Ashalatha Ganesh

Effect of Follicular Fluid Oxidative Stress on Meiotic Spindle Formation in Infertile Women with Polycystic Ovarian Syndrome

Background: The effect of follicular fluid (FF) oxidative stress (OS) on meiotic spindle (MS) formation in oocytes and subsequent outcome in women with polycystic ovarian syndrome (PCOS) are evaluated in this study. Methods: 326 oocytes from 35 PCOS women (group A) and 208 oocytes from 32 women with tubal infertility (group B) were visualized for MS using PolScope. FF was analyzed for OS markers including reactive oxygen species (ROS), lipid peroxidation and total antioxidant capacity (TAC). Group A was further classified into groups A1 and A2, and group B into groups B1 and B2 depending upon the presence or absence of MS, respectively. Results: MS formation was absent in a significantly higher number of oocytes in group A compared to group B (p ≤ 0.05). OS markers were significantly higher in group A compared to group B (p ≤ 0.05). Fertilization rate, number of good quality embryos and clinical pregnancy rates were higher in group B compared to group A, though not statistically significant. FF ROS was significantly higher and TAC significantly lower in groups A2 and B2 compared to groups A1 and B1 (p < 0.001). Conclusion: The presence of MS and oocyte maturation in PCOS women showed a positive correlation with low levels of OS.

Role of Latent Genital Tuberculosis in Repeated IVF Failure in the Indian Clinical Setting

Genital tuberculosis is reported to be a major pelvic factor causing infertility in Indian women and often exists without any apparent signs and symptoms. The role of latent tuberculosis in repeated IVF failure in unexplained infertility is examined. 81 women with unexplained infertility having repeated IVF failure tested for Mycobacterium tuberculosis using PCR, ZN staining and BACTEC-460 culture were selected. Fresh IVF-ET or frozen embryo transfer (FET) was attempted on patients successfully treated with anti-tubercular drugs (ATD). ATD-treated fresh cycles (group A1) and frozen cycles (group B1) were compared to previously failed fresh cycles (group A2) and FET attempts (group B2), respectively. Main outcome measures were gonadotropin required, terminal E2, number of oocytes retrieved, fertilization rate, embryo quality, endometrial thickness and sub-endometrial blood flow (Vmax). Gonadotropin required in group A1 was significantly less as compared to group A2. Number of oocytes retrieved and grade I embryos, endometrial thickness and Vmax were significantly higher in group A1. Endometrial thickness and Vmax were significantly increased in group B1 as compared to B2. The study indicates that latent tuberculosis should be considered in young Indian patients presenting with unexplained infertility with apparently normal pelvic and non-endometrial tubal factors and repeated IVF failure.

Comparison of oral dydrogestrone with progesterone gel and micronized progesterone for luteal support in 1,373 women undergoing in vitro fertilization: a randomized clinical study.

OBJECTIVE To compare the efficacy of oral dydrogesterone with that of micronized vaginal P gel and micronized P capsule for luteal supplementation. DESIGN Prospective, randomized clinical study. SETTING Institute of Reproductive Medicine, Kolkata, India. PATIENT(S) A total of 1,373 infertile women undergoing IVF participated. INTERVENTION(S) Micronized P gel, P capsule, and oral dydrogesterone were administered for luteal support and compared. MAIN OUTCOME MEASURE(S) Demographic profile and pregnancy and miscarriage rates. RESULT(S) The overall pregnancy rate and miscarriage rate were comparable among the three groups. CONCLUSION(S) Oral dydrogesterone seems to be a promising drug for luteal support in woman undergoing IVF.

Congenital Malformations among Babies Born Following Letrozole or Clomiphene for Infertility Treatment

Context Clomiphene citrate (CC) is the first line drug for ovulation induction but because of its peripheral antiestrogenic effect, letrozole was introduced as the 2nd line drug. It lacks the peripheral antiestrogenic effect and is associated with similar or even higher pregnancy rates. Since letrozole is a drug for breast cancer, its use for the purpose of ovulation induction became controversial in the light of studies indicating an increased incidence of congenital malformations. Aims To evaluate and compare the incidence of congenital malformations among offsprings of infertile couples who conceived naturally or with clomiphene citrate or letrozole treatment. Settings and Design A retrospective cohort study done at a tertiary infertility centre. Methods and Material A total of 623 children born to infertile women who conceived naturally or following clomiphene citrate or letrozole treatment were included in this study. Subjects were sorted out from medical files of both mother and newborn and follow up study was done based on the information provided by parents through telephonic conversations. Babies with suspected anomaly were called and examined by specialists for the presence of major and minor congenital malformations. Other outcomes like multiple pregnancy rate and birth weight were also studied. Results Overall, congenital malformations, chromosomal abnormalities were found in 5 out of 171 (2.9%) babies in natural conception group and 5 out of 201 babies in the letrozole group (2.5%) and in 10 of 251 babies in the CC group (3.9%). Conclusions There was no significant difference in the overall rate of congenital malformations among children born to mothers who conceived naturally or after letrozole or CC treatment. Key Messages Congenital malformations have been found to be comparable following natural conception, letrozole and clomiphene citrate. Thus, the undue fear against letrozole may be uncalled for.

Luteal phase estradiol level: a potential predictive marker for successful pregnancy in in vitro fertilization/intracytoplasmic sperm injection

OBJECTIVE To analyze the role of luteal E(2), P, and FSH levels in the prediction of clinical pregnancy in stimulated cycles. DESIGN Prospective study. SETTING Institute of Reproductive Medicine, Salt Lake, Kolkata, India. PATIENT(S) A total of 268 women aged <40 years undergoing their first IVF-ET/intracytoplasmic sperm injection cycles. INTERVENTION(S) Estimation of luteal phase serum E(2), P, and FSH levels on days 0, 7, and 14 of ET. MAIN OUTCOME MEASURE(S) Comparison of luteal phase hormonal profile in pregnant and nonpregnant women and interpretation by discriminant analysis. RESULT(S) A total of 113 pregnancies were achieved (42.16%). Age, basal FSH level, peak E(2) level, number of oocytes retrieved, and number of embryos transferred were comparable between pregnant and nonpregnant women. Day-7 and day-14 luteal E(2) levels were significantly different between the two groups. Progesterone levels were comparable in both groups on days 0 and 14. Levels of FSH showed statistically significant differences between the two groups on days 0, 7, and 14. Classification accuracy for E(2), P, and FSH was 82.1%, 60.8%, and 67.51%, respectively. CONCLUSION(S) Luteal E(2) concentration seems to be a promising marker of successful clinical pregnancy.

Selection of birefringent spermatozoa under Polscope: effect on intracytoplasmic sperm injection outcome

The ideal method for sperm selection during Intracytoplasmic sperm injection (ICSI) is still ill‐defined. Identification of a viable spermatozoon amongst immotile spermatozoa for ICSI often becomes difficult. Ninety‐six ICSI cycles were selected and divided into Group A (azoospermic, n = 58) and Group B (complete asthenozoospermic, n = 38). Oocytes having birefringent meiotic spindle and zona pellucida thickness <20 μm were selected for ICSI. Groups A and B were further divided into A1, A2 and B1, B2, respectively, based on the type of ICSI performed. In Group A1, a motile spermatozoon with normal morphology was injected into a metaphase‐II (M‐II) oocyte. In Group B1, spermatozoon showing coiling of tail following modified hypo‐osmotic swelling test was injected into M‐II oocytes. In Groups A2 and B2, ICSI was performed by injecting a spermatozoan with birefringent head. Pronuclear morphology, fertilisation rate, embryo grading and pregnancy rate were assessed. ICSI outcome measures were better in Group A2 than in Group A1 but were statistically insignificant. However, significantly higher percentage of Z1 and Z2 zygotes, Grade I and Grade II embryos and pregnancy rate were observed in Group B2 as compared to Group B1. Selection of birefringent spermatozoa shows promising results in asthenozoospermic men and men undergoing testicular sperm aspiration or extraction before ICSI.

Live birth in a 50-year-old woman following in vitro fertilization–embryo transfer with autologous oocytes: a rare case report

OBJECTIVE To report a live birth with in vitro fertilization (IVF) from a 50-year-old woman with homologous oocytes. DESIGN Case report. SETTING Referral center. PATIENT(S) A 50-year-old woman. INTERVENTION(S) IVF with fresh embryo transfer (ET). MAIN OUTCOME MEASURE(S) Live birth after IVF. RESULT(S) A 50-year-old woman conceived with her own oocytes by means of IVF. Three fresh embryos were transferred, resulting in a pregnancy and delivery by cesarean section at 35 weeks of a healthy male baby weighing 2,300 g. CONCLUSION(S) Extensive literature search suggests that this is the first case report of live birth in a 50-year-old woman after IVF-ET with her own oocytes. This is a very rare and unusual case that deviates from the norm and therefore warrants attention. In selected cases, assisted reproductive technology might be reasonable to try for a limited number of times with a womans own oocytes.

Endometrial receptivity markers in infertile women stimulated with letrozole compared with clomiphene citrate and natural cycles

Abstract Though various advantages associated with the use of letrozole as an ovulation inducing drug are documented, there is inadequate information regarding its effect on endometrial receptivity. Expression of endometrial receptivity markers including αvβ3 integrin, L-selectin, LIF, and pinopods during the implantation window in infertile women stimulated with letrozole or clomiphene citrate (CC), and spontaneous cycles is investigated. A total of 36 women were included in the study out of which 16 women were diagnosed with primary unexplained infertility and the other 20 women with severe oligoasthenozoospermic/azoospermic male partners. Both groups were sub-divided into three groups; women stimulated with letrozole, or with CC, and the third group was allowed to ovulate spontaneously (natural cycles). Women having natural cycles and not given any drug were considered as controls. Once ovulation was confirmed by ultrasonography, endometrial samples were collected on the seventh day post-ovulation and analyzed. In women with unexplained infertility treated with letrozole and CC, epithelial and stromal expression of αvβ3 integrin, L-selectin, leukemia inhibitory factor (LIF), and pinopod formation was found to be significantly higher as compared to controls. Expression of these receptivity markers was found to be comparable amongst the letrozole, CC, and control groups in women with severe oligoasthenozoospermic/azoospermic male partners. Cell cycle analysis showed similar cell cycle phase fractions on comparing the CC and the letrozole groups. Stimulation with letrozole and CC appears to enhance endometrial receptivity in women with unexplained infertility. However, letrozole and CC did not have any significant effect on the endometrial receptivity markers of women with severe oligoasthenozoospermic/azoospermic male partners.

Assessment of sub-endometrial blood flow parameters following dydrogesterone and micronized vaginal progesterone administration in women with idiopathic recurrent miscarriage: A pilot study

To evaluate differences in uteroplacental blood flow and pregnancy outcome in women with idiopathic recurrent spontaneous miscarriage (IRSM) following administration of micronized vaginal progesterone and oral dydrogesterone.

Analysis of spindle characteristics and embryo quality in mice stimulated with letrozole using Polscope imaging

OBJECTIVE To explore the effect of letrozole as an ovulation inducing agent on oocyte and embryo quality in mice model. DESIGN Prospective study. SETTING Institute of Reproductive Medicine, Kolkata, India. ANIMAL(S) Sixty-nine sexually mature female Swiss Albino mice 6-7 weeks old. INTERVENTION(S) Metaphase II (MII) oocytes from two groups of mice, one group injected with letrozole and the other with rFSH. MAIN OUTCOME MEASURE(S) Number of MII oocytes, number of oocytes with meiotic spindle (MS), different angles of MS relative to the polar body (PB), spindle characteristics, and fertilization outcome. RESULT(S) The MS was present in 84% and 71.2% of the oocytes in the letrozole and rFSH group, respectively. In the letrozole group, 73% of the oocytes had a 0 degrees spindle position compared with 35.7% in the rFSH group. With letrozole, 19% of the oocytes had 0 degrees > MS < 90 degrees and 8% had MS >90 degrees , compared with 46.1% oocytes with 0 degrees > MS < 90 degrees and 18.2% with MS >90 degrees with rFSH. Mean spindle area retardance, spindle dimensions, and 4-8-cell embryo formation rate were significantly higher with letrozole compared with rFSH. Two-cell and blastocyst formation rates were similar in both groups. CONCLUSION(S) Robust birefringent spindles were obtained in mice on superovulation with letrozole. Letrozole does not appear to increase the risk of spindle assembly and preimplantation developmental arrest in mouse oocytes.