Purification, Characterization and Functional Analysis of a Serine Protease Inhibitor from the Pulps of Cicer arietinum L. (Chick Pea)

Serine proteinase inhibitors (SPIs) are present in high amount in legume seeds where they play an important role in plant defence mechanism against pests. In the present study, a serine protease inhibitor has been identified from the seeds of Cicer arietinum (L.) and characterized for its inhibitory potency against trypsin and chymotrypsin. Ammonium sulphate fractionation was executed as an initial step to purify the inhibitor. The fraction which was obtained from 30-60% relative ammonium sulphate saturation exhibited the maximum trypsin inhibition activity against 0.2% casein using radial diffusion method. The 30–60% fraction was further subjected to ion exchange chromatography using 1 mL HiTrap Q HP column. The peak fractions were analyzed for the enzymatic activity and also characterized on 12% SDS PAGE. The results indicated that the flow through fraction has retained a significant proteolytic inhibition towards trypsin with a relative molecular mass of approximately 12-13 kDa. The kinetic results have demonstrated that the purified inhibitor from Cicer arietinum L. not only inhibited trypsin but also chymotrypsin. The Circular Dichroism spectrum analysis of the purified inhibitor has revealed that the secondary structure content is highly composed of random coils which were supported by the reports of other low molecular weight trypsin inhibitors. To conclude, a low molecular weight serine protease inhibitor possessing both trypsin and chymotrypsin inhibition from the seeds of C. arietinum has been purified, characterized and the results are reported

Effect of chemical treatment on germination of nutmeg (Myristica fragrans Routt.) seeds

Studies were undertaken to evaluate the influence of pre-sowing treatment of nutmeg (Myristica fragrans) seeds with various chemicals on the viability of seeds and seedling vigour. Among the various chemicals evaluated, gibberellic acid (100 ppm) was most effective and resulted in significantly higher germination (85%) and vigour index value (3945). &nbsp

Effect of chemical treatment on germination of nutmeg (Myristica fragrans Routt.) seeds

Studies were undertaken to evaluate the influence of pre-sowing treatment of nutmeg (Myristica fragrans) seeds with various chemicals on the viability of seeds and seedling vigour. Among the various chemicals evaluated, gibberellic acid (100 ppm) was most effective and resulted in significantly higher germination (85%) and vigour index value (3945). &nbsp

Design, crystal structure determination, molecular dynamic simulation and MMGBSA calculations of novel p38-alpha MAPK inhibitors for combating Alzheimer's disease

The hallmark of the Alzheimer's disease (AD) is the accumulation of aggregated, misfolded proteins. The cause for this accumulation is increased production of misfolded proteins and impaired clearance of them. Amyloid aggregation and tau hyperphosphorylation are the two proteinopathies which accomplish deprivation of cell and tissue hemostasis during neuropathological process of the AD, as a result of which progressive neuronal degeneration and the loss of cognitive functions. p38 mitogen-activated protein kinase (p38 MAPK) has been implicated in both the events associated with AD: tau protein phosphorylation and inflammation. p38 alpha MAPK pathway is activated by a dual phosphorylation at Thr180 and Tyr182 residues. Clinical and preclinical evidence implicates the stress related kinase p38 alpha MAPK as a potential neurotherapeutic target. Drug design of p38 alpha MAPK inhibitors is mainly focused on small molecules that compete for Adenosine triphosphate in the catalytic site. Here we have carried out the synthesis of phenyl sulfonamide derivatives Sulfo (I) and Sulfo (II). Crystal structures of Sulfo (I) and Sulfo (II) were solved by direct methods using SHELXS-97. Sulfo (I) and Sulfo (II) have R(int)values of 0.0283 and 0.0660, respectively, indicating good quality of crystals and investigated their ability against p38 alpha MAPK. Docking studies revealed that the Sulfo (I) had better binding affinity (-62.24 kcal/mol) as compared to Sulfo (II) and cocrystal having binding affinity of -54.61 kcal/mol and -59.84 kcal/mol, respectively. Molecular dynamics simulation studies of Sulfo (I) and cocrystal of p38 alpha MAPK suggest that during the course of 30 ns simulation run, compound Sulfo (I) attained stability, substantiating the consistency of its binding to p38 alpha MAPK compared to cocrystal. Binding free energy analysis suggests that the compound Sulfo (I) is better than the cocrystal. Thus, this study corroborates the therapeutic potential of synthesized Sulfo (I) in combatting AD

Morphological and Molecular Characterization of Cyclopoid Copepod in and around Coimbatore Lakes Using 18s rRNA Analysis

Published ecological research on the variety and distribution of freshwater planktonic cyclopoid copepods rely on DNA techniques for precise taxonomic identification. Large-scale investigations of plankton identification using molecular techniques are now more practical due to the increased coverage of reference species in the genetic database GenBank and the falling prices for DNA sequencing. Here, we offer a useful morphological and molecular method for identifying  Mesocyclops aspericornis, M. dissimilis, M. pehpiensis, Thermocyclopsdecipiens, and T. hyalinus, which were found in Coimbatore, India, at the Sulur, Singanallur, Ukkadam, and Muthanna lakes. A microscope was used to observe morphological identification. Amplification of 18S rRNA was one technique used for cyclopoids' molecular identification. The evolutionary relationship between 5 species of freshwater cyclopoid copepods is the focus of the current study on molecular phylogenetic analysis. The findings revealed that there was no interpopulation variation in the 18S rRNA molecules of M. dissimilis, M. pehpiensis, T. decipiens, and T. hyalinus. It is intriguing to look at genetic distances within and across taxa to see if a particular group of cyclopoids has diverged, on average, more or less than others. The greatest genetic divergence among cyclopoid taxa was caused by the extreme variance in M. aspericornis, which was more than twice as high as the other cyclopoids. Finally, during the investigation, T. hyalinus was discovered for the first time in the lakes of Coimbatore, India

Purification, Characterization and Functional Analysis of a Serine Protease Inhibitor from the Pulps of <em>Cicer arietinum </em>L. (Chick Pea)

117-124Serine proteinase inhibitors (SPIs) are present in high amount in legume seeds where they play an important role in plant defence mechanism against pests. In the present study, a serine protease inhibitor has been identified from the seeds of&nbsp;Cicer arietinum (L.) and characterized for its inhibitory potency against trypsin and chymotrypsin. Ammonium sulphate fractionation was executed as an initial step to purify the inhibitor. The fraction which was obtained from 30-60% relative ammonium sulphate saturation exhibited the maximum trypsin inhibition activity against 0.2% casein using radial diffusion method. The 30&ndash;60% fraction was further subjected to ion exchange chromatography using 1 mL HiTrap Q HP column. The peak fractions were analyzed for the enzymatic activity and also characterized on 12% SDS PAGE. The results indicated that the flow through fraction has retained a significant proteolytic inhibition towards trypsin with a relative molecular mass of approximately 12-13 kDa. The kinetic results have demonstrated that the purified inhibitor from Cicer arietinum L. not only inhibited trypsin but also chymotrypsin. The Circular Dichroism spectrum analysis of the purified inhibitor has revealed that the secondary structure content is highly composed of random coils which were supported by the reports of other low molecular weight trypsin inhibitors. To conclude, a low molecular weight serine protease inhibitor possessing both trypsin and chymotrypsin inhibition from the seeds of C. arietinum has been purified, characterized and the results are reported

Prediction of Coronary Artery Disease Among Chronic Hepatitis C Virus Infected Patients in Tamilnadu, India

C-reactive protein (CRP) in the form of high sensitivity CRP (hs-CRP) is one of the better extensively used risk marker for the coronary artery disease (CAD) in clinical practice. Increased value of hs-CRP has been related with increased threat for the development of blood pressure, stroke, peripheral arterial disease, temporary ischemic attack and sudden coronary deceased condition. If hs-CRP involves a fundamental role in atherothrombosis, then hs-CRP is the better biomarker proposed to be necessary for improving to detect the risk of coronary events of arterial disease. In this study, the HCV seropositive patients (by Anti HCV-IgG Indirect Elisa) more than 2 years old, hepatic and leukaemia were investigated for extra hepatic manifestation. The Lab request forms with questionnaire were together including CRP, HS-CRP, serum cholesterol and liver function tests were scrutinized. Total of 170 cases (around the Chennai) was taken for the study. In which, 45.3% of cases were observed as hepatitis patients with ALL cases. The other 8% of cases were having liver cirrhosis and rest of the cases was not having cirrhosis. Finally, the study explores the relationship between the HCV seropositive of leukaemia cases, HCV RNA and CRP, hs CRP with cirrhosis. These evaluations point out that HCV infection may amend the chronic inflammatory condition in leukaemia patients. Additionally, the results recommended that screening for hs-CRP should be considered previous to evaluation of cardiovascular risk in HCV patients, because the results may affect the risk scoring of coronary artery disease

Structure and mechanism of action of a novel phosphoglycerate mutase from Bacillus stearothermophilus

Bacillus stearothermophilus phosphoglycerate mutase (PGM), which interconverts 2- and 3–phosphoglyceric acid (PGA), does not require 2,3–diphosphoglyceric acid for activity. However, this enzyme does have an absolute and specific requirement for Mn(2+) ions for catalysis. Here we report the crystal structure of this enzyme complexed with 3PGA and manganese ions to 1.9 Å resolution; this is the first crystal structure of a diphosphoglycerate-independent PGM to be determined. This information, plus the location of the two bound Mn(2+) ions and the 3PGA have allowed formulation of a possible catalytic mechanism for this PGM. In this mechanism Mn(2+) ions facilitate the transfer of the substrate's phosphate group to Ser62 to form a phosphoserine intermediate. In the subsequent phosphotransferase part of the reaction, the phosphate group is transferred from Ser62 to the O2 or O3 positions of the reoriented glycerate to yield the PGA product. Site-directed mutagenesis studies were used to confirm our mechanism and the involvement of specific enzyme residues in Mn(2+) binding and catalysis

Identification of potential PKC inhibitors through pharmacophore designing, 3D-QSAR and molecular dynamics simulations targeting Alzheimer’s disease

<p>Protein kinases are ubiquitously expressed as Serine/Threonine kinases, and play a crucial role in cellular activities. Protein kinases have evolved through stringent regulation mechanisms. Protein kinases are also involved in tauopathy, thus are important targets for developing Anti-Alzheimer’s disease compounds. Structures with an indole scaffold turned out to be potent new leads. With the aim of developing new inhibitors for human protein kinase C, here we report the generation of four point 3D geometric featured pharmacophore model. In order to identify novel and potent PKCθ inhibitors, the pharmacophore model was screened against 80,000,00 compounds from various chemical databases such as., ZINC, SPEC, ASINEX, which resulted in 127 compound hits, and were taken for molecular docking filters (HTVS, XP docking). After in-depth analysis of binding patterns, induced fit docking (flexible) was employed for six compounds along with the cocrystallized inhibitor. Molecular docking study reveals that compound 6F found to be tight binder at the active site of PKCθ as compared to the cocrystal and has occupancy of 90 percentile. MM-GBSA also confirmed the potency of the compound 6F as better than cocrystal. Molecular dynamics results suggest that compound 6F showed good binding stability of active sites residues similar to cocrystal 7G compound. Present study corroborates the pharmacophore-based virtual screening, and finds the compound 6F as a potent Inhibitor of PKC, having therapeutic potential for Alzheimer’s disease. Worldwide, 46.8 million people are believed to be living with Alzheimer’s disease. When elderly population increases rapidly and neurodegenerative burden also increases in parallel, we project the findings from this study will be useful for drug developing efforts targeting Alzheimer’s disease.</p

An Adaptive resistance perturbation based MPPT algorithm for Photovoltaic applications

none5siThis paper proposes a Resistance perturbation based maximum power point tracking (MPPT) with an adaptive control limit algorithm to extract the maximum power from solar photovoltaic (PV) array. This algorithm consists of two main functions, namely 1) resistance perturbation & observation (RP&O) and 2) adaptive resistance control (ARC) limit. The RP&O operates the PV array at maximum power point (MPP), and the ARC limit continuously monitors the resistance of the PV Rpv to determine the operating limit of MPP. The ultimate aim of proposing this algorithm is to reduce the oscillations and improve MPP’s tracking performance for sudden variation in temperature and irradiance conditions. Furthermore, it does not require an expensive pyranometer or temperature sensor to track the MPP of the PV array. This paper also compares the proposed and conventional MPPT algorithm’s performance. Its validation results in both MATLAB/Simulink and experimental studies are presented under constant and sudden changes in irradiance conditions.open1. Maheswaran GUNASEKARAN, Vijayakumar KRISHNASAMY, Sivakumar SELVAM, Dhafer J ALMAKHLES, Norma ANGLANIMaheswaran GUNASEKARAN, 1.; Krishnasamy, Vijayakumar; Selvam, Sivakumar; J ALMAKHLES, Dhafer; Anglani, Norm