Spectrophotometric estimation of nitro tyrosine by azo-coupling reaction

3-Nitrotyrosine has been identified using techniques like immunohistochemistry, HPLC, GCMS to demonstrate as a relevant biomarker in various disorders such as neurological, cardiovascular and others. Estimation of 3-NT by spectrophotometric method has been performed to measure 3-NT over decades. However none of the above methods have been translated to diagnose the diseases, but pathological conditions. Therefore here is an attempt to develop a method to be employed as a diagnostic tool. Current work is focussed on including preparation of 3-NT and further to derivatives using β-Napthol to yield diazotised product. All these intermediates of diazotized products have been characterized by ESI MS and the final colored dye was measured spectrophotometrically which exhibits λ max at 551 nm. The dye has been measured and compared with the direct measurement of 3-NT. The sensitivity of measurement of 3-NT increases by 10 fold, by and large we can estimate 20 μM concentration of 3-NT

Synthesis and biological evaluation of novel pyrazole derivatives as urease inhibitors

Studies on enzyme inhibition remain an important area of pharmaceutical research since these studies have led to the discoveries of drugs useful in a variety of physiological conditions. The enzyme inhibitors can interact with enzymes and block their activity towards natural substrates. Urease inhibitors have recently attracted much attention as potential new anti-ulcer drugs. A series of novel substituted pyrazoles 8(a-j) has been synthesized by diazotization of fluoro chloro aniline (1) and the reaction of the corresponding diazonium salt solution (2) with ethyl cyanoacetate (3) to give the intermediate, ethyl 2-((3-chloro-4-fluorophenyl) diazenyl-2- cyanoacetate (4). The intermediate is then cyclised with chloroacetonitrile (5) using triethyl amine as the base to give the final compound, ethyl 4-amino-1-(3-chloro-4-flurophenyl)-5- cyano-1H-pyrazole-3-carboxylate (6). Nucleophilic substitution group is removed from the final compound and 8(a-j) derivatives have been synthesized. All the synthesized compounds were characterized by physical data (M.P. & TLC) and spectral Data (IR & 1H NMR). The synthesized compounds were evaluated for urease-inhibition activity. Molecular docking studies were carried out for these compounds with the enzyme urease. From the observations it has been noticed that some of the compounds possesses remarkable urease-inhibitory effect

Effect of pH on spectral characteristics of P5C-ninhydrin derivative: Application in the assay of ornithine amino transferase activity from tissue lysate

Currently available method(s) for assaying pyrroline-5-carboxylate (P5C), an important intermediate metabolite of ornithine, proline and glutamate metabolic pathways, are cumbersome or not sensitive enough for microanalysis. The present study involving the synthesis of P5C followed by purity check, molecular mass (amu =113.1) determination by mass spectrometry and spectral characterization of P5C-ninhydrin derivative (λ max: 510 nm) confirmed the authenticity of the preparation. Studies on the effect of pH on spectral characteristics of P5C ninhydrin derivative demonstrated a significant change with respect to λ max (620 nm) and several ∼ 12 fold increase in molar extinction coefficient (ε: 1.96 × 105) in alkaline conditions (pH:7.0–8.0) as compared to the reported Molar ε of 1.65 × 104 at max λ 510 nm in ethanolic solution. The modified method, with the improved sensitivity, is adopted for the assay of ornithine amino transferase activity in WBC’s/platelets lysate(s) from human blood

Discovery of a latent calcineurin inhibitory peptide from its autoinhibitory domain by docking, dynamic simulation, and in vitro methods

Autoinhibitory domain (AID) of calcineurin (CN) was discovered two decades ago. Fewer investigations are reported to find out shortest possible peptide from the AID for CN inhibition. Hence, this study has focused on screening of nearly 150 peptide fragments derived from the AID using in silico method. Therefore, we have employed docking studies, aiming to analyze the best pose of AID-derived peptides on CN active site. We also analyzed binding free energy (ÎG) of docked complex using molecular mechanics/generalized Born surface area (MM/GBSA). MM/GBSA predicts two short peptides P1 and P2 found to be lowest binding free energy. Two peptides exhibit better binding affinity with CN, suggests that the possible candidates for potential CN inhibition. Further, the stability of the docked complex was analyzed using molecular dynamic (MD) simulation. MD study shows that CNA:P2 is the most stable complex than CN A:P1 and CN A:AID. Besides, we have synthesized and purified P1 and P2 peptides over high performance liquid chromatography (HPLC) found to be 90.31 and 98.93 of purity, respectively. In addition, AID peptides were characterized over mass spectral analysis. Peptides were subjected to CN inhibitory assay using malachite green method. Where, P1 and P2 exhibit CN inhibition better than AID. In particular, shortest peptide P2 shows highest inhibitory activity than AID. Enzyme assay reveals CN inhibitory activity of P2 peptide is consistent within silico results. In silico and in vitro, results corroborated each other to confirm short peptide P2 can be used as a potential CN inhibitor. © 2015 Taylor & Franci

Syntheses, characterizations, and antimicrobial activities of binuclear ruthenium(III) complexes containing 2-substituted benzimidazole derivatives

Binuclear ruthenium(III) complexes RuX3L2· nH2O (X=Cl, L=L1, L2, L3; n=1, L4 and L5, X=Br; L=L3), RuX3L 1.52·nH2O (X=Br, L=L1; n=0, L4; n=6 and L5; n=10), and RuX3L 22 (X=Br, L=L2) have been isolated by treatment of hydrated RuX3 (X=Cl/Br) in acetone with 2-(2â²-aminophenylbenzimidazole) (L1), 2-(3â²- aminophenylbenzimidazole) (L2), 2-(3â²-N-salicylidinephenyl) benzimidazole (L3), 2-(3â²-pyridylbenzimidazole) (L 4), and 2-(4â²-pyridylbenzimidazole) (L5) in acetone. The complexes were characterized by elemental analysis, conductivity and magnetic susceptibility measurements, IR, electronic, EPR, and mass spectral studies. The complexes were dimeric; based on analytical and spectral studies, an octahedral geometry was proposed for the complexes. The synthesized complexes were screened against Gram-positive and Gram-negative bacteria and fungi. © 2012 Taylor & Francis

Synthesis, In- vivo and In-silico anti-inflammatory studies of substituted fluoro pyrazole

N-phenyl-5-substituted -aryl-3-p-(fluorophenyl) pyrazoles have been synthesized from cyclization of 4-fluoroacetophenone (1) with various benzaldehydes (2) to give 4-fluorophenylstyrylketone (3) followed by treatment with phenyl hydrazine. The title compounds and their derivatives have been characterized by their elemental and spectral analysis. The newly synthesized compounds are screened for anti-inflammatory activity. All substituted 4-fluorophenylstyrylketones (250mg/kg orally p.o.) possessed anti-inflammatory activity against carrageenan-induced paw oedema in rat. Indomethacin (10mg/kg) was used as standard drug. And all compounds (0.20mM) showed ability to denature bovine serum albumin as observed in vitro inhibition studies. No correlation was found between the anti inflammatory activity and inhibition of bovine serum albumin denaturation. The docking studies were carried out for these compounds against the protein NFκB which is involved in inflammation signal cascade. Some of them showed good activity and molecular binding. Compounds such as 3b, 5b and 5e have exhibited comparative results in both in vivo and in silico studies

In silico and pharmacokinetics of para-nitrophenylphosphate substrate for calcineurin

Calcineurin is a calcium dependent protein ser/thr phosphatase. The assay of this enzyme has been done by using various substrates since it has shown promising evidences to act upon para-nitrophenylphosphate (p-'PP) and many phosphopeptide analogues as well. But none of the methods are as reliable as far sensitivity of the method is concerned. To address the assay method for the Calcineurin, we have isolated and partially purified the enzyme from the bovine brain and employed substrate like p-'PP with two different metal ions (Calcium and Manganese). We have established the binding affinity studies by using enzyme kinetics. The binding affinity of p-'PP against particular metal ion shows the better metal activator and substrate for the enzyme. The affinity in presence of calcium for the substrates p-'PP is found to be Km= 0.48 mM. The affinity in presence of manganese for the substrates p-'PP, is found to be Km= 0.77 mM. At the same time we have used Autodock 3.05 for minimal binding energy calculations for the p-'PP, the 1st cluster which had the maximum number of runs (30) reported mean dock energy of -4.24 kcal / mol. We conclude from the study that p-'PP has more affinity toward the calcineurin in presence of Calcium than Manganese. Copyright© (2010) by the International Society for Research in Science and Technology

Performance of potato (Solanum tuberosum L.) genotypes under hill zone of Karnataka.

An experiment was conducted to study the performance of nine genotypes of potato under hill zone of Karnataka. Among nine genotypes, maximum tuber sprout percentage was observed in Kufri Jyoti (100) followed by TT7011 (98.75) and least percentage was in C-28 (89.16). Kufri Jyoti performed better for most of the characters such as plant height(68.04 cm), number of leaves(42.21), leaf area (154.88 cm2), tuber weight (116.42 g), tuber length (8.20 cm), tuber diameter (5.88 cm), chlorophyll content (1.26 mg/g) and which was found to be significantly superior over other genotypes. Number of tubers per plant (4.43), total tuber yield per plot (18.98 kg), total tuber yield per hectare (26.36 t), marketable yield per plot (10.21 kg), marketable yield per hectare (14.18 t), dry matter (19.88 %), total sugars (2.55 %) was observed in genotype TT7011 compared to the rest of the genotypes. The genotype C-28 recorded highest starch content (24.10 %) followed by TT7007 (22.92 %) and the least starch content (19.05 %) was observed in MCIP-9-1. Genotype TT7011 realized maximum gross returns (283600.00 per ha) and net returns (193457.00 per ha) followed by TT7010 (Gross returns 190200.00 per ha and net returns 100057.00 per ha)

Biosorption of chromium(VI) and lead(II): Role of spirulina platensis in the treatment of industrial effluent

Biosorption is the process of removal of any chemical molecules by the treatment of biological material. Industrialization resulted in the discharge of various toxic heavy metals into water bodies, which poses serious health hazards to humans and animals. In the present study, live Spirulina platensis was used as a biosorbent for the removal of the heavy metals chromium (Cr(VI)) and lead (Pb(II)) from the aqueous samples. S. platensis were cultured in the presence of different concentrations of heavy metals. The growth of the algal cells was found to be decreased by 59 and 36 in media containing 50 ppm Cr(VI) and Pb(II), respectively. To assess the biosorption of heavy metals, at different time intervals, the spent culture media were used to detect Cr(VI) by atomic absorption spectroscopy method and Pb(II) by 4-(2-pyridylazo)resorcinol indicator method. Results suggested that there was a significant uptake of Cr(VI) and Pb(II) from the medium by S. platensis, with corresponding decrease of metals in the medium. When metal salt solutions or industrial effluent samples were passed through the column containing immobilized live S. platensis in calcium alginate beads, the concentration of Cr(VI) was found to be reduced drastically. The present study indicates the application of S. platensis for the bioremediation of heavy metals from the samples obtained from industrial effluents. © 2013 Copyright Taylor and Francis Group, LLC

Correlation and path analysis for yield and yield-related traits of potato (Solanum tuberosum L.) in Karnataka

Correlation and path analyses study on total tuber yield and yield attributing traits of nine CIP advance potato genotypes revealed that total tuber yield per plot had highly significant and positive association with marketable yield per plot, number of tubers per plant, dry matter and number of stems at 60 DAP both at genotypic and phenotypic level indicating the possibility of simultaneous selection for these traits to improve the tuber yield. It revealed that numbers of tubers per plant, marketable yield per plot, number of stems at 60 DAP and tuber weight were the most influencing factors to improve the tuber yield. Thus, these characters deserve greater weightage during selection for yield performance attribute