The development, evaluation, performance, and validation of micro-PCR and extractor for the quantification of HIV-1 &-2 RNA.

HIV infection has been a global public health threat and overall reported ~ 40 million deaths. Acquired immunodeficiency syndrome (AIDS) is attributed to the retroviruses (HIV-1/2), disseminated through various body fluids. The temporal progression of AIDS is in context to the rate of HIV-1 infection, which is twice as protracted in HIV-2 transmission. Q-PCR is the only available method that requires a well-developed lab infrastructure and trained personnel. Micro-PCR, a portable Q-PCR device, was developed by Bigtec Labs, Bangalore, India. It is simple, accurate, fast, and operationalised in remote places where diagnostic services are inaccessible in developing countries. This novel micro-PCR determines HIV-1 and HIV-2 viral load using a TruePrep™ extractor device for RNA isolation. Five ml blood samples were collected at the blood collection centre at AIIMS, New Delhi, India. Samples were screened for serology, and a comparison of HIV-1/2 RNA was done between qPCR and micro-PCR in the samples. The micro-PCR assay of HIV-RNA has compared well with those from real-time PCR (r = 0.99, i < 0.002). Micro-PCR has good inter and intra-assay reproducibility over a wide dynamic range (1.0 × 102-1.0 × 108 IU/ml). The linear dynamic range was 102-108 IU/ml. The clinical and analytical specificity of the assay was comparable, i.e., 100%. Intra-assay and inter-assay coefficients of variation ranged from 1.17% to 3.15% and from 0.02% to 0.46%, respectively. Moreover, due to the robust, simple, and empirical method, the Probit analysis has also been done for qPCR LODs to avoid uncertainties in target recoveries. The micro-PCR is reliable, accurate, and reproducible for early detection of HIV-1 and HIV-2 viral loads simultaneously. Thus, it can easily be used in the field and in remote places where quantification of both HIV-1/2 is not reachable.

Prevalence of sarcopenia and its determinants in people with type 2 diabetes: Experience from a tertiary care hospital in north India.

OBJECTIVES: Changes in skeletal muscle mass and quality are associated with type 2 Diabetes (T2D) and its complications. We evaluated the prevalence of sarcopenia in patients with T2D and its association with various anthropometric and metabolic parameters. METHODS: A total of 229 patients with T2D, ≥20-60 years, were screened for sarcopenia using handgrip strength (HGS) by dynamometer, physical performance test (by Short Physical and chair stand test), and height-adjusted appendicular skeletal muscle index (ASMI) by Dual Energy X-ray Absorptiometry (DXA) applying Asian Working Group on Sarcopenia (AWGS). Multiple logistic regressions were performed to identify the factors associated with sarcopenia. RESULTS: The mean age was 46.2 ± 7.4 years with 55% being women. The prevalence of low HGS, poor physical performance, low ASMI, possible sarcopenia, sarcopenia, and severe sarcopenia was 16.2%, 39.3%, 33%, 43%, 18.8%, and 6.1%, respectively. Age >45 years and use of >2 oral hypoglycaemic agents (OHA's) were risk factors for low HGS (OR:3.51, 95%CI = 1.5-8.3) and low ASMI (OR:2.40, 95%CI = 1.05, 5.49, p-0.04), respectively. Female sex (OR:3.3 1.8-6.1 p < 0.01), age >45 years (OR:2.12, 95% CI = 1.2-3.8 p-0.012) and liver fibrosis (OR: 2.12, 95% CI = 1.01-4.46 p-0.048) were independently associated with poor performance. No association was found with HbA1c, dyslipidaemia, albuminuria, hypertension, or duration of diabetes and sarcopenia. CONCLUSION: Sarcopenia is becoming increasingly recognized as a significant complication in younger individuals with T2D, and poor physical performance plays a vital role in its development. The prevalence of sarcopenia rises with advancing age, underscoring the importance of early intervention to address this condition.

Alterations of oligodendrocyte progenitor cells (OPCs) with survival time in humans following high impact brain trauma.

BACKGROUND: As there is a lack of comprehensive literature regarding the molecular environment of the human brain emphasizing on oligodendrocyte progenitor cells (OPCs) following high impact brain trauma. The protagonist of OPCs post severe traumatic brain injury (sTBI) provides a significant thrust towards estimating time elapsed since trauma as well as developing novel therapeutic approaches. The present study was carried out to study post trauma alterations pertaining to myelin sheath and oligodendrocyte response with survival time. MATERIALS AND METHODS: In the present study, victims (both male and female) of sTBI (n = 64) were recruited and contrasted with age and gender matched controls (n = 12). Post mortem brain samples from corpus callosum and grey white matter interface were collected during autopsy examination. Extent of myelin degradation and response of OPC markers Olig-2 and PDGFR-α were evaluated using immunohistochemistry and qRT-PCR. STATA 14.0 statistical software was used for data analysis with P-value<0.05 considered statistically significant. RESULTS: Timewise qualitative correlation with extent of demyelination performed using LFB-PAS/IHC-MBP, IHC Olig-2 and mRNA expression revealed tendency towards remyelination in both corpus callosum and grey white matter interface. Number of Olig-2 positive cells was significantly higher in sTBI group as compared to control group (P-value: 0.0001). Moreover, mRNA expression studies of Olig-2 showed significant upregulation in sTBI patients. mRNA expression of Olig-2 and PDGFR-α in sTBI patients showed significant variation with respect to survival time (p value:0.0001). CONCLUSION: Detailed assessment of post TBI changes implementing various immunohistochemical and molecular techniques shall potentially reveal intriguing and important inferences in medicolegal practices and neurotherapeutics.

Hyperactive behaviour of growth differentiation factor- 15 (GDF-15) in conjunction with iron trafficking transporters and suppression of Nrf-2 gene in diabetes and metabolic syndrome.

Multiple parallel factors are frequently interrogated with various toxic radicals which are abundantly generated in the liver, heart, and pancreas in stress conditions. They are actively involved in the development of diabetes and metabolic aberrations. However, whether over-activation of GDF-15mRNA and influxes of iron-by-iron trafficking genes are directly suppressing the Nrf-2 gene in patients with diabetes and metabolic aberrations in context with undiagnosed individuals with diabetes and metabolic aberrations? Therefore, we have investigated inter and intra- related Zip8/14 mRNA, GDF-15mRNA, and Nrf-2 mRNA expressions in diabetes and metabolic syndrome as it is expected to be up to 134 million by 2045 in India. We recruited 120 subjects from the Department of Medicine, Endocrinology and Metabolic Clinic, All India Institute of Medical Sciences, New Delhi, India. Various investigations related to anthropometry, nutritional, hematological, biochemical, cytokine, and oxidative stress were measured in diabetes, metabolic syndrome, diabetes with metabolic aberration, and healthy controls. Relative expression of GDF-15, ZIP8, ZIP14, Nrf-2, and housekeeping genes was done in all subjects. Stress-responsive cytokines are highly expressed in patients with metabolic aberration with respect to body weight, IR, waist circumference, and fat mass. IL-1ß, TNF-α, and IL-6 levels were significantly higher in metabolic syndrome, whereas Adiponectin levels were profoundly lower side. MDA levels were significantly raised in diabetes with metabolic syndrome while SOD activities were lowered (p = 0.001). GDF-15 mRNA expression was 1.79-fold upregulated in group III as compared with Group I while 2-threefold down-regulation of Nrf-2 expression was observed in diabetes with metabolic aberration groups. Zip 8 mRNA expressions were downregulated (p = 0.014), and Zip 14 mRNA expressions were upregulated (p = 0.06) in diabetes and metabolic aberrations. The association of GDF-15 and Nrf-2 mRNA expression was found contradictory and highly interlinked with ROS. Zip 8/14mRNA expressions were also dysregulated in diabetes and metabolic-associated complications.

A Combination of Inflammatory and Hematological Markers is Strongly Associated with the Risk of Death in Both Mild and Severe Initial Disease in Unvaccinated Individuals with COVID-19 Infection.

Background: Inflammatory and hematological markers are used extensively for early prognostication and monitoring in COVID-19.We aimed to determine whether routinely prescribed laboratory markers can predict adverse outcome at presentation in COVID-19. Methods: This retrospective observational study was performed on 401 samples collected between July to December 2020 from COVID-19 positive subjects, admitted at All India Institute of Medical Sciences, Delhi, India. Clinical details and laboratory investigations within 3 days of COVID-19 positivity were obtained. Clinical outcomes were noted from patient medical records, till discharge or death. Laboratory parameters, with individually defined cut-offs, were used, either singly or in combination to distinguish survival and death for those having severe and non-severe disease at initial presentation. Findings: Total Leukocyte count, Absolute neutrophil count, Neutrophil to Lymphocyte ratio, C-Reactive Protein (CRP), Interleukin-6 (IL-6), Lactate Dehydrogenase, Ferritin and Lymphocyte to CRP ratio (LCR) were significantly altered at presentation in severe COVID-19 as compared to non-severe cases; and, also in those who died due to COVID-19 compared to those who survived. A combination of four markers, CRP (≥3.9mg/dL); IL-6 (≥45.37pg/ml); Ferritin (≥373ng/mL); 1/LCR ≥0.405 was found to strongly predict mortality in cases with non-severe presentation as also in severe cases. Conclusion and Interpretation: The combination of routinely used markers, CRP, IL-6, Ferritin and 1/LCR can be used to predict adverse outcomes, even in those presenting with mild to moderate disease. This would identify subset of patients who would benefit from closer monitoring than usual for non-severe disease.

Mussel-Inspired Adhesive Hydrogels Based on Laponite-Confined Dopamine Polymerization as a Transdermal Patch.

Transdermal patch for local drug delivery has attained huge attention as an attractive alternative to existing drug delivery techniques as it is painless and user-friendly. However, most adhesive hydrogels either do not have adequate adhesion with the skin or cause discomfort while being removed from the skin surface due to excessive adhesion. To address this challenge, we developed an adhesive hydrogel based on laponite-confined dopamine polymerization as a transdermal patch. Laponite RDS nanoclay was used to control the hydrogel's viscous behavior and dopamine polymerization. The laponite polymerized polydopamine (l-PDA) was incorporated into poly(vinyl alcohol) (PVA) to make the PVA-l-PDA hydrogel. The laponite-confined polymerization improved the hydrogels' water contact angle and adhesion strength. The adhesion strength of the PVA-l-PDA hydrogel was adequate to adhere to the evaluated goat skin, glass, and polypropylene surfaces. Notably, the PVA-l-PDA hydrogel was easy to peel off from the skin. Further, we evaluated the drug release profile in goat skin using lidocaine as a model drug. We observed the controlled release of lidocaine from the PVA-l-PDA hydrogel compared to the PVA-PDA hydrogel. In addition, the nanoclay-confined adhesive hydrogel did not show any cytotoxic effect in fibroblasts. Altogether, PVA-l-PDA hydrogels offer appropriate adhesive strength, toughness, and biocompatibility. Thus, the PVA-l-PDA hydrogel has the potential to be an efficient transdermal patch.

Serum Matrix Metalloproteinase 7 as a Diagnostic and Prognostic Biomarker for Extrahepatic Biliary Atresia.

Background: Differentiation of neonatal cholestasis into neonatal hepatitis (NH) and extrahepatic biliary atresia (EHBA) is essential to formulate the treatment plan; promptness is indispensable for optimal outcomes. The clinical and nonoperative algorithms lack precision; the gold standard investigations (liver biopsy or per-operative cholangiogram) are invasive. There is a need for a noninvasive test which is both, sensitive and specific and has a high likelihood ratio. Aim: To study the (diagnostic) role of matrix metalloproteinase 7 (MMP-7) as a serum biomarker to differentiate between EHBA and NH and evaluate the prognostic significance in EHBA based on its correlation with liver histopathology and serological predictors of liver fibrosis - Aspartate-to-Platelet Ratio Index (APRI) and Fibrosis-4 (FIB-4). Materials and Methods: This was a prospective study conducted upon patients of neonatal cholestasis presenting with acholic stools (n = 46) with equal number of controls (n = 45) with no liver pathology. Observational parametric included disease-specific workup and serum MMP-7 levels (all participants); liver biopsyl and APRI-FIB-4 (EHBA). Results: (Diagnostic) Serum MMP-7 levels were significantly elevated in EHBA (n = 25; 28 ng/mL) as compared to those in NH (n = 21; 1.88 ng/mL) and normal infants (n = 45; 1.2 ng/mL) (P < 0.001 for both). Serum cutoff at 4.99 ng/mL differentiated EHBA-NH with a high sensitivity (96%), specificity (90.5%), and a negative predictive value (95%), with the number needed to misdiagnose being 23. (Prognostic) Inflammatory activity and fibrosis-stage on liver histopathology (METAVIR-and-Ishak scores) correlated with MMP-7 levels. APRI and FIB-4 scores also depicted a strong correlation with each other, age of the patient, and liver fibrosis. Conclusions: MMP-7 has a diagnostic value in differentiating EHBA from NH and may also be used as a prognostic biomarker in the follow-up of these patients. MMP-7 levels in controls may be used as a baseline for future studies.

Characterisation of anaemia amongst school going adolescent girls in rural Haryana, India.

OBJECTIVE: High burden of anaemia exists amongst rural adolescent girls in India. The objective of this study was to characterise anaemia in school going adolescent girls in rural Haryana, India. DESIGN: Linear and multiple logistic regression analysis of data collected prior to an intervention trial was conducted. Participants were classified into anaemic (haemoglobin <12 g/dl) and non-anaemic group and were further classified into deficiencies of Fe, folate or vitamin B12, mixed, anaemia of other causes and inflammation. SETTING: Three schools in Ballabgarh block of Faridabad District, Haryana, India. PARTICIPANTS: One hundered and ninety-eight non-anaemic and 202 anaemic adolescent girls (12-19 years). RESULTS: Anaemic girls had 29·6 % Fe deficiency, 28·1 % folate or vitamin B12 deficiency, 15·8 % mixed deficiency and 9·7 % acute inflammation. Anaemia of other causes was found in 16·8 % of the anaemic participants. Girls with Fe and isolated folate deficiency had 2·5 times and four times higher odds of developing anaemia, respectively, as compared with non-anaemic girls. Fe deficiency with no anaemia was found amongst 11 % non-anaemic girls. Non-anaemic girls had a high prevalence of combined deficiency of folate or vitamin B12 (29·5 %) and acute inflammation (14·4 %). CONCLUSIONS: The current strategy of Fe and folic acid supplementation alone will not suffice for achieving the desired reduction in the prevalence of anaemia as unknown causes and anaemia of inflammation contribute to a substantial proportion of anaemia. Integrating other nutrition-specific components like improving water, sanitation and hygiene practices with the ongoing micronutrient supplementation program will comprehensively tackle anaemia. Unknown causes of anaemia warrant further research.

Platelet Function Analysis by Flowcytometry in Thrombocytopenic Trauma Patients.

Hemostasis is dependent on sufficient quantity and quality of circulating functional platelets. Platelet function in trauma patients with thrombocytopenia and its impact on the clinical outcome is not adequately explored. Whether platelet dysfunction has a role in the pathogenesis of acute traumatic coagulopathy needs to be studied. Blood samples were collected from 70 thrombocytopenic trauma patients before transfusing platelets and assessed for platelet activation and platelet aggregation using flowcytometry. Primary outcome was in-hospital mortality. Platelet dysfunction was identified in 57.1% of thrombocytopenic trauma patients. A weak inverse correlation between percentage of activated platelets and APTT was observed (Spearman coefficient - 0.25, p = 0.03). A sensitivity and specificity of 66.6% was achieved for a cut off of ≤ 6.5% of platelet activation post trauma with ROC-AUC of 0.658 for identifying coagulopathy. No correlation with mortality however was observed (p > 0.05). Platelet dysfunction had a weak association with coagulopathy suggesting limited contribution of platelet dysfunction in pathogenesis of acute traumatic coagulopathy and warrants further research.

Prevalence of elevated alanine aminotransferase levels in adult participants from a community-based study from northern part of India.

Alanine aminotransferase (ALT) is a cytosolic enzyme specific to hepatocytes, and its elevated level in the peripheral blood denotes liver cell injury. Detection of persistently elevated ALT levels during routine health check-up in asymptomatic or symptomatic individuals provides a window of opportunity to explore the causes of liver cell damage and for the timely institution of appropriate treatment. This was a retrospective study using a subset of the data from a previous community-based prospective study done for the estimation of the prevalence of celiac disease (CD) in India,  during which estimation of ALT levels in the blood samples of participants was also carried out. Of the 11,053 individuals (4399 [39.8%] males; mean age 37.9 ± 13.3 years) screened, 6209 consented to provide blood samples for testing for CD. Of these, assessment of serum ALT levels was done in 6083 (2235 [36.7%] males) patients. ALT was elevated above the upper limit of normal (ULN) (> 40 IU/L) in 1246 (20.5%) of the participants and > 1.5 times (> 60 IU/L) in 329 (5.4%) participants. The ALT levels were elevated more frequently in men as compared to women (29.4% vs. 15.3%, p < 0.001). There was a significant positive correlation (Pearson correlation coefficient [r] = 0.25, p < 0.0001) between ALT levels and body mass index (BMI). With increasing age, there was a significant decrease in the proportion of subjects with ALT ≥ 1.5× ULN (p < 0.001). Our results suggest that a high proportion (20.5%) of individuals otherwise considered healthy have values of ALT level in the serum above the "normal" range/cut-off suggesting likely ongoing underlying liver damage. There is a need for measures to evaluate and, if found, treat the underlying cause for the same.

Intravenous Iron sucrose and change in hemoglobin, ferritin, and oxidative stress markers among moderately anemic pregnant women attending a secondary care level Hospital in Northern India.

BACKGROUND: Intravenous iron is associated with oxidative stress, and very few studies have assessed change in oxidative stress markers post infusion. OBJECTIVES: The study aimed to measure the change in levels of hemoglobin (Hb), serum ferritin, and select oxidative stress markers (malondialdehyde [MDA], superoxide dismutase [SOD], and ferric reducing ability of plasma [FRAP]) 4 weeks following the administration of intravenous iron sucrose (IVIS) among moderately anemic pregnant women who were attending a secondary-level health-care facility, Haryana, North India. METHODS: An observational study was conducted (May 2016 to Jan 2018) among pregnant women receiving intravenous iron sucrose i.e., IVIS (300 mg per dose) diluted in 300 mL of normal saline over 20-45 min and were followed up for a period of 4 weeks after the last dose of IVIS (end line). The study outcomes were measured in the levels of Hb, serum ferritin, MDA, SOD, and FRAP from the baseline to the end line. RESULTS: The mean (95% confidence interval) change in the Hb and serum ferritin level 4 weeks after the last dose of IVIS was an increase of 2.5 (2.1-3.0) g/dL (P < 0.001) and 63.0 (44.7-81.3) ng/mL (P < 0.001), respectively. There were no significant changes (baseline to end line) in mean (standard deviation [SD]) MDA level and mean (SD) FRAP level. The mean (SD) SOD level declined significantly (2.2 [0.4] U/mL to 1.6 [0.5] U/mL [P < 0.001]). No life-threatening adverse events were encountered during the study. CONCLUSION: IVIS was well tolerated and effective in treating moderate anemia in pregnancy. Body iron store was replenished following IVIS administration. There was no increase in oxidative stress following IVIS therapy.

Non-immunological biomarkers for assessment of villous abnormalities in patients with celiac disease.

BACKGROUND AND AIM: Demonstration of villous abnormalities is an essential component of diagnosis of celiac disease (CeD) that requires duodenal biopsies. There is a need for non-invasive biomarker(s) that can predict the presence of villous abnormalities. METHODS: Levels of plasma citrulline, plasma intestinal fatty acid binding protein (I-FABP), and serum regenerating gene 1α (Reg1α) were estimated in treatment naïve patients with CeD and controls. The levels of these biomarkers and their cyclical pattern were validated in a predicted model of enteropathy. Optimum diagnostic cut-off values were derived, and the results were further validated in a prospective validation cohort. RESULTS: While level of plasma citrulline was significantly lower, the levels of plasma I-FABP and serum Reg1α were significantly higher in patients with CeD (n = 131) in comparison with healthy (n = 216) and disease controls (n = 133), and their levels reversed after a gluten-free diet (GFD). In the model of predicted enteropathy (n = 70), a sequential decrease and then increase in the level of plasma citrulline was observed; such a sequential change was not observed with I-FABP and Reg1α. The diagnostic accuracy for prediction of presence of villous abnormality was 89% and 78% if citrulline level was  ≤ 30 µM/L and I-FABP levels were ≥ 1100 pg/mL, respectively. The results were validated in a prospective validation cohort (n = 104) with a sensitivity and specificity of 79.5% and 83.1%, respectively, for predicting villous abnormalities of modified Marsh grade > 2 at calculated cut-off values of citrulline and I-FABP. CONCLUSIONS: Plasma citrulline  ≤ 30 µM/L is the most consistent, highly reproducible non-invasive biomarker that can predict the presence of villous abnormality and has the potential for avoiding duodenal biopsies in 78% patients suspected to have CeD.

Silica Release from Silane Cross-Linked Gelatin Based Hybrid Scaffold Affects Cell Proliferation.

Earlier, we had reported the synthesis and characterization of star-shaped poly(d,l-lactide)-b-gelatin (ss-pLG) to improve cell adhesion and proliferation, but the stability of ss-pLG scaffolds remained a persistent issue. Here we show an increase in the stability of ss-pLG using 3-glycidoxypropyl trimethoxysilane (GPTMS) as a covalent cross-linker (h-ss-pLG). The rate of cell proliferation within Hep-G2 cultured h-ss-pLG scaffolds increased until the third day, and afterward it drastically declined. Further, we identified the release of inorganic silica from GPTMS cross-linked h-ss-pLG, which may be associated with the decrease in the rate of HepG2 cell proliferation. However, the cross-linking did not affect red blood cells (RBCs) and they were completely hemocompatible. In addition, our in vivo experiments in female rats showed that the hybrid h-ss-pLG scaffolds were not degraded completely after 4 weeks, as they were covalently cross-linked with silane. These results suggest the significance of the cross-linker selection, which is one of the other key factors, and needs to be considered while designing scaffolds.

Synthesis of 8',11'-dihydrospiro[cyclohexane-1,2'-oxepino[2,3-h] chromen]-4'(3'H)-ones with ring closing metathesis as a key step.

A series of novel hybrid molecular entities incorporating various spiro chromanone scaffolds onto the benzannulated oxepine core moiety were synthesised using allylation, Claisen rearrangement, Kabbe condensation and Ring Closing Metathesis (RCM) as a key step. During the synthesis we found that the nitrogen functionality in the substrate influences significantly the catalyst load due to electronic effects. Several iterations have been carried out to achieve complete conversion to products 6a-6e.

Increased oxidative stress and deficient antioxidant levels may be involved in the pathogenesis of idiopathic recurrent acute pancreatitis.

BACKGROUND: Increased Oxidative Stress (OS) is implicated in the pathogenesis of Chronic Pancreatitis (CP). Whether or not OS contributes to disease progression through the stages of Recurrent Acute Pancreatitis(RAP), to CP is not known. Increased OS, if present in RAP could be an important therapeutic target in preventing progression of RAP to CP. OBJECTIVE: To assess the oxidative stress and antioxidant status in patients with idiopathic RAP. METHODS: 50 consecutive patients with Idiopathic Recurrent Acute Pancreatitis (IRAP) were included. Markers of OS [4-hydroxynonenol (4-HNE), malondialdehyde (MDA) and serum SOD (S-SOD)] and antioxidant status [ferric reducing the ability of plasma (FRAP), Glutathione peroxidase (GPX) and Vitamin C (Vit C)] were measured in quiescent phase and during an episode of pancreatitis. Their levels were compared with those in age and sex matched healthy controls and patients with CP. RESULTS: The mean age of patients with IRAP was 22.2 ± 7.7 years and 39 (78%) were males. Levels of 4-HNE were significantly increased in patients with IRAP compared with healthy controls (3.03 ± 2.35 vs. 2.12 ± 1.29 ng/ml; p = 0.03) and were even higher during an episode of acute pancreatitis (5.21 ± 3.51 ng/ml; p = 0.03). Antioxidant levels were reduced in IRAP compared with healthy controls as measured by FRAP (707.0 ± 144.9 vs. 528.8 ± 120.0 µmol/Fe2+liberated; p = 0.0001) and GPX (1472 ± 375.7 vs. 910.0 ± 558.5 pg/ml; p = 0.001). OS and antioxidant profiles were similar in IRAP and CP with no significant difference. CONCLUSION: OS is increased in patients with IRAP, more so during an acute episode. Antioxidant levels are also reduced suggesting that OS may play a role in the pathogenesis of IRAP and its progression to CP.

Serum FRAP Levels and Pre-eclampsia among Pregnant Women in a Rural Community of Northern India.

INTRODUCTION: There is a balance between the oxidative and the anti-oxidative forces in human body. Some studies document decreased level of anti-oxidant in pre-eclampsia while other studies showed normal level of anti-oxidant in pre-eclampsia and the evidence is equivocal. AIM: The aim of the present study was to assess enzymatic anti-oxidant activities in pre-eclamptic women and compare it with normotensive pregnant women with period of gestation between 28 to 36 weeks. MATERIALS AND METHODS: A community-based cross-sectional survey was conducted (from November 2012-December 2013) at the Ballabgarh Health and Demographic Surveillance System (HDSS) site which was managed by Centre for Community Medicine, All India Institute of Medical Sciences, New Delhi, India. All registered pregnant women with period of gestation between 28 weeks - 36 weeks were eligible for inclusion in the study. All eligible pregnant women were contacted through home visit. A total of 217 pregnant women were enrolled out of which 209 blood samples were collected from pregnant women. About three ml of blood from antecubital vein was drawn without use of tourniquet, under aseptic conditions. It was later analysed for the serum anti-oxidative measures {Malanoaldehyde, Vitamin C, Ferric Reducing Ability of Plasma (FRAP) levels}. Data were entered on Epi-Info version 3.5.4. Data management and analysis was carried out in Stata 11. The means were compared using t-test and p-value stated. Categorical data was analysed using chi-square test. Logistic regression was used and adjusted p-value was stated. RESULTS: A total of 217 pregnant women were eligible for the study and all were enrolled. Out of the 217 blood samples collected, eight samples accidently got destroyed. A total of 28 out of remaining 209 pregnant women (13.4%) had pre-eclampsia. Mean age (SD) was 22.4 (2.3) years, mean height (SD) was 156.6 (6.9) cm, mean weight (SD) was 65.1 (9.7) kg in pre-eclampsia group. In pre-eclampsia group mean serum levels of malanoaldehyde, vitamin C and FRAP was 4.2 (0.26) ng/dl, 0.83 (0.11) mg/dl, 678.0 (35.6) µmol/L. It was 4.5 (0.09) ng/dl,0.88 (0.03) mg/dl, 599.3 (13.3) µmol/L in normotensive pregnant women group respectively. The difference was statistically significant for FRAP levels only. CONCLUSION: Serum anti-oxidant levels are not raised among women with pre-eclampsia.

Structural and Functional Changes in the Tight Junctions of Asymptomatic and Serology-negative First-degree Relatives of Patients With Celiac Disease.

BACKGROUND: Ten to 15% of first-degree relatives (FDRs) of celiac disease (CeD) patients develop CeD. Although intestinal barrier functions (intestinal permeability) are abnormal in the subset of serology-negative FDRs, what leads to the abnormal barrier function is not known. GOALS: To study the ultrastructure and functions of tight junctions in serology-negative FDRs of CeD patients. STUDY: The intestinal permeability was measured in 97 asymptomatic and anti-tissue transglutaminase antibody (anti-tTG Ab)-negative FDRs (using the lactulose mannitol ratio) and in 75 controls. The ultrastructure of tight junctions using transmission electron microscopy, and the expression of key tight junction proteins (claudin-2, claudin-3, occludin, JAM-A, and ZO-1) and zonulin using real-time PCR and immunohistochemistry were assessed in anti-tTG Ab-negative, HLA-DQ2/-DQ8-positive FDRs having normal villi and in disease controls. In addition, the serum zonulin level was measured in 172 anti-tTG Ab-negative FDRs and 198 controls. RESULTS: The intestinal permeability was significantly increased in FDRs than in controls. Ultrastructural abnormalities such as dilatation of the tight junction (P=0.004) and loss of the pentalaminar structure (P=0.001) were more common in FDRs than in disease controls. There was significant underexpression of tight junction proteins ZO-1 (P=0.040) and occludin (P=0.041) in FDRs. There was no significant difference in the serum zonulin level between FDRs and controls (P=0.154). CONCLUSIONS: Even asymptomatic, anti-tTG-Ab-negative FDRs with a normal villous histology have both ultrastructural and functional abnormalities in tight junctions. These findings are indirect evidence of the presence of tight junction abnormalities before the onset of the disease and may have therapeutic implications.

Prevalence of celiac disease among first-degree relatives of Indian celiac disease patients.

BACKGROUND: Celiac disease, once thought to be uncommon in Asia, is now recognized in Asian nations as well. We investigated the prevalence of celiac disease in first-degree relatives of celiac disease patients followed in our centre. METHODS: First-degree relatives were screened prospectively for celiac disease using questionnaire-based interview and anti-tissue transglutaminase antibody. Serology positive first-degree relatives underwent duodenal biopsies. Diagnosis of celiac disease was made based on positive serology and villous abnormality Marsh grade 2 or higher. Human leucocyte antigen DQ2/-DQ8 was also assessed in 127 first-degree relatives. RESULTS: 434 first-degree relatives of 176 celiac disease patients were prospectively recruited; 282 were symptomatic (64.9%), 58 were positive for serology (13.3%). Seroprevalence was higher in female than in males (19% vs 8.5%; p=0.001) and highest in siblings (16.9%) than parents (13.6%) and children (5.9%) of celiac patients (p=0.055); 87.4% first-degree relatives were human leucocyte antigen-DQ2/-DQ8 positive. Overall prevalence of celiac disease was 10.9% amongst first-degree relatives. CONCLUSIONS: The prevalence of celiac disease in first-degree relatives of celiac disease patients was 10.9% in our cohort, and 87% had human leucocyte antigen-DQ2 or -DQ8 haplotype. All first-degree relatives of celiac disease patients should be screen for celiac disease even if asymptomatic or with atypical manifestations.