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Setting Studies have suggested a possible role of antitubercular treatment (ATT) for treating infertility in women with latent or preclinical tubeculosis (TB). Objective This study investigates the effect of latent TB on IVF outcome. Study design This was a prospective cohort study which included women referred for assisted reproductive technique between December 2013 to February 2015. Two hundred forty-five women who consented for an endometrial biopsy for TB DNA PCR (Deoxy-ribo nucleic acid-polymerase chain reaction) in the premenstrual phase were included in this study. Methods Nested TB DNA PCR for IS6110 and MPB64 was performed on endometrial samples of the women. The effect of latent TB (positive endometrial DNA PCR) was estimated in terms of statistical difference in clinical pregnancy rate. Results Endometrial TB DNA PCR was positive for 19 women (7.7%). The clinical pregnancy rate was similar in both groups (28.16 vs. 26.3%; p = 0.5) without ATT. Patients with both positive and negative polymerase chain reaction (PCR) were similar in age, duration and type of infertility, and the infertility diagnosis. Conclusion Positive endometrial TB DNA PCR (latent TB) alone does not require antitubercular drugs for treatment of infertility. TB DNA PCR should not be the sole criterion for diagnosis of genital TB and starting or concluding treatment.

Context: Therapeutic proteins can cause immune responses, which may have clinical implications. Aims: The aim of the study was to assess the immunogenicity of recombinant human follicle-stimulating hormone (r-hFSH), when used for controlled ovarian stimulation (COS). Settings and Design: Prospective, multicenter study conducted at reproductive medicine clinics in India and Vietnam. Materials and Methods: A total of 285 women, aged 20–40 years, undergoing 354 COS cycles for either intrauterine insemination (IUI) or in vitro fertilization (IVF) were studied. The primary outcome measure was the incidence of development of anti-drug antibodies (ADA) and their neutralization potential. Other outcome measures were follicle development, dose and duration of r-hFSH, positive serum pregnancy test, clinical pregnancy, cycle cancellation, and adverse events (AEs). Statistical Analysis Used: A sample size of 250 was planned. Descriptive statistics are presented. Results: Four patients tested positive for ADA after r-hFSH administration at different time points; all of them tested negative, subsequently. None were found to have neutralization potential. The mean dose and duration of r-hFSH were 816 IU and 8.1 days in IUI and 2183 IU and 9.5 days in IVF, respectively. The serum and clinical pregnancy rates were 12.4% and 11.6% in IUI and 32.7% and 29.9% in IVF cycles, respectively. Seven AEs were reported, including two cases of ovarian hyperstimulation syndrome; two AEs were judged to be serious. Conclusions: The tested r-hFSH has very low immunogenic potential and did not lead to the development of neutralizing antibodies. The overall efficacy and safety of the drug were in-line with existing literature data, and no specific clinical impact of immunogenicity could be identified.

Context: A new indigenous recombinant human chorionic gonadotropin (r-hCG) has been developed in India with a comparable pharmacological profile to that of Ovitrelle® (Merck Serono). Aims: This study aims to compare the efficacy and safety of the new r-hCG with that of Ovitrelle for induction of ovulation in women undergoing intrauterine insemination (IUI). Settings and Design: Randomized (2:1), multicenter, open-label, equivalence clinical trial conducted in India. Subjects and Methods: A total of 217 women, aged 20–37 years, undergoing IUI were administered the new r-hCG (test) 250 mcg or Ovitrelle 250 mcg (comparator) after ovarian stimulation with gonadotropins. The ovulation rate was compared as the primary outcome. In addition, pregnancy rates, incidence of adverse events (AEs), and development of immunogenicity were assessed. Statistical Analysis Used: The ovulation and pregnancy rates were compared using Chi-squared test with statistical significance at P < 0.05. Results: With 144 women in the test group and 73 in the comparator group, the ovulation rate (85.4% vs. 78.1%; P = 0.17) and pregnancy rate (serum β hCG test) (11.8% vs. 12.3%; P = 0.91) were similar in both groups. A total of 15 AEs were reported (11 in the test r-hCG group and 4 in the comparator group) in 11 women; none of these were serious, and all were judged to be unrelated to the study drug. No subject developed immunogenic reaction to the test drug. Conclusions: The new preparation of r-hCG was equivalent to the conventional preparation of r-HCG in the induction of ovulation in patients undergoing IUI.

To compare pregnancy, miscarriage and live birth rates and cycle monitoring parameters between Natural Cycle (NC-FET) and Hormone replacement cycle (HRC-FET) in eumenorrhoeic women undergoing vitrified-warmed autologous embryo transfer.Single-centre retrospective cohort study analyzed 173 NC-FET and 507 HRC-FET cycles with transfer of day2/3/5/6 embryos. Natural cycle monitoring occurred with serial ultrasound with the first day of the scan determined by the shortest cycle frequency. Serum progesterone was ordered when ultrasound was ambiguous in ascertaining ovulation. For HRC-FET oral estradiol valerate was used in fixed or escalating doses with maximum daily dose of 12 mg. Transdermal estradiol gel was added when desired endometrial thickness was not achieved. Vaginal progesterone was introduced with Endometrial thickness(ET) = 7 mm. Embryos were transferred after stage-appropriate progesterone exposure. Luteal support was given with vaginal progesterone in NC-FET and vaginal and oral progesterone in HRC-FET. Primary outcome was live-birth-rate. Secondary outcomes were ET, length-of-estrogenic-phase, numbers-of-ultrasoundshormone-monitoring, pregnancymiscarriage rate. The odds ratio for live-birth was adjusted for age, embryo number, previous-live-births, previous-losses, past-negative-ET-cycles, IVF-indication and embryo-developmental-stage. Quantitative variables were compared using unpaired-t-test and qualitative variables with chi-square test. Two tailed p-value 0.05 was considered significant. Binary logistic regression was used to calculate adjusted odds for live-births.The two cohorts were comparable in age, infertility-duration, previous-live-births, previous-losses, past-negative-ET-cycles, IVF-indication and embryo-developmental-stage. Length-of-estrogenic-phase was significantly shorter for NC-FET than HRC-FET 14.32 ± 2.83vs.18.18 ± 4.48; p = 0.0001) as was mean ultrasound-monitoring-scans (2.73 ± 0.95vs. 3.3 ± 1.04; p = 0.0001). Mean-endometrial-thickness (8.75 ± 1.83vs. 8.5 ± 1.25; p = 0.25) and mean-hormonal-tests (1.75 ± 1.28 vs. 1.88 ± 0.69; p = 0.09) did not differ significantly between NC-FET vs HRC-FET. Significantly higher live births took place in NC-FET vs. HRC-FET (87/173 = 50.3%vs.204/507 = 40.2%;p = 0.026). No significant difference was found in pregnancy rate (66.5% vs. 58%; p = 0.058) or in the pregnancy loss rate (24.3%vs30.6%; p = 0.23). The odds ratio for live-births adjusted for relevant variables was 1.48 (1.03-2.13) in NC-FET compared to HRC-FET.NC-FET is a superior method of endometrial preparation compared to HRC-FET in eumenorrhoeic women since it has a shorter estrogenic phase, reduces patient visits to the hospital and improves live birth rates. Future adequately powered studies should look at antenatal and perinatal outcomes, patient satisfaction rates and cost-effectiveness in the two endometrial preparation regimes.

Cervical ectopic pregnancy is an extremely rare form of ectopic pregnancy with potential risk of massive bleeding and associated morbidity. Managing this condition is challenging for clinicians due to the serious risk to patient and dilemma faced in deciding the appropriate management plan. This case report describes the role of uterine artery embolisation in managing a case of cervical ectopic pregnancy with heavy bleeding per vaginum post methotrexate treatment with falling beta human chorionic gonadotropin (HCG) levels. It highlights the unpredictable nature of this condition and need for prompt intervention in an emergency situation.

Follicular aspiration under transvaginal ultrasound guidance is performed as part of assisted reproductive technology (ART) to retrieve oocytes for in vitro fertilization (IVF). However, controversy as to whether follicular flushing following aspiration yields a larger number of oocytes than aspiration only is ongoing. The aim of this study was to assess the safety and efficacy (oocyte recovery rates) of follicular flushing when compared with aspiration only performed in women undergoing ART. The study was conducted at Sir Gangaram Hospital, New Delhi. Retrospective data of 100 females undergoing ART was taken from September 2019 to September 2020. Patients were divided into two groups of 50 each. In first group, patients underwent oocyte retrieval using double lumen needle (flushing technique) and in the second group, single lumen needle (no flushing) were used. Females with age ≤37 years who were having ≤9 follicles, size ≥14 mm during oocyte retrieval when undergoing ARTs. Categorical variables were presented in number and percentage (%) and continuous variables, as mean ± standard deviation and median. Normality of data was tested by Kolmogorov–Smirnov test. If the normality was rejected, then nonparametric test was used. Quantitative variables were compared using Mann–Whitney test. Follicular flushing had significantly better oocyte recovery rates when compared with no flushing (P 0.05) in M2 rates in patients undergoing intracytoplasmic sperm injection (ICSI) when compared in both the groups. There was no significant difference (P > 0.05) in fertilization rates in patients undergoing IVF and ICSI, respectively, in both the groups. Double lumen needle with flushing technique gives better oocyte recovery rates in low responder women undergoing ARTs. Hence, follicular flushing may be considered in such patients for oocyte retrieval.

Introduction: There is a paucity of data evaluating role of endometrial blood flow in frozen thawed embryo transfer cycles (FET cycles) in hormone replacement treatment (HRT) cycles. Objective: To compare pregnancy outcomes in women undergoing ultrasound for endometrial morphology alone to those with endometrial morphology with blood flows in frozen embryo transfer cycles. Patients: Patients aged 23 to 38 years undergoing first two frozen thawed single blastocyst embryo transfer cycles. From August 2019 to January 2020, 163 patients undergoing hormone replacement frozen embryo transfer cycles were enrolled in the study. Patients were divided in two groups. Patients in group A (n = 92) had ultrasonographic evaluation of endometrial thickness and morphology and in group B (n = 71) patients underwent ultrasound for endometrial thickness, morphology along with Doppler blood flow studies. Outcome measures: Primary outcome was clinical pregnancy rate. Secondary outcome measures were pregnancy loss rate, ongoing pregnancy rate, and ectopic pregnancy rate. Results: Clinical pregnancy rate in group A and group B was 54.34% and 47.88%. Clinical pregnancy rate was similar between both the groups (P = 0.779). Ongoing pregnancy rate in group A and B was 48.91% and 40.84%, respectively. Pregnancy loss rate (7–13 wks) was 5.4% and 7.04% in group A and group B, respectively. There was no significant difference in ongoing pregnancy rate and pregnancy loss rate between both the groups (P = 0.756). Conclusion: Additional measurement of doppler studies do not help in improving implantation and thus seems unnecessary in improving pregnancy rates and reducing pregnancy loss rates in FET cycles.

The present study presents a case report of a rare anomaly called accessory cavitated uterine mass (ACUM). The patient, 26 years old, had presented with severe dysmenorrhea which was refractory to the medical treatment. The hysteroscopic examination was normal and laparoscopic examination showed a subserosal bulge, present over anterior and lateral fundal region above the attachment of the round ligament. Chromo-pertubation was performed and dye came freely from both the tubes but not from the cavity of this uterine mass. The chocolate-colour material was drained during excision of the mass. The final histopathology report showed myometrial tissue lined by collection of hemosiderin laden macrophages as well as endometrial cuboidal lining and occasional endometrial glands.

Objective: To study the prevalence of metabolic syndrome (MBS) in Indian women and to see how does it correlate to body mass index (BMI) and polycystic ovarian syndrome (PCOS) in this population. Study Design: Prospective cross-sectional observational study. Setting: Infertility clinic of a tertiary center. Materials and Methods: Two hundred women, 120 with PCOs and 80 age-matched controls were enrolled. The prevalence of MBS was studied in the women with and without and was co related to BMI by further subgrouping as team (BMI 23 kg/m2). The sample size was: team controls-40, obese controls-40, team PCOS-80. Each subject underwent a physical examination and laboratory evaluation for the diagnosis of MBS, which was defined according to the guidelines of National Cholesterol Education Program Adult Treatment Pamel (NCEP ATP III) 2005. INTERVENTION: None. Main Outcome Measures: Main Outcome Measures: Subjects with and without PCOs were compared with each other for the prevalence of MBS, and similarly team subjects were compared with obese subjects. Receiver operator characteristic (ROC) curves were obtained for both the PCOS and non PCOS population separately, co-relating the prevalence of MBS with BMI. These ROC curves were used to establish the cut off values of BMI, which could best predict the risk of MBS. Results: The prevalence of MBS was significantly higher in the women with PCOS, as compared to age-matched controls. Similarly, when BMI was considered, MBS was more prevalent in overweight subjects than in lean subjects with or without PCOS. In subgroup analysis, the presence of PCOS had a lesser impact on the prevalence of MBS as compared to non-PCOS controls with higher BMI. The relative risk of MBS increased as follows: lean controls-1, lean PCOS-2.66, obese controls-5.33, and obese PCOS-6.5. The most appropriate cut-off level of BMI for predicting the risk of MBS in Indian women without PCOS seems to be 23 kg/m 2 , whereas, with PCOS, it was 22.5 kg/m 2 . Conclusion: MBS is more prevalent in women with PCOS. However, obesity is an independent and stronger risk factor for developing MBS. To reduce the risk of MBS and its related long-term health consequences, lifestyle modification is advisable above BMI of 23 kg/m 2 in the normal population and 22.5 kg/m 2 in women with PCOS.

Study question: Oocyte donation is an increasingly popular treatment option for pregnancy and parenthood in women with poor ovarian reserve or advanced age. Indian literature still lacks a formal study focusing on expectations and experiences of oocyte donors. Aim: To assess demography, expectations and experiences of oocyte donors in northern India by an interview-based questionnaire. Material and Methods: This is a prospective cross-sectional study at a tertiary care IVF centre from August 2018 to January 2019. Sample size was 65 oocyte donors found fit and willing for egg donation. Interviews were taken using structured questionnaire at the time of registration and after successful oocyte retrieval. Results: Mean age of donors was 25 years. Most of the donors were housewives (70%) and rest were in a private job. Only 29% of donors were educated above tenth standard. Almost all the donors (96%) were commercial donors introduced by paid agents or friends. Most of the donors had very poor understanding of details of the oocyte retrieval procedure. Only 36% of donors felt that compensation given is adequate, rest expected a higher compensation. Average compensation given to donors was INR 15,000–25,000, which was 5000–10,000 less than their expectations. All donors were satisfied with the procedure and did not feel exploited or anxious or depressed but few complained of discomfort experienced in visits and minimal complications like pain, nausea, and fever. None of the donors developed signs/symptoms of OHSS. Average number of oocytes retrieved per donor was 17. Mean dose of gonadotropins required per donor was 1495 IU. Conclusion: Our findings show that the oocyte donation has an overall good acceptance and satisfaction among the donors. However there is a need to impart more detailed information to donors in order to prevent their possible exploitation.

Introduction: There are limited studies in literature regarding the effect of body mass index (BMI) on clinical pregnancy rate following frozen-thawed embryo transfers. Objective: We aim to study the independent effect of BMI on clinical pregnancy rate in single blastocyst frozen-embryo transfer cycle in Asian women. Material and methods: It is a prospective observational study involving 167 women who underwent single, good quality frozen-blastocyst transfer following a uniform protocol. Stimulation was done by hormone replacement therapy in all cycles. The study population was divided into various cohorts as per the BMI classification for Asian adult population (normal: 18.5–22.9, pre-obese: 23–24.9, obese I: 25–29.9, obese II: ≥30). Results: The positive β-HCG rate was 48.93% in normal BMI women, 55.31% in pre-obese, 50% in obese I, and 38.4% in obese II BMI groups. The clinical pregnancy rate was 36.17% in normal BMI women, 48.93% in pre-obese, 45% in obese I, and 30.07% in obese II BMI groups. The difference between various BMI subgroups as regards to positive β-HCG as well as clinical pregnancy is not statistically significant. Women with BMI ≥ 30 had numerically low clinical pregnancy rate, in comparison to those with lower BMI sub-groups. However, this difference was not statistically significant. Conclusion: This study concludes that BMI did not affect clinical pregnancy rate among women following a uniform protocol, single good quality frozen blastocyst transfer in Asian women. The increased difficulty during transfer for women with higher BMI suggests that body habitus may be responsible for difficult transfers, although this may not translate into a worse clinical pregnancy rate. A study with larger sample size may be needed to confirm these findings.

The outcome of ovulation induction in anovulatory polycystic ovarian syndrome (PCOS) may depend, in part, on the pharmacologic compounds used, but also on individual patient characteristics, such as age, body mass index (BMI), hyper- androgenism, luteinizing hormone (LH), hyper-secretion, anti mullerian hormone (AMH) levels and possibly antral follicle count (AFC) with ovarian volume of these women. There exists a subset of clomiphene citrate (CC) resistant PCOS women who require stimulation of ovulation with high doses of human menopausal gonadotropin (hMG), after not having responded to chronic low dose step up regimes of recombinant follicle stimulating hormone (r-FSH).

Objectives: (a) To establish the cut-off levels for anti-Mullerian hormone (AMH) in a population of Indian women that would determine poor response. (b) To determine which among the three ie.,: age, follicle stimulating hormone (FSH), or AMH, is the better determinant of ovarian reserve. Study Design: Prospective observational study. Setting: In vitro fertilization (IVF) unit of a tertiary hospital. Materials and Methods: The inclusion criterion was all women who presented to the center for in-vitro fertilization/Intracytoplasmic sperm injection (IVF/ICSI). The exclusion criteria were age >45 years, major medical illnesses precluding IVF or pregnancy, FSH more than 20 IU/L, and failure to obtain consent. The interventions including baseline pelvic scan, day 2/3 FSH, luteinizing hormone (LH), estradiol estimations, and AMH measurement on any random day of cycle were done. Subjects underwent IVF according to long agonist or antagonist protocol regimen. Oocyte recovery was correlated with studied variables. The primary outcome measure was the number of oocytes aspirated (OCR). Three categories of ovarian response were defined: poor response, OCR ≤ 3; average response, OCR between 4 and 15; hyperresponse, OCR > 15. Results: Of the 198 patients enrolled, poor, average, and hyperresponse were observed in 23%, 63%, and 14% respectively. Correlation coefficient for AMH with ovarian response was r = 0.591. Area under the curve (AUCs) for poor response for AMH, subject's age, and FSH were 0.768, 0.624, and 0.635, respectively. The discriminatory level of AMH for prediction of absolute poor response was 2 pmoL/l, with 98% specificity and 20% sensitivity. Conclusions: AMH fares better than age and FSH in predicting the overall ovarian response and poor response, though it cannot be the absolute predictor of non-responder status. A level of 2 pmol/l is discriminatory for poor response.

BackgroundThis report describes a patient counseling approach, conservative and surgical management (cervical cerclage) of a dichorionic-diamniotic twin pregnancy, where delivery of the second twin followed the delivery of the first by 104 days.Case presentationA 35-year-old Indian lady, with 10 years of primary infertility, conceived twins, following ovulation induction with gonadotropin therapy. At 19 weeks and 4 days gestation, she aborted a female fetus of 410 gm. Tocolytic therapy was initiated; the placenta remainedin situand the umbilical cord of twin A was ligated high and McDonald's cerclage was placed and prophylactic antibiotics initiated. But since the patient's uterus continued to be irritable and the cerclage appeared to be under tension, at 20 weeks 6 days the stitch was taken out and all conservative treatment stopped for 24 hours, but reinitiated after this period, since the uterus remained quiescent. After 3 weeks of uterine inactivity, at 23 weeks 6 days gestation, McDonald's suture was reapplied following which conservative management was offered after counseling for possible risks associated with maternal sepsis, coagulopathy, need for extended hospitalization and potential for hysterectomy. Reassuring fetal status was observed for twin B without evidence of contractions or chorioamnionitis. A viable male infant (1,800 gm) was delivered vaginally at 34 weeks and 4 days gestation.ConclusionThis report outlines a counseling approach useful for patients with premature delivery of one twin, and presents application of conservative obstetrical management principles for the aftercoming twin, even when delivery interval is extreme.How to cite this articleSingh TA, Majumdar A. Successful Obstetrical Management of Over 100-day Interval between the First and Second Twin Delivery in an Infertility-Treated Patient: Counseling and Management Approach to Extreme Asynchronous Twin. Int J Infertility Fetal Med 2012;3(3): 97-101.

Microdeletions in the azoospermia factor region on the long arm of Y chromosome are associated with spermatogenic failure. There are many markers for the diagnosis of Y chromosome microdeletion analysis, but in routine practice only a limited set of markers can be tested.The objectives of this study were to determine the frequency of Y chromosome microdeletion in idiopathic cases of male infertility in India, to attempt genotype-phenotype correlation, and to evaluate whether markers to be tested for diagnosis of Y chromosome microdeletion should be ethnicity specific.Microdeletions in the Y chromosome were analyzed in 200 infertile males. The six sequence tag site (STS) markers prescribed by the European Academy of Andrology (EAA) were used initially. Patients in whom no deletions were detected by use of these markers were tested by markers selected from other studies from India.The STS markers prescribed by EAA detected deletions in only 6 (3%) of 200 infertile males. However, markers selected from previous Indian studies showed deletions in an additional 15 (7.5%) of infertile males. Overall, Y chromosome microdeletions were observed in 21 (10.5%) of 200 patients. Of these, 13 were cases of azoospermia and 8 were cases of severe oligospermia.The markers prescribed by EAA alone are not suitable for the diagnosis of Y chromosome microdeletions in infertile males. The protocol for identification of Y chromosome microdeletions in cases of nonobstructive azoospermia/severe oligospermia would have to include a different set of STS markers.

Background This report describes a patient counseling approach, conservative and surgical management (cervical cerclage) of a dichorionic-diamniotic twin pregnancy, where delivery of the second twin followed the delivery of the first by 104 days. Case presentation A 35-year-old Indian lady, with 10 years of primary infertility, conceived twins, following ovulation induction with gonadotropin therapy. At 19 weeks and 4 days gestation, she aborted a female fetus of 410 gm. Tocolytic therapy was initiated; the placenta remained in situ and the umbilical cord of twin A was ligated high and McDonald's cerclage was placed and prophylactic antibiotics initiated. But since the patient's uterus continued to be irritable and the cerclage appeared to be under tension, at 20 weeks 6 days the stitch was taken out and all conservative treatment stopped for 24 hours, but reinitiated after this period, since the uterus remained quiescent. After 3 weeks of uterine inactivity, at 23 weeks 6 days gestation, McDonald's suture was reapplied following which conservative management was offered after counseling for possible risks associated with maternal sepsis, coagulopathy, need for extended hospitalization and potential for hysterectomy. Reassuring fetal status was observed for twin B without evidence of contractions or chorioamnionitis. A viable male infant (1800 gm) was delivered vaginally at 34 weeks and 4 days gestation. Conclusion This report outlines a counseling approach useful for patients with premature delivery of one twin, and presents application of conservative obstetrical management principles for the aftercoming twin, even when delivery interval is extreme.

Aims: To study the prevalence of clinical manifestations in obese and lean polycystic ovarian syndrome (PCOS) women and their health hazards. Settings and Design: This prospective study was carried out in a tertiary care infertility clinic from 1.7.2005 till 31.12.2007. Materials and Methods: These women were diagnosed to have PCOS by the European Society of Human Reproduction and Embryology and the American Society of Reproductive Medicine, Rotterdam 2003 criteria. They were further divided into two groups according to their body mass index (BMI): Group A (n = 300), overweight and obese with BMI >23 and Group B (n = 150), normal weight and lean with BMI ≤23. Stastical Analysis and Results: The prevalence of menstrual irregularities [79.2% vs. 44%, P = 0.000, 95% confidence interval (CI) = 0.26-0.44)] and clinical hyperandrogenism (74.2% vs. 50.6%, P = 0.000, 95% CI=0.14-0.32) was signifi cantly higher in the obese group, whereas android central obesity (waist to hip ratio >0.85) was similar in both groups, irrespective of body weight (47.7% vs. 38%, P = 0.056, 95% CI=0.06 to +0.18). Comparative data of various health manifestations in lean vs. obese women with POCS [Table 4]. Of the health risk manifestations, hypertension occurred in both groups with a similar frequency (41% vs. 35.5%, P = 0.261, 95% CI=0.03 to +0.15). Group A showed an increased prevalence of IGT (25% vs. 10%, P = 0.000, 95% CI= 0.13-0.29) and type two diabetes mellitus (11.7% vs. 6%, P = 0.000, 95% CI= 0.13-0.29) as compared with group B. endometrial hyperplasia (EH) also showed an increase prevalence in Group A compared with Group B (5.6% vs. 2%, P = 0.055, 95% CI= 0.01-0.08), although not statistically significant. Conclusion: PCOS emerges as a clinically heterogeneous condition with increased prevalence of health risks such as hypertension, diabetes and EH. Of these, diabetes and EH appear to be more prevalent in the obese, putting them at a greater risk of morbid problems at a much younger age than the lean ones.

To study the impact of vitrification of in vitro matured caprine oocytes on in vitro fertilization (IVF), ovaries were collected from a local abattoir and goat oocytes aspirated. The oocytes were subjected to in vitro maturation (IVM) for 27 h. Twenty-seven hours post IVM oocytes were cryopreserved in a vitrification solution (VS) consisting of propylene glycol (40%, w/v) and trehalose (0.25 mol l−1) in PBS with 4% (w/v) BSA. Some oocytes were exposed to propylene glycol without cryopreservation to check the degree of damage induced by propylene glycol. In the vitrified group, 94.3% of the oocytes were viable for IVF. 16.1% of the oocytes were damaged after thawing. From the vitrified group, 75.8% of the oocytes were subjected to IVF, while for 18 h. Cytological studies following fixing of the IVF oocytes revealed 17.37% of the oocytes showing a pronucleus in the control group, compared to 8.08 and 7.86% in the vitrified and cryoprotectants exposed groups, respectively. This study revealed that goat oocytes can be successfully cryopreserved using a simple vitrification method. However, the fertilizing ability of oocytes was severely affected by both vitrification and exposure to cryoprotectants.

Oocytes were aspirated from goat ovaries, washed in TCM-199 containing 10% estrus goat serum (EGS) and subjected to different variables of vitrification viz. exposure to propanediol before maturation of oocytes but without freezing, to detect the level of damage attributed to propanediol alone. The removal of cumulus cells to mimic damage attributed to osmotic stress during cryoprotectant exposure or vitrification procedures and vitrification with propanediol (40%, w/v) and trehelose (0.25 mol l −1 ) in PBS supplemented with 4% (w/v) BSA. The oocytes (n = 432) after treatment with different variables of vitrification were thawed and cultured for 27 h in TCM-199 with 0.5 g/ml FSH, 100 IU/ml LH, 1g/ml E2 and 10% EGS over a granulose cell monolayer. After 27 h of culture the matured oocytes were separated from the cumulus cells by treating them with PBS containing 0.1% hyaluronidase and by passing them through a fine pipette, fixed in acetic alcohol, stained with aceto orcein and observed under a phase contrast microscope for evidence of maturation. Similarly, oocytes (n = 184) following treatment of different variables of vitrification were thawed, matured and fertilized in a TALP medium. After 18 h of co-incubation with spermatozoa, oocytes were denuded, fixed in acetic alcohol, stained with aceto orcein and observed under phase contrast for evidence of fertilization. Maturation and fertilization rates were 27.5, 58.8, 46.8 and 74.1% and 17.8, 28.0, 40.0 and 54.4% for vitrified, exposed, denuded and control oocytes, respectively. These results indicate vitrification to be detrimental for in vitro maturation and in vitro fertilization of goat oocytes. © 2004 Published by Elsevier B.V.

In the present study the developmental potential of fetal fibroblasts was evaluated using nuclear transfer. Studies were undertaken to workout suitable dc pulse length on electrofusion of fetal goat skin fibroblast cells with enucleated in vitro matured goat oocytes and their capability of forming embryos. Skin fibroblast cells from fetal skin were isolated and cultured in monolayer using RPMI-1640 media with 10% FCS in 5% CO 2 incubator at 38.5±1 °C with 95% humidity. Recipient oocytes recovered from slaughterhouse ovaries were matured in vitro for 22–24 h and enucleated using a Leitz micromanipulator. Serum starved cells, cytochalasin B blocked synchronized cells and growing cells were used for insertion into the perivitelline space of the enucleated oocyte and electrofused with constant ac pulse at 7–10 V, while dc pulses at 250, 300 and 350 V were applied for 5–20 μs followed by culture in a CO 2 incubator to observe proper electrofusion and cleavage. After fusion the embryos were activated with cytochalasin B for 2–4 h, and then were transferred to IVC media (without any supporting cells and growth factors). During the study, it was observed that fetal fibroblast became confluence within 2 days. Proportions of oocytes undergoing zona dissolution did not differ significantly among the three dc pulses during electrofusion after reconstruction of the embryos, and also there was no significant difference for the fusion rates among serum starved cells, cytochalasin B blocked cells and growing cells. It was concluded that around 300 V resulted in better electrofusion, and cytochalasin B blocked synchronized cells and fast growing skin fibroblast cells of goats could be used for nuclear cloning.

Follicles are the site of many important and complex phenomena during preovulatory and ovulatory periods, which lead to the processes of luteinization, steriodogenesis, ovum maturation and ovulation. To understand the steriodogenesis within the follicles during early- and late-luteal phase ovaries, experiments were conducted in goat ovarian follicles of >3 mm in size. Estradiol–17β (E2), progesterone (P4), testosterone (T), follicle stimulating hormone (FSH), luteinizing hormone (LH) and prolactin (PRL) activity were localized in early- and late-luteal phase ovaries using immuno-histochemical techniques. The enzymes involved in steroid synthesis i.e. glucose–6–phosphate dehydrogenase (G–6PD), –3β hydroxy steroid dehydrogenase (–3βHSD) and –17β hydroxysteroid dehydrogenase (–17βHSD) were localized histochemically. Results revealed that during the early luteal phase, follicular theca and granulosa cells showed almost no FSH, LH and PRL activity but was present in the follicular fluid of such follicles. Among the steroid hormones, testosterone was found to be less during the early luteal phase, E2 and P4 were located in the theca and granulosa cells as well as in the follicular fluid. The gonadotrophins and steroid hormones were comparatively more in the larger follicles than in the smaller ones. In the late luteal phase, all the gonadotrophins and steroid hormone concentrations were higher than the early luteal phase. FSH was bound on the granulosa and theca cells of mainly the large follicles. LH and PRL were also observed in the theca cells of such follicles. Even the follicular fluid showed an increased localization of these hormones. All the three steroids were observed in the follicular fluid as well as in theca and granulosa cells in a similar patterns during early- and late-luteal phase, except the concentration, which was higher during the late luteal phase. All the enzymes studied were much higher in follicles of the late luteal phase as compared to the early luteal phase ovaries. It may be concluded from this study that there probably are very few gonadotrophin receptors present in the follicular cells during the early luteal phase and hence less steriodogenesis occurred during the early luteal phase compared to the late luteal phase.

The effect of oxytocin in addition to an prostaglandin (PGF2a) injection, given on day 12 of the oestrous cycle (at the time of fifth and sixth FSH injection) in goats pretreated with human chorionic gonadotrophin (hCG) on day 7 of the oestrous cycle was studied. Sixteen cyclic local nondescript goats were randomly allocated to control (without oxytocin, n=8) and treatment (oxytocin, n=8) superovulation groups. All goats in the control group received a total of 20 NIH FSH S1 units divided into eight doses (4/4, 3/3, 2/2 and 1/1 units) over 4 days (day 10 to 13 of the cycle) and 10 mg dinoprost tromethamine (PGF2a) on day 12. Goats in the treatment group in addition also received 500 IU hCG on day 7 of the oestrous cycle and 50 IU oxytocin intramuscularly with the fifth and sixth FSH injection on day 3 of the superovulatory treatment. Goats in both the groups were administered 500 IU hCG, 10–16 h after the onset of oestrus and naturally serviced at 12 hourly intervals till the end of the oestrus. Laparotomies were performed on all goats on day 6 for assessment of ovarian response and embryo recovery. Blood samples were taken on day 7, (before hCG or any pretreatment), day 10 (before beginning of FSH administration), day 12 (before PGF2a administration), day 0 (day of superovulatory oestrus) and day 6 (day of flushing) for the estimation of the progesterone. There was a significant decrease in the number of corpora lutea/donor in the treatment group compared to the control group (0.8±0.3 vs 5.1±0.9; P

To identify common and varying practice patterns used by in vitro fertilization (IVF) providers from a broadly distributed, worldwide survey. Specific information regarding clinical IVF practices involving the oocyte maturation triggering, oocyte retrieval and embryo transfer was elicited.This is an internet-based questionnaire study of IVF practices throughout the world. We used 26 multiple choice questions regarding common clinical practices. The data reported are weighted based on the number of IVF cycles performed at the specific IVF center, represented by a single respondent.Surveys were completed from 359 centers in 71 countries throughout the world. The most common practice patterns (defined as ≥75 % of IVF cycles) identified included: use of human chorionic gonadotropin (hCG) for trigger with an antagonist protocol, no routine patient monitoring from hCG trigger to oocyte retrieval, timing oocyte retrieval 34-37 h following oocyte maturing trigger, use of a single lumen retrieval needle, no routine tests following oocyte retrieval prior to patient discharge and use of ultrasound assistance with embryo transfer.This is the largest and most diversely represented survey of specific IVF practices addressing oocyte maturation triggers, oocyte retrieval and embryo transfers. Several uniform practice patterns were identified that can be correlated with evidence-based medicine; however, we identified multiple variable practice patterns which is likely the result of the absence of definitive evidence to guide IVF practitioners. The results of this survey allow IVF providers to compare their specific practice patterns with those of a global diverse population of IVF providers.