Adverse effects of prolonged use of pausinystalia yohimbe on sperm and reproductive organs in rats

on semen analysis only reporting sperm after centrifugation. Two to four cryopreservation attempts were recommended to all patients. Patients unable to cryopreserve sperm underwent microsurgical testicular sperm extraction (mTESE). Cryopreserved sperm obtained from ejaculate or mTESE where used in in-vitro-fertilization intracytoplasmic sperm injection (IVF-ICSI) per couples desires. Statistical analysis was performed using were the fisher exact test. RESULTS: Twenty-one (75%) men successfully cryopreserved ejaculated sperm. Seven men (25%) were unable to cryopreserve ejaculated sperm and underwent mTESE. Median age for all men was 35.5 (2951). 85.7% of men who underwent mTESE had a successful extraction with sperm. 1 patient who underwent mTESE was found to have sertoli cell only syndrome and unable to cryopreserve sperm. The couple used donor sperm at time of IVF. Amongst the men who successfully cryopreserved ejaculated sperm, 1 (4.8%) underwent a mTESE after cryopreserved sperm used in IVF-ICSI resulted in no viable pregnancy. He was unable to cryopreserve after that and elected to proceed with mTESE. 28.6% of patients who underwent mTESE had live births using their sperm, however none of the patients who successfully cryopreserved ejaculated sperm had no live births with paternal sperm. There were no significant female factors other than advanced maternal age (>35) in 3 (42.9%) of women in the mTESE group and 8 (38.1%) of women in the ejaculated sperm group. CONCLUSIONS: Management of cryptoozospermic men can be challenging. In our study, men using cryopreserved ejaculated sperm had no live births compared to 28.6% of men who underwent mTESE (p¼ 0.0556). This raises the question of sperm integrity in these patients. It is difficult, if not impossible to assess DNA integrity on these patients due to the small amount of sperm obtained. Our experience with managing these patients highlights a potential need for embracing early surgical intervention and honest discussion, including possible donor sperm assuming maternal factors are not present.

Parameters n Median Age Median Partner age Successful cryopreservation of sperm Live birth rate

Ejaculated sperm

mTESE

21 36 (29-46) 32 (23-44) 75%

7 33 (29-51) 32 (29-46) 85.7%

0%

28.6%

0.0556

References: 1. WHO laboratory manual for the examination and processing of human semen - 5th ed. 2010.

P-545 Wednesday, November 1, 2017 IMPACT OF DNA METHYLATION IN FERTILE AND SUBFERTILE PATIENTS ON SPERM PARAMETERS AND FERTILIZATION RATE. Y. A. Alkhaled, M. M. Laqqan, M. Hammadeh. Obstetrics & Gynecology, IVF Lab, Saarland University, Homburg, Germany. OBJECTIVE: To determine the correlation between changes in sperm DNA methylation levels and sperm parameters and their influence on fertilization rate. DESIGN: A cohort study. A total of 82 human semen samples were collected during the period between March 2016 and September 2016. Samples were obtained from sub-fertile (as a case group), and from fertile males (as a control group). MATERIALS AND METHODS: As a pilot study, twenty samples (10 case and 10 controls) were subjected to Infinium 450K BeadChip arrays, to identify genomic regions that differences in sperm DNA methylation patterns in subfertile compared to fertile males who have idiopathic infertility and underwent intracytoplasmic sperm injection (ICSI). Then tow CpGs were validated by local deep bisulfite sequencing on 82 sperm samples (n ¼ 44 subfertile) and (n ¼ 38 fertile). RESULTS: According to the results of pilot study 15 CpGs have been significantly different in the sperm DNA methylation levels between cases compared to controls group. Only 9 CpGs do not have overlap common

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ASRM Abstracts

annotated SNPs. The results of the validation study for the two CpG (cg19779893, cg19406113) found that a significant variation in the methylation level at 2 CpG (CpG2, CpG3) out of 3 CpGs related to cg19779893 site amplicon (P % 0.03, P % 0.01, respectively) in cases compared to controls. Moreover, Six CpGs from eleven CpG3, CpG5, CpG6*, CpG8, CpG9, and CpG10 related to cg19406113 site amplicon (P % 0.009, P % 0.03, P % 0.02, P % 0.03, P % 0.007, and P % 0.02 respectively) showed significant difference in sperm DNA methylation between cases and controls group. Furthermore, significantly decreased was showed in the sperm concentration, motility, morphology, vitality, viability, and fertilization rate (P % 0.04, P % 0.02, P % 0.04, P % 0.003, P % 0.03, and P % 0.04 respectively) in the case compared to control groups. CONCLUSIONS: The present study found two CpG altered in sperm DNA methylation levels. In addition, a strong association between changes in sperm DNA methylation levels in these CpG sites and sperm parameters. Supported by: University of Saarland, Department of Obstetrics and Gynecology, University of Saarland P-546 Wednesday, November 1, 2017 ADVERSE EFFECTS OF PROLONGED USE OF PAUSINYSTALIA YOHIMBE ON SPERM AND REPRODUCTIVE ORGANS IN RATS. L. C. Ajonuma,a S. A. Bamiro,a S. L. Makanjuola.b aDepartment of Physiology, Lagos State University College of Medicine, Ikeja, Lagos, Nigeria; bDepartment of Pharmacology, Lagos State University College of Medicine, Ikeja, Lagos, Nigeria. OBJECTIVE: Pausinystalia yohimbe (P. yohimbe) was first discovered and used by tribes in West Africa, where it grows wild mostly in the Atlantic evergreen forest throughout West Africa from South-east Nigeria to Congo. P. yohimbe bark extract is commonly used as an aphrodisiac among men and for the treatment of erectile dysfunctions. More recently, due to infertility more men have resorted to using P. yohimbe to improve chances of having baby. However, there are no studies on the effects of P. yohimbe on sperm motility and movement characteristics. This study examined its effects on reproductive organs, sperm production and motility in rats. DESIGN: Basic animal research in a university teaching hospital setting MATERIALS AND METHODS: Fifteen male Sprague Dawley (SD) rats were divided into 3 groups of 5 rats. Group A was the control while groups B and C served as the test groups. Group A received 0.5ml of normal saline daily while groups B and C received orally 150mg/kg and 300mg/kg body weights of aqueous extracts of P. yohimbe respectively for 4 weeks. Serum samples obtained from the rats were assayed for reproductive hormones (Testosterone, Follicle Stimulating Hormone (FSH), Luteinizing Hormone (LH) and Estradiol (E2)). Testes, Seminal Vesicles and the Prostates were removed for histology, while sperm count, motility and vitality were done using sperm from the caudal epididymis and vas deference. RESULTS: Mean percentage change in body weights of groups B and C were significant when compared to control group A. There was significant reduction in sperm motility and concentration in the test groups, but none in sperm vitality. Serum hormonal levels of Testosterone, FSH, LH and E2 were not significantly different from the control. However, there was a significant reduction in testosterone / E2 ratio in the test groups. Degenerative changes were observed in the testes, prostate and seminal vesicles. CONCLUSIONS: Prolonged use of P.yohimbe adversely affects the testes, male accessory glands, sperm concentration and motility. This may lead to reduced reproductive functions and male infertility.

P-547 Wednesday, November 1, 2017 SCREENING FOR GERM CELLS IN SURGICAL SPECIMEN THAT FAIL TO YIELD SPERMATOZOA. C. O’Neill, M. Irani, A. Parrella, Z. Rosenwaks, G. D. Palermo. Reproductive Medicine, Weill Cornell Medicine, New York, NY. OBJECTIVE: To identify the presence of germ cell (GC) maturational stages of non-obstructive azoospermic (NOA) patients where no

Vol. 108, No. 3, Supplement, September 2017