- Email: [email protected]
ICSI for treatment of male factor infertility
cytogenetic results to either normal or aneuploid. The donor’s demographics, cycle parameters and semen parameters were similar for both groups. Aneuploidy was found in 30.0% of abortuses among patients undergoing oocyte donation. No significant increase in fetal aneuploidy rate was noted with increasing paternal age (<40 years ¼ 25.0%, 40–50 years ¼ 38.8%, >50 years ¼ 25.0%). A higher rate of aneuploidy though not statistically significant was seen in the ICSI vs. conventional IVF group (39.1% vs. 22.2% (P 0.20, RR 0.78, 95% CI 0.53–1.15)). We compared the cytogenetic results of the 50 oocyte recipient couples to a group of 23 female patients (mean age 28.7 1.1) who had a missed abortion following IVF using autologous oocytes. There was no statistical difference in sperm characteristics between the egg recipient and IVF patients. There was a significant difference in the male partner age between these two groups (33.7 7.6 vs. 41.5 6.8) (P<0.05). No statistically significant difference in aneuploidy rate was observed between egg recipients and IVF patients younger then 30 years using autologous oocytes (30.0% vs. 26.1% (P 0.66, RR 0.93, 95% CI 0.68, 1.27)). CONCLUSIONS: These data support that paternal age has little effect on aneuploidy rates. The increase in aneuploidy noted with ICSI requires further evaluation. Supported by: None.
for 2005 (1,425 cycles). The live delivery rate for TESE cycles increased from 28.3% to 33.6% (P¼0.042) in couples where sperm was successfully retrieved. While the multiple gestation pregnancy rate decreased from 37% to 31.9% for all IVF cycles, it did not do so for TESE cycles (29.6% to 28.0%, P¼0.737). However, the percentage of cycles resulting in triplet or more infants in the latter group did decline from 5.8% to 2.1% (P¼0.008). CONCLUSIONS: The utilization of both TESE and IVF treatments increased from 1999 to 2005. TESE with IVF/ICSI has improved in its ability to treat male factor infertility over time, with a higher live delivery rate. The overall number of multiple gestation pregnancies associated with TESEdriven IVF cycles however did not significantly change. Supported by: None.
MALE REPRODUCTION AND UROLOGY ABSTRACTS Monday, October 15, 2007 3:00 pm O-56 IDENTIFICATION OF GENE VARIABILITY WITHIN THE CONTRIN GENE OF AZOOSPERMIC PATIENTS. S. Hammoud, D. M. Dunn, B. R. Emery, R. B. Weiss, D. T. Carrell. Andrology and IVF Laboratories, University of Utah School of Medicine, Salt Lake City, UT.
Monday, October 15, 2007 4:45 pm O-55 CHANGES OVER TIME OF TESTICULAR SPERM EXTRACTION (TESE) WITH IVF/ICSI FOR TREATMENT OF MALE FACTOR INFERTILITY. R. Lee, B. Luke, P. S. Li, G. Schattman, M. Goldstein, P. N. Schlegel. James Buchanan Brady Foundation and The Center for Male Reproductive Medicine and Microsurgery, Department of Urology and Cornell Institute for Reproductive Medicine, Weill Medical College of Cornell University, New York, NY; Departments of Obstetrics and Gynecology and Epidemiology and Public Health, University of Miami, Coral Gables, FL; Center for Reproductive Medicine and Infertility, Cornell Institute for Reproductive Medicine, Weill Medical College of Cornell University, New York, NY. OBJECTIVE: We examined the effectiveness of testicular sperm extraction (TESE) with in vitro fertilization and intracytoplasmic sperm injection (IVF/ICSI) over time. DESIGN: TESE with IVF/ICSI enhances the ability to treat male factor infertility. We examined temporal changes in TESE outcomes for all types of azoospermia, using live delivery and multiple gestation pregnancy rates as primary outcome measures. MATERIALS AND METHODS: The Society for Assisted Reproductive Technology (SART) database was queried for TESE patient data in 1999, the earliest year for which robust SART data existed for TESE and in 2005, the most recent year for which SART data was available. The total number of fresh IVF cycles using autologous eggs with ICSI and cervical transfer were recorded, as were the number of cycles for TESE patients in whom sperm was successfully retrieved. The live delivery and multiple gestation pregnancy rates in each year were recorded. All statistical analyses were performed using the c2 test. RESULTS: The total number of IVF cycles increased from 62,991 to 88,422 between 1999 to 2005. The percentage of IVF cycles undergone for male factor infertility however did not change significantly: 36.9% to 36.6% (P¼0.375). Only 1.6% of such cycles used TESE with IVF/ICSI to treat male factor infertility in 1999 (1,029 cycles); this remained unchanged
TABLE. SART TESE/ICSI Outcomes
Category
2005
1999
Total number of cycles % cycles for male factor infertility % cycles for male factor infertility with TESE/ICSI % cycles for male factor infertility with TESE/ICSI resulting in live births % male factor infertility TESE/ICSI live births with multiple infants
88,422 36.6% 1.6%
62,991 36.9% 1.6%
33.6%
28.3%
28.0%
29.6%
FERTILITY & STERILITYÒ
OBJECTIVE: Contrin, a Y-box protein, is expressed in male and female germ cells. In the nucleus, it marks specific mRNAs for cytoplasmic storage and stabilization and suppresses translation. Mice with targeted disruption of the MSY2 gene resulted in spermatogenic arrest and infertility. Our objective is to determine whether single nucleotide polymorphisms (SNPs) in the contrin gene account for azoospermia in infertile males. DESIGN: Cohort study with a control group. MATERIALS AND METHODS: Peripheral blood DNA was extracted from 96 men with Azoospermia following standard procedures. Additional, 96 DNA samples from the Utah Genetic Reference Program (UGRP) were selected for controls based on known paternity. All Contrin primers were optimized and PCR reactions were processed using standard techniques. PCR products were sequenced using an ABI 3700 sequencer in a university core laboratory. Traces were then assembled using the Phrap software and analyzed for significant changes using Phred and Consed analysis utilities. The frequency of SNPs was calculated in patients and controls using SPSS vs. 13. RESULTS: A total of 7 SNPs were identified in patients, of which, 6 were novel. Of greatest interest were two SNPs, one was found in exon 1 of azoospermic patients, but was absent in the UGRP patients. The second SNP was located in intron 1, and was only found in the azoospermic patients. However, both these SNPs occurred at relatively low frequency (Table 1). TABLE. SNPs identified in the Contrin gene
SNPs
Region
UGRP genotype frequencies
cggT/Gggc cggT/Gggc gggcC/Tcc gggcC/Tcc ccC/Aggccc Tttttt-/Tctt gagcC/Ttg gagcC/Ttg Ttgt-/Tcc Ttgt-/Tcc
Exon 1 Exon 1 Exon 1 Exon 1 Exon 1 Intron 1 Intron 6 Intron 6 Intron 6 Intron 6
GG: 0.06 TG: 0.23 CT: 0.409 TT: 0.303 CA: 0 –/T: 0 CT: 0.327 TT: 0.19 –/T: 0.428 TT: 0.142
Azoospermic genotype frequencies GG: 0.054 TG: 0.2054 CT: 0.373 TT: 0.22 CA: 0.0657 –/T: 0.0125 CT: 0.368 TT: 0.09 –/T: 0.356 T/T: 0.095
Function
P-value
Val-Gly Val-Gly Pro-Ser Pro-Ser Pro-Gln
NS NS NS NS 0.01 NS NS NS NS NS
All SNPs except for the first two are novel. CONCLUSIONS: This data indicates that there is some genetic variability in the contrin gene that can contribute to the etiology of azoospermia in men. Further studies will need to evaluate the functionality of the mutation present in the conserved cold shock domain of contrin in the azoospermic patients. Supported by: Thanks to the core lab laboratories for their help.
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for 2005 (1,425 cycles). The live delivery rate for TESE cycles increased from 28.3% to 33.6% (P¼0.042) in couples where sperm was successfully retrieved. While the multiple gestation pregnancy rate decreased from 37% to 31.9% for all IVF cycles, it did not do so for TESE cycles (29.6% to 28.0%, P¼0.737). However, the percentage of cycles resulting in triplet or more infants in the latter group did decline from 5.8% to 2.1% (P¼0.008). CONCLUSIONS: The utilization of both TESE and IVF treatments increased from 1999 to 2005. TESE with IVF/ICSI has improved in its ability to treat male factor infertility over time, with a higher live delivery rate. The overall number of multiple gestation pregnancies associated with TESEdriven IVF cycles however did not significantly change. Supported by: None.
MALE REPRODUCTION AND UROLOGY ABSTRACTS Monday, October 15, 2007 3:00 pm O-56 IDENTIFICATION OF GENE VARIABILITY WITHIN THE CONTRIN GENE OF AZOOSPERMIC PATIENTS. S. Hammoud, D. M. Dunn, B. R. Emery, R. B. Weiss, D. T. Carrell. Andrology and IVF Laboratories, University of Utah School of Medicine, Salt Lake City, UT.
Monday, October 15, 2007 4:45 pm O-55 CHANGES OVER TIME OF TESTICULAR SPERM EXTRACTION (TESE) WITH IVF/ICSI FOR TREATMENT OF MALE FACTOR INFERTILITY. R. Lee, B. Luke, P. S. Li, G. Schattman, M. Goldstein, P. N. Schlegel. James Buchanan Brady Foundation and The Center for Male Reproductive Medicine and Microsurgery, Department of Urology and Cornell Institute for Reproductive Medicine, Weill Medical College of Cornell University, New York, NY; Departments of Obstetrics and Gynecology and Epidemiology and Public Health, University of Miami, Coral Gables, FL; Center for Reproductive Medicine and Infertility, Cornell Institute for Reproductive Medicine, Weill Medical College of Cornell University, New York, NY. OBJECTIVE: We examined the effectiveness of testicular sperm extraction (TESE) with in vitro fertilization and intracytoplasmic sperm injection (IVF/ICSI) over time. DESIGN: TESE with IVF/ICSI enhances the ability to treat male factor infertility. We examined temporal changes in TESE outcomes for all types of azoospermia, using live delivery and multiple gestation pregnancy rates as primary outcome measures. MATERIALS AND METHODS: The Society for Assisted Reproductive Technology (SART) database was queried for TESE patient data in 1999, the earliest year for which robust SART data existed for TESE and in 2005, the most recent year for which SART data was available. The total number of fresh IVF cycles using autologous eggs with ICSI and cervical transfer were recorded, as were the number of cycles for TESE patients in whom sperm was successfully retrieved. The live delivery and multiple gestation pregnancy rates in each year were recorded. All statistical analyses were performed using the c2 test. RESULTS: The total number of IVF cycles increased from 62,991 to 88,422 between 1999 to 2005. The percentage of IVF cycles undergone for male factor infertility however did not change significantly: 36.9% to 36.6% (P¼0.375). Only 1.6% of such cycles used TESE with IVF/ICSI to treat male factor infertility in 1999 (1,029 cycles); this remained unchanged
TABLE. SART TESE/ICSI Outcomes
Category
2005
1999
Total number of cycles % cycles for male factor infertility % cycles for male factor infertility with TESE/ICSI % cycles for male factor infertility with TESE/ICSI resulting in live births % male factor infertility TESE/ICSI live births with multiple infants
88,422 36.6% 1.6%
62,991 36.9% 1.6%
33.6%
28.3%
28.0%
29.6%
FERTILITY & STERILITYÒ
OBJECTIVE: Contrin, a Y-box protein, is expressed in male and female germ cells. In the nucleus, it marks specific mRNAs for cytoplasmic storage and stabilization and suppresses translation. Mice with targeted disruption of the MSY2 gene resulted in spermatogenic arrest and infertility. Our objective is to determine whether single nucleotide polymorphisms (SNPs) in the contrin gene account for azoospermia in infertile males. DESIGN: Cohort study with a control group. MATERIALS AND METHODS: Peripheral blood DNA was extracted from 96 men with Azoospermia following standard procedures. Additional, 96 DNA samples from the Utah Genetic Reference Program (UGRP) were selected for controls based on known paternity. All Contrin primers were optimized and PCR reactions were processed using standard techniques. PCR products were sequenced using an ABI 3700 sequencer in a university core laboratory. Traces were then assembled using the Phrap software and analyzed for significant changes using Phred and Consed analysis utilities. The frequency of SNPs was calculated in patients and controls using SPSS vs. 13. RESULTS: A total of 7 SNPs were identified in patients, of which, 6 were novel. Of greatest interest were two SNPs, one was found in exon 1 of azoospermic patients, but was absent in the UGRP patients. The second SNP was located in intron 1, and was only found in the azoospermic patients. However, both these SNPs occurred at relatively low frequency (Table 1). TABLE. SNPs identified in the Contrin gene
SNPs
Region
UGRP genotype frequencies
cggT/Gggc cggT/Gggc gggcC/Tcc gggcC/Tcc ccC/Aggccc Tttttt-/Tctt gagcC/Ttg gagcC/Ttg Ttgt-/Tcc Ttgt-/Tcc
Exon 1 Exon 1 Exon 1 Exon 1 Exon 1 Intron 1 Intron 6 Intron 6 Intron 6 Intron 6
GG: 0.06 TG: 0.23 CT: 0.409 TT: 0.303 CA: 0 –/T: 0 CT: 0.327 TT: 0.19 –/T: 0.428 TT: 0.142
Azoospermic genotype frequencies GG: 0.054 TG: 0.2054 CT: 0.373 TT: 0.22 CA: 0.0657 –/T: 0.0125 CT: 0.368 TT: 0.09 –/T: 0.356 T/T: 0.095
Function
P-value
Val-Gly Val-Gly Pro-Ser Pro-Ser Pro-Gln
NS NS NS NS 0.01 NS NS NS NS NS
All SNPs except for the first two are novel. CONCLUSIONS: This data indicates that there is some genetic variability in the contrin gene that can contribute to the etiology of azoospermia in men. Further studies will need to evaluate the functionality of the mutation present in the conserved cold shock domain of contrin in the azoospermic patients. Supported by: Thanks to the core lab laboratories for their help.
S21