In Silico characterization of growth hormone from freshwater ornamental fishes: Sequence analysis, molecular modelling and phylogeny

The present investigation includes in Silico sequence analysis, three-dimensional (3D) structure prediction and evolutionary profile of growth hormone (GH) from 14 ornamental freshwater fishes. The analyses were performed using the sequence data of growth hormone gene (gh) and its encoded GH protein. The evolutionary analyses were performed using maximum likelihood (ML) estimate and maximum parsimony (MP) methods. Bootstrap test (1000 replicates) was performed to validate the phylogenetic tree. The tertiary structures of GH were predicted using the comparative modelling method. The suitable template for comparative modeling protein databank (PDB IDs: 1HWG A) has been selected on the basis of basic local alignment search tool (BLASTp) and fast analysis (FASTA) results. The target-template alignment, model building, loop modelling and evaluation have been performed in Modeller 9.10. The tertiary structure of GH is α-helix structure connected by loops, which forms a compressed complex maintained by two disulfide bridges. The resultant 3D models are verified by ERRAT and ProCheck programmes. After fruitful verification, the tertiary structures of GH have been deposited to protein model database (PMDB). Sequence analyses and RNA secondary structure prediction was performed by CLC genomics workbench version 4.0. The computational models of GH could be of use for further evaluation of molecular mechanism of function.Keywords: Growth hormone, in Silico, somatotropin, growth hormone gene (gh) mRNA, freshwater ornamental fis

Arabidopsis accelerated cell death 11, ACD11, is a ceramide-1-phosphate transfer protein and intermediary regulator of phytoceramide levels

The accelerated cell death 11 (acd11) mutant of Arabidopsis provides a genetic model for studying immune response activation and localized cellular suicide that halt pathogen spread during infection in plants. Here, we elucidate ACD11 structure and function and show that acd11 disruption dramatically alters the in vivo balance of sphingolipid mediators that regulate eukaryotic-programmed cell death. In acd11 mutants, normally low ceramide-1- phosphate (C1P) levels become elevated, but the relatively abundant cell death inducer phytoceramide rises acutely. ACD11 exhibits selective intermembrane transfer of C1P and phyto-C1P. Crystal structures establish C1P binding via a surface-localized, phosphate headgroup recognition center connected to an interior hydrophobic pocket that adaptively ensheaths lipid chains via a cleft-like gating mechanism. Point mutation mapping con- firms functional involvement of binding site residues. A p helix (p bulge) near the lipid binding cleft distinguishes apo-ACD11 from other GLTP folds. The global two-layer, a-helically dominated, ‘‘sandwich’’ topology displaying C1P-selective binding identifies ACD11 as the plant prototype of a GLTP fold subfamily

Characterisation and mode of in vitro replication of pea chloroplast OriA sequences

A partially purified replicative system of pea chloroplast that replicates recombinant DNAs containing pea chloroplast origin sequences has been characterised. Polymerisation by this system is very fast and insensitive to chain terminators like dideoxynucleotides, arabinosylcytosine 5'-triphosphate, etc. Both strands of template DNA are synthesized and single-stranded DNA templates undergo more than one round of replication. When sequences of either of the two chloroplast origins of replication (OriA or OriB) are used as templates, the replicative intermediates are found to have sigma structures. Electron microscopic analysis of the sigma structures restricted with various enzymes reveals that the initiation site of in vitro replication maps near the displacement-loop regions where replication initiates also in vivo. Although the observed replication initiation in the OriA recombinant template is chloroplast-DNA-specific, the mode of replication is different from that observed in vivo with intact ctDNA. However, when the template DNA contains both the OriA and OriB sequences, the in vitro replication proceeds in the theta mode, the mode of replication usually observed in vivo

Comparative assessment of severity and prognosis of acute pancreatitis through APACHE II and HAPS predictor models

Background: Acute pancreatitis is one of the leading causes of hospitalization amongst all gastrointestinal disorders with high burden of morbidity and mortality. Predicting the progression of AP in terms of course and outcome to determine suitable management strategy and level of care is challenging. A number of predictor models are developed to predict the severity of acute pancreatitis but they vary in their definitions of severity. HAPS have been proposed as a simple scoring tool for assessment of severity and prognosis of acute pancreatitis. Thus, the aim of present study was to investigate the usefulness of HAPS predictor model against APACHE II model.Methods: Current investigation was a hospital based prospective study conducted on 80 proven cases of acute pancreatitis at K. K. hospital, Uttar Pradesh. The serum amylase and lipase levels of all enrolled patients, were tested and measured at admission, and at 48 and 72 hours post admission. The pancreatitis-specific clinical investigations like; HAPS, APACHE II were calculated and assessed statistically in terms of sensitivity, specificity, positive and negative predictive values and accuracy.Results: The findings of present investigation revealed that amongst the two scoring systems, APACHE II was superior predictor model in terms of sensitivity and specificity for various outcomes like severe acute pancreatitis, hospital stay >7 days and in-hospital mortality. However, HAPS exhibited high specificity for all the outcomes.Conclusions: HAPS can be recommended as a useful tool for early evaluation of acute pancreatitis in patients specifically in primary care settings of developing countries like India