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Find video protocols related to scientific articles indexed in Pubmed.
Correlation of TLR-4, IL-18, Transaminases and Uric Acid in the Patients With Chronic Periodontitis and Healthy Adults.
J. Periodontol.
PUBLISHED: 10-28-2014
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Background: Because of the potential association between periodontal disease and inflammation, the purpose of the present study was to examine the level of toll like receptor-4 (TLR-4), Interleukin-18 (IL-18) and uric acid as a marker of the inflammatory host response along with routine biochemical parameters such as fasting glucose, insulin, total cholesterol, High density lipoprotein (HDL), Low density lipoprotein (LDL), triglycerides, Fasting glucose, Alanine transaminase (AST), Aspartate transaminase (ALT) in the plasma and saliva of healthy individuals and periodontitis patients. We also wanted to check whether the patients with chronic periodontitis exhibit different modulations in salivary and/or plasma concentrations of these parameters compared with clinically healthy individuals. Materials and methods: Both salivary and plasma samples were collected from 40 chronic periodontitis patients and 20 healthy control individuals. TLR-4 and IL-18 measurements were done using commercially available ELISA kits. Total cholesterol, HDL, LDL, triglycerides, Fasting glucose, AST and ALT levels were analyzed on Roche p-800 modular system using the specific kits provided by the manufacturer. Non-parametric tests was used in case of certain parameters for the statistical analyses, because our data did not follow the Gaussian distribution. Results: Significant difference was observed in the plasma and salivary TLR-4 and IL-18 levels along with clinical measurements such as plaque index (PI) and pocket depth (PD) in chronic periodontitis patients (P<0.001). The plasma level of TLR-4 was found to be enhanced from 0.99 to 3.28 in chronic periodontitis patients. Salivary TLR-4 levels also showed slightly higher increase in diseased state (12.44 to 29.97). A significant increase of approx 46% was recorded in the plasma IL-18 level. However, salivary IL-18 level rose upto more than 5 fold in the chronic periodontitis patients compared with the control individuals. The level of plasma uric acid was found to be highly significant compared to control individuals. HDL and triglyceride also showed significant difference (P<0.017 and P<0.025 respectively). Moreover, plasma glucose, total cholesterol, LDL and insulin levels did not show any significant difference. There was only slight increase in the plasma AST and ALT levels between diseased and healthy state (22.55 vs 25.50 and 12.35 vs 15.95 respectively). However, salivary AST and ALT level showed approx 6 fold rise in the chronic periodontitis patients compared with the control individuals. The cross correlation analysis in periodontitis disease group showed significant association of plasma AST, salivary AST and salivary ALT with uric acid level. Conclusion: Based on our study, we believe that TLR-4, IL-18 and uric acid could have a role in the inflammatory pathology of periodontitis. These parameters are suggested to be useful in the prognosis and diagnosis of chronic periodontitis. However, mechanistic association of these parameters with inflammatory pathology of periodontitis patients needs to be further elucidated on a higher number of samples.
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Microbe-specific unconventional T cells induce human neutrophil differentiation into antigen cross-presenting cells.
J. Immunol.
PUBLISHED: 08-27-2014
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The early immune response to microbes is dominated by the recruitment of neutrophils whose primary function is to clear invading pathogens. However, there is emerging evidence that neutrophils play additional effector and regulatory roles. The present study demonstrates that human neutrophils assume Ag cross-presenting functions and suggests a plausible scenario for the local generation of APC-like neutrophils through the mobilization of unconventional T cells in response to microbial metabolites. V?9/V?2 T cells and mucosal-associated invariant T cells are abundant in blood, inflamed tissues, and mucosal barriers. In this study, both human cell types responded rapidly to neutrophils after phagocytosis of Gram-positive and Gram-negative bacteria producing the corresponding ligands, and in turn mediated the differentiation of neutrophils into APCs for both CD4(+) and CD8(+) T cells through secretion of GM-CSF, IFN-?, and TNF-?. In patients with acute sepsis, circulating neutrophils displayed a similar APC-like phenotype and readily processed soluble proteins for cross-presentation of antigenic peptides to CD8(+) T cells, at a time when peripheral V?9/V?2 T cells were highly activated. Our findings indicate that unconventional T cells represent key controllers of neutrophil-driven innate and adaptive responses to a broad range of pathogens.
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Optimization of Storage and Stability of Camel Liver Glutathione S-Transferase.
Prep. Biochem. Biotechnol.
PUBLISHED: 07-19-2014
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Glutathione S-transferases (GSTs) are multifunctional enzymes and play an important role in cellular detoxification. Besides this, GSTs act as cytosolic carrier proteins which bind hydrophobic compounds such as heme, bilirubin, steroids and polycyclic hydrocarbons. GST has great importance in Biotechnology, as it is target for vaccine and drug development and biosensors development for xenobiotics. Moreover, GST tag has been extensively used for protein expression and purification. Until now, biophysical properties of camel liver GST have not been characterized. In the present study we have purified Camel (Camelus dromedarius) liver GST to homogeneity in single step by affinity chromatography with 23.4 fold purification and 60.6% yield. Our results showed that maximal activity of GST was at pH 6.5 and it was stable in pH range of 5 and 10. The optimum temperature was 55°C and the Tm was 57°C. Chemical chaperone, glycerol (3.3 M) was able to protect GST activity and aggregation against thermal denaturation by stabilizing the protein structure at 50 and 57°C, respectively. Whereas, L-arginine (125 mM) did not protect GST against thermal stress. Far-UV CD spectra showed that glycerol protected secondary structure of GST while L-arginine induced conformational changes under thermal stress. In conclusion, our studies on the GST stability suggest that glycerol works as a stabilizer and L-arginine acts as destabilizer.
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Periosteum as a barrier membrane in the treatment of intrabony defect: A new technique.
J Indian Soc Periodontol
PUBLISHED: 07-16-2014
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The purpose of the study was to evaluate the clinical effectiveness of periosteum as a barrier membrane for the treatment of intrabony defects.
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Periodontal regeneration by application of chemical root conditioning to intrabony defects utilizing bioresorbable membrane: a comparative study.
Oral Health Dent Manag
PUBLISHED: 07-03-2014
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To improve and combat the limitations of healing capacity of periodontal tissues, quite a few research methods have evaluated the benefits of GTR inflection by root surface conditioning agents.
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Glycoxidation of biological macromolecules: a critical approach to halt the menace of glycation.
Glycobiology
PUBLISHED: 06-19-2014
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Glycation is the result of covalent bonding of a free amino group of biological macromolecules with a reducing sugar, which results in the formation of a Schiff base that undergoes rearrangement, dehydration and cyclization to form a more stable Amadori product. The final products of nonenzymatic glycation of biomacromolecules like DNA, proteins and lipids are known as advanced glycation end products (AGEs). AGEs may be generated rapidly or over long times stimulated by distinct triggering mechanisms, thereby accounting for their roles in multiple settings and disease states. Both Schiff base and Amadori glycation products generate free radicals resulting in decline of antioxidant defense mechanisms and can damage cellular organelles and enzymes. This critical review primarily focuses on the mechanistic insight of glycation and the most probable route for the formation of glycation products and their therapeutic interventions. Furthermore, the prevention of glycation reaction using therapeutic drugs such as metformin, pyridoxamine and aminoguanidine (AG) are discussed with special emphasis on the novel concept of the bioconjugation of these drugs like, AG with gold nanoparticles (GNPs). At or above 10 mM concentration, AG is found to be toxic and therefore has serious health concerns, and the study warrants doing this novel bioconjugation of AG with GNPs. This approach might increase the efficacy of the AG at a reduced concentration with low or no toxicity. Using the concept of synthesis of GNPs with abovementioned drugs, it is assumed that toxicity of various drugs which are used at high doses can be minimized more effectively.
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Orally active osteoanabolic agent GTDF binds to adiponectin receptors, with a preference for AdipoR1, induces adiponectin-associated signaling, and improves metabolic health in a rodent model of diabetes.
Diabetes
PUBLISHED: 05-21-2014
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Adiponectin is an adipocytokine that signals through plasma membrane-bound adiponectin receptors 1 and 2 (AdipoR1 and -2). Plasma adiponectin depletion is associated with type 2 diabetes, obesity, and cardiovascular diseases. Adiponectin therapy, however, is yet unavailable owing to its large size, complex multimerization, and functional differences of the multimers. We report discovery and characterization of 6-C-?-D-glucopyranosyl-(2S,3S)-(+)-5,7,3',4'-tetrahydroxydihydroflavonol (GTDF) as an orally active adiponectin mimetic. GTDF interacted with both AdipoRs, with a preference for AdipoR1. It induced adiponectin-associated signaling and enhanced glucose uptake and fatty acid oxidation in vitro, which were augmented or abolished by AdipoR1 overexpression or silencing, respectively. GTDF improved metabolic health, characterized by elevated glucose clearance, ?-cell survival, reduced steatohepatitis, browning of white adipose tissue, and improved lipid profile in an AdipoR1-expressing but not an AdipoR1-depleted strain of diabetic mice. The discovery of GTDF as an adiponectin mimetic provides a promising therapeutic tool for the treatment of metabolic diseases.
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Sub-MICs of Carum copticum and Thymus vulgaris influence virulence factors and biofilm formation in Candida spp.
BMC Complement Altern Med
PUBLISHED: 04-23-2014
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Emergence of drug-resistant strains of Candida and inefficiency of conventional antifungal therapy has necessitated the search for alternative and new antifungal agents. Inhibition of virulence and biofilm are the potential drug targets. In this study, the oils of Carum copticum, Thymus vulgaris and their major active compound thymol as revealed by Gas chromatography and gas chromatography-mass spectrometry (GC-GC/MS) analysis were tested for their inhibitory activity against growth to determine sub-MIC values against 27 drug-resistant strains of Candida spp.
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EGFRIndb: epidermal growth factor receptor inhibitor database.
Anticancer Agents Med Chem
PUBLISHED: 03-15-2014
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Aberrant activity of epidermal growth factor receptor (EGFR) family proteins has been found to be associated with a number of human cancers including that of lung and breast. Consequently, the search for EGFR family inhibitors, a well established target of pharmacological and therapeutic value has been ongoing. Therefore, over the years several small molecules, which compete for ATP in the kinase domain have been synthesised and some of them have proved to be effective in attenuating EGFR mediated proliferation. Thus, there exists in literature a vast amount of experimental data on EGFR tyrosine kinase inhibitors. In this paper, we describe a comprehensive database EGFRIndb that contains details of the small molecular inhibitors of EGFR family.
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Enhanced immunoprotective effects by anti-IL-17 antibody translates to improved skeletal parameters under estrogen deficiency compared with anti-RANKL and anti-TNF-? antibodies.
J. Bone Miner. Res.
PUBLISHED: 03-07-2014
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Activated T cell has a key role in the interaction between bone and immune system. T cells produce proinflammatory cytokines, including receptor activator of NF-?B ligand (RANKL), tumor necrosis factor ? (TNF-?), and interleukin 17 (IL-17), all of which augment osteoclastogenesis. RANKL and TNF-? are targeted by inhibitors such as denosumab, a human monoclonal RANKL antibody, and infliximab, which neutralizes TNF-?. IL-17 is also an important mediator of bone loss, and an antibody against IL-17 is undergoing phase II clinical trial for rheumatoid arthritis. Although there are a few studies showing suppression of Th17 cell differentiation and induction of regulatory T cells (Tregs) by infliximab, the effect of denosumab remains poorly understood. In this study, we investigated the effects of anti-TNF-?, anti-RANKL, or anti-IL-17 antibody administration to estrogen-deficient mice on CD4(+) T-cell proliferation, CD28 loss, Th17/Treg balance and B lymphopoesis, and finally, the translation of these immunomodulatory effects on skeletal parameters. Adult Balb/c mice were treated with anti-RANKL/-TNF-?/-IL-17 subcutaneously, twice a week, postovariectomy (Ovx) for 4 weeks. Animals were then autopsied; bone marrow cells were collected for FACS and RNA analysis and serum collected for ELISA. Bones were dissected for static and dynamic histomorphometry studies. We observed that although anti-RANKL and anti-TNF-? therapies had no effect on Ovx-induced CD4(+) T-cell proliferation and B lymphopoesis, anti-IL-17 effectively suppressed both events with concomitant reversal of CD28 loss. Anti-IL-17 antibody reduced proinflammatory cytokine production and induced Tregs. All three antibodies restored trabecular microarchitecture with comparable efficacy; however, cortical bone parameters, bone biomechanical properties, and histomorphometry were best preserved by anti-IL-17 antibody, likely attributable to its inhibitory effect on osteoblast apoptosis and increased number of bone lining cells and Wnt10b expression. Based on the superior immunoprotective effects of anti-IL-17, which appears to translate to a better skeletal preservation, we propose beginning clinical trials using a humanized antibody against IL-17 for treatment of postmenopausal osteoporosis.
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In vitro screening for ?-hydroxy-?-methylglutaryl-CoA reductase inhibitory and antioxidant activity of sequentially extracted fractions of Ficus palmata Forsk.
Biomed Res Int
PUBLISHED: 02-26-2014
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Hypercholesterolemia-induced oxidative stress has been strongly implicated in the pathogenesis of atherosclerosis, which is one of the major causes of mortality worldwide. The current work, for the first time, accounts the antioxidant, genoprotective, antilipoperoxidative, and HMG-CoA reductase (EC 1.1.1.34) inhibitory properties of traditional medicinal plant, Ficus palmata Forsk. Our result showed that among sequentially extracted fractions of Ficus palmata Forsk, FPBA (F. palmata bark aqueous extract) and FPLM (F. palmata leaves methanolic extract) extracts have higher phenolic content and also exhibited significantly more radical scavenging (DPPH and Superoxide) and antioxidant (FRAP) capacity. Moreover, FPBA extract also exhibited significantly higher inhibition of lipid peroxidation assay. Additionally, results showed almost complete and partial protection of oxidatively damaged DNA by these plant extracts when compared to mannitol. Furthermore, our results showed that FPBA extract (IC50 = 9.1 ± 0.61 µg/mL) exhibited noteworthy inhibition of HMG-CoA reductase activity as compared to other extracts, which might suggest its role as cardioprotective agent. In conclusion, results showed that FPBA extract not only possess significant antioxidant and genoprotective property but also is able to attenuate the enzymatic activity of HMG-CoA reductase, which might suggest its role in combating various oxidative stress-related diseases, including atherosclerosis.
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An enzoinformatics study targeting polo-like kinases-1 enzyme: Comparative assessment of anticancer potential of compounds isolated from leaves of Ageratum houstonianum.
Pharmacogn Mag
PUBLISHED: 02-21-2014
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Natural products from plant sources, embracing inherently ample structural diversity than synthetic ones are the major sources of anticancer agents and will constantly play as protagonists for discovering new drugs. Polo-like kinases (PLKs) play a leading role in the ordered execution of mitotic events and 4 mammalian PLK family members have been identified. PLK1 is an attractive target for anticancer drugs in mammalian cells, among the four members of PLKs. The present study expresses the molecular interaction of compounds (1,2-Benzenedicarboxylic acid bis (2 ethylhexyl) ester, squalene, 3,5-bis (1,1-dimethylethyl) phenol, Pentamethyl tetrahydro-5H-chromene, (1,4-Cyclohexylphenyl) ethanone and 6-Vinyl-7-methoxy-2,2-dimethylchromene) isolated from methanolic extract of leaves of Ageratum houstonianum with PLK1 enzyme. Docking between PLK1 and each of these compounds (separately) was performed using "Auto dock 4.2." (1,4-Cyclohexylphenyl) ethanone showed the maximum potential as a promising inhibitor of PLK1 enzyme with reference to ?G (-6.84 kcal/mol) and Ki (9.77 ?M) values. This was sequentially followed by Pentamethyl tetrahydro-5H-chromene (?G = -6.60 kcal/mol; Ki = 14.58 ?M), squalene (?G = -6.17 kcal/mol; Ki = 30.12 ?M), 6-Vinyl-7-methoxy-2,2-dimethylchromene (?G = -5.91 kcal/mol; Ki = 46.68 ?M), 3, 5-bis (1,1-dimethylethyl) phenol (?G = -5.70 kcal/mol; Ki = 66.68 ?M) and 1,2-Benzenedicarboxylic acid bis (2 ethylhexyl) ester (?G = -5.58 kcal/mol; Ki = 80.80 ?M). These results suggest that (1,4-Cyclohexylphenyl) ethanone might be a potent PLK1 inhibitor. Further, in vitro and in vivo rumination are warranted to validate the anticancer potential of (1,4-Cyclohexylphenyl) ethanone.
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Protective Effect of Solanum nigrum Leaves Extract on Immobilization Stress Induced Changes in Rat's Brain.
Evid Based Complement Alternat Med
PUBLISHED: 02-09-2014
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The prophylactic or curative antioxidant efficacy of crude extract and the active constituent of S. nigrum leaves were evaluated in modulating inherent antioxidant system altered due to immobilization stress in rat brain tissues, in terms of measurement of glutathione (GSH), lipid peroxidation (thiobarbituric acid reactive substances, TBARS), and free radical scavenging enzymes activities. Rats were treated with single dose of crude extract of S. nigrum prior to and after 6 h of immobilization stress exposure. Exposure to immobilization stress resulted in a decrease in the brain levels of glutathione, SOD, GST, and catalase, with an increase in thiobarbituric acid reactive substances (TBARS) levels. Treatment of S. nigrum extract and its active constituents to both pre- and poststressed rats resulted in significant modulation in the above mentioned parameters towards their control values with a relative dominance by the latter. Brain is vulnerable to stress induced prooxidant insult due to high levels of fat content. Thus, as a safe herbal medication the S. nigrum leaves extract or its isolated constituents can be used as nutritional supplement for scavenging free radicals generated in the brain due to physical or psychological stress or any neuronal diseases per se.
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Exploring Drug Targets in Isoprenoid Biosynthetic Pathway for Plasmodium falciparum.
Biochem Res Int
PUBLISHED: 02-07-2014
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Emergence of rapid drug resistance to existing antimalarial drugs in Plasmodium falciparum has created the need for prediction of novel targets as well as leads derived from original molecules with improved activity against a validated drug target. The malaria parasite has a plant plastid-like apicoplast. To overcome the problem of falciparum malaria, the metabolic pathways in parasite apicoplast have been used as antimalarial drug targets. Among several pathways in apicoplast, isoprenoid biosynthesis is one of the important pathways for parasite as its multiplication in human erythrocytes requires isoprenoids. Therefore targeting this pathway and exploring leads with improved activity is a highly attractive approach. This report has explored progress towards the study of proteins and inhibitors of isoprenoid biosynthesis pathway. For more comprehensive analysis, antimalarial drug-protein interaction has been covered.
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Influence of pH and temperature on the activity of SnO2-bound ?-amylase: a genotoxicity assessment of SnO2 nanoparticles.
Prep. Biochem. Biotechnol.
PUBLISHED: 02-07-2014
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Immobilization of biologically important molecules on a myriad of nanosized materials has attracted great attention due to their small size, biocompatibility, higher surface-to-volume ratio, and lower toxicity. These properties make nanoparticles (NPs) a superior matrix over bulk material for the immobilization of enzymes and proteins. In the present study, Bacillus amyloliquefaciens ?-amylase was immobilized on SnO2 nanoparticles by a simple adsorption mechanism. Nanoparticle-adsorbed enzyme retained 90% of the original enzyme activity. Thermal stability of nanosupport was investigated by thermogravimetric and differential thermal analysis. Scanning electron microscopic studies showed that NPs have porous structure for the high-yield immobilization of ?-amylase. The genotoxicity of SnO2-NPs was analyzed by pUC(19) plasmid nicking and comet assay and revealed that no remarkable DNA damage occurred in lymphocytes. The pH-optima was found to be the same for both free and SnO2-NPs bound enzyme, while the temperature-optimum for NPs-adsorbed ?-amylase was 5°C higher than its free counterpart. Immobilized enzyme retained more than 70% enzyme activity even after its eight repeated uses.
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Immunogenicity of DNA-advanced glycation end product fashioned through glyoxal and arginine in the presence of Fe(3+): Its potential role in prompt recognition of diabetes mellitus auto-antibodies.
Chem. Biol. Interact.
PUBLISHED: 01-30-2014
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Glyoxal, methylglyoxal and 3-deoxyglucosones are reactive dicarbonyl compounds, which transform free amino groups of proteins and lipoproteins macromolecule into advanced glycation end-products (AGEs). AGEs play a significant role in the pathophysiology of aging and diabetic complications because of their genotoxic effect. Glyoxal also reacts with free amino group of nucleic acids resulting in the formation of DNA-AGEs. The present study reports the genotoxicity and immunogenicity of AGEs formed by Glyoxal-Arginine-Fe(3+) (G-Arg-Fe(3+)) system as a glycating agent. Immunogenicity of native and G-Arg-Fe(3+)-DNA was probed in female rabbits. Immunofluorescence suggests the presence of immune complex deposition in the kidney section of immunized rabbits. Spectroscopic analysis and melting temperature indicates the structural modification in the human DNA. The modified human DNA is found to be highly immunogenic, whereas unmodified form was simply non-immunogenic. This study shows the presence of auto-antibodies against G-Arg-Fe(3+) modified human DNA in the sera of diabetes type 1 and in few cases type 2 patients due to secondary complications of nephropathy. The glyco-oxidative lesions have also been detected in the lymphocyte DNA isolated from patients having type 1 and type 2 diabetes. The results show structural perturbations generating new epitopes in G-Arg-Fe(3+)-DNA rendering it pretty immunogenic.
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Effect of copper oxide nanoparticles on the conformation and activity of ?-galactosidase.
Colloids Surf B Biointerfaces
PUBLISHED: 01-24-2014
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The primary objective of this study is to explore the interaction of ?-galactosidase with copper oxide nanoparticles (CuO NPs). Steady-state absorption, fluorescence and circular dichroism (CD) spectroscopic techniques have been employed to unveil the conformational changes of ?-galactosidase induced by the binding of CuO NPs. Temperature dependent fluorescence quenching results indicates a static quenching mechanism in the present case. The binding thermodynamic parameters delineate the predominant role of H-bonding and van der Waals forces between ?-galactosidase and CuO NPs binding process. The binding was studied by isothermal titration calorimetry (ITC) and the result revealed that the complexation is enthalpy driven, the ?H°<0, ?S°<0 indicates the formation of hydrogen bonds between ?-galactosidase and CuO NPs occurs. Disruption of the native conformation of the protein upon binding with CuO NPs is reflected through a reduced functionality (in terms of hydrolase activity) of the protein CuO NPs conjugate system in comparison to the native protein and CuO NPs exhibited a competitive mode of inhibition. This also supports the general belief that H-bond formation occurs with NPs is associated with a lesser extent of modification in the native structure. Morphological features and size distributions were investigated using transmission electron microscopy (TEM) and dynamic light scattering (DLS). Additionally the considerable increase in the Rh following the addition of CuO NPs accounts for the unfolding of ?-galactosidase. Chemical and thermal unfolding of ?-galactosidase, when carried out in the presence of CuO NPs, also indicated a small perturbation in the protein structure. These alterations in functional activity of nanoparticle bound ?-galactosidase which will have important consequences should be taken into consideration while using nanoparticles for diagnostic and therapeutic purposes.
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Antimicrobial potential of bacteriocin producing Lysinibacillus jx416856 against foodborne bacterial and fungal pathogens, isolated from fruits and vegetable waste.
Anaerobe
PUBLISHED: 01-03-2014
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In this study, antimicrobial potential, some probiotics properties and bacteriocin nature of Lysinibacillus, isolated from fruits and vegetable waste were evaluated. For this, 125 Lactobacillus isolates were tested against foodborne bacterial and fungal pathogens. Among these, an isolated Bacillus spp. showed significant aggregation-co-aggregation probiotics properties and potentially inhibits the foodborne gram positive microbial pathogens such as Staphylococcus aureus, (22 mm ZOI), Staphylococcus epedermidis and Bacillus cereus (18 mm). Phenotypically and molecularly it was identified as Lysinibacillus (NCBI accession no. JX416856) and it was found closest to Lysinibacillus fusiformis, Lysinibacillus sphaericus and Lysinibacillus xylanilyticus. Physico-biochemically, it was found to be negative for amylase, protease, gelatinase, nitrate reductase and urease while positive for catalase. The diagnostic fatty acid was 22;2 (3.51). The growth conditions and bacteriocin activity were found to be optimum with MRS media at pH 7-10, Temperature 35-40 °C and salt tolerance at 1-3%. Eventually its production was optimized with MRS broth at pH 7.6, 37 °C, for 36 h in shaking conditions at the rate of 100 rpm. Active bacteriocin was isolated at 60% ammonium sulfate precipitation. The molecular weight of given bacteriocin was found to be nearly 25-35 kDa by SDS-PAGE. Based on physico- biochemical properties, the isolated bacteriocin was to be categories in class II bacteriocin. The bacteriocin was found to be stable in the range of 4-80 °C temperature, 6-10 pH and even in the presence of surfactant (such as SDS and Tween 80). However, proteases like pepsin and trypsin were found to degrade the bacteriocin. Collectively, the broad spectrum inhibitory potential and physical stability offered the antimicrobial potential to Lysinibacillus, and its relevant bacteriocin might be used as an alternative food preservative or therapeutic agent to control spoilage of different food products.
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Potential Use of ?? T Cell-Based Vaccines in Cancer Immunotherapy.
Front Immunol
PUBLISHED: 01-01-2014
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IMMUNOTHERAPY IS A FAST ADVANCING METHODOLOGY INVOLVING ONE OF TWO APPROACHES: (1) compounds targeting immune checkpoints and (2) cellular immunomodulators. The latter approach is still largely experimental and features in vitro generated, live immune effector cells, or antigen-presenting cells. ?? T cells are known for their efficient in vitro tumor killing activities. Consequently, many laboratories worldwide are currently testing the tumor killing function of ?? T cells in clinical trials. Reported benefits are modest; however, these studies have demonstrated that large ?? T-cell infusions were well tolerated. Here, we discuss the potential of using human ?? T cells not as effector cells but as a novel cellular vaccine for treatment of cancer patients. Antigen-presenting ?? T cells do not require to home to tumor tissues but, instead, need to interact with endogenous, tumor-specific ?? T cells in secondary lymphoid tissues. Newly mobilized effector ?? T cells are then thought to overcome the immune blockade by creating proinflammatory conditions fit for effector T-cell homing to and killing of tumor cells. Immunotherapy may include tumor antigen-loaded ?? T cells alone or in combination with immune checkpoint inhibitors.
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Carum copticum and Thymus vulgaris oils inhibit virulence in Trichophyton rubrum and Aspergillus spp.
Braz. J. Microbiol.
PUBLISHED: 01-01-2014
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Emergence of drug-resistant strains has demanded for alternative means of combating fungal infections. Oils of Carum copticum and Thymus vulgaris have long been used in ethnomedicine for ailments of various fungal infections. Since their activity has not been reported in particular against drug-resistant fungi, this study was aimed to evaluate the effects of oils of C. copticum and T. vulgaris on the growth and virulence of drug-resistant strains of Aspergillus spp. and Trichophyton rubrum. The gas chromatography-mass spectrometry analysis revealed thymol constituting 44.71% and 22.82% of T. vulgaris and C. copticum, respectively. Inhibition of mycelial growth by essential oils was recorded in the order of thymol > T. vulgaris > C. copticum against the tested strains. RBC lysis assay showed no tested oils to be toxic even up to concentration two folds higher than their respective MFCs. Thymol exhibited highest synergy in combination with fluconazole against Aspergillus fumigatus MTCC2550 (FICI value 0.187) and T. rubrum IOA9 (0.156) as determined by checkerboard method. Thymol and T. vulgaris essential oil were equally effective against both the macro and arthroconidia growth (MIC 72 ?g/mL). A > 80% reduction in elastase activity was recorded for A. fumigatus MTCC2550 by C. copticum, T. vulgaris oils and thymol. The effectiveness of these oils against arthroconidia and synergistic interaction of thymol and T. vulgaris with fluconazole can be exploited to potentiate the antifungal effects of fluconazole against drug-resistant strains of T. rubrum and Aspergillus spp.
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Expanded Human Blood-Derived ??T Cells Display Potent Antigen-Presentation Functions.
Front Immunol
PUBLISHED: 01-01-2014
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Cell-based immunotherapy strategies target tumors directly (via cytolytic effector cells) or aim at mobilizing endogenous anti-tumor immunity. The latter approach includes dendritic cells (DC) most frequently in the form of in vitro cultured peripheral blood monocytes-derived DC. Human blood ??T cells are selective for a single class of non-peptide agonists ("phosphoantigens") and develop into potent antigen-presenting cells (APC), termed ??T-APC within 1-3?days of in vitro culture. Availability of large numbers of ??T-APC would be advantageous for use as a novel cellular vaccine. We here report optimal ??T cell expansion (>10(7) cells/ml blood) when peripheral blood mononuclear cells (PBMC) from healthy individuals and melanoma patients were stimulated with zoledronate and then cultured for 14?days in the presence of IL-2 and IL-15, yielding ??T cell cultures of variable purity (77?±?21 and 56?±?26%, respectively). They resembled effector memory ??T (TEM) cells and retained full functionality as assessed by in vitro tumor cell killing as well as secretion of pro-inflammatory cytokines (IFN?, TNF?) and cell proliferation in response to stimulation with phosphoantigens. Importantly, day 14 ??T cells expressed numerous APC-related cell surface markers and, in agreement, displayed potent in vitro APC functions. Day 14 ??T cells from PBMC of patients with cancer were equally effective as their counterparts derived from blood of healthy individuals and triggered potent CD8(+) ??T cell responses following processing and cross-presentation of simple (influenza M1) and complex (tuberculin purified protein derivative) protein antigens. Of note, and in clear contrast to peripheral blood ??T cells, the ability of day 14 ??T cells to trigger antigen-specific ??T cell responses did not depend on re-stimulation. We conclude that day 14 ??T cell cultures provide a convenient source of autologous APC for use in immunotherapy of patients with various cancers.
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A report on biocompounds from palm fossil of India.
Bioinformation
PUBLISHED: 01-01-2014
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The occurrence of a large number of fossil woods having resemblance in anatomical features with the modern palm genus, Phoenix L in Deccan Intertrappean fossil flora of Maastrichtian-Danian age (i. e. Late Cretaceous and Earliest Tertiary (65-67 my)) indicates the most primitive record of date palm. Present discovery of biocompounds from fossil wood of Phoenix collected from Deccan Intertrappean having affinity with the biocompounds known from modern plant further exemplify the earliest documentation of Phoenix in Indian peninsula.
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Hypocholesterolemic activity of monascus fermented product in the absence of monacolins with partial purification for functional food applications.
ScientificWorldJournal
PUBLISHED: 01-01-2014
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Hypercholesterolemia is one of the most common chronic diseases in human. Along with chemical therapy traditional medication is used as hypocholesterolemic remedy, however, with unfavorable side effects. Recently, Monascus fermented product (MFP) has become a popular hypocholesterolemic natural supplement. In the present study, the hypocholesterolemic activity of Monascus purpureus FTC5391 fermented product ethanolic extract (MFPe) was investigated in hypercholesterolemic rats. Results showed that MFPe not only reduced the serum total cholesterol (TC), LDL-C, TG concentration, and TC/HDL-C ratio but also increased the HDL-C. Further, solid phase extraction (SPE) was carried out to obtain the hypocholesterolemic bioactive fraction. The high polar fraction of SPE increased the HDL-C (42%) and decreased the TC (53.3%), LDL-C (47%), and TG (50.7%) levels as well as TC/HDL-C ratio (69.1%) in serum. The GC-MS results of the active fraction revealed two main compounds, isosorbide and erythritol, which act as coronary vasodilator compounds.
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Flower-shaped ZnO nanoparticles synthesized by a novel approach at near-room temperatures with antibacterial and antifungal properties.
Int J Nanomedicine
PUBLISHED: 01-01-2014
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Due to enormous applications of metal oxide nanoparticles in research and health-related applications, metal oxide nanoparticles are increasingly being developed through cheaper and more user-friendly approaches. We have formulated a simple route to synthesize zinc oxide nanoparticles (ZNPs) by a sol-gel method at near-room temperatures 25°C, 35°C, 55°C, and 75°C. The results are analyzed by X-ray diffraction, scanning electron microscopy with energy-dispersive X-ray spectroscopy, and ultraviolet-visible absorption spectroscopy. The effect of different temperature conditions (25°C-75°C) on the particulate sizes (23.7-88.8 nm), pH levels (11.7-11.9), and morphologies (slender needle-broad arrow) of flower-shaped ZNP colonies is studied. A possible mechanism depicting the growth rates at different temperatures and of different facets, mainly towards the <0 0 0 I> and <0 I ? 0> planes of the ZNPs has also been discussed. The values of ?max (293-298 nm) suggest that ZNPs prepared at 55°C are the most effective ultraviolet B absorbers, and that they can be used in sunscreens. Highly significant antimicrobial activity against medically important Gram-positive (Staphylococcus aureus) and Gram-negative (Escherichia coli) bacteria and fungi (Candida albicans) by these ZNPs was also revealed. As S. aureus and C. albicans are responsible for many contagious dermal infections such as abscesses, furuncles, carbuncles, cellulitis, and candidiasis, we can postulate that our fabricated ZNPs may be useful as antimicrobial agents in antiseptic creams and lotions for the treatment of skin diseases.
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Protease characteristics of bacteriocin producing Lysinibacilli, isolated from fruits and vegetable waste.
Bioinformation
PUBLISHED: 01-01-2014
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This study describes the physical stability and optimization of nutrient components for an extracellular protease produced by Bacillus strains isolated from fruits and vegetable waste, Lucknow, India. The isolated proteases could hydrolyze various native proteinaceous substrates such as bovine serum albumin, casein, skim milk, but not the gelatin. The strain JX416854 and isolate 10 yielded maximum protease (831; 703 U/ml) under optimized conditions: Nutrient, Casein broth; pH 7.0; shaking condition 37°C for 36 h. Crude protease exhibited activity over a wide range of pH (6.0-10.0) and found to be stable at (10-70°C), pH stable at 7- 9.0. The significant protease activity was observed with divalent cations Ca2+ and Mg2+ and EDTA. Further, significant blood destaining properties and stabilities with detergents were also observed. Thus, the significant potency and stability of these enzymes indicated their industrial importance and could be an alternative protease for various industrial applications.
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1,8-cineole (eucalyptol) mitigates inflammation in amyloid Beta toxicated PC12 cells: relevance to Alzheimer's disease.
Neurochem. Res.
PUBLISHED: 01-01-2014
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Inflammatory process has a fundamental role in the pathogenesis of Alzheimer's disease and insoluble amyloid beta deposits and neurofibrillary tangles provide the obvious stimuli for inflammation. The present study demonstrate the effect of pretreatment of 1,8-cineole (Cin) on inflammation induced by A?(25-35) in differentiated PC12 cells. The cells were treated with Cin at different doses for 24 h and then replaced by media containing A?(25-35) for another 24 h. The cell viability was decreased in A?(25-35) treated cells which was significantly restored by Cin pretreatment. Cin successfully reduced the mitochondrial membrane potential, ROS and NO levels in A?(25-35) treated cells. Cin also lowered the levels of proinflammatory cytokines TNF-?, IL-1? and IL-6 in A?(25-35) treated cells. Moreover, Cin also succeeded in lowering the expression of NOS-2, COX-2 and NF-?B. This study suggests the protective effects of Cin on inflammation and provides additional evidence for its potential beneficial use in therapy as an anti-inflammatory agent in neurodegenerative disease.
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Metal-free, single-polymer device exhibits resistive memory effect.
ACS Nano
PUBLISHED: 11-14-2013
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All-polymer, write-once-read-many times resistive memory devices have been fabricated on flexible substrates using a single polymer, poly(3,4-ethylenedioxythiophene):polystyrene sulfonate (PEDOT:PSS). Spin-cast or inkjet-printed films of solvent-modified PEDOT:PSS are used as electrodes, while the unmodified or as-is PEDOT:PSS is used as the semiconducting active layer. The all-polymer devices exhibit an irreversible but stable transition from a low resistance state (ON) to a high resistance state (OFF) at low voltages caused by an electric-field-induced morphological rearrangement of PEDOT and PSS at the electrode interface. However, in the metal-PEDOT:PSS-metal devices, we have shown a metal filament formation switching the device from an initial high resistance state (OFF) to the low resistance state (ON). The all-PEDOT:PSS memory device has low write voltages (<3 V), high ON/OFF ratio (>10(3)), good retention characteristics (>10?000 s), and stability in ambient storage (>3 months).
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Effect of high-dose zinc supplementation with oral hypoglycemic agents on glycemic control and inflammation in type-2 diabetic nephropathy patients.
J Nat Sci Biol Med
PUBLISHED: 10-02-2013
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The study aims to evaluate the effect of zinc sulfate on markers of glycemic control, lipid profile and inflammation in type-2 diabetes with microalbuminuria patients.
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Phenyl aldehyde and propanoids exert multiple sites of action towards cell membrane and cell wall targeting ergosterol in Candida albicans.
AMB Express
PUBLISHED: 09-03-2013
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In the present study, two phyto-compounds phenyl aldehyde (cinnamaldehyde) and propanoid (eugenol) were selected to explore their modes of action against Candida albicans. Electron microscopy, flow cytometry and spectroscopic assays were employed to determine the targets of these compounds. Treatment of C. albicans (CA04) with sub-MICs of cinnamaldehyde (50 ?g/mL) and eugenol (200 ?g/mL) indicated multiple sites of action including damages to cell walls, cell membranes, cytoplasmic contents and other membranous structures as observed under electron microscopy. Concentration and time dependent increase in the release of cytoplasmic contents accompanied with change in extracellular K+ concentration was recorded. Exposure of Candida cells at 4?×?MIC of cinnaamldehyde and eugenol resulted in 40.21% and 50.90% dead cells, respectively as revealed by flow cytometry analysis. Treatment of Candida cells by cinnamaldehyde and eugenol at 0.5?×?MIC showed 67.41% and 76.23% reduction in ergosterol biosynthesis, respectively. The binding assays reflected the ability of compounds to bind with the ergosterol. Our findings have suggested that the membrane damaging effects of phenyl aldehyde and propanoids class of compounds is attributed to their ability to inhibit ergosterol biosynthesis and simultaneously binding with ergosterol. Indirect or secondary action of these compounds on cell wall is also expected as revealed by electron microscopic studies.
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Glycation-assisted synthesized gold nanoparticles inhibit growth of bone cancer cells.
Colloids Surf B Biointerfaces
PUBLISHED: 08-31-2013
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This study presents a novel approach to synthesize glycogenic gold nanoparticles (glycogenic GNps) capped with glycated products (Schiffs base, Heyns products, fructosylamine etc.). These glycogenic GNps have been found to be active against human osteosarcoma cell line (Saos-2) with an IC50 of 0.187mM, while the normal human embryonic lung cell line (L-132) remained unaffected up to 1mM concentration. The size of glycogenic GNps can also be controlled by varying the time of incubation of gold solution. Glycation reactions involving a combination of fructose and HSA (Human Serum Albumin) were found to be effective in the reduction of gold to glycogenic GNps whereas glucose in combination with HSA did not result in the reduction of gold. The progress of the reaction was followed using UV-visible spectroscopy and NBT (Nitroblue tetrazolium) assay. The glycogenic GNps were found to be spherical in shape with an average size of 24.3nm, in a stable emulsion. These GNps were characterized using UV-visible spectroscopy, zeta potential analysis, transmission electron microscopy (TEM) and scanning electron microscopy (SEM).
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Study of self-defocusing, reverse saturable absorption and photoluminescence in anthraquinone PMMA nanocomposite film.
Spectrochim Acta A Mol Biomol Spectrosc
PUBLISHED: 08-08-2013
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The nanocomposite film of 1,5-diamino-9,10-anthraquinone in poly methyl methacrylate has been fabricated by following guest-host method. Intense reverse saturable absorption and self-defocusing effect have been investigated by employing Z-scan technique with low power CW laser at 532 nm with different intensities for the dye-polymer composite film. The estimated values of nonlinear absorption coefficient ?, nonlinear refractive index n2 and third order susceptibility ?((3)) of the composite film are of the order of 10(-3) (m/W), 10(-11) (m(2)/W) and 10(-4) (esu), respectively. The dye molecules have been encapsulated uniformly between molecules of polymer as a nanocomposite with average roughness ?7.96 nm as characterized by AFM technique. The nanocomposite film also exhibited strong photoluminescence emission when excited with 532 nm. The second order hyperpolarizability of composite film has also been estimated. The evaluated figure of merit W having a value greater than 1, and the results obtained suggest that the composite film of 1,5-diamino-9,10-anthraquinone-PMMA has potential applications in nonlinear optical devices.
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An In Vitro and Molecular Informatics Study to Evaluate the Antioxidative and ?-hydroxy-?-methylglutaryl-CoA Reductase Inhibitory Property of Ficus virens Ait.
Phytother Res
PUBLISHED: 07-09-2013
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The present study is initially intended to evaluate antioxidant and ?-hydroxy-?-methylglutaryl-CoA reductase (HMGR) inhibitory property of Ficus virens Ait., first by in vitro analyses followed by a corroboratory molecular informatics study. Our results show that of all the sequentially extracted fraction of F. virens bark and leaves extract, F. virens bark methanol extract exhibits strong radical scavenging, antioxidant and oxidative DNA damage protective activity, which is well correlated with its total phenolic content. In addition, F. virens bark methanol extract, which is non-cytotoxic, significantly and non-covalently inhibit the HMGR activity (IC50 ?=?3.45?±?0.45?µg/ml) in comparison with other extracts. The mechanistic aspect of this inhibition activity is authenticated by molecular docking study of bioactive compounds as revealed from gas chromatography-mass spectrometry data, with HMGR. The analysis for the first time indicates that quinic acid (?G: -8.11?kcal/mol) and paravastatin (?G: -8.22?kcal/mol) exhibit almost same binding energy, while other compounds also showed good binding energy, suggesting that quinic acid alone or in combination with other major bioactive compound is probably responsible for HMGR inhibitory property of the extract and plausibly can be used in in vivo system for the management, prevention, and alleviation of hypercholesterolemia as well as hypercholesterolemia-induced oxidative stress. Copyright © 2013 John Wiley & Sons, Ltd.
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Suprapubic Fecal Fistula Due To Richters Inguinal Hernia: A Case Report and Review of Literature.
Iran J Med Sci
PUBLISHED: 07-05-2013
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We encountered a 55-year-old man, who presented with an orifice in the suprapubic region, through which fecal material was discharged. On physical examination, the patient was unstable. After resuscitation, thorough evaluation, and laboratory and imaging investigations, he underwent an exploratory laparotomy. The ileum was entrapped at the deep inguinal ring, and there was some pus in the pelvic cavity. The purulent material was cleaned, and anastomosis was done after the resection of the defective ileal segment. In view of the rarity of this presentation and the paucity of published articles, this case is reported here.
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Analysis of constituents of the eastern Nigeria mistletoe, Loranthus micranthus linn revealed presence of new classes of osteogenic compounds.
J Ethnopharmacol
PUBLISHED: 07-04-2013
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Mistletoe extracts (decoctions) are used traditionally in eastern Nigeria for the management of bone pain, post menopausal syndrome and diabetes amongst several other ailments. While scientific evidence supporting its folkloric use as an antidiabetic agent has been documented, the age-long practice of its use in treatment of post menopausal syndrome has not been scientifically validated. Postmenopausal osteoporosis accounts for one of the prevalent disease conditions in aging population globally. This situation is exacerbated by the lack of osteogenic therapy. In search for plants of Nigerian origin with osteogenic potential, we evaluated eastern Nigerian mistletoe, having ethnotraditional claims of anti-diabetic, anti-hypertensive and anti-cancer activities as well as preventive effect in various post-menopausal syndromes.
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Gene-environment interactions in heavy metal and pesticide carcinogenesis.
Mutat. Res.
PUBLISHED: 07-01-2013
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Cancer is a complex disease involving a sequence of gene-environment interactions. Lifestyle, genetics, dietary factors, and environmental pollutants can increase the risk of cancer. Gene-environment interactions have been studied by a candidate-gene approach focusing on metabolism, DNA repair, and apoptosis. Here, we review the influence of gene-environment interactions in carcinogenesis, with emphasis on heavy metal and pesticide exposures.
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Concentration-dependent antagonistic persuasion of SDS and naphthalene derivatives on the fibrillation of stem bromelain.
Arch. Biochem. Biophys.
PUBLISHED: 06-07-2013
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Sodium dodecyl sulfate, a biological membrane mimetic, can be used to study the conversion of globular proteins into amyloid fibrils in vitro. Using multiple approaches, the effect of SDS was examined on stem bromelain (SB), a widely recognized therapeutic protein. SB is known to exist as a partially folded intermediate at pH 2.0, situation also encountered in the gastrointestinal tract (its site of absorption). In the presence of sub-micellar SDS concentration (500-1000?M), this intermediate was found to exhibit great propensity to form large-sized ?-sheeted aggregates with fibrillar morphology, the hall marks of amyloid structure. We also observed inhibition of fibrillation by two naphthalene-based compounds, ANS and bis-ANS. While bis-ANS significantly inhibited fibril formation at 50?M, ANS did so at relatively higher concentration (400?M). Alcohols, but not salts, were found to weaken the inhibitory action of these compounds suggesting the possible involvement of hydrophobic interactions in their binding to protein. Besides, isothermal titration calorimetry and molecular docking studies suggested that inhibition of fibrillation by these naphthalene derivatives is mediated not just through hydrophobic forces, but also by disruption of ?-? interactions between the aromatic residues together with the inter-polypeptide chain repulsion among negatively charged ANS/bis-ANS bound SB.
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Review of past research and proposed action plan for landfill gas-to-energy applications in India.
Waste Manag Res
PUBLISHED: 06-06-2013
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Open dumps employed for disposal of municipal solid waste (MSW) are generally referred to as landfills and have been traditionally used as the ultimate disposal method in India. The deposition of MSW in open dumps eventually leads to uncontrolled emission of landfill gas (LFG). This article reviews the MSW disposal practices and LFG emissions from landfills in India during the period 1994 to 2011. The worldwide trend of feasibility of LFG to energy recovery projects and recent studies in India indicate a changed perception of landfills as a source of energy. However, facilitating the implementation of LFG to energy involves a number of challenges in terms of technology, developing a standardized framework and availability of financial incentives. The legislative framework for promotion of LFG to energy projects in India has been reviewed and a comprehensive strategy and action plan for gainful LFG recovery is suggested. It is concluded that the market for LFG to energy projects is not mature in India. There are no on-ground case studies to demonstrate the feasibility of LFG to energy applications. Future research therefore should aim at LFG emission modeling studies at regional level and based on the results, pilot studies may be conducted for the potential sites in the country to establish LFG to energy recovery potential from these landfills.
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Use of collagen as a biomaterial: An update.
J Indian Soc Periodontol
PUBLISHED: 06-03-2013
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Biomaterial science is an expanding area, which encompasses a wide range of medical knowledge involving arthroplasty, cochlear implants, heart valves designing, lenses, dental fixation and tissue engineering. Within this context, in vitro cell culture on polymer scaffolds is one of the adopted strategies for tissue creation. It consists of a specific cell line that is seeded onto a particular substrate. This scaffold should provide excellent biocompatibility, controllable biodegradability, appropriate mechanical strength, flexibility as well as the ability to absorb body fluids for delivery of nutrients. Collagen certainly fulfils these demands; therefore, it is often chosen as a biomaterial. Moreover, this protein is abundant in the animal kingdom and plays a vital role in biological functions, such as tissue formation, cell attachment and proliferation.
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Co-expression of Arabidopsis transcription factor, AtMYB12, and soybean isoflavone synthase, GmIFS1, genes in tobacco leads to enhanced biosynthesis of isoflavones and flavonols resulting in osteoprotective activity.
Plant Biotechnol. J.
PUBLISHED: 05-14-2013
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Isoflavones, a group of flavonoids, restricted almost exclusively to family Leguminosae are known to exhibit anticancerous and anti-osteoporotic activities in animal systems and have been a target for metabolic engineering in commonly consumed food crops. Earlier efforts based on the expression of legume isoflavone synthase (IFS) genes in nonlegume plant species led to the limited success in terms of isoflavone content in transgenic tissue due to the limitation of substrate for IFS enzyme. In this work to overcome this limitation, the activation of multiple genes of flavonoid pathway using Arabidopsis transcription factor AtMYB12 has been carried out. We developed transgenic tobacco lines constitutively co-expressing AtMYB12 and GmIFS1 (soybean IFS) genes or independently and carried out their phytochemical and molecular analyses. The leaves of co-expressing transgenic lines were found to have elevated flavonol content along with the accumulation of substantial amount of genistein glycoconjugates being at the highest levels that could be engineered in tobacco leaves till date. Oestrogen-deficient (ovariectomized, Ovx) mice fed with leaf extract from transgenic plant co-expressing AtMYB12 and GmIFS1 but not wild-type extract exhibited significant conservation of trabecular microarchitecture, reduced osteoclast number and expression of osteoclastogenic genes, higher total serum antioxidant levels and increased uterine oestrogenicity compared with Ovx mice treated with vehicle (control). The skeletal effect of the transgenic extract was comparable to oestrogen-treated Ovx mice. Together, our results establish an efficient strategy for successful pathway engineering of isoflavones and other flavonoids in crop plants and provide a direct evidence of improved osteoprotective effect of transgenic plant extract.
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Impact of behavior change communication among pregnant women regarding neonatal care.
Indian J Pediatr
PUBLISHED: 05-06-2013
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To assess the behavior of pregnant women regarding neonatal care. Also to implement and assess impact of Behaviour Change Communication (BCC) package among pregnant women regarding neonatal care.
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Ribosylation of bovine serum albumin induces ROS accumulation and cell death in cancer line (MCF-7).
Eur. Biophys. J.
PUBLISHED: 04-28-2013
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Formation of advanced glycation end products (AGE) is crucially involved in the several pathophysiologies associated with ageing and diabetes, for example arthritis, atherosclerosis, chronic renal insufficiency, Alzheimers disease, nephropathy, neuropathy, and cataracts. Because of devastating effects of AGE and the significance of bovine serum albumin (BSA) as a transport protein, this study was designed to investigate glycation-induced structural modifications in BSA and their functional consequences in breast cancer cell line (MCF-7). We incubated D-ribose with BSA and monitored formation of D-ribose-glycated BSA by observing changes in the intensity of fluorescence at 410 nm. NBT (nitro blue tetrazolium) assay was performed to confirm formation of keto-amine during glycation. Absorbance at 540 nm (fructosamine) increased markedly with time. Furthermore, intrinsic protein and 8-anilino-1-naphthalenesulfonate (ANS) fluorescence revealed marked conformational changes in BSA upon ribosylation. In addition, a fluorescence assay with thioflavin T (ThT) revealed a remarkable increase in fluorescence at 485 nm in the presence of glycated BSA. This suggests that glycation with D-ribose induced aggregation of BSA into amyloid-like deposits. Circular dichroism (CD) study of native and ribosylated BSA revealed molten globule formation in the glycation pathway of BSA. Functional consequences of ribosylated BSA on cancer cell line, MCF-7 was studied by MTT assay and ROS estimation. The results revealed cytotoxicity of ribosylated BSA on MCF-7 cells.
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Experimental and theoretical investigations of nonlinear optical properties of 1,4-Diamino-9,10-Anthraquionone.
Spectrochim Acta A Mol Biomol Spectrosc
PUBLISHED: 04-26-2013
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Nonlinear optical properties of 1,4-Diamino-9,10-Anthraquinone dye in solution at different concentrations are investigated by utilizing single beam Z-scan technique using a low power continuous wave laser (?=532 nm). The anthraquinone dye is found to exhibit self-defocusing and reverse saturable absorption behavior. Effect of concentration on nonlinear refractive index and nonlinear absorption coefficient are also studied. The nonlinear absorption coefficient (?) and nonlinear refractive index (n2) have been evaluated from the open aperture and closed aperture Z-scan data and are found to increase with increase in concentration. The order of magnitude obtained for nonlinear refractive index and nonlinear absorption coefficient are found to be 10(-6) esu and 10(-4) m/W, respectively. The optical limiting behavior and induced self-diffraction patterns are also observed. To have a theoretical insight of nonlinear optical properties of 1,4-Diamino-9,10-Anthraquinone, first hyperpolarizability (?) is also evaluated by using quantum chemical calculations employing DFT method using 6-311 G basis set. The results obtained confirm the nonlinear optical behavior of 1,4-Diamino-9,10-Anthraquinone dye.
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Cancer morbidity in rheumatoid arthritis: role of estrogen metabolites.
Biomed Res Int
PUBLISHED: 04-24-2013
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Estrogen metabolites have been implicated in rheumatoid arthritis (RA) and cancer, although the mechanism remains unestablished. Some estrogen metabolites, which are used for the assessment of cancer risk, play an important role in RA. The pathways by which malignancies associated with RA remain elusive. Possible mechanism involves enzymatic or nonenzymatic oxidation of estrogen into catecholestrogen metabolites through semiquinone and quinone redox cycle to produce free radicals that can cause DNA modifications. Modifications of DNA alter its immunogenicity and trigger various immune responses leading to elevated levels of cancer and RA antibodies. However, the role of different estrogen metabolites as a mediator of immune response cannot be ruled out in various immune-related diseases.
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Developmental exposure to As, Cd, and Pb mixture diminishes skeletal growth and causes osteopenia at maturity via osteoblast and chondrocyte malfunctioning in female rats.
Toxicol. Sci.
PUBLISHED: 04-19-2013
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We studied the effect of metal mixture (MM), comprising As, Cd, and Pb, in developing female rat skeleton from gestation day 5 until postnatal day 60 (P-60). MM resulted in synergistic inhibition in viability and differentiation of osteoblasts in vitro, likely induced by reactive oxygen species. MM, administered at their most frequently occurring concentrations present in the groundwater of India, i.e., As: 0.38 ppm, Pb: 0.22 ppm, and Cd: 0.098 ppm or 10× of the ratio to developing rats, exhibited a synergistic decrease in ex vivo mineralization of bone marrow stromal (osteoprogenitor) cells. MM group showed a dose-dependent attenuation in weight and axial lengths and shortening of tibias at P-60. Furthermore, the growth plate was shortened, which was associated with shorter proliferative and hypertrophic zones, decreased parathyroid hormone-related protein and Indian hedgehog expression in the chondrocytes, reduced primary and secondary spongiosa, and hypomineralized osteoids-a major characteristic of osteomalacia. In addition, compared with the control, MM-treated rats were clearly osteopenic based on bone mineral density, microarchitecture, biomechanical strength, and particularly the biochemical profile, that suggested high turnover bone loss. Finally, in comparison to the control, the fracture-healing ability of MM group was delayed and accompanied by inferior quality of the healed bone. Together, these data demonstrated that the mixture of As, Cd, and Pb induced synergistic toxicity to developing skeleton, thereby diminishing modeling-directed bone accrual, inducing osteopenia and dampening fracture healing.
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Antioxidant and ? -amylase inhibitory property of phyllanthus virgatus L.: an in vitro and molecular interaction study.
Biomed Res Int
PUBLISHED: 04-14-2013
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The present study on Phyllanthus virgatus, known traditionally for its remedial potential, for the first time provides descriptions of the antioxidant and inhibition of ? -amylase enzyme activity first by in vitro analyses, followed by a confirmatory in silico study to create a stronger biochemical rationale. Our results illustrated that P. virgatus methanol extract exhibited strong antioxidant and oxidative DNA damage protective activity than other extracts, which was well correlated with its total phenolic content. In addition, P. virgatus methanol extract strongly inhibited the ? -amylase activity (IC50 33.20 ± 0.556? ? g/mL), in a noncompetitive manner, than acarbose (IC50 76.88 ± 0.277? ? g/mL), which showed competitive inhibition. Moreover, this extract stimulated the glucose uptake activity in 3T3-L1 cells and also showed a good correlation between antioxidant and ? -amylase activities. The molecular docking studies of the major bioactive compounds (9,12-octadecadienoic acid, asarone, 11-octadecenoic acid, and acrylic acid) revealed via GC-MS analysis from this extract mechanistically suggested that the inhibitory property may be due to the synergistic effect of these bioactive compounds. These results provide substantial basis for the future use of P. virgatus methanol extract and its bioactive compound in in vivo system for the treatment and management of diabetes as well as in the related condition of oxidative stress.
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Time resolved spectroscopy and gain studies of Fullerenes C60 and C70.
Spectrochim Acta A Mol Biomol Spectrosc
PUBLISHED: 04-13-2013
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The fluorescence decay time of Fullerenes C60 and C70 in pure form as well as in mixture with Coumarin C440 and Quinizarine dyes are studied. Results indicate that the decay of pure fullerenes is constant throughout the solute concentration and it is also independent of excitation wavelength, whereas in the case of mixture with dyes different behavior is noticed. We have also calculated the Stern-Volmer quenching constant and optical gain of both the fullerenes from which it is found that the optical gain is positive for Fullerene C70 only in a very narrow range of concentration.
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Lipoprotein (a) Levels in Relation to Severity of Coronary Artery Disease in North Indian Patients.
Heart Views
PUBLISHED: 04-13-2013
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Lipoprotein (a) [Lp (a)] is an established risk marker of coronary artery disease which is independent from other risk factors.
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Diversity-oriented synthesis of ?-aminophosphonates: a new class of potential anticancer agents.
Eur J Med Chem
PUBLISHED: 04-01-2013
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A small library of structurally diverse ?-aminophosphonates has been synthesized by reacting alkyl/aryl aldehydes, alkyl/aryl amines and alkyl/aryl phosphites in one-pot catalyzed by Amberlite-IR 120 resin (acidic). All the synthesized ?-aminophosphonates were assayed for their in vitro cytotoxic activities against a panel of five human cancer cell lines including A-549, NCI-H23 (Lung), Colo 320DM (Colon), MG-63 (Bone marrow) and Jurkat (Blood T lymphocytes). Compound 4n having (R)-1-phenylethanamine was found to be the most active amongst all the synthesized ?-aminophosphonates against all the five cancer cell lines, most prominent being against Jurkat cell line with an IC50 value of 4 ?M. Surprisingly, compound 4o having (S)-1-phenylethanamine was found to be devoid of any cytotoxicity. Our finding suggests that these chemical entities could further serve as interesting template for the design of potential anticancer agents.
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Biochemical, immunological and kinetic characterisation of thiol protease inhibitor (cystatin) from liver.
Appl. Biochem. Biotechnol.
PUBLISHED: 03-30-2013
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Regulation of the cysteine protease activity is imperative for proper functioning of the various organ systems. Elevated activities of cysteine proteinases due to impaired regulation by the endogenous cysteine proteinase inhibitors (cystatins) have been linked to liver malignancies. To gain an insight into these regulatory processes, it is essential to purify and characterise the inhibitors, cystatins. Present study was undertaken to purify the inhibitor from the liver. The purification was accomplished in four steps: alkaline treatment, ammonium sulphate fractionation, acetone precipitation and gel filtration column (Sephacryl S-100 HR). The eluted protein exhibited inhibitory activity towards papain, and its purity was further reaffirmed using western blotting and immunodiffusion. The purified inhibitor (liver cystatin (LC)) was stable in the pH range of 6-8 and temperature up to 45 °C. In view of the significance of kinetics parameters for drug delivery, the kinetic parameters of liver cystatin were also determined. LC showed the greatest affinity for papain followed by ficin and bromelain. UV and fluorescence spectroscopy results showed that binding of LC with thiol proteases induced changes in the environment of aromatic residues. Recent advances in the field of proteinase inhibitors have drawn attention to the possible use of this collected knowledge to control pathologies.
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Cancer chemoprevention by polyphenols and their potential application as nanomedicine.
J Environ Sci Health C Environ Carcinog Ecotoxicol Rev
PUBLISHED: 03-29-2013
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Today cancer is a leading cause of death among the developed countries. Its highly complex nature makes it difficult to understand as it entails multiple cellular physiological systems such as cell signaling and apoptosis. The biggest challenges faced by cancer chemoprevention/chemotherapy is maintaining drug circulation and avoiding multidrug resistance. Overall there is modest evidence regarding the protective effects of nutrients from supplements against a number of cancers. Numerous scientific literatures available advocate the use of polyphenols for chemoprevention. Some groups have also suggested use of combination of nutrients in cancer prevention. However, we have yet to obtain the desired results in the line of cancer chemotherapy research. Nanotechnology can play a pivotal role in cancer treatment and prevention. Moreover, nanoparticles can be modified in various ways to prolong circulation, enhance drug localization, increase drug efficacy, and potentially decrease the chances of multidrug resistance. In this communication, we will cover the use of various polyphenols and nutrients in cancer chemoprevention. The application of nanotechnology in this regard will also be included. In view of available reports on the potential of nanoparticles, we suggest their usage along with different combination of nutrients as cancer chemotherapeutic agents.
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Optimization of hexametaphosphate-assisted extraction and functional characterization of palm kernel cake protein.
Food Sci Technol Int
PUBLISHED: 03-23-2013
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Response surface methodology was applied to study the optimization of palm kernel cake protein (PKCP) hexametaphosphate-assisted extraction. The optimum PKCP yield (28.37%) when extracted using 1.50% sodium hexametaphosphate (SHMP) of pH 10, at 50?°C, and the 1:70 (w/v) ratio of cake-to-solvent was significantly (P?
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Ethanol extract of Peperomia pellucida (Piperaceae) promotes fracture healing by an anabolic effect on osteoblasts.
J Ethnopharmacol
PUBLISHED: 03-21-2013
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The whole plant or some part of Peperomia pellucida (L.) HBK is used in some parts of Cameroon as a treatment for fracture healing.
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At very low concentrations known chaotropes act as kosmotropes for the N and B isoforms of human serum albumin.
Biochem. Cell Biol.
PUBLISHED: 03-14-2013
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Very few studies have been done to understand the effect of millimolar concentrations of chaotropes on protein structure. In our previous study we observed that the secondary and tertiary structure of human serum albumin (HSA) increases in the presence of 5 mmol/L urea. Micelle formation in amphoteric detergents increases in the presence of equivalent concentrations of urea. Here, we observed a significant increase in the secondary and tertiary structure of HSA. Interestingly, guanidine hydrochloride, another chaotropic agent, also shows a similar effect. Our results show electrostatic interaction may play a role in neutral to basic transition in HSA. This study further supports the claim that at millimolar concentrations the chaotropes may act as kosmotropes for proteins.
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Greater Skeletal Gains in Ovary Intact Rats at Maturity Are Achieved by Supplementing a Standardized Extract of Butea monosperma Stem Bark that Confers Better Bone Conserving Effect following Ovariectomy and Concurrent Treatment Withdrawal.
Evid Based Complement Alternat Med
PUBLISHED: 03-13-2013
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With a longitudinally designed study, we tested whether an acetone soluble fraction (ASF) from the stem bark of Butea monosperma resulted in maximizing bone gain in rats during growth and maturation and thus protected against osteopenia following ovariectomy (OVx) with concomitant treatment withdrawal. Female rats at weaning were given ASF (100?mg/kg/d) or vehicle for 12 weeks, and baseline skeletal parameters (micro-CT) and total plasma antioxidant status (TAS) were measured. At this stage, one group was OVx and the other group was sham operated. Vehicle group (untreated) after OVx was given E2 or continued with vehicle (OVx control). ASF group after OVx was given vehicle (ASF withdrawn, ASFW). After another 12 weeks, all groups were killed and various skeletal parameters were determined. ASF resulted in substantially better skeletal parameters and higher plasma TAS over control at maturity. Rats treated with ASF before OVx had reduced rates of bone loss compared to OVx control. Twelve weeks after OVx, the ASFW group exhibited better trabecular microarchitectural preservation, bone turnover profiles, increased cortical deposition, and biomechanical strength over the OVx control, and the effects were comparable to OVx + E2 group. ASF supplementation during skeletal growth could maximize bone accrual and could confer increased resistance to post-OVx osteopenia despite treatment withdrawal.
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Benzo(a)pyrene induced structural and functional modifications in lung cystatin.
Environ Monit Assess
PUBLISHED: 02-26-2013
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Cystatins are thiol proteinase inhibitors ubiquitously present in the mammalian body. They serve a protective function to regulate the activities of endogenous proteinases, which may cause uncontrolled proteolysis and damage. In the present study, the effect of benzo(a)pyrene [BaP] on lung cystatin was studied to explore the hazardous effects of environmental pollutant on structural and functional integrity of the protein. The basic binding interaction was studied by UV-absorption, FT-IR, and fluorescence spectroscopy. The enhancement of total protein fluorescence with a red shift of 5 nm suggests structural scratch of lung cystatin by benzo(a)pyrene. Further, ANS binding studies reaffirm the unfolding of the thiol protease inhibitor (GLC-I) after treating with benzo(a)pyrene. The results of FT-IR spectroscopy reflect perturbation of the secondary conformation (alpha-helix to ?-sheet) in goat lung cystatin on interaction with BaP. Finally, functional inactivation of cystatin on association with BaP was checked by its papain inhibitory activity. Benzo(a)pyrene (10 ?M) caused complete inactivation of goat lung cystatin. Benzo(a)pyrene-induced loss of structure and function in the thiol protease inhibitor could provide a caution for lung injury caused by the pollutants and smokers.
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Development of new atom transfer radical polymerization system by iron (III)-metal salts without using any external initiator and reducing agent.
Macromol Rapid Commun
PUBLISHED: 02-23-2013
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Atom transfer radical polymerization (ATRP) catalyzed by high oxidation state metal salts of FeX3 is developed for the first time in the absence of both external initiator and reducing agent. Methyl methacrylate (MMA) and styrene are polymerized successfully using FeX3 /Phosphorous ligands with well-controlled molecular weight distributions (=1.5). The molecular weight of the polymers increases with monomer consumption with the progress of time and the polymerization behaviors show a decent ATRP trend. Activators and initiators are suggested to generate in situ by the addition reaction of MMA and one equivalent of FeX3 . The PMMA synthesized from without-initiator system is characterized by (1) H, (13) C and DEPT (distortionless enhancement by polarization transfer nuclear magnetic resonance) nuclear magnetic resonance spectroscopy. Chain extension and copolymerization experiments prove the livingness of the obtained polymer.
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Homology modeling of LmxMPK4 of Leishmania mexicana and virtual screening of potent inhibitors against it.
Interdiscip Sci
PUBLISHED: 02-17-2013
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Leishmaniasis is one of the most important diseases of mankind. In the life cycle of Leishmania mexicana, two most important developmental stages are observed. In insect vector it is in promastigote form and in mammalian macrophages is the amastigote form. The family of protein kinases are extremely important regulators of many different cellular processes such as transcriptional control, cell cycle development and differentiation, and also draw much attention as possible drug targets for protozaon parasites. Leishmania mexicana mitogen activated protein kinase 4 (LmxMPK4) is essential for proliferation and survival of the parasite promastigote and amastigote forms and is a potential drug target for leishmaniasis. The existing therapy for leishmaniasis is not enough due to host toxicity and drug resistance. The experimental 3D structure of this protein has not yet been determined. In this study, we have used homology modelling techniques to generate the 3D structure of LmxMPK4 and selected effective inhibitors by ZINC database on the basis of structure of berberine alkaloid for molecular docking studies with LmxMPK4. The inhibitors ZINC05999210, ZINC40402312 and ZINC40977377 were found to be more potent for inhibition of leishmaniasis due to the robust binding affinity and strong inhibition constant (Ki) of the protein-ligand interactions. This finding may help to understand the nature of MAP kinase and development of specific anti-leishmanial therapies.
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Improper inhaler technique is associated with poor asthma control and frequent emergency department visits.
Allergy Asthma Clin Immunol
PUBLISHED: 02-09-2013
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Uncontrolled asthma remains a frequent cause of emergency department (ED) visits and hospital admissions. Improper asthma inhaler device use is most likely one of the major causes associated with uncontrolled asthma and frequent ED visits.
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Taurine ameliorates neurobehavioral, neurochemical and immunohistochemical changes in sporadic dementia of Alzheimers type (SDAT) caused by intracerebroventricular streptozotocin in rats.
Neurol. Sci.
PUBLISHED: 01-30-2013
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Oxidative loads in the brain are involved in age related impairments like learning and memory as well as neurodegeneration. Taurine, the most abundant free amino acid in humans has many potential health benefits through its anti-oxidant and anti-inflammatory properties. Therefore, we investigated the neuroprotective potential of taurine on oxidative stress, neuronal loss and memory impairments in streptozotocin model of cognitive impairments in rats. The cognitive impairment was developed by giving single intracerebroventricular (ICV) injection of streptozotocin (STZ) 3 mg/kg body weight bilaterally. An increased latency and path length was observed in ICV-STZ group animals as compared to sham group animals and these were inhibited significantly in STZ group pre-treated with taurine (50 mg/kg body weight orally once daily for 15 days). Moreover, the significantly depleted content of GSH and elevated level of thiobarbituric acid reactive substances (TBARS) in ICV-STZ group animals were protected significantly with pre-treatment of taurine. The activity of antioxidant enzymes, glutathione peroxidase, glutathione reductase, glutathione-S-transferase, catalase, and superoxide dismutase was decreased in STZ group as compared to sham group and pre-treatment of STZ group with taurine has protected their activities significantly. Furthermore, the increased activity of acetylcholine esterase and decreased expression of choline acetyl transferase were attenuated by the pre-treatment of taurine. Taurine also protected the morphology of the hippocampal pyramidal neurons. This study concludes that the prophylactic intervention of taurine may be used to prevent the deterioration of cognitive functions and neurobehavioral activities, often associated with the generation of free radicals.
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Amelioration of cognitive impairment and neurodegeneration by catechin hydrate in rat model of streptozotocin-induced experimental dementia of Alzheimers type.
Neurochem. Int.
PUBLISHED: 01-26-2013
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Alzheimers disease (AD) is a progressive neurodegenerative disorder resulting in cognitive decline and enhancement of oxidative loads in the brain. Flavonoids have been considered to exert human health benefits by anti-oxidant and anti-inflammatory properties. The present study is aimed to elucidate the neuroprotective effect of catechin hydrate (CH), a natural flavanoid with potential antioxidant and anti-inflammatory properties, on intracerebroventricular streptozotocin (ICV-STZ) induced neuronal loss and memory impairment. To test this hypothesis, male Wistar rats were pretreated with CH (10 and 20mg/kgb wt) orally once daily for 21 days and then bilaterally injected with ICV-STZ (3mg/kgb wt), while sham group rats receive the same volume of vehicle. After 2 weeks of ICV-STZ infusion, rats were tested for cognitive performance using Morris water maze (MWM) test and then sacrifice for biochemical and histopathological assays. CH was found to be successful in upregulating the antioxidant status and prevented the memory loss. The expression of choline acetyl transferase (ChAT) was decreased in ICV-STZ group and CH pretreatment increases the expression of ChAT. Moreover, inflammatory mediators like TNF-?, IL-1? levels and expression of iNOS were significantly attenuated by CH pretreatment. The study suggests that CH is effective in preventing memory loss, ameliorating the oxidative stress and might be beneficial for the treatment of sporadic dementia of Alzheimers type (SDAT).
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Attenuation of oxidative damage-associated cognitive decline by Withania somnifera in rat model of streptozotocin-induced cognitive impairment.
Protoplasma
PUBLISHED: 01-07-2013
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Oxidative stress is a critical contributing factor to age-related neurodegenerative disorders. Therefore, the inhibition of oxidative damage, responsible for chronic detrimental neurodegeneration, is an important strategy for neuroprotective therapy. Withania somnifera (WS) extract has been reported to have potent antioxidant and free radical quenching properties in various disease conditions. The present study evaluated the hypothesis that WS extract would reduce oxidative stress-associated neurodegeneration after intracerebroventricular injection of streptozotocin (ICV-STZ) in rats. To test this hypothesis, male Wistar rats were pretreated with WS extract at doses of 100, 200, and 300 mg/kg body weight once daily for 3 weeks. On day 22nd, the rats were infused bilaterally with ICV-STZ injection (3 mg/kg body weight) in normal saline while sham group received only saline. Two weeks after the lesioning, STZ-infused rats showed cognitive impairment in the Morris water maze test. The rats were sacrificed after 3 weeks of the lesioning for the estimation of the contents of lipid peroxidation, reduced glutathione, and activities of glutathione reductase, glutathione peroxidase, and catalase. Pretreatment with WS extract attenuated behavioral, biochemical, and histological alterations significantly in dose-dependent manner in the hippocampus and cerebral cortex of ICV-STZ-infused rats. These results suggest that WS affords a beneficial effect on cognitive deficit by ameliorating oxidative damage induced by streptozotocin in a model of cognitive impairment.
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Different conformation of thiol protease inhibitor during amyloid formation: inhibition by curcumin and quercetin.
J Fluoresc
PUBLISHED: 01-07-2013
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Cystatins are thiol proteinase inhibitors ubiquitously present in mammalian body and serve various important physiological functions. In the present study, we examined the effects of acid denaturation on newly identified thiol protease inhibitors from the lungs of Capra hircus (Goat) with a focus on protein conformational changes and amyloid fibril formation. Acid denaturation as studied by CD (Circular Dichroism) and fluorescence spectroscopy showed that purified inhibitor named GLC (Goat Lung Cystatin) populates three partly unfolded species, a native like state at pH 3.0, a partly unfolded intermediate at pH2.0, and unstructured unfolded state at pH 1.0, from each of which amyloid like fibrils grow as assessed by thioflavin T (ThT) spectroscopy. The result showed, native like structure formed at pH 3.0 is more responsive towards amyloid formation when compare to other conformation of proteins. Morphology of the protein species incubated for amyloid process was observed using transmission electron microscopy (TEM). Moreover, anti-fibrillogenic effects of curcumin and quercetin were analysed using ThT binding assay. Curcumin and quercetin produced a concentration dependent decline inThT fluorescence suggesting deaggregation of the fibrils. When added prior to amyloid fibril initiation 50 ?M curcumin inhibited amyloid aggregation. However, more quercetin is needed to prevent the same extent of fibrillation. Implications for therapeutics in view of polyphenols as essential nutrients are suggested in lung diseases.
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Total Water, Phosphorus Relaxation and Inter-Atomic Organic to Inorganic Interface Are New Determinants of Trabecular Bone Integrity.
PLoS ONE
PUBLISHED: 01-01-2013
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Bone is the living composite biomaterial having unique structural property. Presently, there is a considerable gap in our understanding of bone structure and composition in the native state, particularly with respect to the trabecular bone, which is metabolically more active than cortical bones, and is readily lost in post-menopausal osteoporosis. We used solid-state nuclear magnetic resonance (NMR) to compare trabecular bone structure and composition in the native state between normal, bone loss and bone restoration conditions in rat. Trabecular osteopenia was induced by lactation as well as prolonged estrogen deficiency (bilateral ovariectomy, Ovx). Ovx rats with established osteopenia were administered with PTH (parathyroid hormone, trabecular restoration group), and restoration was allowed to become comparable to sham Ovx (control) group using bone mineral density (BMD) and µCT determinants. We used a technique combining (1)H NMR spectroscopy with (31)P and (13)C to measure various NMR parameters described below. Our results revealed that trabecular bones had diminished total water content, inorganic phosphorus NMR relaxation time (T1) and space between the collagen and inorganic phosphorus in the osteopenic groups compared to control, and these changes were significantly reversed in the bone restoration group. Remarkably, bound water was decreased in both osteopenic and bone restoration groups compared to control. Total water and T1 correlated strongly with trabecular bone density, volume, thickness, connectivity, spacing and resistance to compression. Bound water did not correlate with any of the microarchitectural and compression parameters. We conclude that total water, T1 and atomic space between the crystal and organic surface are altered in the trabecular bones of osteopenic rats, and PTH reverses these parameters. Furthermore, from these data, it appears that total water and T1 could serve as trabecular surrogates of micro-architecture and compression strength.
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Inhibitory effect of metformin and pyridoxamine in the formation of early, intermediate and advanced glycation end-products.
PLoS ONE
PUBLISHED: 01-01-2013
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Non-enzymatic glycation is the addition of free carbonyl group of reducing sugar to the free amino groups of proteins, resulting in the formation of a Schiff base and an Amadori product. Dihydroxyacetone (DHA) is one of the carbonyl species which reacts rapidly with the free amino groups of proteins to form advanced glycation end products (AGEs). The highly reactive dihydroxyacetone phosphate is a derivative of dihydroxyacetone (DHA), and a product of glycolysis, having potential glycating effects to form AGEs. The formation of AGEs results in the generation of free radicals which play an important role in the pathophysiology of aging and diabetic complications. While the formation of DHA-AGEs has been demonstrated previously, no extensive studies have been performed to assess the inhibition of AGE inhibitors at all the three stages of glycation (early, intermediate and late) using metformin (MF) and pyridoxamine (PM) as a novel inhibitor.
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Design, synthesis, evaluation and thermodynamics of 1-substituted pyridylimidazo[1,5-a]pyridine derivatives as cysteine protease inhibitors.
PLoS ONE
PUBLISHED: 01-01-2013
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Targeting papain family cysteine proteases is one of the novel strategies in the development of chemotherapy for a number of diseases. Novel cysteine protease inhibitors derived from 1-pyridylimidazo[1,5-a]pyridine representing pharmacologically important class of compounds are being reported here for the first time. The derivatives were initially designed and screened in silico by molecular docking studies against papain to explore the possible mode of action. The molecular interaction between the compounds and cysteine protease (papain) was found to be very similar to the interactions observed with the respective epoxide inhibitor (E-64c) of papain. Subsequently, compounds were synthesized to validate their efficacy in wet lab experiments. When characterized kinetically, these compounds show their Ki and IC50 values in the range of 13.75 to 99.30 µM and 13.40 to 96.50 µM, respectively. The thermodynamics studies suggest their binding with papain hydrophobically and entropically driven. These inhibitors also inhibit the growth of clinically important different types of Gram positive and Gram negative bacteria having MIC50 values in the range of 0.6-1.4 µg/ml. Based on Lipinskis rule of Five, we also propose these compounds as potent antibacterial prodrugs. The most active antibacterial compound was found to be 1-(2-pyridyl)-3-(2-hydroxyphenyl)imidazo[1,5-a]pyridine (3a).
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Mapping and validation of the major sex-determining region in Nile tilapia (Oreochromis niloticus L.) Using RAD sequencing.
PLoS ONE
PUBLISHED: 01-01-2013
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Sex in Oreochromis niloticus (Nile tilapia) is principally determined by an XX/XY locus but other genetic and environmental factors also influence sex ratio. Restriction Associated DNA (RAD) sequencing was used in two families derived from crossing XY males with females from an isogenic clonal line, in order to identify Single Nucleotide Polymorphisms (SNPs) and map the sex-determining region(s). We constructed a linkage map with 3,802 SNPs, which corresponded to 3,280 informative markers, and identified a major sex-determining region on linkage group 1, explaining nearly 96% of the phenotypic variance. This sex-determining region was mapped in a 2 cM interval, corresponding to approximately 1.2 Mb in the O. niloticus draft genome. In order to validate this, a diverse family (4 families; 96 individuals in total) and population (40 broodstock individuals) test panel were genotyped for five of the SNPs showing the highest association with phenotypic sex. From the expanded data set, SNPs Oni23063 and Oni28137 showed the highest association, which persisted both in the case of family and population data. Across the entire dataset all females were found to be homozygous for these two SNPs. Males were heterozygous, with the exception of five individuals in the population and two in the family dataset. These fish possessed the homozygous genotype expected of females. Progeny sex ratios (over 95% females) from two of the males with the "female" genotype indicated that they were neomales (XX males). Sex reversal induced by elevated temperature during sexual differentiation also resulted in phenotypic males with the "female" genotype. This study narrows down the region containing the main sex-determining locus, and provides genetic markers tightly linked to this locus, with an association that persisted across the population. These markers will be of use in refining the production of genetically male O. niloticus for aquaculture.
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Antibiofilm activity of certain phytocompounds and their synergy with fluconazole against Candida albicans biofilms.
J. Antimicrob. Chemother.
PUBLISHED: 12-13-2011
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The aim of this study was to evaluate four phytocompounds (cinnamaldehyde, citral, eugenol and geraniol) for their in vitro inhibitory activity against pre-formed biofilms of Candida albicans alone or in combination with fluconazole and amphotericin B. These compounds were also tested at subinhibitory concentrations for their ability to inhibit biofilm formation.
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CD4+ T cell-derived novel peptide Thp5 induces interleukin-4 production in CD4+ T cells to direct T helper 2 cell differentiation.
J. Biol. Chem.
PUBLISHED: 11-30-2011
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The differentiation of naïve CD4(+) T cells into T helper 2 (Th2) cells requires production of the cytokine IL-4 in the local microenvironment. It is evident that naïve/quiescently activated CD4(+) T cells produce the IL-4 that drives Th2 cell differentiation. Because early production of IL-4 in naïve T cells leads to preferential Th2 cell differentiation, this process needs to be tightly regulated so as to avoid catastrophic and misdirected Th2 cell differentiation. Here, we show that Thp5, a novel peptide with structural similarity to vasoactive intestinal peptide, regulates production of early IL-4 in newly activated CD4(+) T cells. Induction of IL-4 in CD4(+) T cells by Thp5 is independent of the transcription factor STAT6 but dependent on ERK1/2 signaling. Furthermore, cytokines (IL-12 and TGF-?) that promote the differentiation of Th1 or Th17 cells inhibit Thp5 induction, thus suppressing Th2 cell differentiation. We further showed that Thp5 enhances Th2 responses and exacerbates allergic airway inflammation in mice. Taken together, our findings reveal that early activated CD4(+) T cells produce Thp5, which plays a critical role as a molecular switch in the differentiation of Th cells, biasing the response toward the Th2 cell phenotype.
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Rutin protects dopaminergic neurons from oxidative stress in an animal model of Parkinsons disease.
Neurotox Res
PUBLISHED: 11-29-2011
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This study was undertaken to investigate the neuroprotective effects of rutin (vitamin P) on 6-hydroxydopamine (6-OHDA)-induced Parkinsons disease (PD) in rats. Oxidative stress and inflammation is an important event, play a crucial role in neurodegenerative diseases. Rutin has been shown to have antioxidant and anti-inflammatory actions, and thus was tested for its beneficial effects using 6-OHDA-induced PD rat model. Male Wistar rats were pre-treated with rutin (25 mg/kg bwt, orally) for 3 weeks and subjected to unilateral intrastriatal injection of 6-OHDA (10 ?g in 0.1% ascorbic acid in normal saline). Three weeks after 6-OHDA infusion, rats were tested for neurobehavioral activity, and were killed after 4 weeks of 6-OHDA infusion for the estimation of thiobarbituric acid reactive substances, glutathione, and its dependent enzymes (glutathione peroxidase and glutathione reductase), dopamine (DA) and its metabolite 3,4-dihydroxyphenyl acetic acid. The increase in 6-OHDA-induced rotations and deficits in locomotor activity and motor coordination and decrease in antioxidant level, DA content and its metabolite and increase in the number of dopaminergic D2 receptors in striatum were protected significantly with lesioned group pre-treated with rutin. These findings were further supported by the histopathological and immunohistochemical findings in the substantia nigra that showed that rutin protected neurons from deleterious effects of 6-OHDA. These results suggest that the consumption of rutin, which is novel vitamin, may have the possibility of protective effect against the neurological disorder such as PD.
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