Namita Kattal

Granulocyte-macrophage colony stimulating factor (GM-CSF) and co-culture can affect post-thaw development and apoptosis in cryopreserved embryos

Purpose: The objective of this study was to evaluate the effects of growth factor supplementation and Vero cell co-culture on apoptosis and development of frozen thawed one-cell mouse embryos.Methods: The following treatment regimens were assessed: (a) control medium (b) Vero cell co-culture and (c) growth factor supplemented medium. The individual growth factors tested were: GM-CSF, IGF-I, IGF-II, TNF-α, FGF-4, LIF, TGF-α, TGF-β, IL-6, PDGF and EGF. Blastocyst development and differentiation were monitored. At termination of the experiments, overall blastomere number and apoptosis were assessed using the TUNEL assay.Results: No differences were observed in blastulation and hatching rates. ICM differentiation in thawed embryos was notably improved with either co-culture or growth factor supplementation. The only growth factor significantly modulating apoptosis in thawed embryos was granulocyte-macrophage colony stimulating factor (GM-CSF). GM-CSF enhanced continued cell survival and prevented apoptosis but did not influence overall cell number in developing blastocysts. Vero cell co-culture significantly increased cell number in blastocysts (124±42 vs 100±44 in control; P<0.05). Embryonic apoptosis was higher in the co-cultured embryos. The increased presence of apoptotic cells in blastocysts of high cell number may reflect the regulatory role of apoptosis in balancing ICM: TE ratios.Conclusion: These data indicate that culture conditions can modulate post-thaw embryonic development and apoptosis.

Outcome of intracytoplasmic sperm injection (ICSI) using epididymal and testicular sperm from azoospermic men: the cleveland clinic experience

USING EPIDIDYMAL AND TESTICULAR SPERM FROM AZOOSPERMIC MEN: THE CLEVELAND CLINIC EXPERIENCE Mehmet Oder, Ramadan A Saleh, Namita Kattal, Mohamed A Bedaiwy, Anthony J Thomas, Ashok Agarwal, Cleveland Clinic Foundation Objective: Sperm retrieval from testis and epididymis, combined with ICSI, has significantly enhanced the chances of azoospermic men to father their own genetic offspring. The impact of the sperm origin (epididymal versus testicular) and status (fresh versus frozen-thawed), and the etiology of azoospermia 〈obstructive versus nonobstructive azoospermia (NOA)〉 on sperm reproductive capacity is not well established. In this retrospective analysis, we examined the outcome of ICSI with use of epididymal versus testicular spermatozoa in patients with OA and NOA. Design: Retrospective study Materials/Methods: We collected data of 32 infertile men (26 with OA and 6 with NOA) treated with ICSI. Out of the 26 obstructive cases, 10 were diagnosed with congenital absence of vas deferens, 9 with failed vasectomy reversal, 2 with congenital bilateral ejaculatory duct obstruction and 5 with post-inflammatory obstruction. Female partners underwent a standard protocol of controlled ovarian stimulation. Techniques of MESA and TESE were used for sperm retrieval in OA, whereas TESE was used in NOA cases. Data were collected on normal fertilization and embryo development rates and on pregnancy rates (per patient and per cycle). Results: A total of 52 ICSI cycles were performed including 42 cycles from patients with OA and 10 cycles from NOA. Epididymal sperm was used in 32 cycles (12 cycles with fresh and 20 with frozen-thawed sperm), whereas testicular sperm was used in the remaining 20 cycles (11 cycles with fresh and 9 with frozen sperm). Study parameters are shown in the table. Normal fertilization and pregnancy rates per cycle did not differ significantly (P>0.05) in relation to etiology (obstructive versus non-obstructive), origin (epididymal versus testicular) or type of sperm (fresh versus frozen-thawed). However, normal embryo development rates were significantly lower in NOA versus OA cases (0.0004). Normal embryo development rates and pregnancy rates per patient were significantly lower with frozen testicular sperm compared to frozen epididymal sperm (p = 0.004 & 0.03, respectively). Abortion was noted in 1/12 (8%) cycles with fresh epididymal sperm, 1/20 (5%) cycles with frozen-thawed epididymal sperm and 1/9 (11%) cycles with of frozen-thawed testicular sperm. Conclusions: Our study showed no significant difference in fertilization, embryo development or pregnancy rates with fresh epididymal versus fresh testicular sperm. Also, fertilization and pregnancy rates were comparable for OA versus NOA cases, and for fresh versus frozen-thawed sperm both from epididymis and testis. However, frozen testicular sperm resulted in a lower embryo development rate and a relative increase of abortion rate than frozen epididymal sperm. Simultaneous retrieval of fresh sperm from testis combined with ICSI may help improve the outcome in NOA. Supported by: None

Correlation of nuclear factor kappa B (NFKB) with sperm quality and clinical diagnoses in infertile men

SPERM QUALITY AND CLINICAL DIAGNOSES IN INFERTILE MEN Pavithra Ranganathan, Namita Kattal, Mohamed H Moustafa, Rakesh K Sharma, Anthony J Thomas, Ashok Agarwal, Cleveland Clinic Foundation Objective: NFΚB (p65, p50 and IΚB) plays a major role in regulating apoptosis. Its role in the pathogenesis of male infertility has never been studied. The objectives of our study were to examine 1) the relationship of NFΚB with semen quality (sperm concentration, motility and morphology) and the clinical diagnoses of male infertility, and 2) correlate levels of NFΚB and apoptosis in ejaculated human spermatozoa

Decreased expression of P65, P50 and I kappa B in ejaculated spermatozoa from infertile men

EJACULATED SPERMATOZOA FROM INFERTILE MEN Pavithra Ranganathan, Namita Kattal, Mohamed H Moustafa, Ramadan A Saleh, Rakesh K Sharma, Ashok Agarwal, Cleveland Clinic Foundation Objective: Recent studies indicate high levels of apoptosis in ejaculated spermatozoa from infertile men. Nuclear factor Kappa B (NFΚB) family is an important regulator of apoptosis. It is composed of the sub units p65, p50, and IΚB and is sequestered in the cytoplasm in an inactive form. Recent studies have shown that NFΚB is present in an active form in the nuclei of Sertoli cells in rat testis. However, the activity of NFΚB in ejaculated human spermatozoa has never been investigated. The objective of our study was to examine levels of p65, p50 and IΚBΑ in ejaculated spermatozoa from infertile men as compared to a group of normal sperm donors.

Utilization rate and fertility outcome of cryopreserved sperm from oncological patients: American experience

CRYOPRESERVED SPERM FROM ONCOLOGICAL PATIENTS: AMERICAN EXPERIENCE Pavithra Ranganathan, Ashok Agarwal, Saba Khayal, Namita Kattal, Cleveland Clinic Foundation Objective: The worldwide review of scientific literature reveals a paucity of information regarding the utilization and fertility outcome of cryopreserved semen samples from patients with cancer by assisted reproductive techniques (ART). Our study examined the results of the utilization of cryopreserved semen specimens by oncological patients for reproductive purposes.

Efficacy and compliance of early use vacuum constriction device for erectile dysfunction following radical prostatectomy

CONSTRICTION DEVICE FOR ERECTILE DYSFUNCTION FOLLOWING RADICAL PROSTATECTOMY Rupesh Raina, Ashok Agarwal, Saba Khayal, Pavithra Ranganathan, Namita Kattal, Craig D Zippe, Cleveland Clinic Foundation Objective: To assess the efficacy and compliance of vacuum constriction device (VCD) following radical prostatectomy (RP) and to determiner whether its early use can potentially prevent corporeal fibrosis and expedite recovery of erectile function.

Effects of co-administration of metformin and clomiphine citrate (CC) on hormonal profile and pregnancy rates in non-obese patients with polycystic ovary syndrome (PCOS): results of a clinical trial

CLOMIPHINE CITRATE (CC) ON HORMONAL PROFILE AND PREGNANCY RATES IN NON-OBESE PATIENTS WITH POLYCYSTIC OVARY SYNDROME (PCOS): RESULTS OF A CLINICAL TRIAL Indu Singh, Surya Medi-Tech Hospital & Research Center, Ramadan A Saleh, Namita Kattal, Cleveland Clinic Foundation, Amitabh Hatwal, Surajit Sarkar, Surya Medi-Tech Hospital & Research Center, Ashok Agarwal, Cleveland Clinic Foundation Objective: Insulin resistance, and its compensatory hyperinsulinemia, plays a key role in the pathogenesis of infertility in women with PCOS. Metformin is an insulin-sensitizing agent, which appear to ameliorate the biochemical profile and improve reproductive function in obese patients with PCOS. The objective of this study was to examine the effects of combined metformin and CC treatment on endocrine function and pregnancy rates in nonobese infertile women with PCOS.