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The number of total motile sperm inseminated (TMSI) is predictive of pregnancy by intrauterine insemination (IUI) using sperm from men with spinal cord injury (SCI)
live birth rate was 40%. Sperm retrieval rates were similar (P>0.05) in men with BMI<25, 25-30, and >30 (59%, 57%, and 54% respectively). Mean BMI of men who contributed to pregnancy (27.34.9 kg/m2) was lower (P¼0.02) than for men whose sperm did not contribute to a pregnancy (28.25.4 kg/m2). No man with BMI > 43 (n¼11) contributed to a successful pregnancy even though sperm were found in men with BMI upto 57 kg/m2. On multivariable logistic regression analysis male BMI was the only predictor of successful pregnancy among the variables analyzed. Female age and female BMI did not predict clinical pregnancy outcome in this study. CONCLUSION: Obese men have lower clinical pregnancy rate after microTESE and ICSI compared to men with normal BMI. Men with BMI > 43 did not contribute to any pregnancies despite successful sperm retrieval. This information may be valuable in counseling couples prior to microdissection TESE.
O-296 Wednesday, October 24, 2012 04:30 PM ASSESSMENT OF DNA DAMAGE AND STRICT MORPHOLOGY. M. G. McIntyre, A. Patuszack, P. Shah, T.-C. Hsieh, D. J. Lamb, L. I. Lipshultz. Urology, Baylor College of Medicine, Houston, TX. OBJECTIVE: We sought to determine which patients will most benefit from investigation of DNA damage and strict morphology (SM) in semen samples. DESIGN: This is a retrospective review of a prospectively collected database. MATERIALS AND METHODS: Semen analysis data from 1496 patients obtained between 7/2002-12/2011. Semen analyses were performed using phase contrast microscopy. DNA damage was assessed using a comet assay with R27% damage considered a positive assay. Strict morphology was assessed using Kruger form criteria. Abnormal was considered to be <4% normal forms. Comparisons between DNA damage level and SM data, were performed using Student’s t-test and Spearman’s rank correlation analysis. Cutoffs for SM parameters below which most patients have significant DNA damage were determined using regression analysis. RESULTS: MeanSD age of our cohort was 35.46.3 years, DNA damage 17.115.4%, head defects 91.67.3%, neck defects 4.24.2%, tail defects 2.34.2%, and normal forms 1.91.9%. DNA damage positively correlated with head defects, but negatively correlated with neck and tail defects and normal forms. Significant DNA damage Entire cohort(n¼1479) <4% Normal forms(n¼1249) >4% Normal forms(n¼230) **
% Head % Mid-piece % Tail % Normal defects defects defects sperm .193** .178** -.069
-.173** -.177** .123
-.119** -.098** -.125
-.227** -.200** .037
Correlation is significant at the 0.01 level (2-tailed).
When men were grouped using a cutoff of 4% normal forms, numerous significant correlations were again observed between DNA damage and SM parameters in the <4% group, but not in the >4% group. Regression analysis revealed significant DNA damage at a cutoff of %1.43% normal forms as well as R92% head defects. CONCLUSION: Patients with %1.43% normal sperm or R92% sperm with head defects using SM criteria are at risk for significant DNA damage and should have DNA integrity investigated. Further study is needed to fully understand the relationship between sperm morphology and DNA damage. O-297 Wednesday, October 24, 2012 04:45 PM SEMINAL INFECTION AND ABNORMAL SPERM CHROMATIN PACKAGING. P. Cohen-Bacrie,a M. Cohen-Bacrie,a J. de Mouzon,b S. Belloc,a I. Lichtblau,a V. Napoly.a aART Unit Eylau Unilabs, Paris, France; bService de Gynecologie Obstetrique II et Medecine de la Reproduction, CHU Cochin Port Royal, APHP, Universite Paris Descartes, Paris, France.
S88
ASRM Abstracts
OBJECTIVE: It was recently found that increased seminal leukocytes was associated with significant deleterious effect on sperm concentration, motility, morphology, and DNA fragmentation; bacteriospermia was linked with sperm fragmentation only (Domes 2012). The objective of our study was to determine, in a large subfertile population, the impact of semen infection on chromatin packaging assessed by decondensation. DESIGN: Retrospective study of 4278 sperm cultures performed in a private ART Unit. MATERIALS AND METHODS: We analyzed all cultures performed from January 2006 to December 2012 and associated with sperm DNA decondensation evaluation (aniline blue staining). Most of them had a semen analysis (n¼4192) and a fragmentation measurement (TUNEL assay, n¼3844). Culture was considered positive for bacteria > 3000 colonies, and Mycoplasma R 10000; Bacterial skin flora were excluded (n¼437). Sperm count, total and progressive motility, DNA fragmentation and decondensation were compared for each germ (chi-square, variance analysis). Then, multivariate models were conducted according to the age. P value <0.05 was used. RESULTS: In total, 229 semen were contaminated with bacteria, and 187 with Mycoplasma (mainly Ureaplasma urealyticum). No negative effect of bacteria on semen parameters was found, except an increased sperm DNA fragmentation in cases with Klebsiella (24.6 19.1 vs. 22.5 11.3%, P¼0.03). On the other hand, we observed an increased sperm DNA decondensation with Mycoplasma (19.4 14.1 vs. 17.3 11.2%, P¼0.03). All sperm parameters were significantly impacted by men’s age. However, the detrimental role of Mycoplasma on decondensation remained significant when considering the age (P<0.001). CONCLUSION: Mycoplasma (urealyticum), usually considered as saprophytic, seems to play a deleterious effect on chromatine packaging when R 10000. This could explain its negative impact on sperm fertilizing capacity.
O-298 Wednesday, October 24, 2012 05:00 PM THE NUMBER OF TOTAL MOTILE SPERM INSEMINATED (TMSI) IS PREDICTIVE OF PREGNANCY BY INTRAUTERINE INSEMINATION (IUI) USING SPERM FROM MEN WITH SPINAL CORD INJURY (SCI). A. S. Q. Kathiresan,a E. Ibrahim,b T. Aballa,b G. R. Attia,a C. M. Lynne,c N. L. Brackett.c aDepartment of Obstetrics and Gynecology, University of Miami, Miami, FL; bMiami Project to Cure Paralysis, Miami, FL; cDepartment of Urology, University of Miami, Miami, FL. OBJECTIVE: Due to anejaculation and poor semen quality, most men with SCI require medical assistance to father children. It is recommended that IUI be considered as an option for assisted conception before proceeding to IVF. The purpose of this study was to determine a TMSI threshold predictive of pregnancy for IUI cycles using sperm from men with SCI. DESIGN: Retrospective chart review. MATERIALS AND METHODS: Charts were reviewed of 528 SCI patients participating between 1991 and 2011 in the Male Fertility Research Program. Of these, 57 men participated in 224 IUI cycles after ejaculation occurred by masturbation, penile vibratory stimulation, or electroejaculation. For each IUI cycle, data were collected on male and female age, level of injury, years post injury, method of ejaculation, semen volume, sperm concentration, sperm motility, TMSI, ovarian stimulation protocol, and pregnancy outcome. The Mann-Whitney U test was used to compare continuous variables, and the Chi square test was used to compare nominal variables. Receiver operating characteristic (ROC) curve analysis was used to determine a TMSI threshold predictive of pregnancy. RESULTS: When divided into cycles that resulted in pregnancy versus no pregnancy, there were no significant differences in male or female age, years post injury, level of injury, method of ejaculation, or ovarian stimulation protocol. There were no significant differences in mean semen volume or sperm concentration between groups; however, cycles that resulted in pregnancy had significantly higher sperm motility (32.1 4.7% vs. 20.7 1.3%, P¼0.005) and higher TMSI (35.9 13.9 million vs. 11.9 1.0 million, P¼0.005). ROC analysis indicated that the TMSI threshold predictive of pregnancy was 12.2 million. CONCLUSION: Couples that achieved pregnancy had sperm parameters with significantly higher sperm motility and TMSI. If R 12.2 million total motile sperm are available for insemination, couples with SCI male partners are encouraged to attempt pregnancy by IUI before proceeding to IVF.
Vol. 98, No. 3, Supplement, September 2012
O-296 Wednesday, October 24, 2012 04:30 PM ASSESSMENT OF DNA DAMAGE AND STRICT MORPHOLOGY. M. G. McIntyre, A. Patuszack, P. Shah, T.-C. Hsieh, D. J. Lamb, L. I. Lipshultz. Urology, Baylor College of Medicine, Houston, TX. OBJECTIVE: We sought to determine which patients will most benefit from investigation of DNA damage and strict morphology (SM) in semen samples. DESIGN: This is a retrospective review of a prospectively collected database. MATERIALS AND METHODS: Semen analysis data from 1496 patients obtained between 7/2002-12/2011. Semen analyses were performed using phase contrast microscopy. DNA damage was assessed using a comet assay with R27% damage considered a positive assay. Strict morphology was assessed using Kruger form criteria. Abnormal was considered to be <4% normal forms. Comparisons between DNA damage level and SM data, were performed using Student’s t-test and Spearman’s rank correlation analysis. Cutoffs for SM parameters below which most patients have significant DNA damage were determined using regression analysis. RESULTS: MeanSD age of our cohort was 35.46.3 years, DNA damage 17.115.4%, head defects 91.67.3%, neck defects 4.24.2%, tail defects 2.34.2%, and normal forms 1.91.9%. DNA damage positively correlated with head defects, but negatively correlated with neck and tail defects and normal forms. Significant DNA damage Entire cohort(n¼1479) <4% Normal forms(n¼1249) >4% Normal forms(n¼230) **
% Head % Mid-piece % Tail % Normal defects defects defects sperm .193** .178** -.069
-.173** -.177** .123
-.119** -.098** -.125
-.227** -.200** .037
Correlation is significant at the 0.01 level (2-tailed).
When men were grouped using a cutoff of 4% normal forms, numerous significant correlations were again observed between DNA damage and SM parameters in the <4% group, but not in the >4% group. Regression analysis revealed significant DNA damage at a cutoff of %1.43% normal forms as well as R92% head defects. CONCLUSION: Patients with %1.43% normal sperm or R92% sperm with head defects using SM criteria are at risk for significant DNA damage and should have DNA integrity investigated. Further study is needed to fully understand the relationship between sperm morphology and DNA damage. O-297 Wednesday, October 24, 2012 04:45 PM SEMINAL INFECTION AND ABNORMAL SPERM CHROMATIN PACKAGING. P. Cohen-Bacrie,a M. Cohen-Bacrie,a J. de Mouzon,b S. Belloc,a I. Lichtblau,a V. Napoly.a aART Unit Eylau Unilabs, Paris, France; bService de Gynecologie Obstetrique II et Medecine de la Reproduction, CHU Cochin Port Royal, APHP, Universite Paris Descartes, Paris, France.
S88
ASRM Abstracts
OBJECTIVE: It was recently found that increased seminal leukocytes was associated with significant deleterious effect on sperm concentration, motility, morphology, and DNA fragmentation; bacteriospermia was linked with sperm fragmentation only (Domes 2012). The objective of our study was to determine, in a large subfertile population, the impact of semen infection on chromatin packaging assessed by decondensation. DESIGN: Retrospective study of 4278 sperm cultures performed in a private ART Unit. MATERIALS AND METHODS: We analyzed all cultures performed from January 2006 to December 2012 and associated with sperm DNA decondensation evaluation (aniline blue staining). Most of them had a semen analysis (n¼4192) and a fragmentation measurement (TUNEL assay, n¼3844). Culture was considered positive for bacteria > 3000 colonies, and Mycoplasma R 10000; Bacterial skin flora were excluded (n¼437). Sperm count, total and progressive motility, DNA fragmentation and decondensation were compared for each germ (chi-square, variance analysis). Then, multivariate models were conducted according to the age. P value <0.05 was used. RESULTS: In total, 229 semen were contaminated with bacteria, and 187 with Mycoplasma (mainly Ureaplasma urealyticum). No negative effect of bacteria on semen parameters was found, except an increased sperm DNA fragmentation in cases with Klebsiella (24.6 19.1 vs. 22.5 11.3%, P¼0.03). On the other hand, we observed an increased sperm DNA decondensation with Mycoplasma (19.4 14.1 vs. 17.3 11.2%, P¼0.03). All sperm parameters were significantly impacted by men’s age. However, the detrimental role of Mycoplasma on decondensation remained significant when considering the age (P<0.001). CONCLUSION: Mycoplasma (urealyticum), usually considered as saprophytic, seems to play a deleterious effect on chromatine packaging when R 10000. This could explain its negative impact on sperm fertilizing capacity.
O-298 Wednesday, October 24, 2012 05:00 PM THE NUMBER OF TOTAL MOTILE SPERM INSEMINATED (TMSI) IS PREDICTIVE OF PREGNANCY BY INTRAUTERINE INSEMINATION (IUI) USING SPERM FROM MEN WITH SPINAL CORD INJURY (SCI). A. S. Q. Kathiresan,a E. Ibrahim,b T. Aballa,b G. R. Attia,a C. M. Lynne,c N. L. Brackett.c aDepartment of Obstetrics and Gynecology, University of Miami, Miami, FL; bMiami Project to Cure Paralysis, Miami, FL; cDepartment of Urology, University of Miami, Miami, FL. OBJECTIVE: Due to anejaculation and poor semen quality, most men with SCI require medical assistance to father children. It is recommended that IUI be considered as an option for assisted conception before proceeding to IVF. The purpose of this study was to determine a TMSI threshold predictive of pregnancy for IUI cycles using sperm from men with SCI. DESIGN: Retrospective chart review. MATERIALS AND METHODS: Charts were reviewed of 528 SCI patients participating between 1991 and 2011 in the Male Fertility Research Program. Of these, 57 men participated in 224 IUI cycles after ejaculation occurred by masturbation, penile vibratory stimulation, or electroejaculation. For each IUI cycle, data were collected on male and female age, level of injury, years post injury, method of ejaculation, semen volume, sperm concentration, sperm motility, TMSI, ovarian stimulation protocol, and pregnancy outcome. The Mann-Whitney U test was used to compare continuous variables, and the Chi square test was used to compare nominal variables. Receiver operating characteristic (ROC) curve analysis was used to determine a TMSI threshold predictive of pregnancy. RESULTS: When divided into cycles that resulted in pregnancy versus no pregnancy, there were no significant differences in male or female age, years post injury, level of injury, method of ejaculation, or ovarian stimulation protocol. There were no significant differences in mean semen volume or sperm concentration between groups; however, cycles that resulted in pregnancy had significantly higher sperm motility (32.1 4.7% vs. 20.7 1.3%, P¼0.005) and higher TMSI (35.9 13.9 million vs. 11.9 1.0 million, P¼0.005). ROC analysis indicated that the TMSI threshold predictive of pregnancy was 12.2 million. CONCLUSION: Couples that achieved pregnancy had sperm parameters with significantly higher sperm motility and TMSI. If R 12.2 million total motile sperm are available for insemination, couples with SCI male partners are encouraged to attempt pregnancy by IUI before proceeding to IVF.
Vol. 98, No. 3, Supplement, September 2012