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1.
J Food Sci Technol ; 60(2): 528-537, 2023 Feb.
Article in English | MEDLINE | ID: mdl-36712215

ABSTRACT

Present studies were carried out to find the effect of different multigrain viz., finger millet, foxtail millet and little millet on litti. The multigrain powder was blended in whole wheat flour. Litti composite flour was studied for nutritional, rheological, gluten, sedimentation value, falling number and compared with regular wheat flour. Flaxseed and soybean were blended with barley, besan, spices for inner composite stuffing and samples were evaluated for proximate analysis. Shelf life studies of litti were assessed for 1 month at room temperature 25 ± 2℃ and freezer at 4℃. This research work explores with the aim to have benefits of multifunctional ingredients for the improvement of litti to have a healthy product; and increase the popularity of litti all over India to make it a commercially important product because of the incorporated multifunctional ingredients. The RDA calorie for human can be met with 4-5 litti per day.

2.
Arch Med Sci ; 17(1): 177-188, 2021.
Article in English | MEDLINE | ID: mdl-33488870

ABSTRACT

INTRODUCTION: The role of interferon gamma (IFN-γ) +874 A>T (rs2430561) gene polymorphism has been evaluated in different ethnicities with pulmonary tuberculosis (PTB) infection, and inconsistent results have been reported. In this study, a meta-analysis was performed to determine the precise association between IFN-γ +874 A>T gene polymorphism and PTB susceptibility. MATERIAL AND METHODS: A total of 21 studies comprising 4281 confirmed PTB cases and 5186 healthy controls were included in this meta-analysis by searching the PubMed (Medline), EMBASE, and Google Scholar web-databases. RESULTS: We observed reduced risk of PTB in allelic contrast (T vs. A: p = 0.001; OR = 0.818, 95% CI: 0.723-0.926), homozygous (TT vs. AA: p = 0.017; OR = 0.715, 95% CI: 0.543-0.941), heterozygous (AT vs. AA: p = 0.002; OR = 0.782, 95% CI: 0.667-0.917), dominant (TT+AT vs. AA: p = 0.002; OR = 0.768, 95% CI: 0.652-0.906), and recessive (TT vs. AA+AT: p = 0.042; OR = 0.802, 95% CI: 0.649-0.992) genetic models. In ethnicity-wise subgroup analysis, reduced risk of PTB was found in the Caucasian population. However, we did not find an association with any of the genetic models in the Asian population. CONCLUSIONS: In conclusion, the IFN-γ +874 A>T gene polymorphism is significantly associated with reduced risk of PTB, showing a protective effect in the overall and in the Caucasian population. However, this polymorphism is not associated with PTB risk in the Asian population.

3.
Sci Rep ; 9(1): 6482, 2019 04 24.
Article in English | MEDLINE | ID: mdl-31019210

ABSTRACT

Arterial/venous thrombosis is the major cardiovascular disorder accountable for substantial mortality; and the current demand for antithrombotic agents is extensive. Heparinases depolymerize unfractionated heparin (UFH) for the production of low molecular-weight heparins (LMWHs; used as anticoagulants against thrombosis). A microbial strain of Streptomyces sp. showing antithrombotic activity was isolated from the soil sample collected from north India. The strain was characterized by using 16S rRNA homology technique and identified as Streptomyces variabilis MTCC 12266 capable of producing heparinase enzyme. This is the very first communication reporting Streptomyces genus as the producer of heparinase. It was observed that the production of intracellular heparinase was [63.8 U/mg protein (specific activity)] 1.58 folds higher compared to extracellular heparinase [40.28 U/mg protein]. DEAE-Sephadex A-50 column followed by Sepharose-6B column purification of the crude protein resulted 19.18 folds purified heparinase. SDS-PAGE analysis of heparinase resulted an estimated molecular-weight of 42 kDa. It was also found that intracellular heparinase has the ability to depolymerize heparin to generate LMWHs. Further studies related to the mechanistic action, structural details, and genomics involved in heparinase production from Streptomyces variabilis are warranted for large scale production/purification optimization of heparinase for antithrombotic applications.


Subject(s)
Bacterial Proteins/metabolism , Heparin Lyase/metabolism , Heparin, Low-Molecular-Weight/metabolism , Heparin/metabolism , Streptomyces/metabolism , Bacterial Proteins/isolation & purification , Chromatography/methods , DEAE-Dextran/analogs & derivatives , Electrophoresis, Polyacrylamide Gel , Fibrinolytic Agents/chemistry , Fibrinolytic Agents/metabolism , Heparin/chemistry , Heparin Lyase/isolation & purification , Heparin, Low-Molecular-Weight/chemistry , India , Kinetics , Molecular Weight , Phylogeny , RNA, Ribosomal, 16S/genetics , Sepharose , Soil Microbiology , Streptomyces/classification , Streptomyces/genetics , Substrate Specificity
4.
Microb Pathog ; 130: 1-9, 2019 May.
Article in English | MEDLINE | ID: mdl-30825502

ABSTRACT

Interferon-γ (IFN-γ) plays a crucial role in immunological responses against Mycobacterium tuberculosis (M.tb) infection. The polymorphism at +874 A > T (rs2430561) influences the levels of IFN-γ, which may further influence the susceptibility to extrapulmonary tuberculosis (EPTB). This polymorphism has been investigated with respect to EPTB occurrence in different populations and provided contradictory and conflicting results. This study was performed to meta-statistically analyze the data and draw a more accurate conclusion regarding the association of IFN-γ +874 A > T gene polymorphism and EPTB susceptibility. A quantitative synthesis was executed for the pertinent studies retrieved from online web-databases viz. Google Scholar, PubMed/Medline and EMBASE. The pooled odds ratios (ORs) and confidence intervals (95% CIs) were estimated for all the genetic models by meta-analysis. A total of eight studies were retrieved which included 762 confirmed EPTB cases and 1341 controls. The meta-analysis results revealed reduced association of EPTB in allelic contrast (T vs. A: p = 0.001; OR = 0.668, 95% CI = 0.524 to 0.850), homozygous (TT vs. AA: p = 0.017; OR = 0.450, 95% CI = 0.234 to 0.868), heterozygous (AT vs. AA: p = 0.004; OR = 0.574, 95% CI = 0.395 to 0.835), dominant (TT + AT vs. AA: p = 0.003; OR = 0.536, 95% CI = 0.354 to 0.810) and recessive (TT vs. AA + AT: p = 0.039; OR = 0.662, 95% CI = 0.448 to 0.980) genetic models. Furthermore, re-sampling statistics also revealed reduced risk of EPTB in overall population and Asian subgroup. This meta-analysis concluded that IFN-γ +874 A > T gene polymorphism is meaningfully related with the reduced EPTB risk in overall and Asian population, and further necessitates larger studies to be conducted on this topic in other races.


Subject(s)
Genetic Predisposition to Disease , Interferon-gamma/genetics , Mycobacterium tuberculosis/immunology , Polymorphism, Single Nucleotide , Tuberculosis/genetics , Humans
5.
Front Physiol ; 9: 1793, 2018.
Article in English | MEDLINE | ID: mdl-30618805

ABSTRACT

Angiotensin-converting enzyme (ACE) gene is indispensable for endothelial control and vascular tone regulatory systems, usually affected in Systemic Lupus Erythematosus (SLE). ACE insertion/deletion (I/D) polymorphism may influence the progress of SLE. Earlier studies have investigated this association without any consistency in results. We performed this meta-analysis to evaluate the precise association between ACE I/D polymorphism and SLE susceptibility. The relevant studies were searched until December, 2017 using Medline (PubMed), Google-Scholar and EMBASE search engines. Twenty-five published studies involving 3,308 cases and 4,235 controls were included in this meta-analysis. Statistically significant increased risk was found for allelic (D vs. I: p = 0.007; OR = 1.202, 95% CI = 1.052-1.374), homozygous (DD vs. II: p = 0.025; OR = 1.347, 95% CI = 1.038-1.748), dominant (DD+ID vs. II: p = 0.002; OR = 1.195, 95% CI = 1.070-1.334), and recessive (DD vs. ID+II: p = 0.023; OR = 1.338, 95% CI = 1.042-1.718) genetic models. Subgroup analysis stratified by Asian ethnicity revealed significant risk of SLE in allelic (D vs. I: p = 0.045; OR = 1.238, 95% CI = 1.005-1.525) and marginal risk in dominant (DD+ID vs. II: p = 0.056; OR = 1.192, 95% CI = 0.995-1.428) models; whereas, no association was observed for Caucasian and African population. Publication bias was absent. In conclusion, ACE I/D polymorphism has significant role in overall SLE risk and it can be exploited as a prognostic marker for early SLE predisposition.

6.
Sci Rep ; 7(1): 17003, 2017 12 05.
Article in English | MEDLINE | ID: mdl-29208960

ABSTRACT

The macrophage migration inhibitory factor (MIF) is a cytokine that plays an important role in inhibiting the growth of pathogenic Mycobacterium tuberculosis (M.tb) and regulates immune responses against M.tb pathogen. MIF -173 G > C gene polymorphism may affect immunity in an individual and leads to susceptibility to tuberculosis (TB). A large number of studies have investigated the relevance of this polymorphism with TB risk, but their results were inconclusive. To obtain a precise conclusion, a meta-analysis was performed by retrieving six eligible studies from Google Scholar, PubMed (Medline), and EMBASE online databases. Overall combined analysis suggested increased TB risk between MIF -173 G > C polymorphism and overall risk in four genetic models, i.e., allelic (C vs. G: p = 0.001; OR = 1.517, 95% CI = 1.312 to 1.753), homozygous (CC vs. GG: p = 0.026; OR = 1.874, 95% CI = 1.079 to 3.257), heterozygous (GC vs. GG: p = 0.001; OR = 1.542, 95% CI = 1.273 to 1.868) and dominant model (CC + GC vs. GG: p = 0.001; OR = 1.631, 95% CI = 1.362 to 1.955). Similarly, increased TB risk was observed in subgroup analysis of Asian ethnicity. No publication bias was observed. These results suggested that MIF -173 G > C variant is a significant risk factor for TB in overall and in Asian populations, and can be used as prognostic marker for TB susceptibility.


Subject(s)
Intramolecular Oxidoreductases/genetics , Macrophage Migration-Inhibitory Factors/genetics , Mycobacterium tuberculosis/isolation & purification , Polymorphism, Single Nucleotide , Tuberculosis/genetics , Case-Control Studies , Genetic Predisposition to Disease , Humans , Prognosis , Risk Factors , Tuberculosis/epidemiology , Tuberculosis/microbiology
7.
Ann Clin Lab Sci ; 47(5): 628-637, 2017 Sep.
Article in English | MEDLINE | ID: mdl-29066494

ABSTRACT

GOALS: The involvement of the VDR ApaI gene polymorphism in the development of pulmonary tuberculosis (PTB) has been reported by numerous published studies and yielded inconsistent results. The present meta-analysis evaluated the association of VDR ApaI polymorphism and risk of PTB occurrence. PROCEDURES: PubMed (Medline), EMBASE and Google Scholar web-databases were searched and a meta-analysis was performed by calculating the pooled odds ratios (ORs) and 95% confidence intervals (95% CIs). RESULTS: This meta-analysis included a total of 14 eligible studies comprising of 1958 confirmed PTB cases and 2938 controls. We observed decreased risk of PTB in allelic (a vs. A: p=0.003; OR=0.873, 95% CI=0.798 to 0.955), homozygous (aa vs. AA: p=0.006; OR=0.761, 95% CI=0.626 to 0.924), dominant (aa+Aa vs. AA: p=0.039; OR=0.874, 95% CI=0.769 to 0.993) and recessive (aa vs. AA+Aa: p=0.025; OR=0.819, 95% CI=0.688 to 0.975) genetic models. During subgroup analysis, allele (a vs. A: p=0.005; OR=0.846, 95% CI=0.753 to 0.951), homozygous (aa vs. AA: p=0.002; OR=0.662, 95% CI=0.513 to 0.854) and recessive genetic models (aa vs. AA+Aa: p=0.003; OR=0.709, 95% CI=0.566 to 0.889) demonstrated decreased PTB risk in African population. However, no significant association was observed in Asian population. CONCLUSION: In conclusion, VDR ApaI polymorphism is significantly associated with decreased risk of PTB for in overall and African population, but not in Asians.


Subject(s)
Evidence-Based Medicine , Genetic Predisposition to Disease , Global Health , Immunity, Innate/genetics , Polymorphism, Single Nucleotide , Receptors, Calcitriol/genetics , Tuberculosis, Pulmonary/genetics , Alleles , Asian People , Black People , Gene Frequency , Genetic Association Studies , Humans , Immunogenetic Phenomena , Polymorphism, Genetic , Reproducibility of Results , Tuberculosis, Pulmonary/immunology
8.
Free Radic Res ; 50(8): 861-74, 2016 Aug.
Article in English | MEDLINE | ID: mdl-27150237

ABSTRACT

The progression in lifespan has been associated with elevated intracellular reactive oxygen species (ROS) and oxidative stress level which contributes to development of age related disorders. The discovery of lifespan modulating phytomolecules may promote development of natural therapies against age related afflictions. Acacetin (5,7-dihydroxy-4-methoxyflavone), is a naturally occurring flavonoid known to possess therapeutic properties. To this end, the present study evaluates effect of acacetin (AC) on lifespan, stress and neurotoxicity for the first time by using well-established free living, multicellular Caenorhabditis elegans model system. The 25 µM dose of AC significantly prolonged the mean lifespan of worms by 27.31% in comparison to untreated control and other tested doses of AC. Additionally, AC enhanced stress resistance against oxidative and thermal stress in worms. Furthermore, AC attenuated age related intracellular ROS level, aggregation of age pigment lipofuscin and increased the mean survival in stress hypersensitive mev-1 mutant by 40.5%. AC supplementation also reduced the alpha synuclein aggregation in transgenic worm model of Parkinson's disease. The enhanced stress resistance, lifespan and alleviation of age related pathology can be attributed to increment in stress modulatory enzymes like superoxide dismutase (SOD) and catalase (CAT) level. Altogether the results suggest AC exposure maintains stress level, health span and extends mean lifespan of C. elegans. The longevity promoting and neuromodulatory effects of AC are mediated by up regulation of the stress response genes sod-3 and gst-4. The present finding gives new insights of natural remedies and their future prospects in developing therapeutic interventions for managing age related diseases.


Subject(s)
Aging/drug effects , Flavones/pharmacology , Longevity/drug effects , Oxidative Stress/drug effects , Animals , Caenorhabditis elegans , Reactive Oxygen Species/metabolism
9.
Indian J Exp Biol ; 54(1): 64-71, 2016 Jan.
Article in English | MEDLINE | ID: mdl-26891554

ABSTRACT

Chitinase is one of the important enzymes as it is directly linked to Chitin that has wide applications in industrial, medical and commercial fields for its biocompatibility and biodegradability. Here, we report extracellular chitinase production by Streptomyces violascens NRRL B2700 under submerged fermentation condition. Chitinase production started after 10 h of incubation and reached to maximum level at 72 h of cultivation. Studies on the influence of additional carbon and nitrogen sources on chitinase production revealed that maltose, xylose, fructose, lactose, soybean meal and ammonium nitrate served as good carbon and nitrogen sources to enhance chitinase yield by 1.6 to 6 fold. Medium supplemented with 1% colloidal chitin produced high chitinase concentration (0.1714 U/mg). The enzyme chitinase was purified from the culture broth by 75% ammonium sulphate precipitation, DEAE-cellulose ion-exchange and sephadex G-100 gel filtration. The molecular mass of the purified chitinase was 65 kDa as estimated by SDS-PAGE. The apparent Michaelis constant (K(m)) and the maximum rate (V(max)) of the enzyme for colloidal chitin were 1.556 mg/mL and 2.680 µM/min/mg, respectively suggested high affinity towards-chitin. Possibly, it is the first report on production of chitinase from S. violascens NRRL B2700. The findings were encouraging, especially for cost effective production, and further warrants media and purification optimization studies for enhanced yield.


Subject(s)
Chitinases/isolation & purification , Streptomyces/enzymology , Chitin , Electrophoresis, Polyacrylamide Gel , Hydrogen-Ion Concentration
10.
PLoS One ; 10(9): e0137268, 2015.
Article in English | MEDLINE | ID: mdl-26368924

ABSTRACT

Cholesterol oxidase (COD) is a bi-functional FAD-containing oxidoreductase which catalyzes the oxidation of cholesterol into 4-cholesten-3-one. The wider biological functions and clinical applications of COD have urged the screening, isolation and characterization of newer microbes from diverse habitats as a source of COD and optimization and over-production of COD for various uses. The practicability of statistical/ artificial intelligence techniques, such as response surface methodology (RSM), artificial neural network (ANN) and genetic algorithm (GA) have been tested to optimize the medium composition for the production of COD from novel strain Streptomyces sp. NCIM 5500. All experiments were performed according to the five factor central composite design (CCD) and the generated data was analysed using RSM and ANN. GA was employed to optimize the models generated by RSM and ANN. Based upon the predicted COD concentration, the model developed with ANN was found to be superior to the model developed with RSM. The RSM-GA approach predicted maximum of 6.283 U/mL COD production, whereas the ANN-GA approach predicted a maximum of 9.93 U/mL COD concentration. The optimum concentrations of the medium variables predicted through ANN-GA approach were: 1.431 g/50 mL soybean, 1.389 g/50 mL maltose, 0.029 g/50 mL MgSO4, 0.45 g/50 mL NaCl and 2.235 ml/50 mL glycerol. The experimental COD concentration was concurrent with the GA predicted yield and led to 9.75 U/mL COD production, which was nearly two times higher than the yield (4.2 U/mL) obtained with the un-optimized medium. This is the very first time we are reporting the statistical versus artificial intelligence based modeling and optimization of COD production by Streptomyces sp. NCIM 5500.


Subject(s)
Cholesterol Oxidase/metabolism , Culture Media/chemistry , Streptomyces/growth & development , Algorithms , Artificial Intelligence , Bacterial Proteins/metabolism , Culture Media/pharmacology , Fermentation , Models, Genetic , Neural Networks, Computer , Streptomyces/enzymology
11.
JNMA J Nepal Med Assoc ; 53(199): 180-3, 2015.
Article in English | MEDLINE | ID: mdl-27549501

ABSTRACT

INTRODUCTION: This study aimed to analyze the average length of stay of all inpatients in the department of Orthopaedics and to compare the variations in hospital stay between age, gender, traumatic and non-traumatic co-morbidities and modality of payment. METHODS: This hospital based retrospective descriptive epidemiological study was based on patients discharged from a tertiary level health care center of eastern Nepal. Registry data of 1 year was used to calculate length of stay and analyze the variations. RESULTS: Average length of stay was 10.5 days. It was 10.7 days for males and 10.1 days for females. It was 10.12 days for patients paying themselves for their treatment whereas 14.98 days for patients receiving reimbursement (third party payment). CONCLUSIONS: Average length of stay was more in elderly and patients of trauma (longest in pelvis injury). It was 1.5 times longer for patients receiving reimbursement for treatment.


Subject(s)
Bone Diseases, Infectious , Bone Neoplasms , Length of Stay/statistics & numerical data , Wounds and Injuries , Adult , Female , Hospitals, Teaching , Humans , Insurance, Health, Reimbursement/statistics & numerical data , Male , Middle Aged , Nepal , Reimbursement Mechanisms/statistics & numerical data , Retrospective Studies , Tertiary Care Centers , Young Adult
12.
Biomed Res Int ; 2014: 934351, 2014.
Article in English | MEDLINE | ID: mdl-25250339

ABSTRACT

Gymnema sylvestre R.Br., a pharmacologically important herb vernacularly called Gur-Mar (sugar eliminator), is widely known for its antidiabetic action. This property of the herb has been attributed to the presence of bioactive triterpene glycosides. Although some information regarding pharmacology and phytochemical profiles of the plant are available, no attempts have been made so far to decipher the biosynthetic pathway and key enzymes involved in biosynthesis of steryl glucosides. The present report deals with the identification and catalytic characterization of a glucosyltransferase, catalyzing biosynthesis of steryl glycosides. The full length cDNA (2572 bp) contained an open reading frame of 2106 nucleotides that encoded a 701 amino acid protein, falling into GT-B subfamily of glycosyltransferases. The GsSGT was expressed in Escherichia coli and biochemical characterization of the recombinant enzyme suggested its key role in the biosynthesis of steryl glucosides with catalytic preference for C-3 hydroxyl group of sterols. To our knowledge, this pertains to be the first report on cloning and biochemical characterization of a sterol metabolism gene from G. sylvestre R.Br. catalyzing glucosylation of a variety of sterols of biological origin from diverse organisms such as bacteria, fungi, and plants.


Subject(s)
Escherichia coli/physiology , Glucosides/chemistry , Glucosyltransferases/biosynthesis , Glucosyltransferases/chemistry , Gymnema sylvestre/physiology , Recombinant Proteins/chemistry , Amino Acid Sequence , Catalysis , Cloning, Molecular/methods , Enzyme Activation , Enzyme Stability , Glucosyltransferases/genetics , Molecular Sequence Data , Recombinant Proteins/genetics , Recombinant Proteins/metabolism , Substrate Specificity
13.
Biomed Res Int ; 2014: 830285, 2014.
Article in English | MEDLINE | ID: mdl-24511547

ABSTRACT

Gymnema sylvestre (Asclepiadaceae), popularly known as "gurmar" for its distinct property as sugar destroyer, is a reputed herb in the Ayurvedic system of medicine. The phytoconstituents responsible for sweet suppression activity includes triterpene saponins known as gymnemic acids, gymnemasaponins, and a polypeptide, gurmarin. The herb exhibits a broad range of therapeutic effects as an effective natural remedy for diabetes, besides being used for arthritis, diuretic, anemia, osteoporosis, hypercholesterolemia, cardiopathy, asthma, constipation, microbial infections, indigestion, and anti-inflammatory. G. sylvestre has good prospects in the treatment of diabetes as it shows positive effects on blood sugar homeostasis, controls sugar cravings, and promotes regeneration of pancreas. The herbal extract is used in dietary supplements since it reduces body weight, blood cholesterol, and triglyceride levels and holds great prospects in dietary as well as pharmacological applications. This review explores the transition of a traditional therapeutic to a modern contemporary medication with an overview of phytochemistry and pharmacological activities of the herb and its phytoconstituents.


Subject(s)
Gymnema sylvestre , Plant Extracts , Plants, Medicinal , Animals , Anti-Infective Agents , Antineoplastic Agents , Humans , Hypoglycemic Agents , Medicine, Ayurvedic , Mice , Plant Extracts/chemistry , Plant Extracts/pharmacology , Protective Agents , Rats
14.
J Microbiol ; 51(3): 275-82, 2013 Jun.
Article in English | MEDLINE | ID: mdl-23812806

ABSTRACT

Aeromonas hydrophila is an opportunistic bacterial pathogen that is associated with a number of diseases in fish, amphibians, reptiles, and humans. In fish it causes several disease symptoms including tail and skin rot, and haemorrhagic septicemia; in human it causes soft-tissue wound infection and diarrhoea. The pathogenesis of A. hydrophila is multifactorial, but the mechanism is unknown so far. It is considered to be mediated by expression and secretion of extracellular proteins such as aerolysin, lipase, chitinase, amylase, gelatinase, hemolysins, and enterotoxins. A number of the putative virulence-protein-encoding genes that are present in the genome of A. hydrophila have been targeted by PCR for molecular diagnosis. These significant genes are also targeted for over-production of proteins by cloning and expression methods. In this review, we emphasize recent progress in the cloning, expression, and regulation of putative virulence-protein-encoding genes of A. hydrophila for a better understanding of the pathogenesis and also help to provide effective strategies for control of diseases.


Subject(s)
Aeromonas hydrophila/pathogenicity , Bacterial Proteins/metabolism , Aeromonas hydrophila/genetics , Animals , Bacterial Proteins/genetics , Gram-Negative Bacterial Infections/microbiology , Humans , Virulence/genetics , Virulence/physiology
15.
Biochem Genet ; 48(9-10): 760-78, 2010 Oct.
Article in English | MEDLINE | ID: mdl-20549331

ABSTRACT

The population structure of Labeo calbasu from 11 rivers belonging to the Indus, Ganges, Bhima, Mahanadi, and Godavari basins was investigated using allozyme marker systems. Seven out of 20 allozyme loci (35%) were polymorphic (P < 0.99). Both probability and score tests indicated significant deviation of genotype proportions from Hardy-Weinberg expectations at two loci, XDH* (Mahanadi, Bhima, and Godavari) and G6PDH* (Mahanadi). A pairwise genetic homogeneity test and F (ST) values indicated a low-to-moderate level (0.0515) of genetic structuring in the wild population of L. calbasu. AMOVA analysis also indicated moderate differentiation among the samples from different river basins. Analysis for genetic bottleneck was performed under the infinite allele model. The study revealed nine genetic stocks of L. calbasu from the natural population across Indian rivers. Evidence of genetic bottlenecks in some rivers was also revealed.


Subject(s)
Cyprinidae/genetics , Genetic Variation , Animals , Cyprinidae/classification , Female , Genetic Markers/genetics , Genotype , Isoenzymes/genetics , Male , Phylogeography , Polymorphism, Genetic , Population Density , Rivers
16.
Eur J Med Chem ; 45(4): 1560-71, 2010 Apr.
Article in English | MEDLINE | ID: mdl-20153089

ABSTRACT

Thymidylate synthase (TS) is a crucial enzyme for DNA biosynthesis and many nonclassical lipophilic antifolates targeting this enzyme are quite efficient and encouraging as antitumor drugs. Herein, we report some 3D-QSAR analyses using CoMFA and CoMSIA on quinozoline antifolates in order to have a better understanding of the mechanism of action and structure-activity relationship of these compounds. By applying leave-one-out (LOO) cross-validation study, we obtained cross-validated q(2) value of 0.573 for CoMFA and 0.445 for CoMSIA, while the non-cross-validated r(2) values for them were found to be 0.935 and 0.893, respectively. The models were graphically interpreted using CoMFA and CoMSIA contour plots. The results obtained from this study could be used for rational design of potent inhibitors against thymidylate synthase.


Subject(s)
Folic Acid Antagonists/pharmacology , Models, Molecular , Quinazolines/pharmacology , Thymidylate Synthase/antagonists & inhibitors , Folic Acid Antagonists/chemistry , Quantitative Structure-Activity Relationship , Quinazolines/chemistry
17.
Appl Biochem Biotechnol ; 160(7): 1985-91, 2010 Apr.
Article in English | MEDLINE | ID: mdl-19763901

ABSTRACT

Aerolysin is a significant virulent toxin protein secreted by Aeromonas hydrophila; it produces deep wound infections and hemorrhagic septicemia. The complete aerolysin gene (1,482 bp) was amplified from A. hydrophila. Furthermore, it was cloned and expressed into Escherichia coli BL21(DE3) codon plus RP cells using 0.5 mM IPTG for induction. The protein size was 54 kDa as estimated by SDS-PAGE, and it was purified by Ni-NTA affinity chromatography. Anti-His antibodies were used to characterize the expressed aerolysin by Western blotting and showed hemolytic activity with fish red blood cells. Aerolysin may be used as immunoassays for earlier control of A. hydrophila and is also compatible for vaccination.


Subject(s)
Aeromonas hydrophila/genetics , Bacterial Toxins/genetics , Bacterial Toxins/metabolism , Pore Forming Cytotoxic Proteins/genetics , Pore Forming Cytotoxic Proteins/metabolism , Animals , Bacterial Toxins/isolation & purification , Blotting, Western , Cloning, Molecular , Fishes/blood , Fishes/microbiology , Gene Expression , Hemolysis , Polymerase Chain Reaction , Pore Forming Cytotoxic Proteins/isolation & purification , Recombinant Proteins/genetics , Recombinant Proteins/isolation & purification
18.
Protein Expr Purif ; 65(1): 1-7, 2009 May.
Article in English | MEDLINE | ID: mdl-19136063

ABSTRACT

Hemolysin is a significant toxin secreted by Aeromonas hydrophila, which contributes pathogenicity of fish to humans. The complete ORF of hemolysin gene (1886 bp) was amplified using PCR. It was cloned in TA and sub-cloned in pET28a vector then transformed into Escherichia coli BL21(DE3) codon plus RP cells expressed by the induction with 1.0 mM of IPTG. The expected size of expressed protein was 68.0 kDa estimated by migration in 12% SDS-PAGE. Anti-His monoclonal antibodies were used to substantiate the recombinant protein by Western blotting. The percent similarity between hemolysin of A. hydrophila with other hemolytic toxins revealed that the hemolysin/aerolysin/cytotoxin sequence varied from 99.35 to 50.40%. Homology modeling was used to construct 3-D structure of hemolysin of A. hydrophila with the known crystal 3-D structure (PDB: 1XEZ). This protein can be used for immunoassays and it is suitable for vaccine candidate against A. hydrophila infection.


Subject(s)
Aeromonas hydrophila/genetics , Bacterial Proteins/biosynthesis , Cloning, Molecular , Gene Expression , Hemolysin Proteins/biosynthesis , Models, Molecular , Recombinant Proteins/biosynthesis , Aeromonas hydrophila/chemistry , Aeromonas hydrophila/metabolism , Animals , Bacterial Proteins/chemistry , Bacterial Proteins/genetics , Escherichia coli/genetics , Fish Diseases , Fishes , Hemolysin Proteins/chemistry , Hemolysin Proteins/genetics , Humans , Open Reading Frames/genetics , Protein Structure, Tertiary , Recombinant Proteins/chemistry , Recombinant Proteins/genetics , Structural Homology, Protein
19.
Indian J Microbiol ; 48(4): 453-8, 2008 Dec.
Article in English | MEDLINE | ID: mdl-23100746

ABSTRACT

Aerolysin is a hemolytic toxin encoded by aerolysin gene (1482 bp) that plays a key role in the pathogenesis of Aeromonas hydrophila infection in fish. New speciesspecific primers were designed to amplify 326 bp conserved region of aerolysin gene for A. hydrophila. Twenty-five isolates of A. hydrophila recovered from fish and pond water were studied for detection of aerolysin gene. Aerolysin gene was detected in 85% of the isolates during the study. The designed primers were highly specific and showed no cross reactivity with Escherichia coli, Aeromonas veronii, Vibrio cholerae, Flavobacterium spp., Chyseobacterium spp. and Staphylococcus aureus. The sensitivity limit of primers for detection of aerolysin gene in the genomic DNA of A. hydrophila was 5 pg.

20.
J Integr Bioinform ; 5(1)2008 Jan 24.
Article in English | MEDLINE | ID: mdl-20134056

ABSTRACT

The LexA protein is a transcriptional repressor of the bacterial SOS DNA repair system, which comprises a set of DNA repair and cellular survival genes that are induced in response to DNA damage. Its varied DNA binding motifs have been characterized and reported in the Escherichia coli, Bacillus subtilis, rhizobia family members, marine magnetotactic bacterium, Salmonella typhimurium and recently in Mycobacterium tuberculosis and this motifs information has been used in our theoretical analysis to detect its novel regulated genes in radio-resistant Deinococcus radiodurans genome. This bacterium showed presence of SOS-box like consensus sequence in the upstream sequences of 3166 genes with >60% motif score similarity percentage (MSSP) on both strands. Attempts to identify LexA-binding sites and the composition of the putative SOS regulon in D. radiodurans have been unsuccessful so far. To resolve the problem we performed theoretical analysis with modifications on reported data set of genes related to DNA repair (61 genes), stress response (145 genes) and some unusual predicted operons (21 clusters). Expression of some of the predicted SOS-box regulated operon members then was examined through the previously reported microarray data which confirm the expression of only single predicted operon i.e. DRB0143 (AAA superfamily NTPase related to 5-methylcytosine specific restriction enzyme subunit McrB) and DRB0144 (homolog of the McrC subunit of the McrBC restriction modification system). The methodology involved weight matrix construction through CONSENSUS algorithm using information of conserved upstream sequences of eight known genes including dinB, tagC, lexA, recA, uvrB, yneA of B. subtilis while lexA and recA of D. radiodurans through phylogenetic footprinting method and later detection of similar conserved SOS-box like LexA binding motifs through both RSAT & PoSSuMsearch programs. The resultant DNA consensus sequence had highly conserved 14 bp SOS-box like binding site.


Subject(s)
Bacterial Proteins/metabolism , Deinococcus/metabolism , Repressor Proteins/metabolism , Serine Endopeptidases/metabolism , Algorithms , Animals , Bacterial Proteins/genetics , Base Sequence , Binding Sites/genetics , Consensus Sequence/genetics , DNA Repair , DNA, Bacterial/metabolism , Genome, Bacterial , Molecular Sequence Data , Operon , Protein Binding , Repressor Proteins/genetics , SOS Response, Genetics , Serine Endopeptidases/genetics
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