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Vitamin D: Is there a role in male infertility?
cancellation. This was not found with the CIA. The RIA appears to be a more sensitive technique for evaluation of ovarian reserve. Supported by: None
Monday, October 18, 2004 5:45 P.M. O-108
normal sperm FP of 2 pre-prep. to 3 post-prep. Abnormal sperm FP changes has a tendency to adversely affect IVF fertzn. rates, especially when the sperm becomes prematurely hyperactivated post-prep (FP 4). In this study, we did not show a statistically significant difference in ICSI vs. IVF fertzn rates for both the FP 2 pre-prep to FP 4 post-prep and FP 3 pre-prep to FP 3 post-prep groups possibly due to a limited sample size. However, with increasing sample size we anticipate that there will be an improvement with ICSI in these patients and we strongly suggest ICSI for these patients. Supported by: None
Indications for ICSI versus IVF with sperm based upon forward progression. L. G. Chen, W. A. Caswell, L. Sung, M. Tucker, J. Jenkins, G. San Roman. Reproductive Specialists of New York, Mineola, NY; Reproductive Specialists of New York, Dept. of OBGYN, Winthrop University Hospital, Mineola, NY; Reproductive Specialists of New York, Dept. of OBGYN, Winthrop University Hospital, Dept. of OBGYN, Stony Brook University Hospital, Mineola, NY. OBJECTIVE: To determine whether ICSI or IVF should be recommended for patients with various pre- and post-preparation sperm based upon forward progression. DESIGN: Retrospective, cohort, observational study. MATERIALS AND METHODS: This study was conducted on 509 oocytes retrieved between January 2003 to April 2004. Every patient used had split inseminations between ICSI and IVF (half of oocytes used for ICSI, half for IVF). Inclusion criteria were defined as patients with normal sperm concentration ⱖ20 million/ml, motility ⱖ50% and normal morphology ⱖ 7% (as defined by Kruger’s Strict criteria). Sperm was spun down in Promoter® gradient tubes at 300g for 15 minutes, washed in Sperm Wash Media at 400g for 5 minutes, and resuspended in HTF⫹10% HSA. The samples were incubated at 37°C for about 4 hours until the time of insemination. Mortimer’s guideline for modal sperm progression grading was the standard protocol used; this is a semi-subjective scale that assigns 0 – 4 for forward progression. Pre- and Post- analyses of individual semen specimens were performed by the same technician, so inter-observer variability in interpretation was eliminated. All sperm samples studied fell into 4 categories of sperm forward progression.
RESULTS: Only oocytes that were fertilized normally (2 pronuclei, 2 polar bodies) were considered. Comparable fertilization (fertzn.) results between ICSI and IVF were obtained for sperm that demonstrated normal FP of 2 pre-prep. to 3 post-prep. (66.4% ICSI, 67.6% IVF). Sperm that demonstrated abnormal FP changes had considerably lower IVF fertzn. rates. When sperm FP of 2 pre-prep. became 4 post-prep “hyperactivated”, ICSI fertzn. rate was 68.2% compared to 42.1% in IVF. When FP remained the same at 3, ICSI fertzn. rate was again higher than IVF (71.4% and 56.8% respectively). For pre and post-prep FP of 3 to 4, ICSI fertzn. rate was 71.2% versus 51.6% IVF (pⱕ0.05).
MALE FACTOR: ART Monday, October 18, 2004 2:00 P.M. O-109 Vitamin D: Is there a role in male infertility? S. T. Corbett, J. E. Stern, S. Sundaram, A. K. Nangia. Dartmouth-Hitchcock Medical Center, Lebanon, NH. OBJECTIVE: The role of vitamin D in male infertility has been implicated by several animal studies. Deficiencies of vitamin D have been shown to retard spermatogenesis in animal models. The biological activity of vitamin D is mediated by the vitamin D receptor (VDR). VDR interacts with promoter regions of target genes in several cell types including Sertoli cells. VDR null mutant mice have significant gonadal insufficiencies. The presence of VDR in human reproductive tissue and the role of vitamin D in male infertility have not yet been demonstrated. We investigated the presence of VDR in human sperm. DESIGN: We performed a prospective evaluation of sperm from fertile and infertile men for qualitative and semi-quantitative analysis of VDR expression. MATERIALS AND METHODS: Semen specimens were collected from 11 fertile and 20 infertile men based on their semen parameters as defined by the WHO/Kruger criteria and by the length of time attempting conception. Qualitative analysis for VDR was performed by immunohistochemistry using a monoclonal antibody to human VDR. Immunoprecipitation of equal amounts of total sperm protein lysates followed by immunoblotting with VDR antibodies was performed to semi-quantitate the VDR levels. VDR expression was measured against sperm motility and concentration. RESULTS: Immunohistochemical analysis of semen samples demonstrated the presence of VDR on the mid-piece of sperm. Immunoprecipitation followed by immunoblotting with VDR antibody resulted in 2 possible forms of VDR (50 KDa and 40KDa). Infertile men with sperm concentrations ⬍5 million/cc (n⫽10) showed approximately 3-fold increase in the 50kDa VDR expression versus the fertile controls. Infertile men with concentrations ⬎5 million/cc (n⫽10) had approximately 2-fold increase in the 50kDa VDR expression versus the fertile controls. A similar trend was noted for the 40kDa VDR protein. We also observed a downward trend in VDR expression for patients with low motility, irrespective of fertility status. CONCLUSION: Our results demonstrate the presence of VDR on human sperm for the first time. In addition, we showed VDR expression was inversely related to sperm concentration in infertile men versus fertile controls. Finally we observed a linear trend for motility and VDR expression but these data reflect preliminary results and our studies are ongoing. The mechanism of action of vitamin D through the VDR on sperm has yet to be determined. Our data suggest a role for vitamin D in sperm physiology and male infertility. Supported by: Dartmouth-Hitchcock Medical Center Harmes Scholarship.
Monday, October 18, 2004 2:15 P.M. O-110 CONCLUSION: In good prognosis patients in which male factor is not a contributor to infertility, IVF and ICSI fertzn. rates are very similar for
S44
Abstracts
High, but not moderate, levels of sperm DNA fragmentation are predictive of poor outcome in egg donation cycles. C. Adams, L. Anderson, S. Wood. The Reproductive Sciences Center, La Jolla, CA.
Vol. 82, Suppl. 2, September 2004
Monday, October 18, 2004 5:45 P.M. O-108
normal sperm FP of 2 pre-prep. to 3 post-prep. Abnormal sperm FP changes has a tendency to adversely affect IVF fertzn. rates, especially when the sperm becomes prematurely hyperactivated post-prep (FP 4). In this study, we did not show a statistically significant difference in ICSI vs. IVF fertzn rates for both the FP 2 pre-prep to FP 4 post-prep and FP 3 pre-prep to FP 3 post-prep groups possibly due to a limited sample size. However, with increasing sample size we anticipate that there will be an improvement with ICSI in these patients and we strongly suggest ICSI for these patients. Supported by: None
Indications for ICSI versus IVF with sperm based upon forward progression. L. G. Chen, W. A. Caswell, L. Sung, M. Tucker, J. Jenkins, G. San Roman. Reproductive Specialists of New York, Mineola, NY; Reproductive Specialists of New York, Dept. of OBGYN, Winthrop University Hospital, Mineola, NY; Reproductive Specialists of New York, Dept. of OBGYN, Winthrop University Hospital, Dept. of OBGYN, Stony Brook University Hospital, Mineola, NY. OBJECTIVE: To determine whether ICSI or IVF should be recommended for patients with various pre- and post-preparation sperm based upon forward progression. DESIGN: Retrospective, cohort, observational study. MATERIALS AND METHODS: This study was conducted on 509 oocytes retrieved between January 2003 to April 2004. Every patient used had split inseminations between ICSI and IVF (half of oocytes used for ICSI, half for IVF). Inclusion criteria were defined as patients with normal sperm concentration ⱖ20 million/ml, motility ⱖ50% and normal morphology ⱖ 7% (as defined by Kruger’s Strict criteria). Sperm was spun down in Promoter® gradient tubes at 300g for 15 minutes, washed in Sperm Wash Media at 400g for 5 minutes, and resuspended in HTF⫹10% HSA. The samples were incubated at 37°C for about 4 hours until the time of insemination. Mortimer’s guideline for modal sperm progression grading was the standard protocol used; this is a semi-subjective scale that assigns 0 – 4 for forward progression. Pre- and Post- analyses of individual semen specimens were performed by the same technician, so inter-observer variability in interpretation was eliminated. All sperm samples studied fell into 4 categories of sperm forward progression.
RESULTS: Only oocytes that were fertilized normally (2 pronuclei, 2 polar bodies) were considered. Comparable fertilization (fertzn.) results between ICSI and IVF were obtained for sperm that demonstrated normal FP of 2 pre-prep. to 3 post-prep. (66.4% ICSI, 67.6% IVF). Sperm that demonstrated abnormal FP changes had considerably lower IVF fertzn. rates. When sperm FP of 2 pre-prep. became 4 post-prep “hyperactivated”, ICSI fertzn. rate was 68.2% compared to 42.1% in IVF. When FP remained the same at 3, ICSI fertzn. rate was again higher than IVF (71.4% and 56.8% respectively). For pre and post-prep FP of 3 to 4, ICSI fertzn. rate was 71.2% versus 51.6% IVF (pⱕ0.05).
MALE FACTOR: ART Monday, October 18, 2004 2:00 P.M. O-109 Vitamin D: Is there a role in male infertility? S. T. Corbett, J. E. Stern, S. Sundaram, A. K. Nangia. Dartmouth-Hitchcock Medical Center, Lebanon, NH. OBJECTIVE: The role of vitamin D in male infertility has been implicated by several animal studies. Deficiencies of vitamin D have been shown to retard spermatogenesis in animal models. The biological activity of vitamin D is mediated by the vitamin D receptor (VDR). VDR interacts with promoter regions of target genes in several cell types including Sertoli cells. VDR null mutant mice have significant gonadal insufficiencies. The presence of VDR in human reproductive tissue and the role of vitamin D in male infertility have not yet been demonstrated. We investigated the presence of VDR in human sperm. DESIGN: We performed a prospective evaluation of sperm from fertile and infertile men for qualitative and semi-quantitative analysis of VDR expression. MATERIALS AND METHODS: Semen specimens were collected from 11 fertile and 20 infertile men based on their semen parameters as defined by the WHO/Kruger criteria and by the length of time attempting conception. Qualitative analysis for VDR was performed by immunohistochemistry using a monoclonal antibody to human VDR. Immunoprecipitation of equal amounts of total sperm protein lysates followed by immunoblotting with VDR antibodies was performed to semi-quantitate the VDR levels. VDR expression was measured against sperm motility and concentration. RESULTS: Immunohistochemical analysis of semen samples demonstrated the presence of VDR on the mid-piece of sperm. Immunoprecipitation followed by immunoblotting with VDR antibody resulted in 2 possible forms of VDR (50 KDa and 40KDa). Infertile men with sperm concentrations ⬍5 million/cc (n⫽10) showed approximately 3-fold increase in the 50kDa VDR expression versus the fertile controls. Infertile men with concentrations ⬎5 million/cc (n⫽10) had approximately 2-fold increase in the 50kDa VDR expression versus the fertile controls. A similar trend was noted for the 40kDa VDR protein. We also observed a downward trend in VDR expression for patients with low motility, irrespective of fertility status. CONCLUSION: Our results demonstrate the presence of VDR on human sperm for the first time. In addition, we showed VDR expression was inversely related to sperm concentration in infertile men versus fertile controls. Finally we observed a linear trend for motility and VDR expression but these data reflect preliminary results and our studies are ongoing. The mechanism of action of vitamin D through the VDR on sperm has yet to be determined. Our data suggest a role for vitamin D in sperm physiology and male infertility. Supported by: Dartmouth-Hitchcock Medical Center Harmes Scholarship.
Monday, October 18, 2004 2:15 P.M. O-110 CONCLUSION: In good prognosis patients in which male factor is not a contributor to infertility, IVF and ICSI fertzn. rates are very similar for
S44
Abstracts
High, but not moderate, levels of sperm DNA fragmentation are predictive of poor outcome in egg donation cycles. C. Adams, L. Anderson, S. Wood. The Reproductive Sciences Center, La Jolla, CA.
Vol. 82, Suppl. 2, September 2004