Siddharthanagar, India
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Lavu P.S.R.,Defence Food Research Laboratory DFRL | Mondal B.,Defence Food Research Laboratory DFRL | Ramlal S.,Defence Food Research Laboratory DFRL | Murali H.S.,Defence Food Research Laboratory DFRL | Batra H.V.,Defence Food Research Laboratory DFRL
ACS Combinatorial Science | Year: 2016

This study describes the selection of single-stranded DNA (ssDNA) aptamers against Salmonella enterica serovar Typhimurium using a modified whole cell systematic evolution of ligands by exponential enrichment (whole cell SELEX). For evolving specific aptamers, ten rounds of selection to live Salmonella cells, alternating with negative selection against a cocktail of related pathogens, were performed. The resulting highly enriched oligonucleotide pools were sequenced and clustered into eight groups based on primary sequence homology and predicted secondary structure similarity. Fifteen sequences from different groups were selected for further characterization. The binding affinity and specificity of aptamers were determined by fluorescence binding assays. Aptamers (SAL 28, SAL 11, and SAL 26) with dissociation constants of 195 ± 46, 184 ± 43, and 123 ± 23 nM were used to develop a nanogold-based colorimetric detection method and a sedimentation assay. The former showed a better sensitivity limit of 102 CFU/mL using aptamer SAL 26. This approach should enable further refinement of diagnostic methods for the detection of Salmonella enterica serovar Typhimurium and of other microbial pathogens. © 2016 American Chemical Society.


Ilaiyaraja N.,Defence Food Research Laboratory DFRL | Likhith K.R.,University of Mysore | Sharath Babu G.R.,Defence Food Research Laboratory DFRL | Khanum F.,Defence Food Research Laboratory DFRL
Food Chemistry | Year: 2015

In this study, the extraction conditions for the maximum recovery of polyphenols with high antioxidant activity were optimised by response surface methodology (RSM) in Feronia limonia fruit. The independent variables were viz. concentration of ethanol (X1: 30-70%), incubation temperature (X2: 37-60%) and solvent-to-solid ratio (X3: 20-40%). ANOVA results showed that concentration of ethanol and temperature affected the total polyphenol content (TPC, Y1), DPPH (Y2) and ABTS (Y3) radical scavenging activities significantly (p < 0.05) whereas solvent-to-solid ratio was found to be insignificant. A second-order polynomial model satisfactorily fitted the experimental data with the R2 values of 0.966, 0.946 and 0.955, respectively for the responses Y1, Y2 and Y3 (p < 0.0001), implying a good agreement between the predicted and experimental values. The optimal conditions for the highest yield of TPC (7.21 ± 1.4 g GAE/g) with >80% radical scavenging activities were derived at X1 = 62.7%, X1 = 49.7°C and X3 = 39.4 mL/g. © 2014 Elsevier Ltd. All rights reserved.


Ilaiyaraja N.,Defence Food Research Laboratory DFRL | Khanum F.,Defence Food Research Laboratory DFRL
Journal of Dietary Supplements | Year: 2011

The protective effect of a methanolic extract (ME) of Acorus calamus against alcohol-induced hepatotoxicity and oxidative stress was studied in rats. The in vitro assays using DPPH and ABTS showed a strong antioxidant activity of the extract with the total polyphenolic content of 156 mg/g. Chronic ethanol administration causes an increase in oxidative stress and tissue injury with decreased antioxidant status. In this study, continuous administration of ethanol (7.9 g/kg body weight/day) for a period of 6 weeks resulted in a significant (p < .001) increase in the levels of serum aspartate aminotransferase, serum alanine aminotransferase, alkaline phospahatase, and bilirubin with the decreased level of total antioxidant status. Moreover, the levels of lipid peroxidation markers (malondialdehyde and hydroperoxides) as well as protein carbonyl content were also increased (p < .001), whereas the levels of non-enzymic antioxidants (glutathione, vitamin C, and vitamin E) decreased significantly in the liver tissues of ethanol-administered control rats. Pretreatment of rats with ME at doses of 300 and 600 g/kg body weight before alcohol administration significantly reduced the hepatic marker enzymes, level of lipid peroxidation, and protein oxidation, and increased the enzymatic and non-enzymatic antioxidant levels in liver. These observations were supplemented by histopathological examination of liver sections. Overall, the present study shows that the administration of ME ameliorates the antioxidant status as well as protects against the toxic effects of ethanol in rats, thereby suggesting its use as an effective botanical supplement for hepatoprotection. © 2011 Informa Healthcare USA, Inc.


Wadikar D.D.,Defence Food Research Laboratory DFRL | Premavalli K.S.,Defence Food Research Laboratory DFRL
International Journal of Food Sciences and Nutrition | Year: 2011

The present study aimed at evaluating the effect of appetizer administration on plasma leptin levels of human volunteers. The ginger-based appetizers, namely ginger munch, fruit munch, jeera munch and appetizer drink, developed in the Defence Food Research Laboratory were used for 45 volunteers. Leptin was analyzed using the BioSource enzyme-amplified sensitivity immunoassay kit. The fasting plasma leptin level for men and women ranged between 0.5 and 19.5 ng/ml and between 2 and 36 ng/ml, respectively. The decreased (6-16%) plasma leptin levels after consumption of appetizers indicated their appetizing effect. © 2011 Informa UK, Ltd.


Ilaiyaraja N.,Defence Food Research Laboratory DFRL | Khanum F.,Defence Food Research Laboratory DFRL
International Journal of Integrative Biology | Year: 2010

Protective effect of leaves of Cichorium intybus was studied against paracetamol-induced hepatotoxicity in male Wistar rats. The aqueous extract at the dose of 200 mg and 400 mg/kg b.wt were prefed to the rats for 30 days followed by intoxication with a single overdose of paracetamol (2g/kg b.wt). After 24 hours, all the animals were sacrificed and blood and liver samples were obtained for biochemical analysis. The results shows that paracetamol induces hepatotoxicity with significant increase in serum biochemical markers such as glutamate pyruvic transaminase (SGPT), glutamate oxaloacetate transaminase (SGOT), alkaline phosphatase (ALP) and bilirubin and reduces the activities of hepatic antioxidant enzymes as well. Supplementation of the aqueous extract lowered these changes significantly (p<0.05) as compared to paracetamol control group only at the higher dose of 400 mg/kg b.wt. Prefeeding of the extract also resulted in significant increase (p<0.05) in the decreased activities of hepatic antioxidant enzymes such as catalase, glutathione peroxidase, glutathione reductase, glutamate-s-tranferase, superoxide dismutase (SOD). The present study indicate the hepatoprotective properties of Chicory leaf as observed through the improvement in the antioxidant status, which may be attributed to its antioxidant and radical scavenging properties of the extract. © IJIB, All rights reserved.


Mondal B.,Defence Food Research Laboratory DFRL | Ramlal S.,Defence Food Research Laboratory DFRL | Lavu P.S.R.,Defence Food Research Laboratory DFRL | Murali H.S.,Defence Food Research Laboratory DFRL | Batra H.V.,Defence Food Research Laboratory DFRL
Applied Microbiology and Biotechnology | Year: 2015

Aptamers are synthetic DNA recognition elements which form unique conformations that enable them to bind specifically to their targets. In the present study, an attempt was made to standardize a new modified combinatorial method comprising of Ni-NTA affinity Systematic Evolution of Ligands by Exponential Enrichment (SELEX; based on affinity between His tag protein and Ni-NTA), membrane SELEX (based on immobilization of protein on nitrocellulose membrane), and microtiter plate based SELEX (to monitor affinity and to enrich the selected aptamers) for protein targets. For experimental evaluation, staphylococcal interotoxin B was the molecule chosen. The new combinatorial method enhanced selection ability up to 51.20 % in comparison with individual conventional procedures. Employing this method following six rounds of selection, high-affinity aptamers with very different properties could be obtained with a dissociation constant (Kd) value as low as 34.72 ± 25.09 nM. The optimal aptamers could be employed in fluorescence binding assay, enzyme-linked oligonucleotide assays, and aptamer-based Western blot assay for characterization and detection. These results pave a potential path without using of any robotics for high-throughput generation of aptamers with advantages in terms of rapidity, simplicity, and ease in handling. © 2015, Springer-Verlag Berlin Heidelberg.


PubMed | Defence Food Research Laboratory DFRL
Type: Journal Article | Journal: ACS combinatorial science | Year: 2016

This study describes the selection of single-stranded DNA (ssDNA) aptamers against Salmonella enterica serovar Typhimurium using a modified whole cell systematic evolution of ligands by exponential enrichment (whole cell SELEX). For evolving specific aptamers, ten rounds of selection to live Salmonella cells, alternating with negative selection against a cocktail of related pathogens, were performed. The resulting highly enriched oligonucleotide pools were sequenced and clustered into eight groups based on primary sequence homology and predicted secondary structure similarity. Fifteen sequences from different groups were selected for further characterization. The binding affinity and specificity of aptamers were determined by fluorescence binding assays. Aptamers (SAL 28, SAL 11, and SAL 26) with dissociation constants of 195 46, 184 43, and 123 23 nM were used to develop a nanogold-based colorimetric detection method and a sedimentation assay. The former showed a better sensitivity limit of 10(2) CFU/mL using aptamer SAL 26. This approach should enable further refinement of diagnostic methods for the detection of Salmonella enterica serovar Typhimurium and of other microbial pathogens.


PubMed | Defence Food Research Laboratory DFRL
Type: Journal Article | Journal: Applied microbiology and biotechnology | Year: 2015

Aptamers are synthetic DNA recognition elements which form unique conformations that enable them to bind specifically to their targets. In the present study, an attempt was made to standardize a new modified combinatorial method comprising of Ni-NTA affinity Systematic Evolution of Ligands by Exponential Enrichment (SELEX; based on affinity between His tag protein and Ni-NTA), membrane SELEX (based on immobilization of protein on nitrocellulose membrane), and microtiter plate based SELEX (to monitor affinity and to enrich the selected aptamers) for protein targets. For experimental evaluation, staphylococcal interotoxin B was the molecule chosen. The new combinatorial method enhanced selection ability up to 51.20 % in comparison with individual conventional procedures. Employing this method following six rounds of selection, high-affinity aptamers with very different properties could be obtained with a dissociation constant (K d) value as low as 34.7225.09 nM. The optimal aptamers could be employed in fluorescence binding assay, enzyme-linked oligonucleotide assays, and aptamer-based Western blot assay for characterization and detection. These results pave a potential path without using of any robotics for high-throughput generation of aptamers with advantages in terms of rapidity, simplicity, and ease in handling.


PubMed | Defence Food Research Laboratory DFRL
Type: Journal Article | Journal: Journal of dietary supplements | Year: 2014

Guggulsterone or guggulipid is a steroidal constituent present in the neutral fraction of gum resin of Commiphora mukul, commonly known as guggul. The traditional uses of guggul-resin extract are well documented in the Ayurveda-where it is prescribed to treat a variety of ailments including lipid-related disorders such as obesity and arteriosclerosis. The hypolipidemic activity of the extracts known since ancient times can be traced to the two closely related steroidal ketones, E-guggulsterone and Z-guggulsterone. In this study, we have investigated the dose dependent (100, 200, 400 mg/kg body weight) effect of guggulsterones on appetite regulating hormones [ghrelin, leptin, cholecystokinin (CCK)] and neurotransmitters (serotonin and dopamine), which play a major role in the energy homeostasis and thus influence obesity related factors. We have also studied its effect on food intake, body weight and plasma triglycerides and glucose in rats. Guggulsterones at the dose of 400 mg/kg body weight was able to significantly reduce food intake and limit body weight gain over a period of 15 days. It also significantly decreased the plasma ghrelin, glucose, triglyceride levels and increased plasma leptin, serotonin, dopamine levels, but did not show much effect on CCK levels.


Mithila M.V.,Defence Food Research Laboratory DFRL | Khanum F.,Defence Food Research Laboratory DFRL
Journal of Dietary Supplements | Year: 2014

Guggulsterone or guggulipid is a steroidal constituent present in the neutral fraction of gum resin of Commiphora mukul, commonly known as guggul. The traditional uses of guggul-resin extract are well documented in the Ayurveda-where it is prescribed to treat a variety of ailments including lipid-related disorders such as obesity and arteriosclerosis. The hypolipidemic activity of the extracts known since ancient times can be traced to the two closely related steroidal ketones, E-guggulsterone and Z-guggulsterone. In this study, we have investigated the dose dependent (100, 200, 400 mg/kg body weight) effect of guggulsterones on appetite regulating hormones [ghrelin, leptin, cholecystokinin (CCK)] and neurotransmitters (serotonin and dopamine), which play a major role in the energy homeostasis and thus influence obesity related factors. We have also studied its effect on food intake, body weight and plasma triglycerides and glucose in rats. Guggulsterones at the dose of 400 mg/kg body weight was able to significantly reduce food intake and limit body weight gain over a period of 15 days. It also significantly decreased the plasma ghrelin, glucose, triglyceride levels and increased plasma leptin, serotonin, dopamine levels, but did not show much effect on CCK levels. © 2014 Informa Healthcare USA, Inc.

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