Cloning,expression analysis and In silico characterization of HSP101: a potential player conferring heat stress in Aegilops speltoides (Tausch) Gren

Heat shock protein (HSP101) function as molecular chaperones and confer thermotolerance to plants. In the present investigation, identification, comprehensive expression analysis, phylogeny and protein modelling of HSP101 gene has been done in Aegilops speltoides accession Pau3583. In the present study, we cloned and in silico characterized a HSP101C gene designated as AsHSP101C-Pau3583. AsHSP101C-Pau3583 is 4180 bp long with seven exons and six introns and encoded a polypeptide of 910 amino acids predicted by FGENESH. We have identified 58 SNPs between the AsHSP101C-Pau3583 and reference gene sequence extracted from Ae. speltoides TGAC assembly. Real-time RT-PCR analysis of expression levels of HSP101 gene in two wheat genotypes under heat stress revealed that gene namely HSP101C was up-regulated in Aegilops speltoides acc. Pau3583 by > fourfold in comparison to Triticum aestivum cv. PBW343 under heat stress signifies that it plays a role in conferring heat tolerance. Sequence comparison and phylogenetic analysis of AsHSP101C-Pau3583 with seven wheat homologs Triticum aestivum, Aegilops speltoides (TGAC), Triticum durum cv Cappelli, Triticum durum cv Strongfield, Triticum monococcum, Aegilops tauschii and Triticum urartu showed significant similarities with highly conserved coding regions and functional domains (AAA, AAA + 2, ClpB domains), suggesting the conserved function of HSP101C in different species. The illustration of the protein models of HSP101C in homologs provided information for the ATP-binding motifs within the nucleotide binding domains (NBD), specific for the chaperone activity. These findings are important and identified SNPs could be used for designing markers for ensuring the transfer of AsHSP101C-Pau3583 gene into hexaploid wheat and its role in heat tolerance.Supplementary InformationThe online version contains supplementary material available at 10.1007/s12298-021-01005-2.     

Design and Construction of Aroyl‐Hydrazone Derivatives: Synthesis,Crystal Structure,Molecular Docking and Their Biological Activities

Here, we report the synthesis and characterization of four new aroyl‐hydrazone derivatives L₁ – L₄ , and their structural as well as biological activities have been explored. In addition to docking with bovine serum albumin (BSA) and duplex DNA, the experimental results demonstrate the effective binding of L₁ – L₄ with BSA protein and calf thymus DNA (ct‐DNA) which is in agreement with the docking results. Further biological activities of L₁ – L₄ have been examined through molecular docking with different proteins which are involved in the propagation of viral or cancer diseases. L₁ shows best binding affinity with influenza A virus polymerase PB2 subunit (2VY7) with binding energy ?11.42 kcal/mol and inhibition constant 4.23 nm , whereas L₂ strongly bind with the hepatitis C virus NS5B polymerase (2WCX) with binding energy ?10.47 kcal/mol and inhibition constant 21.06 nm . Ligand L₃ binds strongly with TGF‐beta receptor 1 (3FAA) and L₄ with cancer‐related EphA2 protein kinases (1MQB) with binding energy ?10.61 kcal/mol, ?10.02 kcal/mol and inhibition constant 16.67 nm and 45.41 nm , respectively. The binding energies of L₁ – L₄ are comparable with binding energies of their proven inhibitors. L₁ , L₃ and L₄ can be considered as both 3FAA and 1MQB dual targeting anticancer agents, while L₁ and L₃ are both 2VY7 and 2WCX dual targeting antiviral agents. On the other side, L₂ and L₄ target only one virus related target (2WCX). Furthermore, the geometry optimizations of L₁ – L₄ were performed via density functional theory (DFT). Moreover, all four ligands ( L₁ – L₄ ) were characterized by NMR, FT‐IR, ESI‐MS, elemental analysis and their molecular structures were validated by single crystal X‐ray diffraction studies.     

Apoptolysis: a less understood concept in the pathogenesis of Pemphigus Vulgaris

Pemphigus Vulgaris (PV) is a severe autoimmune disease characterized by supra-basal blisters in the skin and mucous membranes of a wide range of mammals, including humans. It not only affects the skin but also has severe oral manifestations. It has been stated that auto-antibodies are produced, for unknown reasons, which are directed against desmogleins present on the epithelium and thus leads to acantholysis and intraepithelial blistering. But the exact mechanism is still not completely understood. Here we would like to shed light on a new pathologic mechanism i.e., apoptolysis, which emphasizes that apoptotic enzymes contribute to acantholysis development both in terms of molecular events and chronologic sequence. A possible role of apoptolysis has been discussed in purview of PV.

     

Serum levels of IgA in peptic ulcers

A study of the IgA levels in 43 duodenal ulcer (DU) patients and 8 gastric ulcer (GU) patients and their comparison with healthy controls reveals significantly elevated levels of IgA in DU and somewhat lower levels in GU. The levels were also associated with the genotypes of the patients for genetic markers such as ABO blood group, ABH sectetor status, haptoglobin, and alkaline phosphatase enzyme. Nutritional factors, such as vegetarianism, chili consumption, and habits such as smoking and alcoholism also showed variation in the IgA levels. These results indicate the response and role of IgA in the immunological mechanisms involving mucosal protection and autoimmunity in ulceration processes in the stomach.     

Analysis of short-term variation and long-term drift during reagent kit lot change in an NABL accredited clinical biochemistry laboratory

BackgroundKit lot change in clinical biochemistry labs leads to variations in patient results. This study planned to identify variations during 60 reagent lot changes in our laboratory during the period from June 2018 to May 2019.MethodsA statistical analysis was performed to identify the difference between patient samples results variations and QC results. The long term drift was analyzed using a regression test.ResultsThere was a significant difference between the patient and QC results in 16.7% of reagent lot changes. Moreover, the extent of variation in QC results was 3.3%. No long-term drift was seen in three analytes which were studied using regression analysis.ConclusionsOur results showed that, during reagent kit lot change, along with QC material, the patient samples should also be run in order to identify the variation. However, this practice is presently ignored by most of the laboratories. There was no accumulated effect in our laboratory due to reagent kit lot change.