTILIZING POTENTIAL: ADVANCED SEMEN ANALYSIS (ASA) AND HYPERACTIVATION (HA) USING CLEAR CASA FOR LOCAL OR REMOTE PATIENTS. L. Burkman,a,b R. G. George,b F. Gonzalez,c J. A. Shelton,b E. J. Sarcione.a aLifeCell Dx, e-fertility Diagnostics, Buffalo, NY; bGynecology & Obstetrics, School of Medicine, University at Buffalo, Buffalo, NY; cObstetrics & Gynecology, Indiana University School of Medicine, Indianapolis, IN. OBJECTIVE: Many couples despair of knowing the male’s fertilizing potential. Why is the basic SA still heavily utilized? Can ASAwith Clear CASA predict a man’s ability to produce a pregnancy? Classically, a capacitated sperm (CAP) reflects the functions needed to fertilize an egg. HA is the easiest CAP function to evaluate. (See Clear CASA). In 1996, we reported on regression & Sensitivity for HA vs. PREG. Here, we have studied 4 new groups. DESIGN: Four random groups of new couples were seen (1998-2012). These represent > 300 ASA from the two labs. After 9-36 months many patients were called, in a blind fashion, to learn whether a PREG had occurred (natural, IUI, IVF). His ASA report was read, showing the highest HA value. MATERIALS AND METHODS: Collection used F10 medium (diluted), & Clear CASA methods included CAP/HA analysis (IVOS). We adopted an HA cut-off of R 5% (Burkman criteria). We studied those cases which had PREG and HA R 5%. RESULTS: See Table. Original data (1996) showed a mean HA of 10.11.0 (PREG) vs. 4.40.9 (Non-PREG; P< 0.001); 49 had a PREG, and 43 had HA R 5%. Thus, 88% PASSED. In Groups 2, 3, 4, 5, the PASS rates were 88%, 100%, 91%, & 91%. In total, 90.3% received a PASS, leaving 9.7% who failed our testing (P< 0.01, chi-squared). In 17 years, the ASA methods showed no drift. # Couples with PREGNANCY ORIGINAL group new Group 2 new Group 3 new Group 4 new Group 5 ALL 5 groups
49 17 14 11 12 103
How many PASSED our Test 43 of 49 15 of 17 14 of 14 10 of 11 11 of 12 93
% of PREG Couples who PASSED our Test 88% 88% 100% 91% 91.7% 90.3%
CONCLUSION: Rigorous HA predicts a given man’s ability to produce a pregnancy–not a correlation. Collection into medium initiates CAP. PREG indicates that the female issues are okay AND that the sperm were fertile. HA motility is very visual, if CASA is not available. Prediction of PREG by HA & Clear CASA has now reached the next level via remote semen analysis in Japan, and in cities far from Buffalo. Excellent CASA is a very powerful tool within Andrology. P-1008 Thursday, October 17, 2013 DEFINING THE FERTILE MAN WITH CLEAR CASA: PHYSIOLOGIC BASICS FOR HYPERACTIVATION (HA) AND SPERM R. G. George,b CAPACITATION (CAP). B. S. Kim,a,b L. J. Burkman,a,b S. Tambar.b aLifeCell Dx, Buffalo, NY; bGyn/Ob, University at Buffalo, Buffalo, NY. OBJECTIVE: Basic semen analysis has poor predictive capacity; therefore, we have defined Clear CASA in several abstracts. The link between good CASA and predicting a man’s fertilizing potential must rely on handling human sperm in accordance with their inherent physiology. Most simple analyzers today do not offer 60 hz. DESIGN: Test the effect on CAP/HA when basic sperm physiologic functions are altered: a) temperature & CAP. Test the effect on CAP/HA results when resolution is doubled; when sperm are tracked longer (biphasic nature). These basic variations will significantly alter the HA outcome. MATERIALS AND METHODS: Sorting of HA sperm was carried out according to Burkman criteria (curvilinear velocity and amplitude of head movement and linearity. We compared 30 hz vs 60 hz; duration (0.5 sec, 0.67 sec, 1.67 sec). Comparative readings were completed within a 4 minute time span. Data were recorded for ALL HA, and 4 individ-
FERTILITY & STERILITYÒ
ual patterns (Circling, Helical, Thrash, Star). Burkman criteria were used in SORT. RESULTS: Clear CASA: confirmed previous studies that the temperature for sperm capacitation will alter velocity, amplitude and %HA. Comparing 30 hz and 60 hz: % Helical was not changed, but % ALL HA (essentially all 4 patterns) showed a 41% increase at 60 hz (P A, p.R155H; c.3992G>A, p.R1331H; c.5048G>A, p.R1683H) found in four individuals were predicted to be damaging using in-silico prediction models. These variants are highly conserved, and are not present in EVS or 1000 Genomes. All individuals with predicted damaging variants fathered children without reproductive assistance. CONCLUSION: Predicted deleterious missense variants in USP9Y present in an unselected population appear to have no significant impact on male fertility. Cohorts with unbiased ascertainment such as ClinSeqÒ are a useful tool for bioinformatic hypothesis testing. P-1010 Thursday, October 17, 2013 DIFFERENTIAL SPERM LIPIDOMIC PROFILE BETWEEN SPERM SAMPLES ACHIEVING PREGNANCY OR NOT IN INTRACYTOPLASMIC SPERM INJECTION (ICSI) CYCLES. R. Rivera,a N. Sota,a F. Domınguez,b M. Meseguer,a J. Remohı,a N. Garrido.a aInstituto Universitario IVI Valencia, Valencia, Spain; bFundacion IVI, INCLIVA, Paterna, Valencia, Spain. OBJECTIVE: The aim of this work was to define the differences in the sperm lipidomic profile between samples achieving pregnancy (P) using ICSI treatment in comparison with those that failed (F). DESIGN: Descriptive, prospective and non-randomized biomedicine study. MATERIALS AND METHODS: Sperm samples (n¼32) from infertile males undergoing ICSI cycles with normal sperm parameters (WHO 2010), were included in our study, 15 of them failing to achieve pregnancy and 17 who succeeded. Lipids were separated and analyzed by means of Ultra Performance Liquid Chromatography coupled to Mass Spectrometry (UPLC/MS) using methanol (MeOH) and methanol/chloroform (MeOH/ CHCl3) platforms. A total number of 104 different lipidic species were
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