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Correlation of sperm dna damage to outcome of in-vitro-fertilization (IVF) cycles
P-962 Thursday, October 17, 2013 CORRELATION OF SPERM DNA DAMAGE TO OUTCOME OF INVITRO-FERTILIZATION (IVF) CYCLES. N. Malhotra,a L. Chawla,a M. B. Shamsi,b R. Dada.b aART Centre, Department of Obstetrics and Gynecology, All India Institute of Medical Sciences, New Delhi, Delhi, India; b Laboratory for Molecular Reproduction and Genetics, All India Institute of Medical Sciences, New Delhi, Delhi, India. OBJECTIVE: The role of sperm DNA damage in predicting outcome of IVF cycles is controversial. The aim of the study was to assess the relationship between sperm DNA damage and outcome in in-vitro-fertilization (IVF) cycles including fertilization, embryo cleavage, pregnancy and live birth rate. DESIGN: Prospective cross sectional study. MATERIALS AND METHODS: Male partners from 64 couples undergoing IVF for tubal factor infertility underwent semen analysis, seminal fluid Reactive Oxygen Species (ROS) levels and assessment of sperm DNA damage. Couples with male factor, unexplained infertility, endometriosis, PCOS were excluded from the study as these could affect fertilization and embryo cleavage. Sperm DNA damage was assessed in native semen and sperms after density gradient centrifugation using single gel electrophoresis with COMET assay and sperm chromatin dispersion (SCD) at the time of insemination during IVF cycle. The results COMET and SCD were compared between those with (n¼14) and without (n¼50) pregnancy and further correlated with IVF outcome. RESULTS: The mean Sperm DNA fragmentation was found to be 35.7 21.48%.There was significantly higher percentage of DNA damaged sperms from cycles without pregnancy (29.8 vs 15.4%, p¼0.04) in comparison to those with pregnancy on COMET assay. Levels of DNA fragmentation were lower in semen from cycles with pregnancy (14.14 17.12 %) as compared to those without pregnancy (38.99 21.74 %, p¼0.10) as assessed by SCD. Levels of ROS in native (9602.9 vs 864.3 RLU/min/20 million sperm, p¼0.05) and washed semen (2387.5 vs 41453.4 RLU/min/20 million sperm, p¼0.04) were significantly higher in cycles without pregnancy when compared to those with pregnancy There was a non –significant negative correlation between COMET and SCD assays with fertilization rate (r ¼ -0.292, p¼0.10). There was no correlation with cleavage rate, pregnancy or live birth rate. CONCLUSION: Sperm DNA damage as assessed by COMET and SCD assays do not correlate with IVF outcome.
P-963 Thursday, October 17, 2013 FOLLICULAR FLUID TOTAL OXIDANT STATUS IS A DETERMINING FACTOR FOR IVF SUCCESS IN POOR RESPONDERS. B. Demir,a S. Dilbaz,b S. Deveci,a A. F. Tuncel,c a aa B. Dundar, I. Kaplanoglu. Obstetrics and Gynaecology, Etlik Z€ubeyde Hanim Women’s Health Teaching and Research Hospital, Ankara, Turkey; bObstetrics and Gynaecology, Duzce University School of Medicine, Duzce, Turkey; cMedical Biochemistry, Gazi University School of Medicine, Ankara, Turkey. OBJECTIVE: The role of oxidative stress in follicular microenvironment and folliculogenesis has been of great interest in recent years. However, there is paucity of data on IVF success for follicular fluid oxidative and antioxidative status.The aim of this study was to investigate, are there any differences in follicular fluid total oxidant status (TOS), total antioxidant status (TAS) and estradiol using microdose flare-up protocol (MF) and antagonist protocol (A) in poor ovarian responders for ICSI-ET cycles? Secondary, does follicular fluid oxidative status affect IVF success? DESIGN: Seventy patients with poor ovarian response were included in the study. Patients were prospectively randomized into A (n¼35) and MF protocols (n¼35). MATERIALS AND METHODS: Follicular fluid from follicles was collected during egg retrieval. Results of controlled ovarian hyperstimulation, and pregnancy outcome were documented. The impact of TOS and TAS on IVF success was investigated using calculated cut off value. RESULTS: There were no significant differences in follicular fluid TAS, TOS and estradiol level between A and MF protocols. Total gonadotrophin consumption and estradiol concentration on oocyte retrieval day were significantly lower in A protocol compared to MF protocol. Cycle cancellation, fertilization, implantation, and clinical pregnancy rates were comparable between two protocols. In A protocol, TAS was positively correlated with number of oocytes retrieved, number of mature oocytes retrieved, and follicular fluid estradiol level. 6 mmol H202Equiv/L was accepted as cut off value for
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ASRM Abstracts
TOS. Clinical pregnancy rates in relation to the higher cut off value for TOS were significantly higher in both A and MF protocols (TOS <6: 21.6% vs R6: 43.8%). CONCLUSION: Different controlled ovarian hyperstimulation protocols might be have different effects on antioxidative process in the follicular microenvironment. Oxidant status of the follicular fluid might be reflected to oocyte metabolism and a potential marker for predicting IVF success.
P-964 Thursday, October 17, 2013 AN ADJUVANT TEST TO MEASURE MALE GAMETE COMPETENCE. M. J. Smith, Q. V. Neri, Z. Rosenwaks, G. D. Palermo. Reproductive Medicine - CRM, Weill Cornell Medical College, New York, NY. OBJECTIVE: To assess the significance of antioxidant capacity in seminal fluid (SF) of infertile men in relation to semen characteristics and reproductive potential. DESIGN: We assessed total antioxidant capacity (TAC) in SF to identify a relationship with sperm characteristics. We plotted the TAC against abstinence period and semen parameters. To identify the effect of oxidative stress on spermatozoal integrity, DFI and mitochondrial membrane potential (MMP) were also measured. MATERIALS AND METHODS: TAC was assessed using a commercial kit and measured at DOD750. DFI and sperm MMP along with pregnancy outcome were assessed. RESULTS: A total of 42 men (38.05yrs) with and average concentration of 45.827million/ml, with motility of 44.913%, and morphology 3.01%. After choosing a threshold of 1500mM Trolox equivalents, the TAC for normospermic (n¼35) men was 2047 912mM, and OAT men (n¼7), 762176mM. A positive correlation of TAC with semen concentration (r¼0.55; P¼0.0001) and motility (r¼0.485; P¼0.0008) was identified. In a limited number of subjects tested, a lower TAC was associated with compromised MMP and DFI. Couples (n¼19) with recurrent IUI failure (3-5 cycles) had a TAC of 1093322mM in their male partner’s specimen. Six of these couples whose TAC was 889103mM were only able to achieve pregnancies following ICSI. CONCLUSION: Antioxidant capacity of seminal fluid correlated with concentration and motility. Couples whose male partners exhibit compromised antioxidant power are characterized by recurrent IUI failure and achieved pregnancies only through ICSI. Antioxidant capacity of a semen specimen may well serve as a useful addition to a standard semen analysis in order to guide treatment and predict outcome. Supported by: Institutional.
P-965 Thursday, October 17, 2013 ABSTRACT WITHDRAWN P-966 Thursday, October 17, 2013 MODULATION OF OXIDATIVE STRESS DURING IN VITRO FERTILIZATION IN BOVINE: EFFECTS OF MELATONIN ON SPERM FUNCTION AND SUBSEQUENT EMBRYO DEVELOPMENT. R. Sanchez,a C. Cheuqueman,b J. Risopatron,c R. Felmer,d J. Alvarez,e T. Mogas.f aBIOREN-CEBIOR, Department of Preclinical Science, Universidad de La Frontera, Temuco, Araucania, Chile; b BIOREN-CEBIOR, Universidad de La Frontera, Temuco, Araucania, Chile; c BIOREN-CEBIOR, Department of Basic Science, Universidad de La Frontera, Temuco, Araucania, Chile; dDepartment of Agricultural Sciences and Natural Resource, Universidad de La Frontera, Temuco, Araucania, Chile; e Centro Androgen, Androgen, La Coru~na, Spain; fDepartament de Medicina i Cirurgia Animals, Universitat Autonoma de Barcelona, Bellaterra, Barcelona, Spain. OBJECTIVE: This study determined the effects of the addition of different concentrations of melatonin during bovine IVF on subsequent embryo development and sperm functionality. DESIGN: Prospective in vitro study. MATERIALS AND METHODS: Mature in vitro oocytes were fertilized in IVF medium supplemented either with high concentrations (10-5M, 10-4M and 10-3M) or low concentrations (10-8M, 10-7M and 10-6M) of melatonin.
Vol. 100, No. 3, Supplement, September 2013
P-963 Thursday, October 17, 2013 FOLLICULAR FLUID TOTAL OXIDANT STATUS IS A DETERMINING FACTOR FOR IVF SUCCESS IN POOR RESPONDERS. B. Demir,a S. Dilbaz,b S. Deveci,a A. F. Tuncel,c a aa B. Dundar, I. Kaplanoglu. Obstetrics and Gynaecology, Etlik Z€ubeyde Hanim Women’s Health Teaching and Research Hospital, Ankara, Turkey; bObstetrics and Gynaecology, Duzce University School of Medicine, Duzce, Turkey; cMedical Biochemistry, Gazi University School of Medicine, Ankara, Turkey. OBJECTIVE: The role of oxidative stress in follicular microenvironment and folliculogenesis has been of great interest in recent years. However, there is paucity of data on IVF success for follicular fluid oxidative and antioxidative status.The aim of this study was to investigate, are there any differences in follicular fluid total oxidant status (TOS), total antioxidant status (TAS) and estradiol using microdose flare-up protocol (MF) and antagonist protocol (A) in poor ovarian responders for ICSI-ET cycles? Secondary, does follicular fluid oxidative status affect IVF success? DESIGN: Seventy patients with poor ovarian response were included in the study. Patients were prospectively randomized into A (n¼35) and MF protocols (n¼35). MATERIALS AND METHODS: Follicular fluid from follicles was collected during egg retrieval. Results of controlled ovarian hyperstimulation, and pregnancy outcome were documented. The impact of TOS and TAS on IVF success was investigated using calculated cut off value. RESULTS: There were no significant differences in follicular fluid TAS, TOS and estradiol level between A and MF protocols. Total gonadotrophin consumption and estradiol concentration on oocyte retrieval day were significantly lower in A protocol compared to MF protocol. Cycle cancellation, fertilization, implantation, and clinical pregnancy rates were comparable between two protocols. In A protocol, TAS was positively correlated with number of oocytes retrieved, number of mature oocytes retrieved, and follicular fluid estradiol level. 6 mmol H202Equiv/L was accepted as cut off value for
S426
ASRM Abstracts
TOS. Clinical pregnancy rates in relation to the higher cut off value for TOS were significantly higher in both A and MF protocols (TOS <6: 21.6% vs R6: 43.8%). CONCLUSION: Different controlled ovarian hyperstimulation protocols might be have different effects on antioxidative process in the follicular microenvironment. Oxidant status of the follicular fluid might be reflected to oocyte metabolism and a potential marker for predicting IVF success.
P-964 Thursday, October 17, 2013 AN ADJUVANT TEST TO MEASURE MALE GAMETE COMPETENCE. M. J. Smith, Q. V. Neri, Z. Rosenwaks, G. D. Palermo. Reproductive Medicine - CRM, Weill Cornell Medical College, New York, NY. OBJECTIVE: To assess the significance of antioxidant capacity in seminal fluid (SF) of infertile men in relation to semen characteristics and reproductive potential. DESIGN: We assessed total antioxidant capacity (TAC) in SF to identify a relationship with sperm characteristics. We plotted the TAC against abstinence period and semen parameters. To identify the effect of oxidative stress on spermatozoal integrity, DFI and mitochondrial membrane potential (MMP) were also measured. MATERIALS AND METHODS: TAC was assessed using a commercial kit and measured at DOD750. DFI and sperm MMP along with pregnancy outcome were assessed. RESULTS: A total of 42 men (38.05yrs) with and average concentration of 45.827million/ml, with motility of 44.913%, and morphology 3.01%. After choosing a threshold of 1500mM Trolox equivalents, the TAC for normospermic (n¼35) men was 2047 912mM, and OAT men (n¼7), 762176mM. A positive correlation of TAC with semen concentration (r¼0.55; P¼0.0001) and motility (r¼0.485; P¼0.0008) was identified. In a limited number of subjects tested, a lower TAC was associated with compromised MMP and DFI. Couples (n¼19) with recurrent IUI failure (3-5 cycles) had a TAC of 1093322mM in their male partner’s specimen. Six of these couples whose TAC was 889103mM were only able to achieve pregnancies following ICSI. CONCLUSION: Antioxidant capacity of seminal fluid correlated with concentration and motility. Couples whose male partners exhibit compromised antioxidant power are characterized by recurrent IUI failure and achieved pregnancies only through ICSI. Antioxidant capacity of a semen specimen may well serve as a useful addition to a standard semen analysis in order to guide treatment and predict outcome. Supported by: Institutional.
P-965 Thursday, October 17, 2013 ABSTRACT WITHDRAWN P-966 Thursday, October 17, 2013 MODULATION OF OXIDATIVE STRESS DURING IN VITRO FERTILIZATION IN BOVINE: EFFECTS OF MELATONIN ON SPERM FUNCTION AND SUBSEQUENT EMBRYO DEVELOPMENT. R. Sanchez,a C. Cheuqueman,b J. Risopatron,c R. Felmer,d J. Alvarez,e T. Mogas.f aBIOREN-CEBIOR, Department of Preclinical Science, Universidad de La Frontera, Temuco, Araucania, Chile; b BIOREN-CEBIOR, Universidad de La Frontera, Temuco, Araucania, Chile; c BIOREN-CEBIOR, Department of Basic Science, Universidad de La Frontera, Temuco, Araucania, Chile; dDepartment of Agricultural Sciences and Natural Resource, Universidad de La Frontera, Temuco, Araucania, Chile; e Centro Androgen, Androgen, La Coru~na, Spain; fDepartament de Medicina i Cirurgia Animals, Universitat Autonoma de Barcelona, Bellaterra, Barcelona, Spain. OBJECTIVE: This study determined the effects of the addition of different concentrations of melatonin during bovine IVF on subsequent embryo development and sperm functionality. DESIGN: Prospective in vitro study. MATERIALS AND METHODS: Mature in vitro oocytes were fertilized in IVF medium supplemented either with high concentrations (10-5M, 10-4M and 10-3M) or low concentrations (10-8M, 10-7M and 10-6M) of melatonin.
Vol. 100, No. 3, Supplement, September 2013