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Rajalakshmy A.R.,L and crobiology Research Center | Rajalakshmy A.R.,Center for Nanotechnology and Advanced Biomaterials | Malathi J.,L and crobiology Research Center | Madhavan H.N.,L and crobiology Research Center | And 2 more authors.
Cornea | Year: 2015

Purpose: Dry eye is one of the suggested extrahepatic complications associated with hepatitis C virus (HCV) infection. HCV RNA has been detected from the tear fluid of patients with chronic HCV. There has been no literature evidence on the presence of HCV RNA in the tear fluid of patients with dry eye without HCV infection. In this study, tear fluid of patients with dry eye with no HCV infection was screened for the presence of HCV RNA.Methods: Tear fluid was collected from patients with dry eye (n = 36) and healthy controls (n = 20). Real-time polymerase chain reaction was performed to detect HCV RNA in the tear fluid. Anti-HCV enzyme-linked immunosorbent assay, alkaline phosphatase, and alanine aminotransferase tests were performed in the serum samples collected from 15 patients with dry eye.Results: Viral RNA was detected in 58.3% of the patients. Serum samples collected from 15 patients with dry eye were negative for anti-HCV. Alkaline phosphatase levels were elevated in 12 of 15 patients. Alanine aminotransferase levels were normal in all 15 patients. The odds ratio for the presence of HCV RNA in patients with dry eye was 22.4.Conclusions: These results indicate a direct correlation between dry eye and HCV in non-HCV patients. Copyright © 2014 by Lippincott Williams & Wilkins.


Rajalakshmy A.R.,L and crobiology Research Center | Rajalakshmy A.R.,Center for Nanotechnology and Advanced Biomaterials | Malathi J.,L and crobiology Research Center | Madhavan H.N.,L and crobiology Research Center | And 2 more authors.
Molecular Vision | Year: 2014

Purpose: Dry eye condition is an extrahepatic manifestation associated with chronic hepatitis C virus (HCV) infection. Since conjunctival inflammation can contribute to the dry eye condition, in the present study we analyzed the conjunc-tival inflammatory response to HCV core and NS3 proteins.Methods: We used primary human conjunctival fbroblasts for our study. Cytokines were measured with enzyme-linked immunosorbent assay (ELISA). Toll-like receptor (TLR) and cell adhesion molecule gene expression patterns were analyzed with semiquantitative reverse transcription (RT)–PCR. Immunofuorescence staining was performed for the MyD88, nuclear factor-kappa B (NF-kB), and inducible nitric oxide synthase (iNOS) proteins. Nitric oxide (NO) was measured with the Griess assay; terminal deoxynucleotidyl transferase-mediated uridine 5′-triphosphate-biotin nick end labeling (TUNEL) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays were performed for apoptosis and cell viability, respectively.Results: When exposed to the HCV core and NS3 proteins, the conjunctival fbroblasts secreted interleukin-8 (IL-8), IL-6, tumor necrosis factor-alpha (TNF-α), and IL-10 in a dose-dependent manner. Various TLRs were involved in the innate immune response via MyD88 signaling without NF-kB involvement. The gene expression of cell adhesion molecules such as CD44 and ICAM-1 was upregulated, and the cells secreted NO via iNOS. As the sum of these stress responses, the cells underwent apoptosis, which eventually lead to cell death.Conclusions: HCV core and NS3 proteins induced conjunctival inflammation that may form the pathogenesis of dry eye condition. © 2014 Molecular Vision.


Rajalakshmy A.R.,L and crobiology Research Center | Rajalakshmy A.R.,Center for Nanotechnology and Advanced Biomaterials | Malathi J.,L and crobiology Research Center | Madhavan H.N.,L and crobiology Research Center
Journal of Viral Hepatitis | Year: 2016

Neuroinvasion of hepatitis C virus (HCV) is evidenced by recent clinical studies. In this study, serum-derived HCV infection of astrocytes was analysed. Astrocytes were infected with HCV-positive serum, and viral replication was assessed on different days postinfection. RT-PCR was positive for HCV-negative strand on 5th and 7th day postinfection in the HCV-positive serum-infected astrocytes. Real-time RNA count in the cell culture supernatant was steadily increasing from day 3 to day 7. To reconfirm the viral replication, astrocytes were treated with an antiviral before the serum infection, and the antiviral treatment significantly reduced the viral RNA count. Further, the virus-infected cells stained positive for the presence of viral core protein. Electron microscopy revealed the presence of HCV-like particles in the astrocyte cell culture supernatant. In conclusion, serum-derived HCV replicates in human astrocyte cell line SVG. © 2015 John Wiley & Sons Ltd.


Rajalakshmy A.R.,L and crobiology Research Center | Rajalakshmy A.R.,Center for Nanotechnology and Advanced Biomaterials | Malathi J.,L and crobiology Research Center | Madhavan H.N.,L and crobiology Research Center
Experimental Eye Research | Year: 2014

Direct association of dry eye syndrome and hepatitis C virus (HCV) infection is a well established fact. In this context, the current study examines the invitro corneal inflammatory response with respect to HCV core and NS3 antigens. Toll like receptors (TLRs) are pattern recognition receptors which can mediate innate immune response. In the present study, corneal epithelial cells responded to HCV core and NS3 proteins by secreting pro-inflammatory cytokines IL-8, IL-6 and TNF-α via TLR1, TLR2 and TLR6 mediated innate immune response. MyD88/NF-kB signalling was involved in pro-inflammatory cytokine production. Corneal epithelium synthesised nitric oxide (NO) via iNOS during HCV core and NS3 exposure. On later stages of inflammation, cells underwent apoptosis which lead to cell death. SiRNA mediated silencing of TLR1, TLR2 and TLR6 resulted in a significant down regulation of IL-8 and NO. In conclusion, this study indicates that HCV core and NS3 proteins are capable of inducing immune response in corneal epithelium which can potentiate the pathology of HCV associated dry eye condition. Blocking specific TLR response can have therapeutic application in controlling the inflammatory response associated with this dry eye condition. © 2014 Published by Elsevier Ltd.


Ezhilan M.,Center for Nanotechnology and Advanced Biomaterials | Alagesan S.,Center for Nanotechnology and Advanced Biomaterials | Ramachandra B.L.,Center for Nanotechnology and Advanced Biomaterials | Ramachandra B.L.,SASTRA University | And 9 more authors.
Sensor Letters | Year: 2015

Linear sweep voltammetric behaviour of methylglyoxal was investigated using Pt/ZnO flakes/GLO 1 bio-electrode and a new analytical method was developed for the precise and accurate determination of methylglyoxal in grilled chicken sample. This method was based on the electrochemical parameters measured for the increasing concentration of methylglyoxal. The electrochemical parameters calculated from the linear sweep voltammograms served as the dependent variables. The concentrations of methylglyoxal were employed as the independent variables. For each electrochemical parameter, a non-linear calibration equation was framed and its predictive performance was evaluated. The analytical performance of the proposed models was assessed with the aid of relative prediction error, % recovery and root mean square error for cross validation. The best results were obtained with current density versus [methylglyoxal]model (RPE=0.033, Recovery=97.379% and RMSECV = 1.429). This calibrated non-linear model was then applied for the determination of methylglyoxal in grilled chicken samples. The recovery of methylglyoxal in spiked grilled chicken was ranging from 95.895% to 108.191%. The predicted methylglyoxal values were very similar to that of the spiked methylglyoxal values. Copyright © 2015 American Scientific Publishers..

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