Anushree Patil

A cross-sectional study of polycystic ovarian syndrome among adolescent and young girls in Mumbai, India

Introduction: Polycystic ovary disease is a common endocrine condition which is rapidly gaining epidemic proportions. No community based prevalence data is available for this syndrome in India. Materials and Methods: A cross-sectional community-based study was undertaken in a sampled census block of Mumbai to assess the prevalence of polycystic ovarian syndrome (PCOS) among 778 adolescents and young girls aged 15-24 years. Among them, 600 completed all clinical, ultrasonography (USG), and biochemical investigations. Results: The prevalence of PCOS among them was 22.5% by Rotterdam and 10.7% by Androgen Excess Society criteria. Nonobese comprised 71.8% of PCOS diagnosed by Rotterdam criteria. Mild PCOS (oligomenorrhea and polycystic ovaries on USG) was the most common phenotype (52.6%). History of oligomenorrhea had a positive predictive value of 93.3% and negative predictive value of 86.7% to detect a possible case of PCOS. Hyperinsulinemia (serum insulin >15 μlU/mL) was present among 19.2% of diagnosed PCOS cases. Obese girls with PCOS were more hirsute, hypertensive, and had significantly higher mean insulin and 2 h post 75 g glucose levels compared with nonobese PCOS. Conclusion: To our knowledge, this is the first urban community-based study diagnosing PCOS and phenotypes among adolescent and young girls in India. This study demonstrates that PCOS is an emerging disorder during adolescence and screening could provide opportunity to target the group for promoting healthy lifestyles and early interventions to prevent future morbidities.

Methylation analysis of idiopathic recurrent spontaneous miscarriage cases reveals aberrant imprinting at H19 ICR in normozoospermic individuals

OBJECTIVE To study H19 ICR methylation levels in association with sperm parameters routinely analyzed in idiopathic recurrent spontaneous miscarriage cases. DESIGN Case-control study. SETTING Academic research setting. PATIENT(S) Male partners of couples with a history of idiopathic recurrent spontaneous miscarriage (RSM group) and male partners of couples with proven fertility (control group). INTERVENTION(S) None. MAIN OUTCOME MEASURE(S) Paternal age, sperm concentration, motility, chromatin compaction status, morphology, and H19 ICR methylation were assessed in control and idiopathic RSM group participants. RESULT(S) Paternal age and basic semen parameters analyzed did not show any significant difference between the two groups; however H19 ICR methylation levels were reduced significantly in the idiopathic RSM group compared with the control group. CONCLUSION(S) Significant reduction in the H19 ICR methylation without significant difference in the sperm parameters demonstrates aberrant imprinting to be associated with idiopathic RSM.

Surfactant Protein D Inhibits HIV-1 Infection of Target Cells via Interference with gp120-CD4 Interaction and Modulates Pro-Inflammatory Cytokine Production

Surfactant Protein SP-D, a member of the collectin family, is a pattern recognition protein, secreted by mucosal epithelial cells and has an important role in innate immunity against various pathogens. In this study, we confirm that native human SP-D and a recombinant fragment of human SP-D (rhSP-D) bind to gp120 of HIV-1 and significantly inhibit viral replication in vitro in a calcium and dose-dependent manner. We show, for the first time, that SP-D and rhSP-D act as potent inhibitors of HIV-1 entry in to target cells and block the interaction between CD4 and gp120 in a dose-dependent manner. The rhSP-D-mediated inhibition of viral replication was examined using three clinical isolates of HIV-1 and three target cells: Jurkat T cells, U937 monocytic cells and PBMCs. HIV-1 induced cytokine storm in the three target cells was significantly suppressed by rhSP-D. Phosphorylation of key kinases p38, Erk1/2 and AKT, which contribute to HIV-1 induced immune activation, was significantly reduced in vitro in the presence of rhSP-D. Notably, anti-HIV-1 activity of rhSP-D was retained in the presence of biological fluids such as cervico-vaginal lavage and seminal plasma. Our study illustrates the multi-faceted role of human SP-D against HIV-1 and potential of rhSP-D for immunotherapy to inhibit viral entry and immune activation in acute HIV infection.

Surfactant protein D induces immune quiescence and apoptosis of mitogen-activated peripheral blood mononuclear cells

Surfactant protein D (SP-D) is an integral molecule of the innate immunity secreted by epithelial cells lining the mucosal surfaces. The C-type lectin domain of SP-D performs pattern recognition functions while it binds to putative receptors on immune cells to modify cellular functions. Activation of immune cells and increased serum SP-D is observed in a range of patho-physiological conditions including infections. We speculated if SP-D can modulate systemic immune response via direct interaction with activated PBMCs. In this study, we examined interaction of a recombinant fragment of human SP-D (rhSP-D) on PHA-activated PBMCs. We report a significant downregulation of activation receptors such as TLR2, TLR4, CD11c and CD69 upon rhSP-D treatment. rhSP-D inhibited production of Th1 (TNF-α and IFN-γ) and Th17 (IL-17A) cytokines along with IL-6. Interestingly, levels of IL-2, Th2 (IL-4) and regulatory (IL-10 and TGF-β) cytokines remained unaltered. Analysis of co-stimulatory CD28 and co-inhibitory CTLA4 receptors along with their ligands CD80 and CD86 revealed a selective up-regulation of CTLA4 in the lymphocyte subset. rhSP-D induced apoptosis in the activated but not in non-activated lymphocytes. Blockade of CTLA4 inhibited rhSP-D mediated apoptosis of activated lymphocytes, confirming involvement of CTLA4. We conclude that SP-D restores immune homeostasis. It regulates expression of immunomodulatory receptors and cytokines, which is followed by induction of apoptosis in activated lymphocytes. These findings suggest a critical role of SP-D in immune surveillance against activated immune cells.

PON1 polymorphisms are associated with polycystic ovary syndrome susceptibility, related traits, and PON1 activity in Indian women with the syndrome

OBJECTIVE To investigate the association of paraoxonase 1 (PON1) polymorphisms (L55M and Q192R) with polycystic ovary syndrome (PCOS) susceptibility and its related traits in Indian women. DESIGN Case-control study. SETTING Academic research institute, infertility, and endocrinology clinics. PATIENT(S) Controls (n = 326), women with PCOS (n = 482). INTERVENTION(S) None. MAIN OUTCOME MEASURE(S) Genotypic and allelic frequency distribution, genotype-phenotype association, different PON1 activities (lactonase, arylesterase, and paraoxonase). RESULT(S) The genotypic and allelic frequency distributions of the L55M polymorphism were significantly different between lean controls and lean women with PCOS, and this polymorphism reduced the risk of PCOS development in lean but not in obese Indian women. Furthermore, this polymorphism was significantly associated with decreased 2-hour glucose, apolipoprotein B, free and bioavailable T, and free androgen index concurrent with increased sex hormone-binding globulin (SHBG) and FSH levels only in lean women with PCOS. However, Q192R polymorphism showed comparable genotypic frequency distribution between controls and women with PCOS. PON1 lactonase and arylesterase activities were significantly decreased in women with PCOS compared with controls. PON1 polymorphisms were shown to influence its activities. CONCLUSION(S) Our study showed that L55M, but not Q192R, polymorphism is significantly associated with reduced PCOS susceptibility only in lean women and also impacts glucose metabolism, lipid parameters, and hyperandrogenemia in them. Our study therefore suggests the possibility of differential genetic pathophysiology of PCOS between lean and obese women.

Monocyte Proteomics Reveals Involvement of Phosphorylated HSP27 in the Pathogenesis of Osteoporosis

Peripheral monocytes, precursors of osteoclasts, have emerged as important candidates for identifying proteins relevant to osteoporosis, a condition characterized by low Bone Mineral Density (BMD) and increased susceptibility for fractures. We employed 4-plex iTRAQ (isobaric tags for relative and absolute quantification) coupled with LC-MS/MS (liquid chromatography coupled with tandem mass spectrometry) to identify differentially expressed monocyte proteins from premenopausal and postmenopausal women with low versus high BMD. Of 1801 proteins identified, 45 were differentially abundant in low versus high BMD, with heat shock protein 27 (HSP27) distinctly upregulated in low BMD condition in both premenopausal and postmenopausal categories. Validation in individual samples (n = 80) using intracellular ELISA confirmed that total HSP27 (tHSP27) as well as phosphorylated HSP27 (pHSP27) was elevated in low BMD condition in both categories (P < 0.05). Further, using transwell assays, pHSP27, when placed in the upper chamber, could increase monocyte migration (P < 0.0001) and this was additive in combination with RANKL (receptor activator of NFkB ligand) placed in the lower chamber (P = 0.05). Effect of pHSP27 in monocyte migration towards bone milieu can result in increased osteoclast formation and thus contribute to pathogenesis of osteoporosis. Overall, this study reveals for the first time a novel link between monocyte HSP27 and BMD.

Serum levels of phosphorylated heat shock protein 27 (pHSP27) are associated with bone mineral density in pre- & postmenopausal women: A pilot study

Background & objectives: Phosphorylated heat shock protein 27 (pHSP27) has been implicated in the pathogenesis of osteoporosis. Oxidative stress and proinflammatory cytokines, which are known to be involved in aetiology of osteoporosis, can trigger HSP27 phosphorylation. Since pHSP27 is present in circulation, it was hypothesized that serum pHSP27 would be elevated in low bone mineral density (BMD) condition and might serve as an indicator of osteoporosis/osteopenia. Hence, the aim of this study was to examine serum levels of pHSP27 in relation with BMD in pre- and postmenopausal women. Methods: Premenopausal (30 to 40 yr) and postmenopausal (50 to 60 yr) women having either low BMD (osteopenia/osteoporosis) or high BMD were selected (n=80) from a prospective cohort (n=200). Serum levels of pHSP27; along with levels of oestradiol, malondialdehyde, total antioxidant capacity, interleukin (IL)-1, IL-6, tumour necrosis factor - alpha, (TNF-α), c-telopeptide fragments of collagen type I (CTX-1) and osteocalcin were estimated. Results: The serum levels of pHSP27 were significantly elevated in low BMD groups in premenopausal and postmenopausal categories (P<0.05). It also exhibited a significant odds ratio (OR) to differentiate between low and high BMD in both premenopausal (OR=1.734, P=0.013) and postmenopausal (OR=1.463, P=0.042) categories. Additionally, area under the curve to predict low BMD was non-significantly higher for pHSP27 than CTX-1 in premenopausal and postmenopausal categories. Interpretation & conclusions: This study highlights a novel relation between serum pHSP27 and BMD in Indian women however, these findings need to be confirmed in larger studies.

Preterm birth among pregnancies conceived by assisted reproduction techniques in Mumbai, Maharashtra, India

Background: Preterm births are an enormous global problem on families, medical system and economy. The rates of preterm birth are increasing and one of the contributors is growing use of Assisted Reproduction Techniques (ART) leading to multifetal gestations. Some risk factors for preterm birth are specific to women who conceive by ART. Since there is limited data from India, this pilot study was undertaken to assess the magnitude of preterm birth among pregnancies conceived by ART and to study the contributing factors. Methods: Clinic based descriptive cohort study through eight ART clinics in Mumbai for one year. Data was collected using an in-depth questionnaire on socio-demographic characteristics, medical history, ART details, course and complications during pregnancy, mode of delivery, pregnancy outcome, risk factors related to preterm birth and neonatal outcome. Complete details of 113 participants who completed the study were analyzed. Results: Study showed high incidence of preterm birth (76.23%) among women conceived with ART. Multiple gestations were observed in 45.1%. Pregnancy related complications like heterotrophic pregnancy (3%), pre eclampsia (15%) and gestational diabetes (11%) were high. Incidence of caesarean section was very high (98%). Neonatal outcome was good with 98% live births and only 2 still births. Conclusions: Present study highlights that preterm birth, multiple pregnancies, pregnancy related complications like preeclampsia, gestational diabetes and caesarean sections are very high among women conceived by ART. With growing use of ART there is an urgent need to develop a National ART Surveillance system in India like the one in Centre for Disease Control Atlanta to get complete data on the pregnancy course and outcomes of ART conceptions. Efforts to limit the number of embryos transferred should be strengthened to prevent multiple births.

Capacity assessment of district health system in india on services for prevention and management of infertility

Background: Infertility is a neglected service component in the public health-care system in India. Objectives: This study aims to assess the availability and practices on prevention and management services for infertility in the district health system. Methodology: A cross-sectional survey of selected health facilities and the staff from 12 district hospitals (DHs), 24 community health centers (CHCs), 48 primary health centers (PHCs), and 48 subcenters was conducted using qualitative and quantitative methods. Interviewed staff included 26 gynecologists; 91 medical officers; 91 auxiliary nurse midwife; 67 laboratory technicians; and 84 accredited social health activist workers. Results: The findings indicate that adequate staff was in place at more than 70% of health facilities, but none of the staff had received any in-service training on infertility management. Most of the DHs had basic infrastructural and diagnostic facilities. However, the majority of the CHCs and PHCs had inadequate physical and diagnostic facilities related to infertility management. Semen examination service was not available at 94% of PHCs and 79% of CHCs. Advanced laboratory services were available in <42% at DHs and 8% at CHCs. Diagnostic laparoscopy and hysteroscopy were available in 25% and 8% of DHs, respectively. Ovulation induction with clomiphene was practiced at 83% and with gonadotropins at 33% of DHs. Conclusion: The district health infrastructure in India has a potential to provide basic services for infertility. With some policy decisions, resource inputs and capacity strengthening, it is possible to provide advanced services for infertility in the district health system.

PON1 promoter polymorphisms contribute to PCOS susceptibility and phenotypic outcomes in Indian women

Polycystic ovary syndrome is a common endocrinopathy characterized by anovulatory infertility, hyperandrogenism, insulin resistance and oxidative stress, which predisposes affected women to reproductive and cardiometabolic complications in later life. We have investigated the association of PON1 promoter polymorphisms with PCOS susceptibility, PON1 activity and its related traits in Indian women. The genotypic and allelic frequency distribution of only -907G/C polymorphism in PON1 promoter showed significant difference between non-hyperandrogenic control and PCOS women, and was significantly associated with reduced susceptibility to PCOS, considering the recessive model. PON1 lactonase and arylesterase activities were also significantly decreased in women with PCOS compared to controls. Further, PON1 promoter polymorphisms were linked to altered insulin and testosterone levels in hyperandrogenic and non-hyperandrogenic women with PCOS. This study highlights PON1 as an important candidate gene influencing genetic pathophysiology of PCOS.