In vitro analysis of material micro-leakage in pit and fissure sealants.

A preferable choice of material offers superior resistance against micro-leakage for clinical applications in preventing dental caries in pits and fissures is of interest. A total of 45 extracted human premolars were cleaned, stored in a saline solution, and randomly divided into three groups, each intended for treatment with one of the sealants: Fuzi VII, ClinPro, and Embrace Wetbond. The application of the sealants followed the manufacturers' instructions strictly. The teeth were subjected to thermal cycling to simulate oral conditions. Marginal micro-leakage was then assessed by dye penetration method using a 0.5% methylene blue dye. Teeth were sectioned, and dye penetration was measured under a stereomicroscope. The results showed that all the tested materials exhibited some degree of micro-leakage. Within the limitations of this In vitro study, it was concluded that Embrace Wetbond exhibited superior performance in terms of minimizing marginal micro-leakage among the tested pit and fissure sealants.

Genome-wide screens identify SEL1L as an intracellular rheostat controlling collagen turnover.

Accumulating evidence has implicated impaired extracellular matrix (ECM) clearance as a key factor in fibrotic disease. Despite decades of research elucidating the effectors of ECM clearance, relatively little is understood regarding the upstream regulation of this process. Collagen is the most abundant constituent of normal and fibrotic ECM in mammalian tissues. Its catabolism occurs through extracellular proteolysis and cell-mediated uptake of collagen fragments for intracellular degradation. Given the paucity of information regarding the regulation of this latter process, here we execute unbiased genome-wide screens to understand the molecular underpinnings of cell-mediated collagen clearance. Using this approach, we discover a mechanism through which collagen biosynthesis is sensed by cells internally and directly regulates clearance of extracellular collagen. The sensing mechanism appears to be dependent on endoplasmic reticulum-resident protein SEL1L and occurs via a noncanonical function of this protein. This pathway functions as a homeostatic negative feedback loop that limits collagen accumulation in tissues. In human fibrotic lung disease, the induction of this collagen clearance pathway by collagen synthesis is impaired, thereby contributing to the pathological accumulation of collagen in lung tissue. Thus, we describe cell-autonomous, rheostatic collagen clearance as an important pathway of tissue homeostasis.

Genome-wide screens identify SEL1L as an intracellular rheostat controlling collagen turnover.

Accumulating evidence has implicated impaired extracellular matrix (ECM) clearance as a key factor in fibrotic disease. Despite decades of research elucidating the effectors of ECM clearance, relatively little is understood regarding the upstream regulation of this process. Collagen is the most abundant constituent of normal and fibrotic ECM in mammalian tissues. Its catabolism occurs through extracellular proteolysis and cell-mediated uptake of collagen fragments for intracellular degradation. Given the paucity of information regarding the regulation of this latter process, we executed unbiased genome-wide screens to understand the molecular underpinnings of cell-mediated collagen clearance. Using this approach, we discovered a previously unappreciated mechanism through which collagen biosynthesis is sensed by cells internally and directly regulates clearance of extracellular collagen. The sensing mechanism is dependent on endoplasmic reticulum-resident protein SEL1L and occurs via a noncanonical function of SEL1L. This pathway functions as a homeostatic negative feedback loop that limits collagen accumulation in tissues. In human fibrotic lung disease, the induction of this collagen clearance pathway by collagen synthesis is impaired, thereby contributing to the pathological accumulation of collagen in lung tissue. Thus cell-autonomous, rheostatic collagen clearance is a previously unidentified pathway of tissue homeostasis.

Genetic and Functional Characterization of HIV-1 Vpu from HIV-1-Infected North Indian Population.

Acquired immunodeficiency syndrome is a pandemic disease due to increased variability in causative agent in global distribution; it is attributed to various complications in developing the vaccine, namely, error-prone reverse transcriptase, rapid replication, and high recombination rate. Vpu downmodulates CD4 in infected cells, and it targets the newly synthesized CD4 molecules from the endoplasmic reticulum. The aim of this study was to identify the level of genetic changes in the Vpu gene from HIV-1-infected North Indian individuals and determine the functional relevance with respect to the CD4 downregulation potential of this protein. Genomic DNA was isolated from peripheral blood mononuclear cells, and the Vpu gene was polymerase chain reaction amplified with specific primers followed by cloning, sequencing, and sequence analyses using bioinformatic tools for predicting HIV-1 subtypes, recombination events, conservation of domains, and phosphorylation sites. Among all Vpu variants, three of the variants having serine substitution (serine-52 and serine-56 conversion to isoleucine; S52I and S56I) had lost their functional ß-TrcP binding motif. However, the specific determinants for CD4 (V20, W22, S23) and BST-2 (A11, A15, I17, and A19) binding remained highly conserved. The data obtained with Vpu mutants recommend that the serine residue substitutions in cytoplasmic domain distress the CD4 downregulation activity of Vpu. These events are likely to have implications for viral pathogenesis and vaccine formulations.

IL-6: An endogenous activator of MMP-9 in preterm birth.

Preterm birth or PTB (<37 weeks) is a heterogeneous phenotype with numerous biological pathways. The lack of explanation due to complex pathways, inconsistent observations indicates the need for employing the role of genetic determinants to anticipate the danger of PTB. In this present study, we investigated the possible gene-gene interaction of five SNPs with PTB and its association with total MMP-9 levels. A total of 510 recruitments (250 terms and preterm each) were made and were genotyped by Restriction Fragment length polymorphism (RFLP). Generalized Multifactor Dimensionality Reduction (GMDR) method was carried out for determining gene-gene interaction. ANOVA and t-test were used to identify the association of IL-6 polymorphism with PTB alone and correspondingly with PTB and low birth weight infants (i.e. < 2500 kg). The combination of IL-6 and MMP-9 and MMP-1, MMP-8 and MMP-9 polymorphism was selected through GMDR analysis concerning mothers with preterm and term birth (accuracy 0.5921 and 0.8030 with Cross-Validation Consistency (CVC) 10/10 respectively). Increased expression of MMP-9 was reported in cases in those mothers carrying IL-6 G allele, which was profoundly associated with PTB independently. IL-6 polymorphisms showed synergistic effects in terms of increased total MMP-9 levels in the present study.

Effect of Exposure of Plastic Infant Feeding Bottle Leached Water on Biochemical, Morphological and Oxidative Stress Parameters in Rats.

Bisphenol A (BPA) is leached out from plastic infant feeding bottles that are filled with warm milk/water due to high temperatures, exposing the infants to BPA. The present study aims to understand the effects of ingestion of BPA leached from plastic infant feeding bottle and delineate the underlying mechanisms in rats. In this study, adult rats of Wistar strain were divided into 3 groups. In the first group, the rats consumed normal food and tap water ad libitum. In the second group, the rats ingested BPA (20 µg/kg bodyweight/day, orally). In the third group, the rats drank water leached from plastic infant feeding bottles. After 30days, tests involving biochemical parameters, histopathological examination, and oxidative stress enzyme markers were performed, and the levels of BPA in plastic-leached water were estimated by HPLC analysis. There were significant biochemical changes in the form of increased alkaline phosphatase (ALP), creatine kinase-muscle/brain (CK-MB), and lactate dehydrogenase (LDH) levels in both treated groups as compared to control group, accompanied by structural damage to the vital organs, and lipid peroxidation, glutathione reductase, and catalase activity were also high in the treated groups. Further, the BPA concentration in plastic leached water was estimated to be 0.1 ± 0.02 µg/mL.

Role of MMP-1, MMP-8 and MMP-9 gene polymorphisms in preterm birth.

Novel approaches to preterm births are underway building upon our prior discoveries and probing into unknown discovery pathways. The recent findings showed a high affinity of MMP-9 in serum and its polymorphisms for preterm birth. This study, which is a hospital-based case-control study, aims to investigate the association of MMP-1, MMP-8 and MMP-9 polymorphisms, and levels of MMP-9 in preterm birth. Increased level of MMP-9 was reported in cases as compared to control. The significant association of MMP-9 (-1562) CT (P = 0.001; OR = 1.44(CI = 0.97-2.14)) and TTgenotype (P = 0.05;OR = 2.6 (CI = 1.46-4.69)) were reported in preterm birth. Our findings suggest that the MMP-9 plays an important role in contributing preterm labour and this can be used as a diagnostic tool during pregnancy.

Association of TLR4 and TNF-a Gene Polymorphisms and TLR4 mRNA Levels in Preterm Birth in a Northern Indian Population.

OBJECTIVE: To assess the association of TLR4 (rs4986790 and rs4986791) and TNF-a (rs1800629) genes polymorphisms and TLR4mRNA levels with preterm birth. METHODS: Hospital-based case-control study on women of Caucasoid morphological subtype ethnicity in Northern India. Inclusion criteria for cases: women aged between 18-40 years with preterm birth (<37 weeks gestation), and for controls: women who delivered a term neonate consecutive to an enrolled case. Three polymorphisms TLR4 (Asp299 Gly, Thr399 Ill) and TNF-á (-308G/A) and TLR4 mRNA levels were compared between cases and controls. RESULTS: From 2012-2015, 559 cases and 559 controls were recruited. TLR4 mRNA levels were found to be higher (P<0.001) in cases [(0.7 (0.04)] than in controls [(0.5 (0.04)]. No association was found between TLR4 Asp299 Gly, TLR4 Thr399 Ill and TNF-a (-308G/A) with preterm birth. CONCLUSIONS: Increased TLR4 mRNA levels seem to be associated with preterm birth, and can be investigated further as a potential biomarker for identifying women at risk.

Interleukin 1 receptor antagonist (IL1RA) gene polymorphism and levels associated with adverse outcome in severe community-acquired pneumonia in children: A hospital-based study in India.

BACKGROUND: High morbidity and mortality due to community-acquired pneumonia (CAP) is seen in children under 5 years of age in India. Besides identified risk factors for CAP, there may be a phenotype-genotype association with cytokines, resulting in enhanced inflammatory response resulting in the adverse outcome (AO), namely complications and death. AIM: To assess the association of IL1RA gene polymorphism on serum levels of IL1RA and with AO in children under 5 years of age hospitalized with WHO-defined severe CAP. METHOD: A prospective cohort study with nested case-control design conducted in a tertiary care teaching hospital after obtaining institutional ethical approval. Included were children between 2 and 59 months of age hospitalized with WHO-defined severe CAP with consistent radiological abnormalities. Excluded were those with suspected or proven cystic fibrosis, pulmonary tuberculosis, malignancy, immunodeficiency, and congenital heart disease. Polymerase chain reaction (PCR) was used to analyze the Variable Number of Tandem Repeats (VNTRs) of IL1RA gene polymorphism and ELISA test to detect serum levels of IL1RA. RESULTS: From 2014 to 2016, of 420 screened cases, 350 were eligible and included, of which 132 (37.7%) had no complication and 218 (62.3%) had AO, which included complications like empyema, pyopneumothorax, acute respiratory distress syndrome (ARDS), and septic shock of these 24 (6.9%) expired. Higher risk of AO was seen in A2A2 genotype (OR 11.18, p 0.0001) and lower in A1A1 genotype (OR 0.18, P < 0.0001). Serum IL1RA (ng/mL) was statistically significantly elevated in CAP with AO (2.55 ± 1.44) versus uncomplicated (0.87 ± 0.52) (P < 0.0001). CONCLUSION: In IL1RA gene, A1A1 genotype was associated with lower risk and A2A2 genotype with increased the risk of AO. Higher serum levels of IL1RA were found in A2A2 genotype indicating possibly enhanced inflammatory response resulting in AO of CAP.

Association of IL-10 Gene Polymorphism (-819C > T, -592C > A and -1082G > A) with Preterm Birth.

OBJECTIVE: To analyze the association of IL-10 gene and its polymorphisms with preterm birth (PTB). METHODS: Five hundred and fifty nine women with term birth and 559 with preterm birth were recruited from Lucknow, India. Genetic association analysis was conducted between cases and controls. Subjects recruited as cases were women (aged between 18-40 y) with singleton delivery before 37 wk of gestation and controls were with delivery after or on 37 wk. The genotyping was performed for rs1800871, rs1800872 and rs1800896 for assessing the allelic distribution, haplotypic association and linkage disequilibrium analysis. IL-10mRNA levels were evaluated by real time quantitative polymerase chain reaction (PCR) method. RESULTS: The risk of PTB was found significant in women carrying IL-10 (-1082) GA genotype [OR=1.72(1.7-2.5), p=0.006]. The haplotypic analysis of studied polymorphisms for rs1800871, rs1800872 and rs1800896 depicted the association of ATA (p=0.02) and ATC (p=0.01) haplotypes with PTB. The IL-10 mRNA levels were significantly lower in cases (p=0.05). CONCLUSIONS: IL-10 marks a protective impact in the inflammatory pathway of PTB.

Interplay of cytokines in preterm birth.

Preterm infants (i.e., born before <37 wk of gestation) are at increased risk of morbidity and mortality and long-term disabilities. Global prevalence of preterm birth (PTB) varies from 5 to 18 per cent. There are multiple aetiological causes and factors associated with PTB. Intrapartum infections are conventionally associated with PTB. However, maternal genotype modulates response to these infections. This review highlights the association of cytokine gene polymorphisms and their levels with PTB. Varying PTB rates across the different ethnic groups may be as a result of genetically mediated varying cytokines response to infections. Studies on genetic variations in tumour necrosis factor-alpha, interleukin-1 alpha (IL-1α), IL-1ß, IL-6, IL-10 and toll-like receptor-4 genes and their association with PTB, have been reviewed. No single polymorphism of the studied genes was found to be associated with PTB. However, increased maternal levels of IL-1ß and IL-6 and low levels of IL-10 have been found to be associated with PTB.

Cholesterol-lowering drug, in combination with chromium chloride, induces early apoptotic signals in intracellular L. donovani amastigotes, leading to death.

Leishmania establishes a successful parasitism by evading both oxidative and non-oxidative killing pathways, and its drug resistance against the currently available therapeutics demands for a safe and cheap drug. Since the parasite synthesizes ergosterol instead of cholesterol, using the same biochemical pathway and enzymes, an inhibitor of HMG-CoA-Reductase, Lovastatin, has been tried for its anti-Leishmanial effect. Lovastatin, being an inhibitor of HMG-CoA-Reductase, inhibits infection by cholesterol depletion, while chromium chloride complexes, at their higher concentrations, are reported to exhibit cytotoxicity. In intracellular amastigotes, cytotoxicity has been checked by assessing various manifestation of cell death, viz. DNA fragmentation, AnnexinV-FITC binding and JC-1 fluorescence ratio. Release of hydrogen peroxide (HPO) and nitric oxide (NO) has been assessed in live cell. Lovastatin and CrCl3.6H2O in combination has appeared to be ineffective on promastigotes but has induced cytotoxic effect on the intracellular amastigotes through up-regulation of cellular signalling mechanisms. CrCl 3.6H2O stimulates generation of NO, leading to reduction of the number of intracellular amastigote, while Lovastatin shows HPO-mediated killing of the same, keeping the host cell unaffected. This novel therapeutic approach, involving two known safe compounds in suboptimal doses, may resolve human visceral Leishmaniasis.