Constitutive expression and discovery of antimicrobial peptides in Zygogramma bicolorata (Coleoptera: Chrysomelidae).

The expression, identification, and discovery of less toxic antimicrobial peptides (AMPs) are significant in managing infectious pathogens. AMPs triggered in response to the immune system have evolved to defend against pathogens and wounding. The protein composition of Zygogramma bicolorata hemolymph is of diagnostic importance as the open circulatory systems of the insects involve signaling through hemolymph. They have conserved many ancestral vertebrate genes that may help better understand the evolution of innate immunity. The present work describes the isolation, purification, identification, and bioinformatics analysis of AMPs from the immunized hemolymph of Z. bicolorata. Thirty-nine peptides were isolated from reverse-phase high-performance liquid chromatography and sequenced via mass spectrometry analysis. The immunization process recorded a threefold higher protein concentration in immunized hemolymph when compared with nonimmunized one. For the first time, the proteomic study on Z. bicolorata hemolymph unveils the three novel proteins in the family Chrysomelidae with no homology in the database, indicating its novelty and the expression of the rest of 36 well-known proteins, including heat-shock, immune, structural, signaling proteins, and others speak for its method validity. Combining the expression of novel AMPs, detoxifying enzymes, hemolytic, and cytotoxic assays, and this work can elucidate new pathways to immune response mechanisms. Its molecular basis also holds the potential applicability in the future drug development process against pathogenic fungi such as Aspergillus niger and Candida albicans.

Metabolism of cisplatin in the organs of Rattus norvegicus: role of Glutathione S-transferase P1.

Glutathione S-transferases (GSTs) play an important role in the biotransformation of endogenous compounds and xenobiotics as well as in the metabolic inactivation of pharmacologically active substances, including anticancer drugs. Using cisplatin as the prototype drug, we investigated if any correlation exists between GSH levels, GSTs/GSTP1 activity and the fate of cisplatin in different organs of Rattus norvegicus. GSH-cisplatin complex was prepared, purified by anion-exchange chromatography and subjected to mass spectroscopic analysis which confirmed the structure to be diglutathione-monoplatinum (diglutathionylplatinum). Purified diglutathionylplatinum was used to quantify metabolite formed in different tissue homogenates. Specific GSTP1 activity was found to be highest in kidneys, which correlated positively with the levels of metabolite formed in renal tissues. Altogether, our results showed that cisplatin metabolism in different organs of rats correlated positively with specific GSTP1 activities and this enzyme may be a critical determinant of extent of cellular uptake or retention of cisplatin in renal and liver tissues.