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Reddy V.,National Institute of Mental Health and Neuro Sciences | Ravi V.,National Institute of Mental Health and Neuro Sciences | Desai A.,National Institute of Mental Health and Neuro Sciences | Parida M.,Defence Research and Development Establishment DRDE | And 2 more authors.
Journal of Medical Virology | Year: 2012

Chikungunya fever a re-emerging infection with expanding geographical boundaries, can mimic symptoms of other infections like dengue, malaria which makes the definitive diagnosis of the infection important. The present study compares the utility of four laboratory diagnostic methods viz. IgM capture ELISA, an in house reverse transcription PCR for the diagnosis of Chikungunya fever, TaqMan real-time PCR, and a one step reverse transcription-loop mediated isothermal amplification assay (RT-LAMP). Out of the 70 serum samples tested, 29 (41%) were positive for Chikungunya IgM antibody by ELISA and 50 (71%) samples were positive by one of the three molecular assays. CHIKV specific nucleic acid was detected in 33/70 (47%) by reverse transcription PCR, 46/70 (66%) by TaqMan real-time PCR, and 43/70 (62%) by RT-LAMP assay. A majority of the samples (62/70; 89%) were positive by at least one of the four assays used in the study. The molecular assays were more sensitive for diagnosis in the early stages of illness (2-5 days post onset) when antibodies were not detectable. In the later stages of illness, the IgM ELISA is a more sensitive diagnostic test. In conclusion we recommend that the IgM ELISA be used as an initial screening test followed one of the molecular assays in samples that are collected in the early phase of illness and negative for CHIKV IgM antibodies. Such as approach would enable rapid confirmation of the diagnosis and implementation of public health measures especially during outbreaks. © 2012 Wiley Periodicals, Inc.


Jarouliya U.,Jiwaji University | Anish Zacharia J.,Jiwaji University | Kumar P.,Defence Research and Development Establishment DRDE | Bisen P.S.,Jiwaji University | And 2 more authors.
Indian Journal of Medical Research | Year: 2012

Background & objectives: Diabetes mellitus is a metabolic disorder characterized by hyperglycaemia. Several natural products have been isolated and identified to restore the complications of diabetes. Spirulina maxima is naturally occurring fresh water cyanobacterium, enriched with proteins and essential nutrients. The aim of the study was to determine whether S. maxima could serve as a therapeutic agent to correct metabolic abnormalities induced by excessive fructose administration in Wistar rats. Methods: Oral administration of 10 per cent fructose solution to Wistar rats (n=5 in each group) for 30 days resulted in hyperglycaemia and hyperlipidaemia. Aqueous suspension of S. maxima (5 or 10%) was also administered orally once daily for 30 days. The therapeutic potential of the preparation with reference to metformin (500 mg/kg) was assessed by monitoring various biochemical parameters at 10 day intervals during the course of therapy and at the end of 30 days S. maxima administration. Results: Significant (P<0.001) reductions in blood glucose, lipid profile (triglycerides, cholesterol and LDL, VLDL) and liver function markers (SGPT and SGOT) were recorded along with elevated level of HDL-C at the end of 30 days therapy of 5 or 10 per cent S. maxima aquous extract. Co-administration of S. maxima extract (5 or 10% aqueous) with 10 per cent fructose solution offered a significant protection against fructose induced metabolic abnormalities in Wistar rats. Interpretation & conclusions: The present findings showed that S. maxima exhibited anti-hyperglycaemic, anti-hyperlipidaemic and hepatoprotective activity in rats fed with fructose. Further studies are needed to understand the mechanisms.


Kumar J.S.,Defence Research and Development Establishment DRDE | Khan M.,Defence Research and Development Establishment DRDE | Gupta G.,Canadian Department of National Defence | Bhoopati M.,Canadian Department of National Defence | And 2 more authors.
Viral Immunology | Year: 2012

The resurgence of Chikungunya (CHIK) virus in the form of an explosive, unprecedented epidemic with high virulence and unusual numbers of fatalities has created an immense public health concern in recent years. In the absence of an effective vaccine and specific antiviral therapy, early accurate diagnosis is essential for the best patient management. The present study describes the production and characterization of high-affinity and selective monoclonal antibodies (Mabs) against recombinant E2 protein (rE2) of the CHIK virus. The reactivity of Mabs for rE2 protein was demonstrated using ELISA. The specificity of the generated Mabs for rE2 was demonstrated by Western blot and indirect immunofluorescence. The application of this CHIK virus E2-specific monoclonal antibody in early clinical diagnosis was demonstrated by various analytical methods, such as immunoblotting, indirect immunofluorescence assay (IFA), and antigen-capture ELISA (AC-ELISA), for the detection as well as the identification of the novel ECSA genotypes of CHIK virus. These findings suggest that the high-affinity E2-specific monoclonal antibodies reported in this study will be useful for early clinical diagnosis and epidemiological studies of CHIK virus in developing countries. © 2012, Mary Ann Liebert, Inc.


Dhanwani R.,Defence Research and Development Establishment DRDE | Dhanwani R.,University of Minnesota | Khan M.,Defence Research and Development Establishment DRDE | Lomash V.,Defence Research and Development Establishment DRDE | And 3 more authors.
PLoS ONE | Year: 2014

While a number of studies have documented the persistent presence of chikungunya virus (CHIKV) in muscle tissue with primary fibroblast as the preferable cell target, little is known regarding the alterations that take place in muscle tissue in response to CHIKV infection. Hence, in the present study a permissive mouse model of CHIKV infection was established and characterized in order to understand the pathophysiology of the disease. The two dimensional electrophoresis of muscle proteome performed for differential analysis indicated a drastic reprogramming of the proteins from various classes like stress, inflammation, cytoskeletal, energy and lipid metabolism. The roles of the affected proteins were explained in relation to virus induced myopathy which was further supported by the histopathological and behavioural experiments proving the lack of hind limb coordination and other loco-motor abnormalities in the infected mice. Also, the level of various proinflammatory mediators like IL-6, MCP-1, Rantes and TNF-α was significantly elevated in muscles of infected mice. Altogether this comprehensive study of characterizing CHIKV induced mouse myopathy provides many potential targets for further evaluation and biomarker study. © 2014 Dhanwani et al.


Srivastava A.,Indian Institute of Toxicology Research | Yadav S.,Indian Institute of Toxicology Research | Sharma A.,Indian Institute of Toxicology Research | Dwivedi U.,University of Lucknow | And 2 more authors.
Xenobiotica | Year: 2012

Freshly prepared peripheral blood lymphocytes (PBL) are known to express cytochrome P450s (CYPs) and glutathione S-transferases (GSTs) involved in the bioactivation and detoxification of organic components of diesel exhaust particles (DEPs). To validate that blood lymphocyte expression profiles could be used as a biomarker to predict exposure to vehicular emissions, similarities in the alterations in the mRNA expression of CYPs and GSTs were studied in PBL and lungs of rats exposed to DEPs. Adult male Wistar rats were treated transtracheally with different doses of DEPs (3.75- or 7.5- or 15- or 30-mg/kg b.wt.). The animals were anaesthetized after 24h and blood was drawn and lungs were taken out and processed. DEP produced a similar pattern of increase in the mRNA expression of CYPs (CYP1A1, 1A2, 1B1, 2E1), associated arylhydrocarbon receptor (Ahr) and arylhydrocarbon nuclear translocator (Arnt) and GSTs (GSTPi, GSTM1 and GSTM2) at all the doses in lungs and PBL. The protein expression of CYP1A1/1A2 and 2E1 and catalytic activity of CYPs and GSTs also showed a similar pattern of increase in blood lymphocyte and in lungs isolated from DEP treated rats. Our data indicating similarities in the alterations in the expression of carcinogen metabolizing CYPs and GSTs in PBL with the lung enzymes suggests the suitability of using expression profiles of blood lymphocyte CYPs and GSTs as a biomarker to predict exposure to vehicular emissions. © 2012 Informa UK, Ltd.


Sharma S.,Defence Research and Development Establishment DRDE | Parida M.,Defence Research and Development Establishment DRDE | Shukla J.,Defence Research and Development Establishment DRDE | Rao P.V.L.,Defence Research and Development Establishment DRDE
Virology Journal | Year: 2011

Background: The H1N1pandemic virus is a newly emergent human influenza A virus that is closely related to a number of currently circulating pig viruses in the 'classic North American' and 'Eurasian' swine influenza virus lineages and thus referred as S-OIV. Since the first reports of the virus in humans in April 2009, H1N1 virus has spread to 168 countries and overseas territories. India also witnessed severe H1N1 pandemic virus epidemic with considerable morbidity and mortality in different parts starting from May 2009. Findings. The suspected swine flu outbreak from Gwalior India during October- December 2009 was confirmed through S-OIV HA gene specific RT-LAMP and real time RT-PCR. Positive samples through CDC real time and Lamp assay were further processed for isolation of the virus. Full HA gene sequencing of the H1N1 isolates of Gwalior, India revealed 99% homology with California and other circulating novel swine flu viruses. Three major changes were observed at nucleotide level, while two major amino acid shifts were observed at the position C9W and I30M corresponding to the ORF with prototype strain. The HA gene sequence phylogeny revealed the circulation of two genetically distinct lineages belonging to Clade VII and Clade I of S-OIV. Conclusions: Our findings also supported the earlier report about circulation of mixed genogroups of S-OIV in India. Therefore continuous monitoring of the genetic makeup of this newly emergent virus is essential to understand its evolution within the country. © 2011 Sharma et al; licensee BioMed Central Ltd.


Maurya C.K.,Defence Research and Development Establishment DRDE | Gupta P.K.,Defence Research and Development Establishment DRDE
Tetrahedron Letters | Year: 2015

Abstract Sodium metal stabilized in silica gel (Na-SG) has been reported as a powerful reducing agent. Being less pyrophoric than free metal, it has been used to carry out a variety of organic transformations including desulfonation, Birch reduction, Bouveault-Blanc Ester Reduction, etc. In this Letter, we describe a newer method involving its use for decarbamoylation of primary carbamates to the corresponding alcohols/phenols. In comparison with the reported methods, the present method requires milder reaction conditions and proceeds at room temperature. The reaction worked in solvents like THF and acetonitrile, the later giving better results in terms of reaction profiles. The process has been extended for decarbamoylation of a variety of carbamates. © 2015 Elsevier Ltd. All rights reserved.


Dhanwani R.,Defence Research and Development Establishment DRDE | Khan M.,Defence Research and Development Establishment DRDE | Alam S.I.,Defence Research and Development Establishment DRDE | Rao P.V.L.,Defence Research and Development Establishment DRDE | Parida M.,Defence Research and Development Establishment DRDE
Proteomics | Year: 2011

Chikungunya infection is a major disease of public health concern. The recurrent outbreaks of this viral disease and its progressive evolution demands a potential strategy to understand major aspects of its pathogenesis. Unlike other alphaviruses, Chikungunya virus (CHIKV) pathogenesis is poorly understood. In every consecutive outbreak, some new symptoms associated with virulence and disease manifestations are being reported such as neurological implication, increased severity and enhanced vector competence. In order to unravel the mechanism of the disease process, proteomic analysis was performed to evaluate the host response in CHIKV-infected mice tissues. Comparative analysis of the multiple gels representing the particular tissue extract from mock and CHIKV-infected tissues revealed a drastic reprogramming of physiological conditions through 35 and 15 differentially expressed proteins belonging to different classes such as stress, inflammation, apoptosis, urea cycle, energy metabolism, etc. from liver and brain, respectively. Based on the alterations obtained in the CHIKV mouse model, most of the aspects of CHIKV infection such as disease severity, neurological complications, disease susceptibility and immunocompetence could be defined. This is the first report unravelling the complicated pathways involved in the mechanism of Chikungunya disease pathogenesis employing proteomic approach. © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.


Kumar J.S.,Defence Research and Development Establishment DRDE | Saxena D.,Defence Research and Development Establishment DRDE | Parida M.,Defence Research and Development Establishment DRDE
Molecular and Cellular Probes | Year: 2014

The recent outbreaks of West Nile Virus (WNV) in the Northeastern American continents and other regions of the world have made it essential to develop an efficient protocol for surveillance of WN virus. Nucleic acid based techniques like, RT-PCR have the advantage of sensitivity, specificity and rapidity. A one step single tube Env gene specific real-time RT-PCR was developed for early and reliable clinical diagnosis of WNV infection in clinical samples. The applicability of this assay for clinical diagnosis was validated with 105 suspected acute-phase serum and plasma samples from the recent epidemic of mysterious fever in Tamil Nadu, India in 2009-10. The comparative evaluation revealed the higher sensitivity of real-time RT-PCR assay by picking up 4 additional samples with low copy number of template in comparison to conventional RT-PCR. All the real-time positive samples further confirmed by CDC reported TaqMan real-time RT-PCR and quantitative real-time RT-PCR assays for the simultaneous detection of WNV lineage 1 and 2 strains. The quantitation of the viral load samples was done using a standard curve. These findings demonstrated that the assay has the potential usefulness for clinical diagnosis due to detection and quantification of WNV in acute-phase patient serum samples. © 2014 Elsevier Ltd.


Maurya C.K.,Defence Research and Development Establishment DRDE | Mazumder A.,Defence Research and Development Establishment DRDE | Kumar A.,Defence Research and Development Establishment DRDE | Gupta P.K.,Defence Research and Development Establishment DRDE
Synlett | Year: 2016

We report an efficient procedure for the synthesis of symmetrical disulfanes from organic thiocyanates in the presence of sodium in silica gel at room temperature. By avoiding the use of foul-smelling thiols, the present protocol provides an attractive alternative to existing methods for the preparation of symmetrical disulfanes. © Georg Thieme Verlag Stuttgart New York - Synlett 2016.

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