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1H NMR based metabolomic profiling in seminal plasma of asthenozoospermic men: a pilot study
RESULTS: Opiorphin treatment resulted in a 1.2 fold change in motility in men with asthenozoospermia (p¼0.01) bringing 40% of cases into the normal motility range. Conversely, opiorphin did not affect sperm motility in the control group. CONCLUSION: Overall, these exciting preliminary data suggest that the endogenous NEP inhibitor, opiorphin, may offer potential benefit for the treatment of idiopathic male infertility characterized by asthenozoospermia. Supported by: This work was partly Supported by NIH/NIDDK (Grant# DK087872) awarded to KPD. P-634 Wednesday, October 22, 2014 1H NMR BASED METABOLOMIC PROFILING IN SEMINAL PLASMA OF ASTHENOZOOSPERMIC MEN: A PILOT STUDY. S. Singh,a E. Subramani,b R. Chattopadhyay,a S. Yasmin,a K. Chowdhury,b B. Chakravarty.a aReproductive Medicine, Institute of Reproductive Medicine, Kolkata, West Bengal, India; bSchool of Medical Science and Technology, Indian Institute of Technology, Kharagpur, West Bengal, India. OBJECTIVE: Sperm motility is usually evaluated in research and infertility clinics for assessing the male fertility. However, there is a need to identify metabolite markers for the improved diagnosis and treatment of male infertility.[1] The present pilot study, therefore, aims to identify differentially expressed metabolites in seminal plasma of asthenozoospermic men using proton NMR based metabolomics. DESIGN: Asthenozoospermic semen samples (n¼19) were randomly collected from subjects (28–40 years) reporting at the Institute of Reproductive Medicine, Salt Lake City, Kolkata for male factor infertility treatment. Proven fertile men (n¼19) whose partners had delivered healthy babies during the last six months without assisted reproductive technology were considered as controls. MATERIALS AND METHODS: Proton NMR spectra of seminal plasma were recorded using 700 MHz Bruker AvanceAV III spectrometer. Following phased and baseline correction of all NMR[2] spectra using Mest ReNova, spectral binning of the data was performed. The acquired data were analyzed using multivariate principal component analysis, partial least-squaresdiscriminant analysis, and orthogonal projection to latent structure with discriminant analysis and metabolites were identified. RESULTS: A significant alteration in metabolites including lactate, alanine, choline, glycerophosphocholine, tyrosine, histidine, phenylalanine and isoleucine was observed in seminal plasma of asthenozoospermic men as compared with controls. CONCLUSION: The metabolic changes in seminal plasma of asthenozoospermic men produced a distinct pattern which helps in differentiating asthenozoospermia from controls. The altered endogenous metabolites in the seminal plasma of patients showed glycolysis intermediates, amino acids, and molecules related to lipid catabolism. These findings suggest that the metabolomic profiling may contribute for the diagnosis and treatment of infertility. However, this study warrants further investigation with large sample size. P-635 Wednesday, October 22, 2014 SEMEN VARIATION IN A POPULATION OF FERTILE CANDIDATES FOR SPERM BANK. C. Borghi, A. Nabel, F. Aguirre, S. Papier, M. A. Barros, C. Alvarez Sedo. CEGYR - Genetics and Reproductive Medicine, Capital Federal, Buenos Aires, Argentina. OBJECTIVE: The seminal parameters related to male fertility have been studied for many years by the World Health Organization (WHO), in their last manual (2010) many of the values suffered some important changes in relation to the 1999 version. The aim of this study was to compare the seminal parameters of fertile candidates for sperm banking with the current WHO values. DESIGN: Prospective blind study. MATERIALS AND METHODS: Since one year, our institution is recruiting men for creating a sperm bank. After passing the medical and psychological evaluation, a total of 80 candidates were considered for the evaluation of a semen sample. The evaluation was conducted using the guidelines of the WHO (2010) and blinded to the personal history of each candidate. Mainly, we proceeded to measure: volume, concentration / mL, progressive motility, vitality and sperm morphology. DNA fragmentation (TUNEL), is not considered as an essential parameter by the WHO, but it was also evaluated. Subsequently, we proceeded to check out the medical
FERTILITY & STERILITYÒ
history each candidate, stating that 28 were fertile, but only 20 were fertile in a period not exceeding two years, so it was decided to take this group of men for the analysis. RESULTS: Considering the average value of each seminal parameter, all of them were over the threshold established by WHO (table 1). However, some candidates showed values under the cut-off for specific parameters. In that sense, 25% of fertile men had abnormal volume values, 35% for concentration, 20% for progressive motility and 10% for vitality. Sperm morphology was the only parameter that didn’t had an abnormal value. There were not candidates who had two or more altered parameters. The average of DNA fragmention levels was 8.43.9.
Volume Concentration Progressive motility Vitality Morphology DNA fragmentation
Candidates
WHO (2010)
2.21.6 49.127.2 36.511.0 71.112.1 8.53.4 8.43.9
1.5 39 32 58 4
CONCLUSION: In our experience, the seminal parameters of fertile men had normal average values according to WHO, but some individual cases may be below the cutoff values. The sperm concentration, is the seminal parameter that suffered more variation between the candidates, and morphology seems to be the most stable parameter. Supported by: CEGYR Foundation.
P-636 Wednesday, October 22, 2014 OUTCOME OF INTRACYTOPLASMIC SPERM INJECTION USING FRESH AND CRYOPRESERVED-THAWED TESTICULAR SPERMATOZOA IN 83 AZOOSPERMIC MEN WITH KLINEFELTER SYNDROME. K. Vicdan,a C. Akarsu,a E. S€ozen,a B. Buluc¸,a A. Vicdan,b K. Biberoglu.c aPrivate Ankara IVF Center, Ankara, Turkey; b Department of Genetics, Ankara University Medical School, Ankara, Turkey; cDepartment of Obcstetric and Gynecology, Gazi University Medical School, Ankara, Turkey. OBJECTIVE: To report the outcome of intracytoplasmic sperm injection (ICSI) using fresh or cryopreserved-thawed testicular spermatozoa in patients with Klinefelter syndrome. DESIGN: Retrospective clinical study. MATERIALS AND METHODS: Medical records of 83 azoospermic men with Klinefelter syndrome who underwent 88 TESE procedures between 2003-2013 were reviewed. The clinical parameters for predicting sperm recovery, the sperm retreival, pregnancy and live birth rates of ICSI cycles were evaluated. RESULTS: Seventy seven out of 83 azoospermic men had classic and the remaining 6 had mosaic Klinefelter syndrome. A total of 88 TESE procedures were performed of which conventional method consisting of multiple tissue biopsies and microsurgery were applied in 48 and 40 of the cases, respectively. Spermatozoa were found in 35 men with a retrieval rate of 39.7 %. The age, volume of testes, serum FSH and testosterone levels, presence of mosaicism, the method of sperm retreival and smoking were not found to be predictive in obtaining testicular spermatozoa. A total of 41 embryo transfer cycles were carried out using fresh testicular spermatozoa in 30, cryopreserved-thawed spermatozoa in 10 and frozen thawed embryo replacement in one. Twenty two clinical pregnancies were established, including 14 singleton, 5 twin, 2 triplet and 1 quadriplet gestation (53.6 %). Fifteen women delivered singleton and 7 women twin fetuses. In total, while 21 of the newborns (15 female and 6 male) were healthy, 5 newborns died following delivery due to various reasons. The outcome of three fetuses was unknown because the mothers were lost to follow-up. Karyotype analysis were available in 12 of the healthy newborns and all were normal. CONCLUSION: Testicular sperm extraction (TESE) and ICSI provide high sperm recovery and pregnancy rates in infertile couples with azoospermia due to Klinefelter syndrome. There is no clinical, epidemiological or laboratory finding that predicts sperm retrieval in TESE procedures.
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FERTILITY & STERILITYÒ
history each candidate, stating that 28 were fertile, but only 20 were fertile in a period not exceeding two years, so it was decided to take this group of men for the analysis. RESULTS: Considering the average value of each seminal parameter, all of them were over the threshold established by WHO (table 1). However, some candidates showed values under the cut-off for specific parameters. In that sense, 25% of fertile men had abnormal volume values, 35% for concentration, 20% for progressive motility and 10% for vitality. Sperm morphology was the only parameter that didn’t had an abnormal value. There were not candidates who had two or more altered parameters. The average of DNA fragmention levels was 8.43.9.
Volume Concentration Progressive motility Vitality Morphology DNA fragmentation
Candidates
WHO (2010)
2.21.6 49.127.2 36.511.0 71.112.1 8.53.4 8.43.9
1.5 39 32 58 4
CONCLUSION: In our experience, the seminal parameters of fertile men had normal average values according to WHO, but some individual cases may be below the cutoff values. The sperm concentration, is the seminal parameter that suffered more variation between the candidates, and morphology seems to be the most stable parameter. Supported by: CEGYR Foundation.
P-636 Wednesday, October 22, 2014 OUTCOME OF INTRACYTOPLASMIC SPERM INJECTION USING FRESH AND CRYOPRESERVED-THAWED TESTICULAR SPERMATOZOA IN 83 AZOOSPERMIC MEN WITH KLINEFELTER SYNDROME. K. Vicdan,a C. Akarsu,a E. S€ozen,a B. Buluc¸,a A. Vicdan,b K. Biberoglu.c aPrivate Ankara IVF Center, Ankara, Turkey; b Department of Genetics, Ankara University Medical School, Ankara, Turkey; cDepartment of Obcstetric and Gynecology, Gazi University Medical School, Ankara, Turkey. OBJECTIVE: To report the outcome of intracytoplasmic sperm injection (ICSI) using fresh or cryopreserved-thawed testicular spermatozoa in patients with Klinefelter syndrome. DESIGN: Retrospective clinical study. MATERIALS AND METHODS: Medical records of 83 azoospermic men with Klinefelter syndrome who underwent 88 TESE procedures between 2003-2013 were reviewed. The clinical parameters for predicting sperm recovery, the sperm retreival, pregnancy and live birth rates of ICSI cycles were evaluated. RESULTS: Seventy seven out of 83 azoospermic men had classic and the remaining 6 had mosaic Klinefelter syndrome. A total of 88 TESE procedures were performed of which conventional method consisting of multiple tissue biopsies and microsurgery were applied in 48 and 40 of the cases, respectively. Spermatozoa were found in 35 men with a retrieval rate of 39.7 %. The age, volume of testes, serum FSH and testosterone levels, presence of mosaicism, the method of sperm retreival and smoking were not found to be predictive in obtaining testicular spermatozoa. A total of 41 embryo transfer cycles were carried out using fresh testicular spermatozoa in 30, cryopreserved-thawed spermatozoa in 10 and frozen thawed embryo replacement in one. Twenty two clinical pregnancies were established, including 14 singleton, 5 twin, 2 triplet and 1 quadriplet gestation (53.6 %). Fifteen women delivered singleton and 7 women twin fetuses. In total, while 21 of the newborns (15 female and 6 male) were healthy, 5 newborns died following delivery due to various reasons. The outcome of three fetuses was unknown because the mothers were lost to follow-up. Karyotype analysis were available in 12 of the healthy newborns and all were normal. CONCLUSION: Testicular sperm extraction (TESE) and ICSI provide high sperm recovery and pregnancy rates in infertile couples with azoospermia due to Klinefelter syndrome. There is no clinical, epidemiological or laboratory finding that predicts sperm retrieval in TESE procedures.
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