- Email: [email protected]
P-994
CONCLUSION: Since the 90% layer of the density gradient preparation is known to produce the best sperm quality, these data may appear paradoxical. However, previous data has indicated that the P1/P2 ratio of fertile sperm donors is lower than that of the general population, and similar to the ratio found in the 90% layer. Therefore, this study may indicate a potentially important difference in the processing of protamines and histones in populations of sperm within a semen sample. Supported by: None
SPERM PREPARATION P-992 SEMINAL VISCOSITY: IT’S EFFECTS ON ROUTINE SEMEN ANALYSIS OUTCOME. P. M. Zavos, S. Koundouros, J. R. CorreaPe´rez, K. Kaskar, P. N. Zavos. Andrology Institute of America, Lexington, KY; Genesis Infertility Clinic, Nicosia, Cyprus; Infertility & IVF Center, Town & Country, MO; Kentucky Center for Reproductive Medicine & IVF, Lexington, KY. OBJECTIVE: Seminal hyperviscosity was shown to be associated with male infertility. However, specimens with higher viscosity and liquefaction difficulties may present additional challenges to those attempting to measure their seminal and spermatogenic profile during the performance of routine semen analysis (RSA). The aim of the study was to investigate the accuracy and possible variations that could exist during the performance of RSA between liquefied and non-liquefied seminal specimens. DESIGN: Prospective controlled study. MATERIALS AND METHODS: Seminal specimens (n⫽66) obtained from sixty-six patients with semen viscosity disorders attending the Andrology Institute of America between November 2004 and April 2005 were included in this study. Semen samples were analyzed by the same technician under blind conditions, using a Makler counting chamber within one hour of collection. Sperm concentration and percentage of motile sperm were determined according to WHO criteria. Five repeated measurements were made from each specimen prior to liquefaction and then liquefied using the Viscolytic System (VLS), a commercially available product (ZDL, Inc., Lexington, KY USA). Five repeated measurements were carried out again following liquefaction. The liquefied samples were used as their own controls (non-liquefied) and the results were compiled and compared between the two treatments. RESULTS: Mean seminal profiles (⫾SD) of the non-liquefied and liquefied specimens are shown in Table 1.The mean sperm count was lower (p⬍0.05) in the non-liquefied group of specimens than the liquefied specimens with significantly higher degrees of variation as evidenced by the standard deviations (14.7 vs 5.0 respectively). The overall seminal measurements continued to remain consistently lower in the non-liquefied than the liquefied specimens. Furthermore, statistically significant differences (p⬍0.01) were noted in the TFSF values (an inclusive term) between the non-liquefied and liquefied specimens (25.9⫾12.6 vs 47.2⫾3.5x106), respectively. Interestingly, the non-liquified specimens were clinically classified as subfertile when compared to those following liquefaction.
P-993 SUCCESSFUL MODIFIED SPERM WASHES IN SEVERE OLIGOASTHENOZOOSPERMIC MALES INFECTED WITH HUMAN IMMUNODEFICENCY AND HEPATITIS C VIRUSES. N. Garrido, J. A. Martinez-Conejero, J. Remohı´, A. Pellicer, M. Meseguer. Instituto Valenciano de Infertilidad, Universidad de Valencia, Valencia, Spain; Fundacion IVI, Universidad de Valencia, Valencia, Spain. OBJECTIVE: HIV/HCV positive males can father children safely with sperm wash to remove sperm viral particles, nested polymerase chain reaction (nPCR) techniques to test viral presence and followed by intracytoplasmic sperm injection (ICSI) procedures to conceive. Nevertheless, a small percentage of these males present severely impaired spermatogenesis, and current sperm wash protocols make impossible to recover motile spermatozoa after the procedure. We hypothesized about the possibility of adapting the washing protocols for these samples. Our aim was to determine if less rigorous sperm washing methods still yield negative HIV and HCV specimens, tested by nested PCR, to be employed in assisted reproduction treatments (ART). DESIGN: Prospective study. MATERIALS AND METHODS: Eight semen samples were obtained from 7 males with HIV and/or HCV, attending Instituto Universitario IVI for ART procedures, where the total motile sperm count was below 2 million. All the samples were processed in an exclusive laboratory, following strict safety guidelines of viral handling, and semen parameters were analyzed (World Health Organization criteria). Ejaculates were diluted 1:1 (vol:vol) with Human Tubal Fluid Medium. Then they were centrifuged at 400g 10 min, and the supernatants were carefully discarded. This procedure was repeated twice, and the resulting cells were resuspended into a final volume of 0.5-1 ml. One half was immediately submerged into liquid nitrogen for PCR determinations, and the other half was kept frozen after the negative PCR result was obtained. Nucleic acids were extracted using the NuclisenseR method following manufacturer instructions. Extracted samples were used to detect genes from the gag and pol region for HIV and NCR region for HCV by nested DNA amplification. For HIV also RNA reverse transcription before PCR was conducted. Controls for extraction and amplification steps were added. In all the samples a consistent result was obtained. RESULTS: Viral acquisition was, for the HIV infected males (6 of them also co-infected with HCV, 64.3%), 3 ex-addictions to parenteral drugs (50%), 3 heterosexual transmissions (50%). The mean duration of the HIV disease was 9.3 years (range 1-20). Regarding treatments, 2 (28.6%) of them were treatment-free, while the remaining were receiving multiple antiretroviral drug combinations. HIV blood load of the unique patient with detectable levels was 699 IU/ml, while 5 patients (83.3%) showed negative viral blood load. Peripheral blood CD4 levels median was 223.6 ranging from (99-525). Sperm concentrations ranged from 0.1 to 1.75 millions of motile sperm/ml before the sperm wash, and all of them were lower than 0.5 after the sperm wash. We found that no ejaculate treated in this way resulted positive after the washing procedure neither for HIV RNA, HIV DNA, nor HCV RNA, when needed, therefore concluding its usefulness in these sperm samples with severe oligoasthenozoospermia. CONCLUSION: Given the negative results in viral detection after modified sperm wash, although other semen components remain within the sample, we can conclude that these males should be offered ART. This clinical procedure would be only possible if the molecular confirmation of viral absence after modified sperm washes is successful and motile sperm are found, regardless sperm quality. If these results are maintained after increasing the sample size or they are just occasional, remain to be elucidated in longer series. Supported by: None
P-994 CONCLUSION: Seminal hyperviscosity during the performance of RSA can interfere with the proper sampling process during the performance of a RSA. These variations in the sampling process have shown to produce inaccurate data with large variations that could result in the misdiagnosis of the male and subsequently affect his ability to receive the proper treatment during the couples infertility work up. It is therefore recommended that specimens should be liquefied prior to the performance of RSA. Supported by: None.
S502
Abstracts
COMPARATIVE STUDY OF VARIOUS SEMEN PREPARATION TECHNIQUES IN IUI AND THEIR EFFECT ON PREGNANCY RATE. K. D. Nayar, P. Sehgal, A. Tiwari. Akanksha IVF Centre, Mata Chanan Devi Hospital, New Delhi, India. OBJECTIVE: To evaluate the efficacy of different semen preparation methods & their impact on pregnancy rate in IUI cycles. DESIGN: Retrospective study from Jan 2005 to September 2005.
Vol. 86, Suppl 2, September 2006
MATERIALS AND METHODS: A total of 166 couples were allocated to IUI treatment cycles with three different semen preparation techniques i.e. simple washing , swim up and density gradient.. The inclusion criteria was set according to age less than 38 years, having normal fallopian tubes and ovaries. The semen criteria were set according to WHO standards & patients with mild male factors were also included in the study. The ejaculate of male partner must be analysed twice before insemination.All patients underwent ovarian hyperstimulation protocol using clomiphene alone or clomiphene with gonadotrophins. The swim up technique allows motile sperms to swim out of the ejaculate into the salt solution, therby seperating them from seminal plasma, immotile sperms, extraneuos cells and debris.However since seperation is based on motility alone, the swim-up technique does not remove morphologically abnormal sperms or chromatin damaged sperms. WHO manual suggest Earls Balaced salt solution or commercially available sperm washing media for swimup. However since ionic balance, pH and osmolality are crucial in preventing sperm hyperactivation, an optimised medium was used to maintain the sperm in a fertile state. In density gradient technique liquefied semen layer is layered on a discontinuous gradient consisting of two or more layers of different densities.During centrifugation, motile sperms move through the layer of colloid, thus becoming seperated from dead or immotile sperm, cellular debris and seminal plasma. Bacteria viruses and macropahages also remain in the upper layers.Careful removal of gradient layers after centrifugation allows sperm pellet to be recovered without contamination. RESULTS: The study group were divided into 80 in swimup. 56 in density gradient,30 patients in simple washing group.The mean age of females in swimup 32.08⫾ 2.3, density gradient 32.23⫾3 years and simple washing was 31.80⫾2 years respectively. The total number of IUI cycles were 116 in swim-up, 75 in density gradient and 35 in simple washing group. There were 21 clinical pregnancies in swim-up, 6 in density gradient, 2 in simple washing. There was one spontaneous abortion each in swimup & density group. The pre & post wash semen charateristics with number of total motile sperms are illustrated ,presented as mean ⫾ SD.
MATERIALS AND METHODS: Infertile couples who were undergoing treatment with superovulation and IUI during 2004 to 2006 were included in the study (n⫽120). Indications for superovulation and IUI were idiopathic infertility (84%) or male factor infertility (16%). Women were superovulated with exogenous gonadotropins or clomiphene. All women had a hysterosalpingogram to document tubal patency. Exclusion criteria included severe oligospermia and tubal factor infertility. In gonadotropin cycles, hCG was given when there were at least two follicles ⱖ 18mm and insemination was performed 24-34 hours later. In clomid cycles, insemination was done when there were one or more follicles ⱖ20mm in size. If there was no LH surge by ovulation predictor kit, 10,000 units of human chorionic gonadotropin was given prior to insemination. In the IUI cycles performed before July, 2005 (group I), total pellet (all particulates of semen) recovered by 2 centrifugation- 2 wash treatment method was used for sperm preparation. After July 2005, motile sperm recovered by standard gradient column (Isolate, Irvine Scientific) with 1 wash treatment was used for all the IUI cycles (group II). Hepes-HTF supplemented with 5% HSA was used as wash medium in both groups. Total sperm recovered in each method was inseminated in a fixed volume (0.5 ml) by a single physician using same catheter. Patient population managed by a single physician was included in the study to minimize clinical variances. We compared the pregnancy rates between group I and group II. Statistical analysis was done by student t test and Chi-square analysis and the results are reported as mean ⫾ SE. RESULTS: The clinical characteristics of the women in both groups were similar with regards to age and duration of infertility. The pre wash sperm count and motility were identical (count 55 ⫾5 and 50⫾1 106/ml (⫾ SE) and motility 50 ⫾ 1% and 55⫾2% in the respective groups). The post wash motility was significantly (P ⬍ 0.001) higher in group II (79 ⫾ 2%) compared to group I (56 ⫾ 2%). The total number of motile sperm used in the insemination in group I and group II were similar (30⫾2 and 25⫾2 x106/ ml). The pregnancy rate per insemination cycle was 10 % for group I and 20% for group II. (P ⬍ 0.01). Cumulative pregnancy rate per patient was 19% in group I and 38% for group II (P ⬍ 0.03) CONCLUSION: These results indicate that 1) sperm preparation using gradient column gives better success rate with IUI than the conventional pellet wash method, 2) the higher pregnancy rate is apparently due to the higher post wash sperm motility seen with the gradient method, and 3) lower pregnancy rate with the pellet wash method could be due to lower sperm motility, and the presence of higher number of non-spermatozoal particles (round cells, epithelial cells, other debris) in the post wash sample. Supported by: None
P-996 RELATIONSHIP AMONG HBA, SPERM MOTILITY, KRUGER MORPHOLOGY WITH FERTILIZATION RATE AND PREGNANCY OUTCOME IN PATIENTS TREATED WITH IVF OR ICSI. T. Wu, D. Dasig, M. Dangcil, W. Shen. Kaiser Permanente Center for Reproductive Health, Fremont, CA. CONCLUSION: The swimup group has higher pregnancy rates and total cost of IUI is also less compared to gradient. It is a good standard technique, widely used for normal semen samples in IUI.The gradient method is useful for poor quality samples. Simple washing is approximately equal in cost to swim-up and has a role in extremely low sperm densities. Choosing an individualised Semen preparation technique gives the couple a modrate chance of pregnancy with IUI. Supported by: None P-995 SPERM WASH BY GRADIENT COLUMN TECHNIQUE IMPROVES IUI SUCCESS RATE. A. Hossain, C. Osuamkpe, D. Castle, M. Nagamani. Univ. of Texas Medical Branch, Galveston, TX. OBJECTIVE: Despite considerable development in infertility treatment, pregnancy by intrauterine insemination (IUI) remains low compared to that of in vitro fertilization (IVF). However, superovulation and IUI remains a cost-effective treatment for infertile couples. The purpose of our study was to investigate the effect of two different methods of sperm preparation on IUI success rate. DESIGN: Retrospective study of IUI cycles in a tertiary care academic institution.
FERTILITY & STERILITY威
OBJECTIVE: Semen analysis with Kruger morphology is widely used to assess male fertility potential. More recently hyaluronan-binding protein has been established as a biomarker for mature, functional sperm. Consequently the HBA assay (Sperm-Hyaluronan Binding assay) has been developed to differentiate functional integrity and fertilization potential of sperm cells. We investigated the relationships among HBA score, sperm motility, and Kruger morphology with fertilization rate in patients undergoing in vitro fertilization (IVF) with standard insemination or intracytoplasmic sperm injection (ICSI). DESIGN: Prospective comparative data analysis. MATERIALS AND METHODS: The Andrology and Embryology laboratories at the Kaiser Permanente Center for Reproductive Health performed HBA assay in conjunction with a standard semen analysis with Kruger morphology for patients undergoing IVF. The decision to perform standard IVF or ICSI was made by the primary physicians and the patients. Sperm HBA score, sperm motility and Kruger morphology before and after density gradient preparation were compared and correlated with fertilization rate of oocytes inseminated with standard insemination or ICSI. Linear regression analysis was performed. RESULTS: We studied prospectively 31 cases in which 20 cases were treated with ICSI, 7 cases were treated with IVF, and 4 cases were performed split IVF/ICSI. In the ICSI treatment group, the fertilization rate had no correlation with sperm motility, Kruger morphology, or HBA score (r ⫽ -0.15,
S503
SPERM PREPARATION P-992 SEMINAL VISCOSITY: IT’S EFFECTS ON ROUTINE SEMEN ANALYSIS OUTCOME. P. M. Zavos, S. Koundouros, J. R. CorreaPe´rez, K. Kaskar, P. N. Zavos. Andrology Institute of America, Lexington, KY; Genesis Infertility Clinic, Nicosia, Cyprus; Infertility & IVF Center, Town & Country, MO; Kentucky Center for Reproductive Medicine & IVF, Lexington, KY. OBJECTIVE: Seminal hyperviscosity was shown to be associated with male infertility. However, specimens with higher viscosity and liquefaction difficulties may present additional challenges to those attempting to measure their seminal and spermatogenic profile during the performance of routine semen analysis (RSA). The aim of the study was to investigate the accuracy and possible variations that could exist during the performance of RSA between liquefied and non-liquefied seminal specimens. DESIGN: Prospective controlled study. MATERIALS AND METHODS: Seminal specimens (n⫽66) obtained from sixty-six patients with semen viscosity disorders attending the Andrology Institute of America between November 2004 and April 2005 were included in this study. Semen samples were analyzed by the same technician under blind conditions, using a Makler counting chamber within one hour of collection. Sperm concentration and percentage of motile sperm were determined according to WHO criteria. Five repeated measurements were made from each specimen prior to liquefaction and then liquefied using the Viscolytic System (VLS), a commercially available product (ZDL, Inc., Lexington, KY USA). Five repeated measurements were carried out again following liquefaction. The liquefied samples were used as their own controls (non-liquefied) and the results were compiled and compared between the two treatments. RESULTS: Mean seminal profiles (⫾SD) of the non-liquefied and liquefied specimens are shown in Table 1.The mean sperm count was lower (p⬍0.05) in the non-liquefied group of specimens than the liquefied specimens with significantly higher degrees of variation as evidenced by the standard deviations (14.7 vs 5.0 respectively). The overall seminal measurements continued to remain consistently lower in the non-liquefied than the liquefied specimens. Furthermore, statistically significant differences (p⬍0.01) were noted in the TFSF values (an inclusive term) between the non-liquefied and liquefied specimens (25.9⫾12.6 vs 47.2⫾3.5x106), respectively. Interestingly, the non-liquified specimens were clinically classified as subfertile when compared to those following liquefaction.
P-993 SUCCESSFUL MODIFIED SPERM WASHES IN SEVERE OLIGOASTHENOZOOSPERMIC MALES INFECTED WITH HUMAN IMMUNODEFICENCY AND HEPATITIS C VIRUSES. N. Garrido, J. A. Martinez-Conejero, J. Remohı´, A. Pellicer, M. Meseguer. Instituto Valenciano de Infertilidad, Universidad de Valencia, Valencia, Spain; Fundacion IVI, Universidad de Valencia, Valencia, Spain. OBJECTIVE: HIV/HCV positive males can father children safely with sperm wash to remove sperm viral particles, nested polymerase chain reaction (nPCR) techniques to test viral presence and followed by intracytoplasmic sperm injection (ICSI) procedures to conceive. Nevertheless, a small percentage of these males present severely impaired spermatogenesis, and current sperm wash protocols make impossible to recover motile spermatozoa after the procedure. We hypothesized about the possibility of adapting the washing protocols for these samples. Our aim was to determine if less rigorous sperm washing methods still yield negative HIV and HCV specimens, tested by nested PCR, to be employed in assisted reproduction treatments (ART). DESIGN: Prospective study. MATERIALS AND METHODS: Eight semen samples were obtained from 7 males with HIV and/or HCV, attending Instituto Universitario IVI for ART procedures, where the total motile sperm count was below 2 million. All the samples were processed in an exclusive laboratory, following strict safety guidelines of viral handling, and semen parameters were analyzed (World Health Organization criteria). Ejaculates were diluted 1:1 (vol:vol) with Human Tubal Fluid Medium. Then they were centrifuged at 400g 10 min, and the supernatants were carefully discarded. This procedure was repeated twice, and the resulting cells were resuspended into a final volume of 0.5-1 ml. One half was immediately submerged into liquid nitrogen for PCR determinations, and the other half was kept frozen after the negative PCR result was obtained. Nucleic acids were extracted using the NuclisenseR method following manufacturer instructions. Extracted samples were used to detect genes from the gag and pol region for HIV and NCR region for HCV by nested DNA amplification. For HIV also RNA reverse transcription before PCR was conducted. Controls for extraction and amplification steps were added. In all the samples a consistent result was obtained. RESULTS: Viral acquisition was, for the HIV infected males (6 of them also co-infected with HCV, 64.3%), 3 ex-addictions to parenteral drugs (50%), 3 heterosexual transmissions (50%). The mean duration of the HIV disease was 9.3 years (range 1-20). Regarding treatments, 2 (28.6%) of them were treatment-free, while the remaining were receiving multiple antiretroviral drug combinations. HIV blood load of the unique patient with detectable levels was 699 IU/ml, while 5 patients (83.3%) showed negative viral blood load. Peripheral blood CD4 levels median was 223.6 ranging from (99-525). Sperm concentrations ranged from 0.1 to 1.75 millions of motile sperm/ml before the sperm wash, and all of them were lower than 0.5 after the sperm wash. We found that no ejaculate treated in this way resulted positive after the washing procedure neither for HIV RNA, HIV DNA, nor HCV RNA, when needed, therefore concluding its usefulness in these sperm samples with severe oligoasthenozoospermia. CONCLUSION: Given the negative results in viral detection after modified sperm wash, although other semen components remain within the sample, we can conclude that these males should be offered ART. This clinical procedure would be only possible if the molecular confirmation of viral absence after modified sperm washes is successful and motile sperm are found, regardless sperm quality. If these results are maintained after increasing the sample size or they are just occasional, remain to be elucidated in longer series. Supported by: None
P-994 CONCLUSION: Seminal hyperviscosity during the performance of RSA can interfere with the proper sampling process during the performance of a RSA. These variations in the sampling process have shown to produce inaccurate data with large variations that could result in the misdiagnosis of the male and subsequently affect his ability to receive the proper treatment during the couples infertility work up. It is therefore recommended that specimens should be liquefied prior to the performance of RSA. Supported by: None.
S502
Abstracts
COMPARATIVE STUDY OF VARIOUS SEMEN PREPARATION TECHNIQUES IN IUI AND THEIR EFFECT ON PREGNANCY RATE. K. D. Nayar, P. Sehgal, A. Tiwari. Akanksha IVF Centre, Mata Chanan Devi Hospital, New Delhi, India. OBJECTIVE: To evaluate the efficacy of different semen preparation methods & their impact on pregnancy rate in IUI cycles. DESIGN: Retrospective study from Jan 2005 to September 2005.
Vol. 86, Suppl 2, September 2006
MATERIALS AND METHODS: A total of 166 couples were allocated to IUI treatment cycles with three different semen preparation techniques i.e. simple washing , swim up and density gradient.. The inclusion criteria was set according to age less than 38 years, having normal fallopian tubes and ovaries. The semen criteria were set according to WHO standards & patients with mild male factors were also included in the study. The ejaculate of male partner must be analysed twice before insemination.All patients underwent ovarian hyperstimulation protocol using clomiphene alone or clomiphene with gonadotrophins. The swim up technique allows motile sperms to swim out of the ejaculate into the salt solution, therby seperating them from seminal plasma, immotile sperms, extraneuos cells and debris.However since seperation is based on motility alone, the swim-up technique does not remove morphologically abnormal sperms or chromatin damaged sperms. WHO manual suggest Earls Balaced salt solution or commercially available sperm washing media for swimup. However since ionic balance, pH and osmolality are crucial in preventing sperm hyperactivation, an optimised medium was used to maintain the sperm in a fertile state. In density gradient technique liquefied semen layer is layered on a discontinuous gradient consisting of two or more layers of different densities.During centrifugation, motile sperms move through the layer of colloid, thus becoming seperated from dead or immotile sperm, cellular debris and seminal plasma. Bacteria viruses and macropahages also remain in the upper layers.Careful removal of gradient layers after centrifugation allows sperm pellet to be recovered without contamination. RESULTS: The study group were divided into 80 in swimup. 56 in density gradient,30 patients in simple washing group.The mean age of females in swimup 32.08⫾ 2.3, density gradient 32.23⫾3 years and simple washing was 31.80⫾2 years respectively. The total number of IUI cycles were 116 in swim-up, 75 in density gradient and 35 in simple washing group. There were 21 clinical pregnancies in swim-up, 6 in density gradient, 2 in simple washing. There was one spontaneous abortion each in swimup & density group. The pre & post wash semen charateristics with number of total motile sperms are illustrated ,presented as mean ⫾ SD.
MATERIALS AND METHODS: Infertile couples who were undergoing treatment with superovulation and IUI during 2004 to 2006 were included in the study (n⫽120). Indications for superovulation and IUI were idiopathic infertility (84%) or male factor infertility (16%). Women were superovulated with exogenous gonadotropins or clomiphene. All women had a hysterosalpingogram to document tubal patency. Exclusion criteria included severe oligospermia and tubal factor infertility. In gonadotropin cycles, hCG was given when there were at least two follicles ⱖ 18mm and insemination was performed 24-34 hours later. In clomid cycles, insemination was done when there were one or more follicles ⱖ20mm in size. If there was no LH surge by ovulation predictor kit, 10,000 units of human chorionic gonadotropin was given prior to insemination. In the IUI cycles performed before July, 2005 (group I), total pellet (all particulates of semen) recovered by 2 centrifugation- 2 wash treatment method was used for sperm preparation. After July 2005, motile sperm recovered by standard gradient column (Isolate, Irvine Scientific) with 1 wash treatment was used for all the IUI cycles (group II). Hepes-HTF supplemented with 5% HSA was used as wash medium in both groups. Total sperm recovered in each method was inseminated in a fixed volume (0.5 ml) by a single physician using same catheter. Patient population managed by a single physician was included in the study to minimize clinical variances. We compared the pregnancy rates between group I and group II. Statistical analysis was done by student t test and Chi-square analysis and the results are reported as mean ⫾ SE. RESULTS: The clinical characteristics of the women in both groups were similar with regards to age and duration of infertility. The pre wash sperm count and motility were identical (count 55 ⫾5 and 50⫾1 106/ml (⫾ SE) and motility 50 ⫾ 1% and 55⫾2% in the respective groups). The post wash motility was significantly (P ⬍ 0.001) higher in group II (79 ⫾ 2%) compared to group I (56 ⫾ 2%). The total number of motile sperm used in the insemination in group I and group II were similar (30⫾2 and 25⫾2 x106/ ml). The pregnancy rate per insemination cycle was 10 % for group I and 20% for group II. (P ⬍ 0.01). Cumulative pregnancy rate per patient was 19% in group I and 38% for group II (P ⬍ 0.03) CONCLUSION: These results indicate that 1) sperm preparation using gradient column gives better success rate with IUI than the conventional pellet wash method, 2) the higher pregnancy rate is apparently due to the higher post wash sperm motility seen with the gradient method, and 3) lower pregnancy rate with the pellet wash method could be due to lower sperm motility, and the presence of higher number of non-spermatozoal particles (round cells, epithelial cells, other debris) in the post wash sample. Supported by: None
P-996 RELATIONSHIP AMONG HBA, SPERM MOTILITY, KRUGER MORPHOLOGY WITH FERTILIZATION RATE AND PREGNANCY OUTCOME IN PATIENTS TREATED WITH IVF OR ICSI. T. Wu, D. Dasig, M. Dangcil, W. Shen. Kaiser Permanente Center for Reproductive Health, Fremont, CA. CONCLUSION: The swimup group has higher pregnancy rates and total cost of IUI is also less compared to gradient. It is a good standard technique, widely used for normal semen samples in IUI.The gradient method is useful for poor quality samples. Simple washing is approximately equal in cost to swim-up and has a role in extremely low sperm densities. Choosing an individualised Semen preparation technique gives the couple a modrate chance of pregnancy with IUI. Supported by: None P-995 SPERM WASH BY GRADIENT COLUMN TECHNIQUE IMPROVES IUI SUCCESS RATE. A. Hossain, C. Osuamkpe, D. Castle, M. Nagamani. Univ. of Texas Medical Branch, Galveston, TX. OBJECTIVE: Despite considerable development in infertility treatment, pregnancy by intrauterine insemination (IUI) remains low compared to that of in vitro fertilization (IVF). However, superovulation and IUI remains a cost-effective treatment for infertile couples. The purpose of our study was to investigate the effect of two different methods of sperm preparation on IUI success rate. DESIGN: Retrospective study of IUI cycles in a tertiary care academic institution.
FERTILITY & STERILITY威
OBJECTIVE: Semen analysis with Kruger morphology is widely used to assess male fertility potential. More recently hyaluronan-binding protein has been established as a biomarker for mature, functional sperm. Consequently the HBA assay (Sperm-Hyaluronan Binding assay) has been developed to differentiate functional integrity and fertilization potential of sperm cells. We investigated the relationships among HBA score, sperm motility, and Kruger morphology with fertilization rate in patients undergoing in vitro fertilization (IVF) with standard insemination or intracytoplasmic sperm injection (ICSI). DESIGN: Prospective comparative data analysis. MATERIALS AND METHODS: The Andrology and Embryology laboratories at the Kaiser Permanente Center for Reproductive Health performed HBA assay in conjunction with a standard semen analysis with Kruger morphology for patients undergoing IVF. The decision to perform standard IVF or ICSI was made by the primary physicians and the patients. Sperm HBA score, sperm motility and Kruger morphology before and after density gradient preparation were compared and correlated with fertilization rate of oocytes inseminated with standard insemination or ICSI. Linear regression analysis was performed. RESULTS: We studied prospectively 31 cases in which 20 cases were treated with ICSI, 7 cases were treated with IVF, and 4 cases were performed split IVF/ICSI. In the ICSI treatment group, the fertilization rate had no correlation with sperm motility, Kruger morphology, or HBA score (r ⫽ -0.15,
S503