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To compare fertilization, cleavage, and pregnancy outcome using pentoxifylline and a hypoosmotic swelling (HOS) test to select viable spermatozoa from testicular biopsy specimens. Open, comparative, prospective study. G.S. Medical College and Fertility clinic, Mumbai, India. A total of 50 couples enrolled for infertility treatment having a male factor indication of nonobstructive azoospermia. Assessment of viable spermatozoa using pentoxifylline and using an HOS test from a population of nonmotile spermatozoa obtained from testicular biopsies. Comparison of fertilization, cleavage, and clinical pregnancy rates using viable sperms recovered using pentoxifylline and an HOS test. Viable spermatozoa were obtained in both the study groups. Significantly higher fertilization rates (pentoxifylline 62.05% vs. HOS 41.07%) and clinical pregnancy rates (pentoxifylline 32% vs. HOS 16%) were observed. There was no significant difference in cleavage rates among both groups. We found that obtaining viable spermatozoa using pentoxifylline was more effective in terms of fertilization and pregnancies than obtaining it through an HOS test. Copyright © 2011 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

Desai S.S.,National Health Research Institute | Achrekar S.K.,National Health Research Institute | Paranjape S.R.,National Health Research Institute | Desai S.K.,Fertility Clinic and Center | And 2 more authors.
Reproductive BioMedicine Online | Year: 2013

During an IVF protocol, exogenous FSH is administered to women for ovulation induction. The ovarian response to gonadotrophin stimulation is variable and unpredictable in these women. The FSHR is the most studied gene in relation to ovarian response. The association of a FSHR gene polymorphism at position 680 (p.Asn680Ser) with ovarian response has been well documented. Recently, a polymorphism at position-29 in the 50-untranslated region of FSHR (g.-29G>A) has been reported to be associated with poor ovarian response and reduced FSHR expression. The present study evaluated the combined effect of the polymorphisms at positions-29 and 680 of FSHR with type of ovarian response and receptor expression. The two FSHR gene polymorphisms together formed four discrete haplotypes and nine allelic combinations. Various clinical parameters revealed that 75% of the subjects with A/A-Asn/Asn genotype were poor ovarian responders (odds ratio 7.92; P = 0.009). The relative FSHR mRNA expression in granulosa cells indicated that subjects with A/A-Asn/Asn genotype express significantly lower level of FSHR as compared to the subjects with G/G-Asn/Ser genotype (P = 0.029). These results indicate that A/A-Asn/Asn genotype could be used as a potential marker to predict poor ovarian response. © 2013, Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

Desai S.S.,National Health Research Institute | Achrekar S.K.,National Health Research Institute | Pathak B.R.,National Health Research Institute | Desai S.K.,Fertility Clinic and Center | And 3 more authors.
Journal of Clinical Endocrinology and Metabolism | Year: 2011

Context: Polymorphisms of the FSHR gene are associated with variable ovarian response to FSH stimulation in subjects undergoing in vitro fertilization (IVF) treatment. The type of ovarian response is correlated with the level of FSH receptor (FSHR) expression on granulosa cells. Objective:Weinvestigated whether the polymorphism at position -29 in the promoter of theFSHR gene may contribute in altered receptor expression. Design and patients: FSHR polymorphism at position -29 was studied in 100 subjects undergoing IVF treatment. Association of this polymorphism with level of FSHR expression was retrospectively analyzed. Setting: The study was conducted at an academic research institute and private IVF clinic. Methods: The genotype at position -29 of the FSHR gene was studied in IVF subjects by PCRrestriction fragment length polymorphism. Total RNA and protein was extracted from granulosa cells. The relative FSHR mRNA expression was carried out by real-time PCR. The receptor protein expression was evaluated by Western blot and confocal microscopy. Results:Theclinicalandendocrinological parameters revealed that almost72%of subjects with the AA genotype at position -29 of FSHR gene were poor ovarian responders (odds ratio 8.63, 95% confidential interval 1.84-45.79; P = 0.001). The lower cleavage intensity predicted by in silico analysis for A allele as compared with the G allele suggest the difference in the DNA-protein binding affinity. The relative expression of FSHR at mRNA and protein level was significantly reduced in subjects with AA genotype as compared with the GG genotype. Conclusion: Poor ovarian response observed in subjects with the AA genotype at position -29 of the FSHR gene is due to reduced receptor expression. Copyright © 2011 by The Endocrine Society.

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