Spermatozoa from azoospermic males can be retrieved from either the epididymis or the testis, depending on the type of azoospermia, using different surgical methods such as percutaneous epididymal sperm aspiration (PESA), testicular sperm aspiration (TESA), testicular sperm extraction (TESE), and microsurgical testicular sperm extraction (micro- TESE). After collecting the epididymal fluid or testicular tissue, laboratory techniques are used to remove contaminants, cellular debris, noxious microorganisms, and red blood cells. Processed spermatozoa may be used for intracytoplasmic sperm injection or eventually be cryopreserved. However, spermatozoa collected from either the epididymis or the testis are often compromised and more fragile than ejaculated ones. Therefore, sperm processing techniques should be used with great caution to avoid jeopardizing the sperm fertilizing potential in treatment cycles. In this review, we describe the current methods for processing surgically-retrieved specimens, either fresh or frozen- thawed, and provide the tips and pitfalls for facilitating the handling of such specimens. In addition, we present the available laboratory tools to aid in the identification of viable immotile spermatozoa to be used in conjunction with assisted reproductive techniques. Review of the literature was carried out using PubMed and Science Direct search engines.

Development of ART has great benefit for millions of couples all over the world and with falling fertility rate there are a growing numbers of children born with the help of ART, it is important to investigate potential risks to these children. IVF-ICSI pregnancies are associated with higher risk for multiple gestation, preterm labor and low birth weight. It is an area of great conflict and interest that whether ART is associated with increased congenital malformations or not. So, this article reviews the data and evidences linking ART to congenital malformations.

Context: Recurrent miscarriages, the loss of three or more consecutive intrauterine pregnancies before 20 weeks of gestation with the same partner, affect 1%-1.5% of the pregnant population. The inadequate secretion of progesterone in early pregnancy has been proposed as a cause of recurrent miscarriages. Aims: The aim was to investigate the efficacy of progesterone supplementation in patients with unexplained recurrent miscarriages. Settings And Design: This was a 9-year cohort study of women with otherwise unexplained recurrent miscarriages who attended a recurrent miscarriage clinic in a tertiary care university hospital. Subjects and Methods: Women with at least three unexplained recurrent miscarriages were included in the study. They were divided into three groups according to their initial and 48-h repeat progesterone levels. For women with inadequate endogenous progesterone secretion, natural progesterone vaginal pessaries 400 mg 12-hourly were offered until 12 weeks gestation. Statistical Analysis: Proportions and 95% confidence intervals calculated for categorical variables and the chi-square test were used to show statistical significance. Medians and ranges were calculated for noncontinuous variables. Results: Pregnancy cycles (n = 203) were analyzed to examine the miscarriage rate following progesterone supplementation. Overall live birth and miscarriage rates were 63% and 36%, respectively. When analyzed by the number of previous miscarriages there was a reduction in the miscarriage rate following progesterone supplementation in women with 4 previous miscarriages when compared with historical data. Conclusions: Progesterone supplementation may have beneficial effects in women with otherwise unexplained recurrent miscarriages.

Background: Oocyte donation is an invaluable therapy for couples with impending or complete ovarian failure. In addition, oocyte donation affords a scientific opportunity to study the unique biologic participation of the uterus in the process of human embryo implantation. Aim: To identify the recipient variables that may have a significant impact on pregnancy outcome in order to optimize results of an oocyte donation program. Design and Settings: A prospective study conducted from March 1, 2010 to March 31, 2011 at a private tertiary care IVF Clinic. Materials and methods A total of 270 recipients resulting in embryo transfer as a result of oocyte donation were enrolled. Clinical and Ongoing pregnancy rates, Implantation rates were calculated according to different age groups, Endometrial thickness, Indication, Day and number of embryos transferred. Data was evaluated as chi square analyses with comparative significance determined at P <.05. Results: In recipients less than 40 years of age, higher ongoing pregnancy and implantation rates (41.9% and 24.6%) were seen as compared to recipients above 40 years (21.8% and 13.18%) respectively. Pregnancy and implantation rates increased with increasing endometrial thickness but the difference was not statistically significant. A higher ongoing pregnancy rate (40.9% vs.28.8%) and implantation rate (23% vs.19.6%) was demonstrated with Day 3 embryo transfer compared to Day 2 transfer. Conclusion: A declining endometrial receptivity may result in lower implantation and pregnancy rates in recipients above the age of 40 years, more pronounced after the age of 45 years. An endometrial thickness of >8 mm is considered ideal before transfer. Transfer of two selected embryos on day 3 yields a favorable pregnancy outcome with reduced multiple pregnancy rates. Recipient's age above 45 years has negative impact on pregnancy outcome whereas embryo transfers on Day 3 yields better pregnancy

Aim: To correlate the ease or difficulty of embryo transfer and blood at catheter tip with pregnancy rate when embryo transfer (ET) was performed by the same operator using soft catheter. Materials and Methods: A retrospective analysis of 342 patients who underwent in vitro fertilization or ICSI cycle from January 2008 to December 2010 in a single centre was done. The type of transfer was divided into two groups: 'easy' or 'difficult'. Transfer was considered difficult when additional instrumentation was required or firmer catheter was used or required changing of catheter. Patients undergoing cryo-preserved ET were excluded from the study. Results: On the day of transfer in 284 (83%) patients, ET was easy and difficulty was encountered in 58 (17%) patients. Blood at catheter was seen in 101 (29.53%) patients. In the group of 58 difficult transfers, 10 pregnancies resulted with a clinical pregnancy rate of 17.2%, while 67 pregnancies resulted in 284 cycles of easy transfer with clinical pregnancy rate of 23.6% (P value = 0.045). While no significant difference was seen in pregnancies with blood on outer catheter and blood less transfer, there was significant reduction in pregnancy rate when blood was present on catheter tip compared to bloodless transfer (13.3% v/s 24.1; P value = 0.032). Conclusion: Reduction in clinical pregnancy rate is seen with difficult ETs, more when blood is present at the catheter tip.

Objective: To compare Letrozole (5 mg) and clomiphene citrate (100 mg) as first line ovulation induction drug in infertile PCOS women. Study Design: Prospective Randomised trial. Setting: A Tertiary level infertility centre. Patients: 103 infertile PCOS women Intervention(S): Treatment naοve infertile PCOS women were randomised to treatment with 5 mg letrozole (51 patients) or 100 mg clomiphene citrate (52 patients) daily starting day 2 to day 6 of menstrual cycle. Timed intercourse or Intra Uterine Insemination (IUI) was advised 24 to 36 hours after Human Chorionic Gonadotropin (HCG) injection. Main Outcome Measures: Ovulation rate, mono or multi follicular rate, days to ovulation, endometrial thickness, serum progesterone, serum estrogen, pregnancy rate, miscarriage rate. Results: The mean age, Body Mass Index (BMI), duration of infertility in both Clomiphene Citrate (CC) and Letrozole groups were similar.Ovulation rate was 73.08% in letrozole group and 60.78% in CC, which was not statistically significant (P=0.398). There was no statistically significant difference between Endometrial thickness (CC 7.61 ±1.96, Let 7.65 ± 2.10), Sr E ₂ on day of HCG (CC 178.3 ± 94.15, Let 162.09 ± 73.24), Days to ovulation (CC 14.2 ± 3.41; Let 13.13 ± 2.99) and Sr P ₄ on D ₂₁ (CC 10.58 ± 6.65; Let 11.86 ± 6.51). Monofolliculo genesis (CC 54.84, Let 79.49 %, P=0.027) and Pregnancy rate (CC 7.84%, Let 21.56% P=0.0125) were statistically significantly higher in letrozole group. Conclusion: Our study shows that letrozole has excellent pregnancy rates compared to clomiphene citrate. Letrozole should be considered at par with clomiphene citrate as first line drug for ovulation induction in infertile PCOS women.

Aims: The purpose of this study was to identify the differentially expressed genes (DEG) in human testis and also evaluate the relationship between human testis, human Embryonic Stem Cells (hESC), mouse testis and mouse ESCs (mESC). Settings and Design: It is a prospective analysis designed computationally. Methods and Material: The microarray data for human testis, hESCs, mouse testis and mESCs were obtained from NCBI-GEO and analyzed for identification of DEGs. The results were then compared with mouse testis and extended to ESCs. Statistical Analysis Used: Data was analyzed in R using various Bioconductor packages. To identify DEGs, 2-fold cut-off and a False Discovery Rate (FDR) below 0.01 criterions was used. Results: A total 2868 transcripts (DEGs) were found to be significantly up-regulated and 2011 transcripts significantly down-regulated in human testis compared to other normal tissues. Of the up-regulated transcripts, 232 transcripts were grouped as unclassified i.e. had unknown annotations at the time of analysis. Gene Ontology (GO) based functional annotation of testis specific DEGs indicate that most of the DEGs (~80%) are involved in various metabolic processes. Pathway analysis shows over-representation of Ubiquitin-mediated proteolysis pathway. A core group of 67 transcripts were found to be common among human testis, mouse testis, hESCs and mESCs. Conclusions: Testis seems to be metabolically very active relative to other normal tissues as indicated by functional annotation. The comparison of human and mouse testis shows conserved functions and pathways involved in both species. Large numbers of genes were found conserved between testis and ESCs suggesting very close expression level relationship between reproductive organs and complex phenomenon such as dedifferentiation and reprogramming.

Aim: This study aims to compare the efficacy of AFC and AMH, as markers for ovarian reserve. Materials and Methods: 75 patients with PCO (polycystic ovaries) undergoing IVF were randomized with 75 non-PCO patients. On day 3, volume of ovary was acquired, ovarian volume was defined by VOCAL, and Sono AVC was used to count the number of antral follicles. Sum total of antral follicles in both ovaries was taken as total antral follicle count (AFC). AMH was measured on the same day. Long agonist protocol with recombinant FSH (rFSH) was used for IVF stimulation till at least two follicles of 18 mm were seen. hCG 10,000 iu was given and ovum pick up was done after 34-35 h. Primary end point was number of follicles >12 mm seen on day of hCG. Final end point was number of ova retrieved on ovum pick up. Correlation of AFC and AMH was checked for both end points and with each other. Results: Correlation of AFC and follicles >12 mm on day of hCG in PCO group is 0.56 and non-PCO group is 0.63, ¹ and for AMH and follicles >12 mm on day of hCG in PCO group is 0.42 and non-PCO group is 0.47. Correlation of AFC with number of ova retrieved on OPU in PCO group is 0.44 and for non-PCO group is 0.50. The value for AMH is 0.39 in PCO and 0.43 for non-PCO group. Comparing correlation of AFC and AMH for primary end point in PCO group has 'z' value 1.11(onetailed significance 0.1335, twotailed significance 0.267) and in non-PCO group comparison shows a 'z' value of 1.39 (one tailed significance 0.0823, two-tailed significance 0.1645). Therefore in both groups, AFC and AMH correlates with total number of follicles >12 mm on day of hCG, but both AFC and AMH have independent significance. Comparing correlation of AFC and AMH with number of ova retrieved on OPU, in non-PCO group has 'z' value of 0.54(one tailed 0.2946, two-tailed 0. 5892). In PCO group, this comparison shows, 'z' value of 0.36(one tailed 0.3594, two tailed 0.7188). Conclusion: AFC and AMH correlate with each other but have independent significance for estimating follicles >12 mm on day of hCG. For ova retrieved on OPU, in PCO group, AMH and AFC have similar significance. In non-PCO, AMH may increase accuracy of estimation of number of ova retrieved on OPU over AFC, but difference is not significant. AFC only may suffice for estimation of number of ova retrieved on OPU for all practical purposes.

Objectives: To evaluate whether three daily doses of GnRH agonist (Inj. Lupride 1 mg SC) administered 6 days after oocyte retrieval increases ongoing pregnancy rates following embryo transfer (ET) in cycles stimulated with the long GnRH agonist protocol. Settings And Design: Prospective randomized controlled study in a tertiary care center. Materials and Methods: Four hundred and twenty six women undergoing ET following controlled ovarian stimulation with a long GnRH agonist protocol were included. In addition to routine luteal-phase support (LPS) with progesterone, women were randomized to receive three 1 mg doses of Lupride 6 days after oocyte retrieval. Computer-generated randomization was done on the day of ET. Ongoing pregnancy rate beyond 20 ^th week of gestation was the primary outcome measure. The trial was powered to detect a 13% absolute increase from an assumed 27% ongoing pregnancy rate in the control group, with an alpha error level of 0.05 and a beta error level of 0.2. Results: There were 59 (27.69%) ongoing pregnancies in the GnRHa group, and 56 (26.29%) in the control group ( P = 0.827). Implantation, clinical pregnancy and multiple pregnancy rates were likewise similar in the GnRHa and placebo groups. Conclusions: Three 1 mg doses of Lupride administration 6 days after oocyte retrieval in the long protocol cycles does not result in an increase in ongoing pregnancy rates.

Cystic adenomyosis is a rare form of adenomyosis mostly seen in middle aged women.We report two cases of cystic adenomyosis in juvenile patients, which simulate uterine malformation and presented as a diagnostic dilemma. The first patient initially was diagnosed as uterus bicornis with a hematometra in obstructed rudimentary horn while the second patient was diagnosed as broad ligament fibroid. Surgical exploration by laparoscopic approach confirmed the diagnosis and excision of the cystic mass relieved the symptoms of the patients.

Despite advances in diagnostic techniques, approximately 10,000 babies with β-thalassemia major are born annually in India. Pre-implantation genetic diagnosis (PGD), an alternative to prenatal diagnosis, helps in negative selection of affected embryos prior to implantation. Hereby, we report the first successful β-thalassemia PGD pregnancy in an Indian carrier couple. β-Thalassemia mutation analysis by Amplification-Refractory Mutation Sequence (ARMS)-polymerase chain reaction (PCR) in the parents, followed by PGD for β-thalassemia mutation in embryos in two consequent in vitro fertilization (IVF) cycles, with transfer for three β-thalassemia minor embryos, resulted in singleton successful pregnancy, the results of which were confirmed on prenatal diagnosis. With advances in assisted reproductive techniques and molecular diagnosis, PGD for monogenic diseases is feasible in high-risk couples. The methodology in the current study included two rounds of PCR using fluorescently labeled primers, fragment analysis using the ABI 3100 nucleotide sequencer and the GeneMapper software, purification, and concentration of PCR product, which enabled distinct clear peaks making the analysis and interpretation non-ambiguous.

This paper presents a rare anatomical variation in the form of accessory fallopian tube on right side. The duplication of fallopian tube was observed in a 34-year-old female during routine undergraduate dissection in our department. Fallopian tube is the part of uterus that carries the ovum from the ovary to the uterus. Accessory fallopian tube is the congenital anomaly attached to the ampullary part of main tube. This accessory tube is common site of pyosalpinx, hydrosalpinx, cystic swelling and torsion. The ovum released by the ovary may also be captured by the blind accessory tube leading to infertility or ectopic pregnancy. Hence, all patients of infertility or pelvic inflammatory disease should be screened to rule out the presence of accessory fallopian tube and if encountered should be removed.

Mullerian duct anomalies, though rare, can be a treatable cause of pelvic pain and infertility. Various complex Mullerian duct anomalies may exist with combination of features of more than one class. Since there are no precise clinical or imaging criteria to enable specific categorisation, there is ambiguous classification of these anomalies by various radiologists and clinicians. A young female presented with complaints of chronic pelvic pain, primary amenorrhoea and infertility. The patient was evaluated by sonography and Magnetic Resonance Imaging and diagnosed as case of complex mullerian duct anomaly, a unicornuate uterus with cervical dysgenesis and cavitated, noncommunicating, rudimentary right horn. The findings were confirmed on laprohysteroscopy and the patient underwent hystertectomy. There should be an integrated clinico-radiological classification scheme and familiarity with rare and complex anomalies for appropriate diagnosis and management of complex Mullerian duct anomalies.