- Email: [email protected]
Association of sperm DNA fragmentation measured by TUNEL with sperm maturation measured by DNA cytometry
DESIGN: All patients undergoing sextant site-specific biopsies, i.e. testis mapping, from 2004-2008 were retrospectively reviewed. MATERIALS AND METHODS: 114 patients with nonobstructive azoospermia were evaluated for testis volume, hormonal parameters, chromosomal abnormalities by karyotype and y-chromosomal deletions and testis pathology. Each patient was further categorized by their major, and if present, minor testis pathology. Testicular histolopathologic subtypes included sertoli-cell only, early maturation arrest, late maturation arrest, hypospermatogenesis, and normal. RESULTS: 42 men (36.8%) with nonobstructive azoospermia were found to have purely SCO histology. An additional 20 men (17.5%) were found to have predominantly SCO pathology in a mixed histologic pattern. Suspected causes for SCO included Klinefelter’s, bilateral intra-abdominal testes, testicular cancer, radiation therapy, chemotherapy, and the use of anabolic steroids. Men with pure SCO pathology were characterized by high FSH values (mean 29.3), small testicular volume (12.9 mL), low percentage of y-chromosome deletions (2.7%) and karyotypic abnormalities (5.6%), and low probability of testicular sperm extraction on conventional testis mapping was (7%). CONCLUSIONS: Sertoli-cell only is a very common histopathologic diagnosis in men with nonobstructive azoospermia. They can be characterized by high FSH and small testicular volume. In our series, few men had chromosomal abnormalities. There were no characteristics in men with SCO that predicted successful retrieval of sperm during testis mapping. Since patients with SCO have a low probability of having sperm extracted on conventional testis biopsy, Urologists may consider extended testicular mapping or micro testicular sperm extraction instead. Supported by: None.
P-623 EARLY VERSUS LATE MATURATION ARREST: INCIDENCE AND CLINICAL SIGNIFICANCE IN TESTICULAR FAILURE. J. W. Weedin, D. M. Fenig, J. N. Mills, L. I. Lipshultz. Urology, Baylor College of Medicine, Houston, TX. OBJECTIVE: There is a paucity of data in the literature characterizing the stages of maturation arrest (MA). We hypothesize that patients with earlier MA have a higher serum FSH reflecting increased sertoli cell/testicular damage and that the probability of mature sperm extraction increases at later stages of MA. DESIGN: All patients undergoing sextant site-specific testis biopsies, i.e. testis mapping, at a single institution from 2004-2008 were retrospectively reviewed. MATERIALS AND METHODS: 114 patients with nonobstructive azoospermia (NOA) and 23 patients with severe oligospermia (density < 10,000 sperm/mL) were categorized by their major, and if present, minor histopathology patterns. These categories consisted of sertoli-cell only (SCO), early maturation arrest (EMA), late maturation arrest (LMA), hypospermatogenesis (H) and normal. EMA was characterized by maturation arrest up to the level of the primary spermatocyte and LMA up to the level of the spermatid. Each group was analyzed with respect to serum hormone concentration, testicular volume, chromosomal abnormalities (karyotype, Y-deletions) and presence of sperm at the time of testicular mapping. Statistical analysis was carried out by paired T-test. RESULTS: 42 (30.9%) patients were identified with MA. 15 patients had concomitant SCO histology. Pathology was organized according to primary and secondary components (Table 1). Mean FSH of pure EMA versus pure LMA was 16.2 vs. 6.8 (p ¼ 0.037). FSH correlated with testis pathology (Table 1) but not with the probability of obtaining sperm at the time of biopsy (p ¼ 0.32). The frequency of successful sperm extraction during testis mapping increased with later stages of MA (SCO/mixed MA ¼ 26.7%, EMA ¼ 14.3%, mixed MA ¼ 37.5%, LMA ¼ 46.1%). Y-chromosome deletions were detected in 35.7% of patients with EMA, 14.2% with LMA, and 2.1% of all other patients. TABLE 1. Histopathologic categories
N FSH % pts with sperm extracted
SCO
SCO/ EMA
SCO/ LMA
EMA
EMA/ LMA
LMA/ EMA
LMA
42 29.3 7.1
8 24.2 25
7 25.3 28.6
7 16.2 14.2
4 11 50
4 5.3 25
13 6.8 46.2
FERTILITY & STERILITYÒ
CONCLUSIONS: MA is a common pattern in NOA that is frequently associated with other histopathology. In patients with pure MA, FSH is higher in patients with EMA and tends to decrease and approach normal with later stages of spermatogenesis present. We also report that a higher percentage of patients with EMA have Y-chromosome microdeletions. Although FSH did not correlate with the probability of sperm extraction, the probability of sperm found on testis mapping was higher with late maturation arrest. Supported by: None.
P-624 HANDHELD INTRA-OPERATIVE DOPPLER FLOW TO LOCALIZE THE PRESENCE OF SPERM DURING TESTIS BIOPSY IN AZOOSPERMIC MEN. S. J. Parekattil, M. S. Cohen, J. W. Vieweg. Urology, University of Florida, Gainesville, FL. OBJECTIVE: Testis biopsy is a diagnostic procedure performed in men with possible non-obstructive azoospermia (NOA). Recent studies have illustrated that it is likely to find active spermatogenesis in areas with better blood supply within the testicle. These studies utilized detailed color Doppler ultrasonography and needle guidance techniques to localize possible areas of spermatogenesis within the testicle. Our goal was to assess the efficacy of simple percutaneous hand-held Doppler blood flow measurements of the testicle at the time of biopsy to localize areas of spermatogenesis. DESIGN: Retrospective case series review and blinded analysis. MATERIALS AND METHODS: Review of 4 patients who underwent testis biopsy (3 to 4 biopsies from each testicle – 14 total biopsies) performed for NOA from Nov’07 to Mar’08. Percutaneous hand-held Doppler (Vascular TechnologyÔ, Nashua, NH) blood flow measurements were taken from 12 different regions of each testicle (only one testicle scanned on each patient – the testicle that was larger was chosen). The surgeon then obtained 3 to 4 random testicular biopsies (blinded to the Doppler analysis) from that testicle. The findings from the biopsies were then compared to the pre-biopsy Doppler mapping to assess if the Doppler readings had any predictive value in detecting spermatogenesis. RESULTS: The Doppler mapping identified three distinct types of blood flow waveforms within the testicle – high, medium and low. The medium amplitude waveform provided the highest predictive value in correctly locating spermatogenesis (sperm found at that biopsy location). The detection of this waveform was 79% accurate in predicting the location of sperm at the time of biopsy (area under the ROC of 0.8). This waveform may correlate to intermediary arteries within the testicle and thus identify areas with good blood supply. CONCLUSIONS: Our preliminary evaluation of hand-held Doppler flow mapping of the testicle at the time of biopsy appears to have some promise in detecting areas of spermatogenesis. Further evaluation and testing will reveal its potential in this role. Supported by: Research equipment support from VTI Technology.
P-625 ASSOCIATION OF SPERM DNA FRAGMENTATION MEASURED BY TUNEL WITH SPERM MATURATION MEASURED BY DNA CYTOMETRY. R. Mahfouz, R. Sharma, A. Agarwal. Center for Reproductive Medicine, Cleveland Clinic, Cleveland, OH; Cleveland Clinic, Cleveland, OH. OBJECTIVE: The terminal uridine deoxynucleotidyl transferase (dUTP) nick end labeling (TUNEL) measures single and double DNA strand breaks. DNA cytometry using propidium iodide (PI) can provide information on sperm chromatin maturation/ condensation status based on the fluorescent intensity of PI. The aim of our study was to evaluate the relationship between sperm DNA fragmentation as measured by TUNEL assay with chromatin maturation/ condensation by DNA cytometry. DESIGN: Prospective study. MATERIALS AND METHODS: 117 semen samples were prepared for TUNEL assay using the Apoptosis Detection Kit (APO-Direct, BD
S317
Bioscience). PI/ RNase were added after the FITC-dUTP reaction for DNA staining according to manufacturer’s instructions. Flowcytometry was used and data acquisition was done for FITC (FL1) as well as PI (FL2-area). RESULTS: The percentage of TUNELþve sperm were positively correlated with percentage of immature (r ¼ 0.36, p ¼ <0.001) and subhaploid sperm (r ¼ 0.53, p ¼ <0.001). TUNELþve spermatozoa showed a negative correlation with percentage of mature (haploid) sperm (r ¼-0.55, p ¼<0.001). % haploid mature sperm were positively correlated with TUNEL-ve percentage (r ¼ 0.7, p <0.001). There was a significant negative interdependence between percentage of haploid sperm and subhaploid sperm (r ¼ 0.76, p <0.001) and between haploid and % of immature sperm (r ¼ 0.44, p <0.001). CONCLUSIONS: Low DNA fragmentation detected by TUNEL assay was associated with sperm maturation/ condensation detected by DNA cytometry. We recommend evaluating semen samples by both assays for better understanding of the underlying pathology i.e. sperm immaturity, apoptosis and/ or oxidative stress associated with abnormal spermatozoa. Supported by: None.
P-626 ENHANCEMENT OF SPERM FERITILIZATION CAPABILITY BY PEROXISOME PROLIFERATION-ACTIVATED RECEPTOR (PPAR) g AGONIST. I. C. Wun, J.-C. Huang. OBGYN, University of Texas Medical School at Houston, Houston, TX. OBJECTIVE: PPAR g is among the nuclear receptors that belong to a superfamily pertaining to energy integration, along with lipid and glucose metabolism. Aquila et al (2006) first reported that PPAR g was expressed in human spermatozoa and enhanced sperm capacitation, acrosome reaction, and motility. This study implements a mouse model to examine the effect of Troglitzaone (a PPAR gamma agonist) upon fertilization. DESIGN: A prospective study. MATERIALS AND METHODS: Spermatozoa were collected from the vas deferens, while the oocytes were obtained from superovulated mice. A Hamilton Thorn semen analyzer was used for the sperm kinetic study. The definition of hyperactivation was a curvilinear velocity greater than 135 m/ s, a linearity of less than 5.0, and lateral head displacement amplitude greater than 6.5 m. A triple-stain technique was used to observe for acrosomal reaction occurrence (Balbot and Chacon, 1981). The insemination concentration was 10,000 motile sperm/ml with Troglitazone from 0.3 mM to 30 mM. The method of least squares was used for kinetic parameter analysis. The acrosomal stain was analyzed by an approach of a generalized estimate equation linked with a logit transformation. The trend of the dose-fertilization relationship was analyzed by the Cochran-Mentel-Haenzel test. Specific Troglitazone concentrations were compared to the control by using the chi-square test.
Fertilization Rate
Item
Sperm Activity Attribute
1 mM vs. 3 mM vs. 10 mM vs. 30 mM vs. Acrosome control control control control Motility Hyperactivation Reaction
Analysis p <0.05 p <0.005 p <0.005 p <0.05
p <0.05 p <0.05
p <0.05
RESULTS: 1. Overall motility increased by Troglitazone treatment. Sperm hyperactivation significantly increased both overall and at 90 minutes Troglitazone treatment. 2. Acrosome reaction increased significantly due to Troglitazone treatment. 3. The fertilization rate showed a marginal trend (p¼0.0527) along with the Troglitazone dosage. Troglitazone at 1 mM or higher significantly increased the fertilization rate. CONCLUSIONS: Troglitazone significantly increases sperm motility, hyperactivation, and acrosome reaction. These activated parameters correspond to the spermatic ability to fertilize eggs. The study demonstrated that expression of functional PPAR g serves to enhance the spermatic ability to fertilize eggs. Aquila, S. et al. J Cell Physiol; 209: 977-86, 2006. Supported by: None.
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Abstracts
P-627 PREVALENCE OF SECONDARY INFERTILITY FOLLOWING VARICOCELECTOMY. P. Salehi, H. Abbasi, K. Nouri-mahdavi, M. H. Nasr Esfahani, M. Zargham, M. Yazdani. Urology, Isfahan University of Medical Sciences, Isfahan, Islamic Republic of Iran; Isfahan University of Medical Sciences, Isfahan, Islamic Republic of Iran; Isfahan Fertility and Infertility Center, Isfahan, Islamic Republic of Iran. OBJECTIVE: To determine the prevalence of secondary infertility following varicocelectomy. DESIGN: This stuty is a cross-sectional research which is performed in isfahan fertility and infertility center on 96 men with a history of primary infertility who had undergone microsurgical varicocelectomy. MATERIALS AND METHODS: Ninety-six men with a history of primary infertility who had fathered children following microsurgical varicocelectomy and wished to have another child were followed after the spouses’ first pregnancy to determine the number of couples who had achieved a second pregnancy. RESULTS: The mean duration follow-up was 3 years (range 1-5). Secondary infertility was observed in 72 (75%), with only 24 couples (25%) achieving a second pregnancy. Secondary infertility was directly related to patient age, duration of prevaricocelectomy infertility, and time to first pregnancy (P<0.05). CONCLUSIONS: It seems that varicocelectomy is associated with unexpectedly high rates of secondary infertility. Supported by: None.
P-628 MICRODISSECTION TESTICULAR SPERM EXTRACTION (MICROTESE) AND INTRACYTOPLASMIC SPERM INJECTION (ICSI) IN THE TREATMENT OF PERSISTENT POSTCHEMOTHERAPY AZOOSPERMIA (PPCA): LESSONS LEARNED OVER 12 YEARS. P. J. Stahl, E. J. Nejat, G. D. Palermo, L. L. Veeck, Z. Rosenwaks, P. N. Schlegel. Department of Urology, Weill Cornell Medical College, New York, NY; Department of Obstetrics & Gynecology, Weill Cornell Medical College, New York, NY; Center for Reproductive Medicine & Infertility, Weill Cornell Medical College, New York, NY. OBJECTIVE: In 2001 we published our initial experience with microTESE and ICSI for the treatment of PPCA. The objective of the present study was to report our updated outcomes in this population to provide prognostically helpful information to physicians and affected couples. DESIGN: We queried our database of men who consecutively underwent microTESE at our institution from 1995-2007 and identified all patients who had previously received cytotoxic chemotherapy for retrospective analysis. MATERIALS AND METHODS: Oncologic data, pretreatment hormone profiles, testicular histology, and outcomes of microTESE-ICSI were reviewed. MicroTESE was performed by a single surgeon. Most patients without preoperatively available testicular histology underwent random testicular biopsy at the time of microTESE. Histology was classified by the most advanced spermatogenic pattern present. ICSI was performed using fresh spermatozoa. Embryos were transferred into the uterine cavity on the third day after microinjection. Clinical pregnancy was established by transvaginal ultrasonographic detection of a fetal heartbeat. RESULTS: 53 patients underwent microTESE-ICSI. Lymphoma was the most common malignancy (n¼25, 47%), followed by leukemia (n¼9, 17%), sarcoma (n¼7, 13%), and testicular cancer (n¼6, 11%). Mean patient and female partner ages were 34.8 years (range 22-54) and 32.5 years (range 21-43). The mean male serum follicle-stimulating hormone level was 22.4 mIU/mL (range 3.3-62.7). Testicular histology was available in 46 patients, 40 of whom had Sertoli cell only (SCO) pattern (87%) and 6 of whom had hypospermatogenesis (HS) (13%). MicroTESE was successful in 28 of 64 attempts (43% retrieval rate, mean 1.2 attempts per patient). Retrieval rates were significantly higher in men with HS (100%) than in men with SCO (33%) (p¼ .003, Fisher exact test). Clinical pregnancies following successful sperm retrieval were achieved in 15 of 28 couples (54%), which resulted in 6 live deliveries (21%). Four boys and 4 girls were delivered.
Vol. 90, Suppl 1, September 2008
P-623 EARLY VERSUS LATE MATURATION ARREST: INCIDENCE AND CLINICAL SIGNIFICANCE IN TESTICULAR FAILURE. J. W. Weedin, D. M. Fenig, J. N. Mills, L. I. Lipshultz. Urology, Baylor College of Medicine, Houston, TX. OBJECTIVE: There is a paucity of data in the literature characterizing the stages of maturation arrest (MA). We hypothesize that patients with earlier MA have a higher serum FSH reflecting increased sertoli cell/testicular damage and that the probability of mature sperm extraction increases at later stages of MA. DESIGN: All patients undergoing sextant site-specific testis biopsies, i.e. testis mapping, at a single institution from 2004-2008 were retrospectively reviewed. MATERIALS AND METHODS: 114 patients with nonobstructive azoospermia (NOA) and 23 patients with severe oligospermia (density < 10,000 sperm/mL) were categorized by their major, and if present, minor histopathology patterns. These categories consisted of sertoli-cell only (SCO), early maturation arrest (EMA), late maturation arrest (LMA), hypospermatogenesis (H) and normal. EMA was characterized by maturation arrest up to the level of the primary spermatocyte and LMA up to the level of the spermatid. Each group was analyzed with respect to serum hormone concentration, testicular volume, chromosomal abnormalities (karyotype, Y-deletions) and presence of sperm at the time of testicular mapping. Statistical analysis was carried out by paired T-test. RESULTS: 42 (30.9%) patients were identified with MA. 15 patients had concomitant SCO histology. Pathology was organized according to primary and secondary components (Table 1). Mean FSH of pure EMA versus pure LMA was 16.2 vs. 6.8 (p ¼ 0.037). FSH correlated with testis pathology (Table 1) but not with the probability of obtaining sperm at the time of biopsy (p ¼ 0.32). The frequency of successful sperm extraction during testis mapping increased with later stages of MA (SCO/mixed MA ¼ 26.7%, EMA ¼ 14.3%, mixed MA ¼ 37.5%, LMA ¼ 46.1%). Y-chromosome deletions were detected in 35.7% of patients with EMA, 14.2% with LMA, and 2.1% of all other patients. TABLE 1. Histopathologic categories
N FSH % pts with sperm extracted
SCO
SCO/ EMA
SCO/ LMA
EMA
EMA/ LMA
LMA/ EMA
LMA
42 29.3 7.1
8 24.2 25
7 25.3 28.6
7 16.2 14.2
4 11 50
4 5.3 25
13 6.8 46.2
FERTILITY & STERILITYÒ
CONCLUSIONS: MA is a common pattern in NOA that is frequently associated with other histopathology. In patients with pure MA, FSH is higher in patients with EMA and tends to decrease and approach normal with later stages of spermatogenesis present. We also report that a higher percentage of patients with EMA have Y-chromosome microdeletions. Although FSH did not correlate with the probability of sperm extraction, the probability of sperm found on testis mapping was higher with late maturation arrest. Supported by: None.
P-624 HANDHELD INTRA-OPERATIVE DOPPLER FLOW TO LOCALIZE THE PRESENCE OF SPERM DURING TESTIS BIOPSY IN AZOOSPERMIC MEN. S. J. Parekattil, M. S. Cohen, J. W. Vieweg. Urology, University of Florida, Gainesville, FL. OBJECTIVE: Testis biopsy is a diagnostic procedure performed in men with possible non-obstructive azoospermia (NOA). Recent studies have illustrated that it is likely to find active spermatogenesis in areas with better blood supply within the testicle. These studies utilized detailed color Doppler ultrasonography and needle guidance techniques to localize possible areas of spermatogenesis within the testicle. Our goal was to assess the efficacy of simple percutaneous hand-held Doppler blood flow measurements of the testicle at the time of biopsy to localize areas of spermatogenesis. DESIGN: Retrospective case series review and blinded analysis. MATERIALS AND METHODS: Review of 4 patients who underwent testis biopsy (3 to 4 biopsies from each testicle – 14 total biopsies) performed for NOA from Nov’07 to Mar’08. Percutaneous hand-held Doppler (Vascular TechnologyÔ, Nashua, NH) blood flow measurements were taken from 12 different regions of each testicle (only one testicle scanned on each patient – the testicle that was larger was chosen). The surgeon then obtained 3 to 4 random testicular biopsies (blinded to the Doppler analysis) from that testicle. The findings from the biopsies were then compared to the pre-biopsy Doppler mapping to assess if the Doppler readings had any predictive value in detecting spermatogenesis. RESULTS: The Doppler mapping identified three distinct types of blood flow waveforms within the testicle – high, medium and low. The medium amplitude waveform provided the highest predictive value in correctly locating spermatogenesis (sperm found at that biopsy location). The detection of this waveform was 79% accurate in predicting the location of sperm at the time of biopsy (area under the ROC of 0.8). This waveform may correlate to intermediary arteries within the testicle and thus identify areas with good blood supply. CONCLUSIONS: Our preliminary evaluation of hand-held Doppler flow mapping of the testicle at the time of biopsy appears to have some promise in detecting areas of spermatogenesis. Further evaluation and testing will reveal its potential in this role. Supported by: Research equipment support from VTI Technology.
P-625 ASSOCIATION OF SPERM DNA FRAGMENTATION MEASURED BY TUNEL WITH SPERM MATURATION MEASURED BY DNA CYTOMETRY. R. Mahfouz, R. Sharma, A. Agarwal. Center for Reproductive Medicine, Cleveland Clinic, Cleveland, OH; Cleveland Clinic, Cleveland, OH. OBJECTIVE: The terminal uridine deoxynucleotidyl transferase (dUTP) nick end labeling (TUNEL) measures single and double DNA strand breaks. DNA cytometry using propidium iodide (PI) can provide information on sperm chromatin maturation/ condensation status based on the fluorescent intensity of PI. The aim of our study was to evaluate the relationship between sperm DNA fragmentation as measured by TUNEL assay with chromatin maturation/ condensation by DNA cytometry. DESIGN: Prospective study. MATERIALS AND METHODS: 117 semen samples were prepared for TUNEL assay using the Apoptosis Detection Kit (APO-Direct, BD
S317
Bioscience). PI/ RNase were added after the FITC-dUTP reaction for DNA staining according to manufacturer’s instructions. Flowcytometry was used and data acquisition was done for FITC (FL1) as well as PI (FL2-area). RESULTS: The percentage of TUNELþve sperm were positively correlated with percentage of immature (r ¼ 0.36, p ¼ <0.001) and subhaploid sperm (r ¼ 0.53, p ¼ <0.001). TUNELþve spermatozoa showed a negative correlation with percentage of mature (haploid) sperm (r ¼-0.55, p ¼<0.001). % haploid mature sperm were positively correlated with TUNEL-ve percentage (r ¼ 0.7, p <0.001). There was a significant negative interdependence between percentage of haploid sperm and subhaploid sperm (r ¼ 0.76, p <0.001) and between haploid and % of immature sperm (r ¼ 0.44, p <0.001). CONCLUSIONS: Low DNA fragmentation detected by TUNEL assay was associated with sperm maturation/ condensation detected by DNA cytometry. We recommend evaluating semen samples by both assays for better understanding of the underlying pathology i.e. sperm immaturity, apoptosis and/ or oxidative stress associated with abnormal spermatozoa. Supported by: None.
P-626 ENHANCEMENT OF SPERM FERITILIZATION CAPABILITY BY PEROXISOME PROLIFERATION-ACTIVATED RECEPTOR (PPAR) g AGONIST. I. C. Wun, J.-C. Huang. OBGYN, University of Texas Medical School at Houston, Houston, TX. OBJECTIVE: PPAR g is among the nuclear receptors that belong to a superfamily pertaining to energy integration, along with lipid and glucose metabolism. Aquila et al (2006) first reported that PPAR g was expressed in human spermatozoa and enhanced sperm capacitation, acrosome reaction, and motility. This study implements a mouse model to examine the effect of Troglitzaone (a PPAR gamma agonist) upon fertilization. DESIGN: A prospective study. MATERIALS AND METHODS: Spermatozoa were collected from the vas deferens, while the oocytes were obtained from superovulated mice. A Hamilton Thorn semen analyzer was used for the sperm kinetic study. The definition of hyperactivation was a curvilinear velocity greater than 135 m/ s, a linearity of less than 5.0, and lateral head displacement amplitude greater than 6.5 m. A triple-stain technique was used to observe for acrosomal reaction occurrence (Balbot and Chacon, 1981). The insemination concentration was 10,000 motile sperm/ml with Troglitazone from 0.3 mM to 30 mM. The method of least squares was used for kinetic parameter analysis. The acrosomal stain was analyzed by an approach of a generalized estimate equation linked with a logit transformation. The trend of the dose-fertilization relationship was analyzed by the Cochran-Mentel-Haenzel test. Specific Troglitazone concentrations were compared to the control by using the chi-square test.
Fertilization Rate
Item
Sperm Activity Attribute
1 mM vs. 3 mM vs. 10 mM vs. 30 mM vs. Acrosome control control control control Motility Hyperactivation Reaction
Analysis p <0.05 p <0.005 p <0.005 p <0.05
p <0.05 p <0.05
p <0.05
RESULTS: 1. Overall motility increased by Troglitazone treatment. Sperm hyperactivation significantly increased both overall and at 90 minutes Troglitazone treatment. 2. Acrosome reaction increased significantly due to Troglitazone treatment. 3. The fertilization rate showed a marginal trend (p¼0.0527) along with the Troglitazone dosage. Troglitazone at 1 mM or higher significantly increased the fertilization rate. CONCLUSIONS: Troglitazone significantly increases sperm motility, hyperactivation, and acrosome reaction. These activated parameters correspond to the spermatic ability to fertilize eggs. The study demonstrated that expression of functional PPAR g serves to enhance the spermatic ability to fertilize eggs. Aquila, S. et al. J Cell Physiol; 209: 977-86, 2006. Supported by: None.
S318
Abstracts
P-627 PREVALENCE OF SECONDARY INFERTILITY FOLLOWING VARICOCELECTOMY. P. Salehi, H. Abbasi, K. Nouri-mahdavi, M. H. Nasr Esfahani, M. Zargham, M. Yazdani. Urology, Isfahan University of Medical Sciences, Isfahan, Islamic Republic of Iran; Isfahan University of Medical Sciences, Isfahan, Islamic Republic of Iran; Isfahan Fertility and Infertility Center, Isfahan, Islamic Republic of Iran. OBJECTIVE: To determine the prevalence of secondary infertility following varicocelectomy. DESIGN: This stuty is a cross-sectional research which is performed in isfahan fertility and infertility center on 96 men with a history of primary infertility who had undergone microsurgical varicocelectomy. MATERIALS AND METHODS: Ninety-six men with a history of primary infertility who had fathered children following microsurgical varicocelectomy and wished to have another child were followed after the spouses’ first pregnancy to determine the number of couples who had achieved a second pregnancy. RESULTS: The mean duration follow-up was 3 years (range 1-5). Secondary infertility was observed in 72 (75%), with only 24 couples (25%) achieving a second pregnancy. Secondary infertility was directly related to patient age, duration of prevaricocelectomy infertility, and time to first pregnancy (P<0.05). CONCLUSIONS: It seems that varicocelectomy is associated with unexpectedly high rates of secondary infertility. Supported by: None.
P-628 MICRODISSECTION TESTICULAR SPERM EXTRACTION (MICROTESE) AND INTRACYTOPLASMIC SPERM INJECTION (ICSI) IN THE TREATMENT OF PERSISTENT POSTCHEMOTHERAPY AZOOSPERMIA (PPCA): LESSONS LEARNED OVER 12 YEARS. P. J. Stahl, E. J. Nejat, G. D. Palermo, L. L. Veeck, Z. Rosenwaks, P. N. Schlegel. Department of Urology, Weill Cornell Medical College, New York, NY; Department of Obstetrics & Gynecology, Weill Cornell Medical College, New York, NY; Center for Reproductive Medicine & Infertility, Weill Cornell Medical College, New York, NY. OBJECTIVE: In 2001 we published our initial experience with microTESE and ICSI for the treatment of PPCA. The objective of the present study was to report our updated outcomes in this population to provide prognostically helpful information to physicians and affected couples. DESIGN: We queried our database of men who consecutively underwent microTESE at our institution from 1995-2007 and identified all patients who had previously received cytotoxic chemotherapy for retrospective analysis. MATERIALS AND METHODS: Oncologic data, pretreatment hormone profiles, testicular histology, and outcomes of microTESE-ICSI were reviewed. MicroTESE was performed by a single surgeon. Most patients without preoperatively available testicular histology underwent random testicular biopsy at the time of microTESE. Histology was classified by the most advanced spermatogenic pattern present. ICSI was performed using fresh spermatozoa. Embryos were transferred into the uterine cavity on the third day after microinjection. Clinical pregnancy was established by transvaginal ultrasonographic detection of a fetal heartbeat. RESULTS: 53 patients underwent microTESE-ICSI. Lymphoma was the most common malignancy (n¼25, 47%), followed by leukemia (n¼9, 17%), sarcoma (n¼7, 13%), and testicular cancer (n¼6, 11%). Mean patient and female partner ages were 34.8 years (range 22-54) and 32.5 years (range 21-43). The mean male serum follicle-stimulating hormone level was 22.4 mIU/mL (range 3.3-62.7). Testicular histology was available in 46 patients, 40 of whom had Sertoli cell only (SCO) pattern (87%) and 6 of whom had hypospermatogenesis (HS) (13%). MicroTESE was successful in 28 of 64 attempts (43% retrieval rate, mean 1.2 attempts per patient). Retrieval rates were significantly higher in men with HS (100%) than in men with SCO (33%) (p¼ .003, Fisher exact test). Clinical pregnancies following successful sperm retrieval were achieved in 15 of 28 couples (54%), which resulted in 6 live deliveries (21%). Four boys and 4 girls were delivered.
Vol. 90, Suppl 1, September 2008