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Endometriosis is a common benign gynecological disease, characterized by proliferation of functional endometrial glands and stroma outside the uterine cavity. The present study involves investigation of alterations in the serum proteome of endometriosis patients compared to healthy controls using 2DE and 2D-DIGE combined with MALDI TOF/TOF-MS. Comparison of serum proteome of endometriosis patients and healthy subjects revealed 25 significant differentially expressed proteins. Gene ontology and network analysis, performed using PANTHER, DAVID, WebGestalt and STRING, revealed that the differentially expressed proteins are majorly involved in response to stimulus, immune system, metabolic, localization and cellular processes. For serum diagnostic marker identification, several robust statistical screening procedures were applied to identify the set of the most significant proteins responsible for successful diagnosis of different endometriosis stages. Partial least squares (PLS) based marker selection tool and orthogonal partial least squares-discriminant analysis (OPLS-DA) were used to identify the most significant proteins for disease prediction. Western blotting validation in a separate cohort of patients revealed that haptoglobin (HP), Ig kappa chain C region (IGKC), alpha-1B-glycoprotein (A1BG) can be considered effective serum protein markers for the diagnosis of Stage II, III and IV endometriosis. For diagnosis of Stage I, only IGKC and HP seemed promising.Globally, about 12 in 100 women of reproductive age are diagnosed with endometriosis. The pathogenesis of the disease still remains unclear, leading to non-specific therapeutic approaches for disease management. Moreover, there is a delay of 8-12years in correct diagnosis after the initial onset of symptoms leading to a considerable impact on the woman's lifestyle. Also, the gold standard for diagnosis of endometriosis, laparoscopy, is an invasive procedure. The value of a noninvasive or semi-invasive diagnostic test for endometriosis with easily accessible fluids such as plasma, serum, urine, and saliva is, therefore, rightfully recognized. The present study is expected to considerably improve the understanding of the disease pathogenesis along with improved diagnostics and therapeutic approaches leading to better management of the disease.

Though the role of reactive oxygen species (ROS) in female infertility has been a subject of rigorous research worldwide, there is inadequate information on the cut-off value of ROS in the oocyte microenvironment beyond which ART outcome may be adversely affected. An upper ROS level in follicular fluid (FF) samples of women undergoing IVF beyond which good quality embryo formation is unlikely, is established. ROS, lipid peroxidation and total antioxidant capacity were estimated. The upper cut-off ROS level beyond which viable embryo formation is not favorable was found to be approximately 107 cps/400 microl FF. This level, determined in women with tubal factor infertility, was further validated in women with endometriosis and PCOS and correlated with fertilization and pregnancy rate and embryo quality. Summarizing, a threshold level in FF has been established for the first time beyond which ROS may be considered toxic for viable embryo formation and pregnancy outcome.

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° spindle position compared with 35.7% in the rFSH group. With letrozole, 19% of the oocytes had 0° > MS 90°, compared with 46.1% oocytes with 0° > MS 90° 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.

Objective: To evaluate the significance of an estrogen-upregulated 64.0-kDa human uterine fluid (hUF) glycoprotein in relation to promotion of fertility. Design: Prospective study. Setting: Department of Assisted Reproductive Biology Unit, Institute of Reproductive Medicine, Kolkata. India. Patient(s): Sixty-three women with unexplained infertility having normal ovulatory cycle with normal endocrine profile and 18 parous women. Intervention(s): Women either received no stimulation (n = 35) or were stimulated with clomiphene citrate (CC) alone (100 mg/day from day 3 to day 7; n = 56) or in combination with pure FSH (75 IU on day 3 and day 8; n = 54) and were subjected to IUI. Parous women (n = 18) receiving no treatment served as control and were followed-up for spontaneous ovulation. Main Outcome Measure(s): Uterine fluid protein profile, relative intensity of 64.0-kDa protein, number of mature follicles, endometrial thickness, and pregnancy. Result(s): Expression of the 64.0-kDa protein exhibited positive correlation with prevailing estradiol levels, but the degree of the protein response to estrogen was comparatively blunted in the CC-incorporated cycles. Endometrial thickness and pregnancy outcomes correlated positively with the expression of the protein. Conclusion(s): The 64.0-kDa hUF protein perhaps plays a role in endometrial receptivity to support pregnancy. Failure of pregnancy despite documented ovulation in CC-stimulated cycles may be due to its attenuating effects on expression of the protein.

pregnancy rate and LBR in non-donor sperm, but this association is not observed in donor sperm; this lack of association in donor group may be due to small sample size. Furthermore, our data suggest that male factor infertility in cycles using non-donor sperm (despite high sperm counts) continues to compromise pregnancy outcomes as compared to donor sperm. References: 1: Freour T, Jean M, Mirallie S, Langlois ML, Dubourdieu S, Barriere P. Predictive value of CASA parameters in IUI with frozen donor sperm. Int J Androl. 2009 Oct;32(5):498-504. 2: Marshburn PB, McIntire D, Carr BR, Byrd W. Spermatozoal characteristics from fresh and frozen donor semen and their correlation with fertility outcome after intrauterine insemination. Fertil Steril. 1992 Jul;58(1):17986. 3: ByrdW, BradshawK, Carr B, Edman C, Odom J, AckermanG. A prospective randomized study of pregnancy rates following intrauterine and intracervical insemination using frozen donor sperm. Fertil Steril. 1990 Mar;53(3):521-7. 4: Tan O, Ha T, Carr BR, Nakonezny P, Doody KM, Doody KJ. Predictive value of postwashed total progressively motile sperm count using CASA estimates in 6871 non-donor intrauterine insemination cycles. J Assist Reprod Genet. 2014 Sep;31(9):1147-53.

Objective To surgically reconstruct rare Mullerian duct anomalies with varying degrees of bladder exstrophy and congenital absence of ampullary portion of fallopian tube. Design Case report. Setting Institute of Reproductive Medicine, Salt Lake, Kolkata, India. Patient(s) Two cases of anteriorly displaced vagina with stenotic introitus, divergent pubic rami, and varying degrees of bladder exstrophy (group A) and three cases of congenital absence of ampullary portion of the fallopian tube with intact but blind fimbria attached to mesoovarium (group B). Intervention(s) Introitoplasty and tubal anastomosis were performed for groups A and B, respectively. Main Outcome Measure(s) Description and treatment of two types of patients with rare Mullerian anomalies. Result(s) Three spontaneous pregnancies and viable deliveries, two in group A and one in group B, were achieved after surgical reconstruction. Conclusion(s) Assisted reproductive technologies usually provide a solution to infertility in rare Mullerian anomalies; reconstructive surgical procedures specific to the type of anatomical defect may also be considered for spontaneous pregnancy with viable deliveries.

Objective To report two cases of advanced pelvic endometriosis, both with deep rectal involvement and one with extensive cervical involvement appearing like a carcinoma, where both patients achieved spontaneous pregnancy and delivered viable babies. Design Case report. Setting Institute of Reproductive Medicine. Patient(s) Two nulliparous patients, one with extensive cervical and rectal and other with deep rectal endometriosis and primary infertility. Intervention(s) Medical management. Main Outcome Measure(s) Description and treatment of two patients with advanced pelvic endometriosis and pregnancy. Result(s) Delivery of viable babies in both cases. Conclusion Prolonged medical treatment may have helped to arrest the progression of the disease by restoring anatomic proximity leading to spontaneous pregnancy in both cases.

Objective: To evaluate the efficacy of prolonged gonadotropin-releasing hormone-agonist (GnRH-a) hormonal suppression on IVF-ET outcome in the patients with moderate/severe grades of endometriosis (AFS grades III & IV). A retrospective analysis was done on 117 cycles of 54 patients undergoing IVF-ET treatment for endometriosis-associated infertility. Materials/Methods: Diagnosis and staging of endometriosis were confirmed by laparoscope only in 33 women, while laparoscope was followed by laparotomy in 21 cases. The patients were randomly distributed into 2 group-I (n � 25) received GnRH-a hormonal suppression for a period of 6 months followed by IVF-ET. Patients failing to achieve pregnancy in the first cycle were subjected to IVF-ET in subsequent cycles, maximum for 3 cycles (total 53 cycles), using conventional protocol of controlled ovarian hyperstimulation. The remaining 29 patients (group-II) had conventional ‘long’ protocol GnRH-a down regulation. In this group, like in group-I, IVF-ET was performed maximum for 3 cycles (total 64 cycles). Treatment outcome was assessed in terms of ovarian response, rates of clinical pregnancy/cycle, cumulative pregnancy and pregnancy wastage. The prolonged GnRH-a suppressed group resulted in significantly (p � 0.01) increased the rates of oocyte retrieval (7.9 � 1.5) and transferable embryos per cycle (3.9 � 1.3) in comparison to those of the conventionally stimulated group (oocytes: 4.9 � 1.5; transferable embryo: 2.8 � 0.8). The pregnancy rate per cycle was statistically comparable between the groups (group-I: 15% vs. group-II: 9%), but in group-I, cumulative pregnancy rate after 3 cycles was significantly (p � 0.05) higher (group-I: 52% vs. group-II: 27%) and rate of miscarriage was lower (group-I: 12% vs. group-II: 33%). In moderate and severe grades of endometriosis-associated infertility, prolonged GnRH-a hormonal suppression is an effective regime for improving cumulative pregnancy and take home baby rates following IVF-ET.

Klinefelter syndrome (47,XXY) with male phenotype is the most common sex chromosomal abnormality. It is believed that SRY (sex determining region on Y) is the major gene necessary to induce the undifferentiated bipotential gonadal primordium to develop as testis. Recently, several SRY-boxrelated (SOX) genes have been identified on autosomes. Mutations in the SRY or SOX gene have been implicated in sex reversal. We report an individual having 47,XXY chromosome constitution, normal SRY, SOX9, and ZFY genes and yet with a female phenotype. A 15-year-old phenotypic female has well developed breasts; prominent labioscrotal folds; an enlarged clitoris; feminine pubic hair distribution; urethra in the centre of the vestibule; a urogenital sinus 5 cm deep; a uterus; prepubertal in shape and size, palpable per rectum; and a cervix which could not be palpated. Ultrasonography of the abdomen revealed normal shape of urinary bladder with smooth and regular walls. Uterus was small in size, with normal shape and outline. Myometrium was homologous, cavity was empty, midline echo was normal. Laparoscopic assessment revealed a normal uterus. Right ovary was normal in size and shape with a cyst and a normal Fallopian tube. Left ovary was not seen as it was hidden beneath an epithelial fold. Chromosome analysis showed 47,XXY karyotype in all 100 metaphases analysed. Hormone profiles showed average female range of corticotrophin, T3, T4, thyrotrophin, and testosterone. Progesterone and prolactin were higher when compared with normal female levels. Histology of the right gonad showed features compatible with ovarian stroma. No RESEARCH LETTERS