Shotgun proteomics based on peptide fractionation by using liquid chromatography has become the common procedure for proteomic studies, although in the very beginning of the field, protein separation by using electrophoresis was the main tool. Nonetheless, during the last two decades, the electrophoretic techniques for peptide mixtures fractionation have evolved as a result of relevant technological improvements. We also proposed the combination of sodium dodecyl sulfate polyacrylamide gel electrophoresis for protein fractionation and sodium dodecyl sulfate free polyacrylamide gel electrophoresis for peptide separation as a novel procedure for proteomic studies. Here, we present an optimized device for sodium dodecyl sulfate free polyacrylamide gel electrophoresis improving peptide recoveries respect to the established electrophoretic technique off gel electrophoresis meanwhile conserving the excellent resolution described for the former technique in slab gel based systems. The device simultaneously allows the separation and the collection of fractionated peptides in solution.
The new antigen Bm95 from the cattle tick Boophilus microplus was recently isolated, cloned and expressed in the methylotrophic yeast Pichia pastoris. The recombinant protein has shown to induce protection in cattle against infestations of B. microplus under controlled and production conditions. In this paper we report the production and large-scale purification of the Bm95 protein, following a simple and cost-effective process. The antigen was obtained highly aggregated, forming particles ranging from 26 to 30 nm and with purity higher than 80%. The process yield was 0.55 g of pure Bm95 protein per liter of culture. The 98% of the primary structure of the recombinant protein was verified by mass spectrometry. Three amino acid changes in comparison with the sequence deduced from cDNA were detected by LC-MS/MS. The antigen was also obtained N-glycosylated, as previously reported for heterologous protein expression in P. pastoris.
In an attempt to find possible targets for benzimidazole action in muscle-stage larvae of Trichinella spiralis, the effects of mebendazole and thiabendazole were tested in vivo by oral treatment of infested mice and in vitro by including these anthelmintics in an adequate maintenance medium containing decapsulated larvae. The effects of the anthelmintics on succinate dehydrogenase and fumarate reductase activities, measured in the mitochondrial fraction obtained from the in vivo- or in vitro-treated larvae showed that only thiabendazole causes significant inhibition of fumarate reductase activity. On the other hand, measurements of free glucose, glycogen reserves and soluble protein in the treated larvae indicate that in vivo, mebendazole and thiabendazole clearly diminish free glucose levels, although in vitro only mebendazole produces the same diminution. Both the glycogen and protein contents of the larvae remained unchanged after treatment in vivo or in vitro. The importance of these findings with regard to a possible site of action for mebendazole and thiabendazole is discussed.
El incremento de las autolesiones en la adolescencia ha sido progresivo en las últimas décadas, con un incremento exponencial en los 10 últimos años. Además, en la actualidad se ha detectado un incremento notorio de las autolesiones en la población adolescente, en parte, relacionado con el estrés psicológico y distanciamiento social causado por la pandemia por COVID-19. El objetivo general es elaborar un documento basado en la evidencia científica que permita favorecer una mejor evaluación y manejo clínico de las autolesiones en la adolescencia.
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