02664nas a2200301   4500008004100000022001400041245023700055210006900292260009500361300001000456490000700466520151000473653001101983653001601994653001702010653001002027653001402037653002002051653002202071653002702093653002402120100001602144700001502160700001702175700002802192700001602220856012602236       2006                        eng d  a0048-357500aFlufenoxuron, an acylurea insect growth regulator, alters development of Tribolium castaneum (Herbst) (Coleoptera :tenebrionidae) by modulating levels of chitin, soluble protein content, and HSP70 and p34(cdc2) in the larval tissues0 aFlufenoxuron an acylurea insect growth regulator alters developm  a525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USAbACADEMIC PRESS INC ELSEVIER SCIENCEcJUN  a84-900 v853 a<p>The effect of sublethal concentrations, 0.00141% (LC20), 0.00251% (LC30), and 0.0036% (LC40) of a dispersible formulation of an acylurea insect growth regulator, flufenoxuron (Cascade) on certain biochemical parameters in the larvae of Tribolium castaneum was investigated. When neonates were fed on diet treated with sublethal concentrations for 24 It, it was observed that at all concentrations tested, there was a significant reduction in chitin content on the 15th day of development. Total soluble protein content at LC20 and LC30 decreased with increasing age of the larvae. At LC20 and LC40 concentrations there was a progressive increase in the protein: chitin ratio as a function of increase in age of the larvae. SDS-PAGE analysis of the larval tissue extracts indicated gross quantitative changes in some of the protein bands (MW 50-97 kDa). Western blot analysis revealed significant increase in the level of HSP70 in the extracts of larvae fed on LC30 treated diet, on the 7th and 10th day of development in the decreasing order. Conversely, a significant decrease in the hyper-phosphorylated form of p34(cdc2) kinase due to flufenoxuron treatment indicating modulation of cell cycle regulation was observed. Thus, sublethal concentrations of flufenoxuron alter expression of developmentally regulated proteins, HSP70 and p34(cdc2) and chitin formation in a stage-specific manner thereby resulting developmental abnormalities in T castaneum. (c) 2005 Elsevier Inc. All rights reserved.</p>10achitin10aDevelopment10aflufenoxuron10aHSP7010ap34(cdc2)10aphosphotyrosine10asublethal effects10atotal soluble proteins10aTribolium castaneum1 aSalokhe, S.1 aSarkar, A.1 aKulkarni, A.1 aMukherjee, Samindra, N.1 aPal, J., K.  uhttps://library.ncl.res.in/content/flufenoxuron-acylurea-insect-growth-regulator-alters-development-tribolium-castaneum-002224nas a2200205   4500008004100000245011400041210006900155260000800224300001200232490000700244520150600251100001701757700001801774700002101792700002201813700001601835700001701851700002501868856012501893       2016                        eng d00aMolecular cloning and in silico studies of physiologically significant trehalase from drosophila melanogaster0 aMolecular cloning and in silico studies of physiologically signi  cNOV  a282-2920 v923 aTrehalase, a physiologically important glycosidase is known for its crucial role in insect glycometabolism and stress recovery. The present study describes the molecular cloning of a gene fragment, encoding the catalytically active trehalase from Drosophila melanogaster (DmTre) and its heterologous expression in Escherichia coli. The 1275 bp gene was overexpressed in two different vectors viz., pET28a and pCOLD TF and investigated for variable soluble expression, purification and activity of the recombinant enzyme with optimum pH and temperature of enzyme as 6 and 55 degrees C, respectively. The sequence was characterized in silico by subjecting it to homology search, multiple sequence alignment and phylogenetic tree construction revealing its identity to other trehalases which belong to glycoside hydrolase family 37. The deduced amino acid sequence and modeled 3D structure of DmTre possessed all features of trehalase superfamily, including signature motifs and catalytic domain. The active site pocket of recombinant DmTre was compared with the crystal structure of E. coli trehalase identifying Glu424 and Asp226 as the putative catalytic residues. Additionally, enzyme-substrate docking suggests possible involvement of other residues in the catalysis along with Asp226. The present study holds significance in understanding the structural aspects of Drosophila trehalase in spite of unavailabilty of eukaryotic trehalase crystal structure. (C) 2016 Elsevier B.V. All rights reserved.1 aShukla, Ekta1 aThorat, Leena1 aBhavnani, Varsha1 aBendre, Ameya, D.1 aPal, J., K.1 aNath, B., B.1 aGaikwad, Sushama, M.  uhttps://library.ncl.res.in/content/molecular-cloning-and-silico-studies-physiologically-significant-trehalase-drosophila