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There has been a paradign shift in the status of immunoassays. There used to be a time where immunoassays had a very narrow role in clinical medicine, but that is not the case in today's world. Immunoassays have taken a central role in helping us better understand and treat human diseases. The literature around anti-ovarian antibodies (AOA) immunoassay testings have been conflicting. Researchers challenged the specificity of the reported assays, but a systematic study was never elucidated on what/who the trouble maker was in rendering these tests so nonspecific. Attempts were made by our group in Mumbai, India, to throw light on the culprit behind the nonspecificity casative factor in the immunoassays and a method to overcome this was reported and published. This review highlights the stories back five and a half decades to date, to demonstrate where the status of AOA testing was, is and will be.

Diagnosis of female genital tuberculosis (FGTB) remains elusive due to the paucibacillary nature of the disease. We evaluated if analysis of inflammatory pathways of endometrial tissue could establish a better diagnosis of FGTB.One hundred and four infertile women suspected of having GTB or having been treated for GTB in the past, underwent endometrial biopsies for diagnosis and Gene Inflammatory Pathways analysis at our center between 2018-2020. Diagnosis of FGTB was based on acid-fast bacilli culture, immunocytochemistry, nested-polymerase chain reaction, histopathological examination, TB GeneXpert, or combinations thereof. Gene expression profiles were also analyzed.Based on diagnostic tests of 104 women, 44 (42%) were considered TB-positive, 35 (34%) TB-negative, and 25 (24%) TB-negative after TB treatment in the past. Inflammatory pathways were significantly upregulated in TB-positive women versus TB-negative (41% vs. 6%; p = .0005), and in women who were TB-negative after TB treatment in the past versus TB-negative (never treated for TB in the past) (38% vs. 6%; p = .0037). Two-hundred seventy-one genes were upregulated, and 61 genes were downregulated in TB-positive women versus those who were TB-negative. Differentially expressed genes were mapped to various interlinked inflammatory signaling pathways, including mitogen-activated protein kinase (MAPK), Natural Killer (NK) cells, nuclear factor kappa-B (NF-kB), tumor necrosis factor (TNF), and Toll-like receptors (TLR) signaling.Inflammatory pathways and gene expression profiles add to the diagnostic tools to identify TB-positive women at an early stage. The results from this study are still experimental and large multi-centric studies are suggested before their recommendation in routine clinical practice.

We here report a case of a 23-year-old female from Mumbai, Maharashtra, India who was detected to carry the α chain variant Hb J-Norfolk [HBA2: c.173G>A (or HBA1]. She had no clinical symptoms and was referred to us for routine investigations and screening. An abnormal peak was detected on both high performance liquid chromatography (HPLC) and capillary electrophoresis (CE) with a fast-moving band on cellulose acetate electrophoresis. There is no detailed study on the HPLC and CE pattern of this hemoglobin (Hb) variant, and therefore, this study will help in detecting and avoiding missing these variants during routine investigations and population screening. This is the first report of this variant in the Indian population.

The technique of pre-implantation genetic diagnosis (PGD) by fluorescence in situ hybridization (FISH) in cases of repeated miscarriages due to parental balanced inversions and translocations is relatively new in India. In a couple with a history of recurrent miscarriages and implantation failures, karyotyping done in three laboratories showed that the husband had an insertion or inversion of chromosome 12. Hence, they were referred to us for PGD. The anomaly turned out to be more complex. A pre-PGD workup using a series of FISH probes on metaphases accompanied by reflex FISH was required to characterize the anomaly. For subsequent PGD, single blastomeres were biopsied from seven embryos obtained by intracytoplasmic sperm injection. FISH analysis had to be carried out using ten probes in four rounds. On pre-PGD workup for inversion 12 by FISH, an additional anomaly of a cryptic translocation between 9qter and 12qter was detected in the husband. His complex karyotype according to the detailed ISCN nomenclature was therefore 46,XY,t(9;12)(9pter→9q34.1::12q24.2→12qter),der(12)inv(12)(12pter→12p11.2::12q24.2→12p11.2::9q34.1→9qter). After PGD, the normal and balanced embryos transferred, resulted in the birth of healthy twins conceived in the first cycle itself. Therefore, a pre-PGD workup is important and needs reflex FISH in the event of an unexpected cytogenetic anomaly. PGD will need the analysis of additional chromosomes on the same cell by FISH in such cases. An experienced in vitro fertilization and Genetics team is essential for success. This is the first report of PGD by FISH for an inversion coupled with a cryptic translocation from India.

Antibodies to multiple ovarian antigens have been proposed as markers of ovarian autoimmunity. The role of ovarian autoantibodies has been widely discussed in the pathophysiology of premature ovarian failure and unexplained infertility, but the autoantigens are yet to be identified. Three immunodominant ovarian autoantigens, α-actinin 4 (αACTN4), heat shock 70 protein 5 (HSPA5) and β-actin (ACTB), have been identified using anti-ovarian antibody-positive sera from women with idiopathic premature ovarian failure ( n =50) and women undergoing IVF ( n =695), using mass spectrometry. These autoantigens were subsequently validated using Western blot, immunohistochemistry and enzyme-linked immunosorbent assay. These autoantigens are localized to different components of the ovary such as the ooplasm of the oocyte, theca, granulosa, corpus luteum and zona pellucida. All the above antigens were found to be expressed in the ooplasm throughout follicular development. All the autoantigens are expressed specifically in the oocyte except αACTN4. The three autoantigens could contribute to the array of biomarkers to be used for developing specific and sensitive tests for diagnosis of women at risk of premature ovarian failure and IVF failure due to ovarian autoimmunity and could give an insight into the molecular mechanisms involved in the pathophysiology of these conditions. Anti-ovarian antibodies (AOA) have been reported in women with premature ovarian failure and unexplained infertility. Among women with infertility, those with evidence of ovarian autoimmunity appear to have poorer IVF–embryo transfer outcomes. Diagnosis of an autoimmune mechanism in these pathologies has relied for a long time on the detection of AOA. Little is known about the precise ovarian antigenic targets in terms of its molecular and cellular identities that are recognized by the antibodies and immune cells in autoimmune diseases of the ovary. In the present study, we observed that 31% of the total women recruited under an IVF–embryo transfer programme (group I) and 46% of women with premature ovarian failure (group II) tested positive for AOA. Three immunodominant ovarian autoantigens, namely non-muscle α-actinin 4, heat shock 70 protein 5 and cytoplasmic β-actin, were identified using mass spectrometry and validated and characterized using AOA-positive sera from women from both groups. Further investigation of the identified targets could give us an insight into the molecular mechanism involved in the pathophysiology of human ovarian autoimmunity.

A couple with a history of five early miscarriages due to a balanced reciprocal translocation t(6;19) (p22;q13.4) in the wife, was referred for preimplantation genetic diagnosis (PGD) by fluorescence in situ hybridization (FISH). After oocyte retrieval, nine embryos were obtained. One cell from each embryo was biopsied, fixed and subjected to FISH using centromere and subtelomere probes for chromosomes 6 and 19. Five embryos which had unbalanced translocations, were not transferred. A balanced translocation or absence of translocation was seen in three embryos, which were transferred. One embryo had an anucleate cell and subsequently arrested. A pregnancy was achieved in the first intracytoplasmic sperm injection cycle itself and resulted in the birth of a healthy baby. This is the first baby after PGD for a reciprocal translocation in India. In 2010, the team of the authors reported the first successful pregnancy after PGD for a Robertsonian translocation and a normal child was born.

KEYWORDS Prader-Willi. Angelman. Williams. DiGeorge. Cytogenetics. Fluorescence. Autism ABSTRACT Fluorescence in situ hybridization (FISH) has facilitated the detection of microdeletions seen in Prader- Willi/Angelman (PW/AS), Williams and DiGeorge syndromes. Out of 374 suspected cases tested at Jaslok Hospital in the past 5 years, 73 were positive, including 29 cases of Angelman, 16 of Prader-Willi, 24 of Williams and 4 of DiGeorge syndrome. Male preponderance was seen, mainly in Williams syndrome. The mechanisms causing Prader- Willi and Angelman syndrome include microdeletions, intragenic mutations, uniparental disomy and imprinting defects, though FISH can only detect microdeletions. Metaphase FISH helped to detect 1 case each with deletion of the control (PML) signal and duplication of the critical PW/AS region, which are associated with autism. One suspected case of Prader-Willi syndrome had a Robertsonian translocation t(14;15)(q10;q10) which led to a deletion of a major part of the SNRPN region in 10% cells, resulting in low-grade mosaicism. Another FISH-positive case was due to a reciprocal translocation t(2;15)(q37;q11), where loss of critical genes at the breakpoint on chromosome 15 caused the Prader-Willi phenotype. FISH in a child with an Angelman phenotype showed no microdeletion, though Trisomy 15 was seen in 1 metaphase suggesting uniparental disomy due to trisomy rescue. A known polymorphism in the form of an additional tiny green signal on chromosome 14 was observed in 17 of 284 (6%) cases studied for Prader-Willi/Angelman syndrome. Another inherited polymorphism was seen in 5 cases, where one control signal was very small. Prenatal diagnosis was carried out with normal results, in 12 women with a previously affected child.

Preimplantation genetic testing (PGT) is an early form of prenatal genetic diagnosis where abnormal embryos are identified, thereby allowing transfer of genetically normal embryos. This technology has become an integral part of Assisted Reproductive Technology (ART) procedures. Initial experiments with animals as early as 1890 and those in the mid and later part of the last century paved the forward path of ART and PGT. This review article covers the evolution of PGT and is a pointer toward current and fast-evolving technology, allowing scientists and doctors to better comprehend human reproduction, and ensure healthy pregnancy outcomes.

Serum anti-ovarian antibodies (AOAs) have been shown in autoimmune premature ovarian failure and in vitro fertilization-embryo transfer (IVF-ET) cases. The specificity of assays detecting these antibodies has been questioned. Researchers have used several techniques (e.g., ELISA and indirect immunofluorescence). Few have reported on the non-specificity and the type of molecular and cellular targets. We reported earlier on the presence of naturally occurring anti-albumin antibodies as the likely factor for non-specificity. Having developed a novel blocking recipe, we show substantial elimination of this non-specificity. With these standardized tests, we hereby report multiple targets at protein and histological levels. In our study group, 15 of 50 (30%) patients with premature ovarian failure and 13 of 50 (26%) IVF-ET patients showed the presence of AOAs. Western blotting showed a large number of patients making AOAs to a 90-kDa protein, followed by 97- and 120-kDa proteins. Histochemically, it was evident that the sera of these patients predominantly react with the oocyte; other somatic cellular targets are also involved. The specific non-invasive test developed by us was found to be useful because it could carry out a reliable diagnosis of an autoimmune etiology that would be very helpful to select patients in whom immune-modulating therapy could be recommended, which in turn may restore ovarian function and fertility.

Objective To evaluate the effect of the use of cumulus-aided embryo transfer on pregnancy rates. To study the proximity of expanded cumulus cells to the developing embryo. To document by light microscopy the anchoring of day 3 to day 4 embryos by the expanded cumulus cells. To demonstrate by transmission electron microscopy the cellular activity of the expanded cumulus cells. To evaluate the expression of growth factors (vascular endothelial growth factor, interleukin-6, insulin-like growth factor I) that are secreted by the cumulus cells. Design A comparative study of a group of women undergoing cumulus coculture and cumulus-aided embryo transfer, with those who underwent cumulus coculture but did not undergo cumulus-aided embryo transfer. The endpoint was the achievement of pregnancy. Setting Department of Infertility Management and Assisted Reproduction, Jaslok Hospital and Research Centre, Mumbai, India. Patient(s) Five hundred seventeen women undergoing treatment for infertility using intracytoplasmic sperm injection and embryo transfer and fulfilling set criteria. To validate our initial results, we conducted a similar study on 208 women where randomization was performed. Intervention(s) Embryos were cocultured with the patient's own cumulus cells and were transferred into the uterus with approximately 30 μL of the expanded cumulus cells. Main Outcome Measure(s) Pregnancy, implantation, and multiple gestation rates. Result(s) Our study demonstrated a significant increase in the implantation rate in the study group (group A) of 25.6% versus 14.5% in the control group (group B) and a significant increase in the pregnancy rate in the study group (group A) of 47.6% versus 34% achieved in the control group (group B). Although the incidence of multiple gestation was similar (38.6% in the study group and 32.9% in the control group), the higher-order multiple gestation rate was significantly more in the study group as compared with the control group (18.1% vs. 2.4%). Similar pregnancy and implantation rates were observed in the randomized study. Conclusion(s) This study demonstrates the efficacy of cumulus-aided embryo transfer, using autologous cumulus cells. It indicates a significant increase in implantation and pregnancy rates. The results suggest that cumulus cells play an important role in embryonic development, and that they may provide a mechanism to improve embryo–uterine adhesion by physical proximity, and by secreting cytokines and growth factors required to aid the implantation process.

Polymorphic variants on chromosomes are considered 'normal', as heterochromatin has no coding potential and nucleolar organizing regions (NOR) contain genes coding for rRNA. Variants have been reported in infertility and recurrent abortions. With refined molecular techniques, genes for fertility and viability are now thought to reside in heterochromatin. DNA sequence analysis of human chromosome 9 has shown that it is highly structurally polymorphic, with many intrachromosomal and interchromosomal duplications, and contains the largest autosomal block of heterochromatin. Transcriptional activation of constitutive heterochromatic domains of the human genome in response to environmental stress was reported recently. Heat shock triggers the assembly of nuclear stress bodies on the pericentromeric heterochromatin of human chromosomes including chromosome 9. These are characterized by an epigenetic status typical of euchromatic regions. On acrocentric chromosomes, NOR-associated protein count and morphology was reported to separate benign and malignant melanocytic lesions. Hence all variants may not be 'normal'. The present study of karyotyping 842 individuals attending an IVF clinic with primary infertility or repeated miscarriages, showed polymorphic variants in 28.82% of males and 17.19% of females, which was quite high. It is suggested that variants should not be ignored by cytogeneticists. Screening prospective gamete donors for chromosome variants may help enhance the success of IVF.

KEYWORDS Male infertility; ICSI; semen analysis; PGD; AZF deletions; azoospermia; OAT ABSTRACT Male partners of infertile couples are known to frequently have abnormal semen parameters. Some of these cases are due to underlying genetic factors such as Y chromosome microdeletions, an abnormal karyotype or cystic fibrosis mutations. Y chromosome microdeletions generally cannot be detected by karyotyping. At our clinic we undertook a study of male partners of infertile couples to determine the frequency and common loci of Y chromosome microdeletions in India, using the PCR technique. We studied 100 patients mainly having azoospermia (AZ) or oligoasthenoteratozoospermia (OAT). Multiplex PCR analysis for 18 loci on the Y chromosome was carried out using commercially available kit (Promega Version 1.1). Y chromosome microdeletions were observed in 12/100 (12%) patients including 8/27 (29.63%) with azoospermia, 3/56 (5.35%) with oligoasthenoteratozoospermia and 1/ 7 (14.28%) with only asthenoteratozoospermia. All loci of the DAZ gene were deleted along with DYS237 and DYS236 from AZFd in 5/27 (18.52%) azoospermic males studied. The most commonly deleted loci were DYS240 in 11/12 (91.67%) and DYS219 in 7/12 (58.33%) patients with microdeletions. The use of ICSI in such patients can lead to transmission of Y chromosome microdeletions and subsequent infertility from father to son. Hence screening for Y chromosome microdeletions will help in the proper counseling and management of couples with male factor infertility.

Fluorescence in situ hybridization (FISH) is a rapid reliable technique in molecular cytogenetics. It supplements conventional karyotyping by providing additional information in certain cases. A large number of cells are available for quantitative analysis by FISH, as even interphase nuclei can be studied. It helps in detection of minimal residual disease and disease recurrence, as a very small percentage of abnormal cells can also be identified. The FISH probes used in this study were for the detection of the BCR/ABL fusion or t(9;22) in chronic myeloid leukemia (CML), PML/RARA fusion or t(15;17) in acute promyelocytic leukemia (APML) and determination of the XX/XY ratio in sex mismatched bone marrow transplantation. One hundred and fifty eight heparinized bone marrow or leukemic blood samples referred by consultants were analysed using the above mentioned FISH probes. These included samples of patients who underwent bone marrow transplantation. Karyotyping was carried out where indicated. Cases where FISH provided additional information or a different interpretation to karyotype analysis have been described.

Sjögren’s syndrome (SS) is a chronic slowly progressive autoimmune disorder characterized by symptoms of oral and ocular dryness, exocrine dysfunction, and lymphocytic infiltration of exocrine glands. Multiple myeloma (MM) is a bone-marrow-based malignant neoplasm of plasma cells associated with serum/urine monoclonal paraproteins and lytic skeletal lesions. There have been very few reported cases of MM, who had SS as the first presentation. We report a case of a woman diagnosed with Sjögren’s syndrome, who was later suspected to have multiple myeloma on serum protein electrophoresis. Fluorescencein situhybridization (FISH) was carried out to check for deletions of loci 13q14.3, ATM, p53, and IGH (14q32) rearrangements on a bone marrow aspirate. Monosomy 13 was observed in 49% of cells, and a rearrangement at the IGH locus was seen in 42% of cells. To determine the partner chromosome associated with the IGH rearrangement, further FISH tests were set up for t(4;14)(p16;q32) followed by t(14;16)(q32;q22) on fresh slides. The test was negative for t(4;14) but positive for t(14;16) in 27% of cells. This confirmed the diagnosis of MM. We report the first case from India, having an association of Sjögren’s syndrome with multiple myeloma, which showed t(14;16) and monosomy 13 by FISH analysis.

Objective To assess sperm characteristics and fertility before and after varicocelectomy using computer-assisted semen analysis (CASA). Design Preoperative and postoperative sperm parameters of infertile men with varicocele were analyzed statistically and the outcome of pregnancy was determined. Setting Department of Infertility Management and Assisted Reproduction, Jaslok Hospital and Research Centre, Bombay. Patients Semen samples of 49 men with varicocele-related infertility were analyzed using CASA. Interventions Sperm parameters of 26 men who underwent varicocelectomy were evaluated 3, 6, and 9 months postoperatively. Conception, occurring either naturally or therapeutically, was recorded. Main Outcome Measures Evaluation of improvement of sperm parameters after surgery. Results Sperm count, motility parameters, curvilinear velocity, straightline velocity, lateral head displacement, and normal morphology were significantly lower in men with varicocele. Postoperatively, there was significant improvement in count, motility, and normal morphology, with a decrease in proportion of acrosome-deficient heads and tapering forms. After varicocelectomy, 46.2% of the men had normal semen parameters, with the overall pregnancy rate being 50%. Conclusions Computer-assisted semen analysis provides the potential for accurate quantitative evaluation of semen in men with varicocele. Varicocelectomy results in improvement in semen quality with pregnancy rates of 50%.

Although the factors that are involved in the regulation of blastocyst implantation are incompletely understood, increasing evidence suggests that growth factors and cytokines play a crucial role in the maternal–fetal interaction during embryonic implantation. It is known that the endometrium, preimplantation embryo, and the cumulus cells are a source of these cytokines and growth factors and their corresponding receptors at the developmental stage and also during the point of implantation. A better understanding of these factors during embryonic development and implantation could lead to improved in vitro culture conditions and enhance the outcome of human in vitro fertilization. In this chapter, we discuss the benefits of using cumulus cells in an embryo coculture.

To establish importance of anti-ovarian antibodies (AOA) testing in infertile women.A clinical reproductive outcome comparative study between two groups of women undergoing IVF-ET. Group 1 consists of women tested positive for AOA, put on corticosteroid therapy, reverted to AOA negative and then taken up for IVF-ET. Group 2 were seronegative for AOA.Major urban infertility reference centre and National research institute.Five hundred seventy infertile women enrolled for IVF-ET.AOA testing, corticosteroid therapy and IVF-ET/ICSI.Comparable clinical outcome and significance of AOA testing established.AOA positive serum samples were sent periodically to re-investigate presence of AOA after corticosteroid therapy and women turned AOA negative were taken up for IVF-ET. Of the 70/138 women in group 1 who were treated with corticosteroids and turned seronegative for AOA, 22/70 were poor responders and needed donor oocyte-recipient cycles. Results demonstrated that fertilization and clinical pregnancy rates between both groups are comparable. Nevertheless, it is also observed that there is poor response to stimulation protocol, smaller number of oocytes retrieved and more spontaneous abortions in group 1 women. Hence not all outcomes following the treatment are comparable between the two groups. Usefulness of the test was established in two case studies.AOA testing could be included in the battery of tests investigating and treating infertility.

Objective To study the association of chromosomal polymorphic variations with infertility and subfertility. Design A comparative case-controlled association study using cytogenetic techniques to compare the frequency of chromosomal variations in infertile individuals versus fertile controls. Setting Department of Infertility Management and Assisted Reproduction, Jaslok Hospital and Research Centre, Mumbai, India. Patient(s) 760 infertile individuals and 555 fertile controls. Intervention(s) ICSI, IUI, karyotyping, inverted 4′,6-diamidino-2-phenylindole (DAPI), CBG banding. Main Outcome Measure(s) Frequency of chromosomal polymorphic variations in infertile individuals undergoing infertility treatment versus fertile individuals. Result(s) A highly statistically significant increase in the frequency of total chromosomal variants in infertile women (28.31% vs. 15.16%) and infertile men (58.68% vs. 32.55%) was observed. The frequency of 9qh+ was statistically significantly increased in women with primary infertility (16.22% vs. 6.41%) and in men with severe male factor infertility (14.69% vs. 4.25%). A highly statistically significant increase in the frequency of Yqh+ was observed in men whose wives had a bad obstetric history (30.20% vs. 12.74%). Conclusion(s) The statistically significantly higher incidence of heterochromatic variations found in infertile individuals stresses on the need to evaluate their role in infertility and subfertility. Potential epigenetic, genetic, and chromosomal modifications could be associated with certain complex disorders such as infertility and bad obstetric history.

Fanconi Anemia (FA) is a rare autosomal recessive disorder associated with pancytopenia, sponta- neous chromosome instability and a variety of congenital anomalies. Hypersensitivity to bifunctional alkylating or DNA crosslinking agents like Mitomycin C (MMC), Diepoxybutane (DEB) and Nitrogen Mustard (HN2) is used as a differential diagnostic test. A variable phenotype and age of onset of anemia make diagnosis difficult in some cases. We report a case of Fanconi anemia detected by the MMC stress test in a 2 year old boy, operated for bilateral syndactyly and contractures of fingers. He had a bifid thumb on the left hand and bilateral triphalangeal thumbs. There was no history of consanguinity or malformations, though a maternal uncle had a bifid thumb. USG in a subsequent pregnancy showed bony anomalies like scoliosis, talipes, contractures and radial aplasia, consistent with FA. The parents opted for termination. An early diagnosis of FA in a non-manifesting child would provide more time to ex- plore different treatment options, since a delay in diagno- sis could have serious consequences.

Intracytoplasmic Sperm Injection (ICSI) of- fers the real prospect of genetic parenthood for men with profound oligozoospermia and azoospermia. However, it may result in transgenerational transmission of genetic de- fect, which substantially increases the recurrence risk of infertility in the offspring of couples treated with ICSI. Re- cent developments in the field of genetics and assisted re- production have led to emergence of preimplantation ge- netic diagnosis (PGD). PGD helps in the negative selection of the aneuploid and abnormal embryos. For PGD, the em- bryo is biopsied and 1-2 blastomeres are removed from the 6-10 cell embryo. For establishing the diagnosis, the ge- netic analysis is carried out using FISH (for structural and numerical chromosomal anomalies) or PCR (for single gene disorders). Only the normal embryos are transferred back to the uterus, thus ensuring a normal pregnancy. Spare or arrested embryos obtained following ICSI were used for the study. FISH was performed on 5 embryos using CEP X spectrum green probes. PCR was done for cystic fibrosis common mutation ∆ F508 and for β-thalassaemia IVS1→ 5 and 619 bp deletion using blood as positive control. Mosa- icism was noted in two of the five embryos using FISH. The blastomeres were found to be normal for cystic fibro- sis and β-thalassaemia. PGD has an important role in help- ing patients to avoid the risk of transmission of genetic de- fect or abnormalities and also helps in avoiding repeated medical termination of pregnancy.

Objectives: The purpose of this study was to assess the coping mechanisms in patients presenting for in-vitro fertilization (IVF). Methods: We evaluated thirty consecutive couples presenting for in-vitro-fertilization. All couples were interviewed individually at first, and then together, using a semi- structured interview technique. Psychiatric diagnoses were made using the Diagnostic and Statistical Manual-IV (DSM-IV) criteria. Coping mechanisms used by the individuals were assessed using the Mechanisms of Coping Scale (MOCS). Other instruments used were Hamilton Depression Rating Scale (HAM-D-17), Hamilton Anxiety Rating Scale (HAM-A), Brief Psychiatric Rating Scale (BPRS), Self-Rating Symptom Scale (SRSS), and Eysenck Personality Inventory (EPI). Results: The mean age of the sixty patients was 32.3 ± 5.2 years. Fatalism was the commonest factor on the mechanisms of coping scale. Analysis of variance (ANOVA) across all factors of the MOCS for demographic factors showed that men used problem-solving mechanisms significantly more often than women ( F = 3.0, df = 1, 58, p < 0.05). ANOVA across coping factors on stressors with post-hoc tests of significance revealed that individuals facing social stress used fatalism significantly more often than other coping mechanisms, while those facing career stress used problem-solving significantly more often than other coping mechanisms ( F = 5.6, df = 1, 58, p < 0.05 and F = 3.04, df = 1,58, p < 0.01 respectively). ANOVA across coping factors on HAM-D-17 scores revealed that individuals who used fatalism had significantly higher HAM-D-17 scores compared to those who did not ( F = 4.4, df = 1, 58, p < 0.05). ANOVA across coping factors on HAM-A scores revealed that individuals who used escape-avoidance had significantly lower HAM-A scores than those who did not ( F = 4.3, df = 1, 58, p < 0.05). ANOVA across coping factors on SRSS scores revealed that individuals who used passivity or fatalistic coping mechanisms had significantly higher scores on SRSS than who did not ( F = 4.6, df = 1, 58, p < 0.05 and F = 3.5, df = 1, 58, p < 0.05). Conclusions: Differential patterns of coping were found among the sixty individuals presenting for IVF and were associated with a variety of factors including gender, education, stressors, and levels of depression, anxiety, and overall psychopathology. Efforts to recognize and recruit the coping mechanisms of infertile individuals are likely to enhance their ability to participate effectively in treatment.

Objective: To study the effect of tuberculosis, a common infectious disease in the Indian subcontinent, on the female pelvic factor and its subsequent effect on female fertility. Design: Retrospective case studies. Setting: Department of Infertility Management and Assisted Reproduction, Jaslok Hospital and Research Centre, Bombay, India. Patient(s): Three hundred women, between the ages of 25 and 35 years, with tubal factor as a cause of their infertile state. Result(s): One hundred seventeen women with a tubal factor were found to have tuberculosis as the cause of tubal blockage. On laparoscopy, 49.5% were found to have simple tubal blockage, 15.3% showed tubo-ovarian masses, and 23.9% had a frozen pelvis. Seventy-five percent complained of menstrual irregularities, thus indicating endometrial involvement; 25.6% of these women underwent an IVF procedure. The pregnancy rate after IVF-ET was 16.6% per transfer. Conclusion(s): This study highlights the fact that tuberculosis, a chronic infectious disease, is one of the major etiologic factors of female tubal infertility, especially on the Indian subcontinent.

expression pattern of specific genes important for fetal growth and nutrition. Materials and Methods: Blood samples from children conceived after ART (n = 54) and from spontaneously conceived children (n = 29) were collected after informed consent in PAXGene Blood RNA Tubes. The children were part of an existing cohort from the UK, followed since birth. Blood samples were stored at 20oC and transferred frozen to Belgium. RNA was extracted for reverse transcription and Real-Time PCR analysis using gene expression assays for DNA methyltransferase 1 (DNMT1), Glucocorticoid receptor (NR3C1) and Peroxisome Proliferator Activated Receptor Alpha (PPARA) and Hypoxanthine Phosphoribosyltransferase 1 (HPRT1) as an endogenous control. Statistical analysis was done using the Student T test. Results: Gene expression levels were normalised against the endogenous control HPRT1 and relatively quantified using the DDCt method. The mean blood mRNA levels of DNMT1, NR3C1 and PPARA in the ART children show a 0.95, 1.26 and 1.35 fold change respectively compared to those of the children from spontaneous conception (p = 0.95, p = 0.73 and p = 0.54 respectively). Discussion and Conclusion: No significant difference was observed in mRNA expression levels of DNMT1, NR3C1 and PPARA between children conceived after ART and children spontaneously conceived, reassuring the epigenetic safety of ART methods with respect to the studied genes. Nevertheless, no definitive conclusions can be drawn without testing other gene panels of imprinted and non-imprinted genes.

We are reporting on a possible association between multinucleated spermatozoa and polyploidy after in vitro fertilization. The semen from the male partner of a couple with 40% polyploidy on three in vitro fertilization attempts was examined by light and transmission electron microscopy. In that sample 10% to 12% of spermatozoa demonstrated conjoined or double heads. The significance of these findings is discussed.

This study used CA125, carcinoembryonic antigen (CEA), and alpha-fetoprotein (AFP) to classify ovarian cysts by measuring the levels of the three antigens; this information was useful when fluid obtained through laparoscopic puncture of ovarian cysts was submitted for cytologic examination from patients for whom tissue was unavailable for classification. We studied 136 consecutive cyst fluids (108 benign, 28 malignant) and correlated the findings with the tissue diagnosis. All three antigens were very low (CEA, less than 0.5 ng/ml; CA125, 55-2,143 mu/ml; and AFP, less than 4.8 ng/ml) in follicular and lutein cysts. Markedly elevated CA125 (296-1,950,000 mu/ml) and low CEA (0.5-220 ng/ml) and AFP (less than 4.8 ng/ml) levels were seen in patients with serous neoplasms, both benign and malignant. Elevated CEA (greater than 600 ng/ml) and CA125 (56-65,330 mu/ml) levels were seen in primary mucinous cystadenoma and cystadenocarcinoma. Two patients with colonic carcinoma metastatic to the ovary had an elevated CEA (greater than 600 ng/ml) and a normal CA125. Only one patient, with a malignant teratoma, had an elevated AFP. The adjunctive use of CEA and CA125 is recommended for the classification of ovarian cysts.