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Pre-ART strict sperm morphology assessment and IVF and ICSI outcomes.
matin structure assay (SCSA) to determine the percentage of cells outside the main population (COMP␣t) with abnormal chromatin structure. Results: Leukocytospermia was associated with an abnormality in one or more of the standard semen parameters in 93% (15/16) of cases. Sperm motility (mean ⫾ SD) was significantly lower in the leukocytospermic group (37.9 ⫾ 15.2) compared to the non-leukocytospermic group (47.7 ⫾ 15.3%; P ⬍ 0.04) and the donor group (66.6 ⫾ 10%; P ⬍ 0.001). Patients with leukocytospermia had poor sperm morphology (16.9 ⫾ 9.1%) compared to the donor group (36.7 ⫾ 6.1%; P ⫽ 0.0003). Abnormal sperm morphology was observed in 88% of leukocytospermic samples versus 47% of non-leukocytospermic samples. ROS levels [median (25%, 75% interquartile range)] were significantly higher in the leukocytospermic group [177.9 (31.5, 430.1)], compared to the non-leukocytospermic group [2.7 (0.5, 12.2)] (P ⬍ 0.0001) and the donor group [0.4 (0.1, 2.5)] (P ⬍ 0.0001). Significantly higher levels of %COMP␣t (mean ⫾ SD) were observed in the leukocytospermic group (28.8 ⫾ 9.3) versus non-leukocytospermic group (22.6 ⫾ 10.8; P ⬍ 0.02), and the donors (15.8 ⫾ 6.8; P ⫽ 0.002). A COMP␣t value of ⱖ27% was seen in 62% of leukocytospermic samples compared to 25% in non-leukocytospermic samples. A significant negative correlation was seen between leukocyte concentration in semen and sperm motility (r ⫽ 0.31, P ⫽ 0.01) and sperm morphology (r ⫽ 0.28, P ⫽ 0.03). Leukocytospermia was positively correlated with high ROS levels (r ⫽ 0.7, P ⬍0.0001) and %COMP␣t values (r ⫽ 0.32, P ⫽ 0.01). Conclusions: Our results show a strong correlation between leukocytospermia and poor semen quality, oxidative stress and increased sperm nuclear DNA damage. COMP␣t values ⱖ 27% have been recently reported to be an indicator of negative pregnancy outcome in assisted reproduction. Therefore, infertile men with leukocytospermia should be counseled concerning the potential negative effects of increased sperm DNA damage in their pregnancy outcome. Supported by: Research was supported by the Cleveland Clinic Foundation.
P-120 Testicular volume and hormone levels in varicocele patients (fertile and infertile) and fertile controls without varicoceles. F. F. Pasqualotto, J. Hallak, L. M. Rossi-Ferragut, A. M. Lucon, P. M. Go´es, S. Arap. Univ of Sa˜o Paulo Medical Sch, Sa˜o Paulo, Brazil; Fertility Ctr for Human Reproduction, Sa˜o Paulo, Brazil. Objective: Testicular alterations associated with the varicocele are well documented. Also, altered testicular steroidogenesis as a mechanism by which the varicocele exerts its deleterious effects on spermatogenesis remains not clear. We evaluated the levels of luteinizing hormone (LH), follicle-stimulating hormone (FSH), total testosterone, and testicular volume in fertile and infertile men with varicoceles, and fertile controls without varicoceles. Design: Prospective study. Materials/Methods: The records of 27 infertile patients with varicocele and 70 controls with and without varicoceles were assessed from January 1999 to November 2000. In all patients serum hormone levels as well as testicular volume assessed with calipers were evaluated. Patient’s were divided into three groups: group A (fertile men with varicocele; n ⫽ 21), group B (infertile men with varicocele; n ⫽ 27), and group C (fertile men without varicoceles; n ⫽ 49). Results: No differences were seen in the mean age and testosterone levels across the three groups (P ⬎ 0.05). FSH levels were significantly higher in group B (7.8 ⫾ 7.6) compared to groups A (2.95 ⫾ 1.38) and C (3.43 ⫾ 2) (P ⬍ 0.05). Also LH levels were higher in group B (6.8 ⫾ 5.4) compared to patients in groups A (3.2 ⫾ 1.9) and C (3.4 ⫾ 1.74) (P ⬍ 0.05). The right testicle was smaller in group B (18.9 ⫾ 9) compared to group A (24.4 ⫾ 8.6) and C (25.6 ⫾ 8.5) (P ⬍ 0.05). Also, the left testicle was smaller in group B (16.6 ⫾ 8.9) compared to groups A (21. ⫾ 7.8) and C (23.4 ⫾ 8.3) (P ⬍ 0.05). Conclusions: Infertile patients with varicocele have higher levels of FSH and LH and smaller testes compared to controls with and without varicoceles. Supported by: Grant from the University of Sa˜o Paulo Medical School.
FERTILITY & STERILITY威
P-121 Pre-ART strict sperm morphology assessment and IVF and ICSI outcomes. T. G. Schuster, G. D. Smith, G. M. Christman, Y. R. Smith, D. I. Lebovic, D. A. Ohl. Univ of Michigan, Ann Arbor, MI. Objective: To assess if pre-procedure strict morphology assessment is predictive of in-vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) outcome. Design: Retrospective review of semen analyses, fertilization rates, embryo quality and pregnancy outcomes in conventional IVF and ICSI. Materials/Methods: During preparation for assisted reproductive technology (ART), the male partner’s semen was evaluated using the strict morphology (SM) system of Kruger. Standard IVF protocol with superovulation and ultrasound-guided retrieval of oocytes was used in female partners. Approximately 4 – 6 hours after retrieval, oocytes were incubated with 100,000 sperm/ml or directly injected with isolated sperm with ICSI. Oocytes were examined the next day for evidence of fertilization and followed until transfer at 72 hours post-oocyte collection. Embryos were graded for blastomere number and given a score of A–D based on degree of fragmentation and blastomere irregularities. The best quality embryos (usually 3) were transferred on day 3 of incubation. Pre-ART SM values were compared to rates of fertilization using simple regression. Morphology values were compared between nominal groups based on embryo quality, pregnancy, and miscarriage with unpaired t-tests. Results: Seventy-two cases of conventional IVF and 41 cases of ICSI were analyzed. Average number of oocytes inseminated was 12 with a fertilization rate of 82%. Average number of embryos transferred was 3 (range 0 – 6). Thirty-five patients had positive pregnancy tests (31%) of which 24 had live births (21%). Semen analyses for the ICSI group had lower sperm motility (32 ⫾ 16% vs. 50 ⫾ 11%, p ⬍ 0.0001), lower total motile sperm count (69 ⫾ 103 vs. 216 ⫾ 235 million, p ⫽ 0.007), and lower normal morphology (8.4 ⫾ 6.1% vs. 12.2 ⫾ 5.6%, p ⫽ 0.001) than the IVF group. There was no correlation between pre-ART SM values and any parameter measured, in either the IVF or ICSI groups. Specifically, SM had no effect on oocyte fertilization rate, grade of embryo development, pregnancy rate, or miscarriage rate (p 0.05 for all parameters). The results were not different in the IVF and ICSI subgroups. Conclusions: While selection bias of poorer sperm specimens for ICSI may be present, pre-operative SM assessment, within our treatment groups, did not predict fertilization rates, embryo development, or pregnancy outcome for either IVF or ICSI. Supported by: Internal funding.
P-122 Antisperm antibodies in spinal cord injury patients. R. L. Pagani, D. M. Nudell, D. J. Cozzolino, D. J. Lamb, L. I. Lipshultz. Baylor Coll of Medicine, Houston, TX. Objective: Several etiologies for decreased semen quality in spinal cord injury (SCI) patients have been proposed. Among them, some studies have shown increased levels of antisperm antibodies (ASA) in this patient population. We compared the incidence of ASA in the seminal fluid from a large group of SCI patients to that from men with unexplained infertility and to normal sperm donors. Design: A retrospective clinical study of spinal cord injury patients tested for antisperm antibodies. Materials/Methods: 86 SCI patients, 100 infertile patients (excluding those following vasectomy reversal or known infection) and 14 sperm donors were tested. Seminal fluid ASA were detected using Immunobead Assay. The test was considered positive when greater than 20% of the motile sperm present were bound to immunoglobulins. Results: Only 3 out of 86 (3.4%) SCI patients were positive for ASA in their seminal fluid. In the infertile population, 6% had ASA while, as expected, the sperm donors were all negative for ASA. Conclusions: The etiology of poor semen quality in SCI patients remains unclear and may be multifactorial. We have recently shown, for example, that SCI patients have a significant reduction in sperm penetrating ability. Although some SCI patients in this study demonstrated ASA, there appeared to be no difference when compared to patients with unexplained infertility. Supported by: Supported in part by NIH HD30155 to LIL and DJL.
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P-120 Testicular volume and hormone levels in varicocele patients (fertile and infertile) and fertile controls without varicoceles. F. F. Pasqualotto, J. Hallak, L. M. Rossi-Ferragut, A. M. Lucon, P. M. Go´es, S. Arap. Univ of Sa˜o Paulo Medical Sch, Sa˜o Paulo, Brazil; Fertility Ctr for Human Reproduction, Sa˜o Paulo, Brazil. Objective: Testicular alterations associated with the varicocele are well documented. Also, altered testicular steroidogenesis as a mechanism by which the varicocele exerts its deleterious effects on spermatogenesis remains not clear. We evaluated the levels of luteinizing hormone (LH), follicle-stimulating hormone (FSH), total testosterone, and testicular volume in fertile and infertile men with varicoceles, and fertile controls without varicoceles. Design: Prospective study. Materials/Methods: The records of 27 infertile patients with varicocele and 70 controls with and without varicoceles were assessed from January 1999 to November 2000. In all patients serum hormone levels as well as testicular volume assessed with calipers were evaluated. Patient’s were divided into three groups: group A (fertile men with varicocele; n ⫽ 21), group B (infertile men with varicocele; n ⫽ 27), and group C (fertile men without varicoceles; n ⫽ 49). Results: No differences were seen in the mean age and testosterone levels across the three groups (P ⬎ 0.05). FSH levels were significantly higher in group B (7.8 ⫾ 7.6) compared to groups A (2.95 ⫾ 1.38) and C (3.43 ⫾ 2) (P ⬍ 0.05). Also LH levels were higher in group B (6.8 ⫾ 5.4) compared to patients in groups A (3.2 ⫾ 1.9) and C (3.4 ⫾ 1.74) (P ⬍ 0.05). The right testicle was smaller in group B (18.9 ⫾ 9) compared to group A (24.4 ⫾ 8.6) and C (25.6 ⫾ 8.5) (P ⬍ 0.05). Also, the left testicle was smaller in group B (16.6 ⫾ 8.9) compared to groups A (21. ⫾ 7.8) and C (23.4 ⫾ 8.3) (P ⬍ 0.05). Conclusions: Infertile patients with varicocele have higher levels of FSH and LH and smaller testes compared to controls with and without varicoceles. Supported by: Grant from the University of Sa˜o Paulo Medical School.
FERTILITY & STERILITY威
P-121 Pre-ART strict sperm morphology assessment and IVF and ICSI outcomes. T. G. Schuster, G. D. Smith, G. M. Christman, Y. R. Smith, D. I. Lebovic, D. A. Ohl. Univ of Michigan, Ann Arbor, MI. Objective: To assess if pre-procedure strict morphology assessment is predictive of in-vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) outcome. Design: Retrospective review of semen analyses, fertilization rates, embryo quality and pregnancy outcomes in conventional IVF and ICSI. Materials/Methods: During preparation for assisted reproductive technology (ART), the male partner’s semen was evaluated using the strict morphology (SM) system of Kruger. Standard IVF protocol with superovulation and ultrasound-guided retrieval of oocytes was used in female partners. Approximately 4 – 6 hours after retrieval, oocytes were incubated with 100,000 sperm/ml or directly injected with isolated sperm with ICSI. Oocytes were examined the next day for evidence of fertilization and followed until transfer at 72 hours post-oocyte collection. Embryos were graded for blastomere number and given a score of A–D based on degree of fragmentation and blastomere irregularities. The best quality embryos (usually 3) were transferred on day 3 of incubation. Pre-ART SM values were compared to rates of fertilization using simple regression. Morphology values were compared between nominal groups based on embryo quality, pregnancy, and miscarriage with unpaired t-tests. Results: Seventy-two cases of conventional IVF and 41 cases of ICSI were analyzed. Average number of oocytes inseminated was 12 with a fertilization rate of 82%. Average number of embryos transferred was 3 (range 0 – 6). Thirty-five patients had positive pregnancy tests (31%) of which 24 had live births (21%). Semen analyses for the ICSI group had lower sperm motility (32 ⫾ 16% vs. 50 ⫾ 11%, p ⬍ 0.0001), lower total motile sperm count (69 ⫾ 103 vs. 216 ⫾ 235 million, p ⫽ 0.007), and lower normal morphology (8.4 ⫾ 6.1% vs. 12.2 ⫾ 5.6%, p ⫽ 0.001) than the IVF group. There was no correlation between pre-ART SM values and any parameter measured, in either the IVF or ICSI groups. Specifically, SM had no effect on oocyte fertilization rate, grade of embryo development, pregnancy rate, or miscarriage rate (p 0.05 for all parameters). The results were not different in the IVF and ICSI subgroups. Conclusions: While selection bias of poorer sperm specimens for ICSI may be present, pre-operative SM assessment, within our treatment groups, did not predict fertilization rates, embryo development, or pregnancy outcome for either IVF or ICSI. Supported by: Internal funding.
P-122 Antisperm antibodies in spinal cord injury patients. R. L. Pagani, D. M. Nudell, D. J. Cozzolino, D. J. Lamb, L. I. Lipshultz. Baylor Coll of Medicine, Houston, TX. Objective: Several etiologies for decreased semen quality in spinal cord injury (SCI) patients have been proposed. Among them, some studies have shown increased levels of antisperm antibodies (ASA) in this patient population. We compared the incidence of ASA in the seminal fluid from a large group of SCI patients to that from men with unexplained infertility and to normal sperm donors. Design: A retrospective clinical study of spinal cord injury patients tested for antisperm antibodies. Materials/Methods: 86 SCI patients, 100 infertile patients (excluding those following vasectomy reversal or known infection) and 14 sperm donors were tested. Seminal fluid ASA were detected using Immunobead Assay. The test was considered positive when greater than 20% of the motile sperm present were bound to immunoglobulins. Results: Only 3 out of 86 (3.4%) SCI patients were positive for ASA in their seminal fluid. In the infertile population, 6% had ASA while, as expected, the sperm donors were all negative for ASA. Conclusions: The etiology of poor semen quality in SCI patients remains unclear and may be multifactorial. We have recently shown, for example, that SCI patients have a significant reduction in sperm penetrating ability. Although some SCI patients in this study demonstrated ASA, there appeared to be no difference when compared to patients with unexplained infertility. Supported by: Supported in part by NIH HD30155 to LIL and DJL.
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