A new strategy for RNA isolation from eukaryotic cells using arginine affinity chromatography

The relevance of RNA in many biological functions has been recognized, broadening the scope of RNA research activities, from basic to applied sciences, also aiming the translation to clinical fields. The preparation and purification of RNA is a critical step for further application, since the quality of the template is crucial to ensure reproducibility and biological relevance. Therefore, the establishment of new tools that allows the isolation of pure RNA with high quality is of particular importance. New chromatographic strategies for RNA purification were considered, exploiting affinity interactions between amino acids and nucleic acids. In the present study, a single arginine-affinity chromatography step was employed for the purification of RNA from a total eukaryotic nucleic acid extract, thus eliminating several steps compared with current RNA isolation procedures. The application of this process resulted in a high RNA recovery yield of 96 ± 17% and the quality control analysis revealed a high integrity (28S:18S ratio = 1.96) in RNA preparations as well as a good purity, demonstrated by the scarce detection of proteins and the reduction on genomic DNA contamination to residual concentrations. Furthermore, the performance of the new RNA isolation method was tested regarding the applicability of the isolated RNA in modern molecular biology techniques. Hence, this new affinity approach will simplify the isolation and purification of RNA, which can bring great improvements in biomedical investigation.info:eu-repo/semantics/publishedVersio

Analysis of Mercury Grouping in Soil in the Foz of the Tapajós River, Amazonia-Brazil

The region of Santarém, in the state of Pará, was chosen to carry out this research because it is at the center of discussions of mercury contamination in the Amazon. The objective of this paper is the analysis of Hg and Methyl-Hg speciation in soil. The methodological procedures to determine the total mercury and methylHg were carried out by means of samples, which were submitted to acid digestion and determinations of Hg performed by Atomic Absorption Spectrophotometry with cold vapor generation. The evaluation of the concentrations of mercury in the sampled and depth points was performed with the aid of cluster analysis, using the Hierarchical clustering method. The results obtained indicate that the total mercury in soil showed very high values in the analysis of total mercury with granulometry for fine fraction and gross fraction, indicating an urgent mobilization and political action in the municipal, state and federal spheres regarding the exposure of the environment environment to the mercury detected

Adjusting the Capital Index (ICS), to the Population of Santarém-Pa, It’s Capacity for Mobilization and Political Action the Environmental Exposure to Mercury

Until the mid-1980s, the scientific literature focused on the environmental issue, particularly in the Brazilian Amazon, attributed the mercurial contamination, the anthropic action, especially the mining activities. The region of Santarém, in the state of Pará, was chosen to conduct this research because it is at the center of the discursions of mercury contamination in the Amazon. The objective of this study is to analyze the perception of risk to mercury contamination in the population of Santarém, with the application of Integrated Questionnaires to Measure the Social Capital Index - QI-MCS proposed by the World Bank. The methodological procedures were based on the application of structured questionnaires in open and closed questions, through individual interviews, which addressed several aspects related to groups and networks; trust and solidarity; collective action and cooperation; information and communication; cohesion and social inclusion; authority or capacity building and political action; mercury contamination. In order to determine the ICS, variables that express interpersonal relationships among community members were considered as well as their participation in participatory and cooperative associations. It is concluded that the risk is underestimated by the local population, since it does not have enough information regarding the mercurial contamination and actions taken by the State to prevent this type of accident

Advances in Membrane-Bound Catechol-O-Methyltransferase Stability Achieved Using a New Ionic Liquid-Based Storage Formulation

Funding: The authors acknowledge the support from FEDER funds through the POCI—COMPETE 2020—Operational Program Competitiveness and Internationalization in Axis I—Strengthening research, technological development, and innovation (Project POCI-01-0145-FEDER-007491) and National Funds (Project UID/Multi/00709/2013) LA/P/0006/2020, financed by national funds through the Portuguese Foundation for Science and Technology/MCTES (PIDDAC). This work was also supported by the Applied Molecular Biosciences Unit UCIBIO (UIDB/04378/2020 and UIDP/04378/2020) and the Associate Laboratory Institute for Health and Bioeconomy—i4HB (project LA/P/0140/2020) which are financed by National Funds from FCT/MCTES. Researchers also acknowledge funding by FEDER through COMPETE 2020—Programa Operacional Compet-itividade e Internacionalização (POCI), Augusto Q. Pedro research contract CEEC-IND/02599/2020 under the Scientific Stimulus Publisher Copyright: © 2022 by the authors. Licensee MDPI, Basel, Switzerland.Membrane-bound catechol-O-methyltransferase (MBCOMT), present in the brain and involved in the main pathway of the catechol neurotransmitter deactivation, is linked to several types of human dementia, which are relevant pharmacological targets for new potent and nontoxic inhibitors that have been developed, particularly for Parkinson’s disease treatment. However, the inexistence of an MBCOMT 3D-structure presents a blockage in new drugs’ design and clinical studies due to its instability. The enzyme has a clear tendency to lose its biological activity in a short period of time. To avoid the enzyme sequestering into a non-native state during the downstream processing, a multi-component buffer plays a major role, with the addition of additives such as cysteine, glycerol, and trehalose showing promising results towards minimizing hMBCOMT damage and enhancing its stability. In addition, ionic liquids, due to their virtually unlimited choices for cation/anion paring, are potential protein stabilizers for the process and storage buffers. Screening experiments were designed to evaluate the effect of distinct cation/anion ILs interaction in hMBCOMT enzymatic activity. The ionic liquids: choline glutamate [Ch][Glu], choline dihydrogen phosphate ([Ch][DHP]), choline chloride ([Ch]Cl), 1-dodecyl-3-methylimidazolium chloride ([C12mim]Cl), and 1-butyl-3-methylimidazolium chloride ([C4mim]Cl) were supplemented to hMBCOMT lysates in a concentration from 5 to 500 mM. A major potential stabilizing effect was obtained using [Ch][DHP] (10 and 50 mM). From the DoE 146% of hMBCOMT activity recovery was obtained with [Ch][DHP] optimal conditions (7.5 mM) at −80◦ C during 32.4 h. These results are of crucial importance for further drug development once the enzyme can be stabilized for longer periods of time.publishersversionpublishe

Aroma precursors of grapes: contribution of variety and vineyard traning system to port wine aroma

In order to evaluate the influence of the vine training system in the composition in terpenes, norisoprenoids and their precursors (carotenoids and glycosidic molecules), these compounds were quantified in grapes and Port wines belonging to two varieties: Touriga Nacional and Touriga Franca from Douro Region. The double cordon LYS 2/3 system was compared with the traditional method, the vertical shoot positioning VSP system. Touriga Nacional grapes and wines have higher contents in carotenoids, free and glycosylated fractions of terpenic and norisoprenoids compounds than Touriga Franca. In general, LYS 2/3 training system produced grapes and wines with high content in glycosylated fractions of terpenes and nor-isoprenoids, being more appropriate to the production of Port wines, which are submitted to an aging process

Características físico-químicas, microbiológicas e aceitabilidade de iogurte de leite de búfala para merenda escolar.

bitstream/item/63158/1/Oriental-Doc109.pd

Manejo de mosca branca Bemisia argentifolii bellows & perring no algodoeiro.

bitstream/CNPA-2009-09/22585/1/CIRTEC40.pd

Arquitetura genômica de genes que flanqueiam o regulador LaeA em Trichoderma spp.

Ferramentas de bioinformática e sequenciamento de genoma cada vez mais eficientes e com menor custo tem contribuído com a identificação de um vasto número de grupos de genes relacionados a síntese metabólica, os BGCs. A identificação de compostos bioativos crípticos por meio da estratégia de mineração não é suficiente para obtenção de metabolitos secundários de interesse, uma vez que, fatores epigenéticos podem influenciar na expressão de genes crípticos. O regulador LaeA é considerado um fator importante para desbloquear a expressão de metabólitos crípticos, já constatado em vários fungos. Este trabalho teve como objetivo realizar análise de sintenia das sequências dos genes que flanqueiam o LaeA em 16 espécies de Trichoderma e analisar a correlação filogenética dos isolados, com intuito de verificar a conservação gênica da região