Hipótesis alternativas sobre los beneficios de los fermentados sobre la microbiota intestinal

Cartas al editorRevisión por paresRevisón por pare

FAST, a method based on split-GFP for the detection in solution of proteins synthesized in cell-free expression systems

ackground: Depression and anxiety are two of the most prevalent and disabling mental disorders worldwide, both in the general population and in outpatient clinical settings. Objective: This study aimed to analyze the psychometric properties of the Patient Health Questionnaire-4 (PHQ-4) based on network analysis metrics. Methods: A total of 911 Paraguayans (23.71% women and 76.29% men; mean age 31.25 years, SD = 10.63), selected by non-probabilistic convenience sampling, participated in the study. Network analysis was used to evaluate the internal structure, reliability, and measurement invariance between men and women. Results: The results revealed that the PHQ-4 is a unidimensional measure through Exploratory Graph Analysis (EGA). Reliability, through structural consistency, identified that 100% of the time, only a single dimension was obtained, and all items remained stable, as they were always replicated within the empirical dimension. The unidimensional structure has shown evidence of configural invariance; therefore, the network structure functioned equally among the different sex groups. Conclusion: The PHQ-4 presented optimal preliminary evidence of validity based on its internal structure, reliability, and invariance between sexes. Therefore, it may be useful as an accurate and brief measure of anxiety and depressive symptoms in the Paraguayan context.Horizon 2020 Framework Programm

An in vitro protocol to assay mRNA translation inhibitors using the fluorescent assembly of the split-GFP

Here, we present an in vitro protocol to assay mRNA translation inhibitors using the fluorescent assembly of split-GFP for translation test (FAST), based on the small fragment GFP11 binding to GFP1-10fast. We detail the expression and purification of the GFP1-10fast protein, DNA template amplification, in vitro GFP11-tagged CspA synthesis, FAST detection of the GFP11-tagged protein, and optional recovery of the fluorescent complex. In vitro synthesis of GFP11 maximizes the molar yield of synthesized proteins, providing enhanced sensitivity to test translation inhibitors. For complete details on the use and execution of this protocol, please refer to Pham et al.

Новые возможности в изучении этапа инициации трансляции методами микротермофореза и дифференциальной сканирующей флуориметрии

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The Cryo-EM Structure of a Complete 30S Translation Initiation Complex from Escherichia coli

Formation of the 30S initiation complex (30S IC) is an important checkpoint in regulation of gene expression. The selection of mRNA, correct start codon, and the initiator fMet-tRNAfMet requires the presence of three initiation factors (IF1, IF2, IF3) of which IF3 and IF1 control the fidelity of the process, while IF2 recruits fMet-tRNAfMet. Here we present a cryo-EM reconstruction of the complete 30S IC, containing mRNA, fMet-tRNAfMet, IF1, IF2, and IF3. In the 30S IC, IF2 contacts IF1, the 30S subunit shoulder, and the CCA end of fMet-tRNAfMet, which occupies a novel P/I position (P/I1). The N-terminal domain of IF3 contacts the tRNA, whereas the C-terminal domain is bound to the platform of the 30S subunit. Binding of initiation factors and fMet-tRNAfMet induces a rotation of the head relative to the body of the 30S subunit, which is likely to prevail through 50S subunit joining until GTP hydrolysis and dissociation of IF2 take place. The structure provides insights into the mechanism of mRNA selection during translation initiation

Mechanistics of the binding, function and\ud dissociation of ribosomal ligands during translation\ud initiation analysed by fast kinetics\ud

Initiation is the most regulated and rate limiting step of mRNA translation. In prokaryotes translation begins with an initial interaction between 30S ribosomal subunit and initiation factors (IF1, IF2, IF3), fMet-tRNAfMet and mRNA to eventually yield a 30S initiation complex (30S IC) which subsequently interacts with the 50S subunit to produce a 70S initiation complex (70S IC). Although biochemical and\ud structural properties of most of these interactions have been extensively characterized in the past, there are fundamental aspects of the kinetics of initiation\ud factors association to and dissociation from the ribosomal subunits which have not been extensively and/or directly studied and which therefore remain somewhat\ud controversial. In the present work, I present a direct study of the kinetics of binding and release of IF1 and IF3 using florescent reporter groups on each ribosomal ligand\ud involved in translation initiation (Chapter 1).\ud The functional implications and properties of the nucleotide binding domain of IF2 have been investigated. It is shown that translation initiation may be regulated by\ud switching the IF2 conformation from an active GTP-bound form to an inactive ppGppbound form. Thus, the guanine nucleotide binding domain confers upon IF2 the\ud properties of a metabolic sensor and might explain its evolutionary conservation (Chapter 2).\ud \ud \ud \u

Smartphones encuentran moleculas inteligentes: eBioPhy lleva diagnóstico rápido a zonas remotas

eBioPhy is a diagnostic platform that uses biochemical and biophysical principles together with informatic and communication tools to probe the presence of pathogens in biological samples. The platform aims to bring real-time diagnostics to remote locations where health services are rare and it is based in two main principles: 1) The recognition of pathogens using fluorescently and chemically modified molecules, smart molecules. 2) Data collection and analysis using smartphone capabilities.Proyecto financiado por el FINCyT y el Gobierno de Canadá (Grand Challenges Canada, Bold Idea with Bog impact). Se inicia inica en Octubre 2014 y finaliza en Abril 2016. La instituciones particpantes son Universidad Peruana de Ciencias Aplicadas - UPC y el Instituto de Investigación Nutricional - IIN. Lima, Per

Herramientas Bioinformáticas - BL11 - 202301

DESCRIPCIÓN: El curso Herramientas Bioinformáticas presenta al estudiante los conceptos básicos de bioinformática y los instruye en el uso de herramientas computacionales que les permitirán analizar diferentes tipos de datos biológicos. El estudiante aprenderá a usar diferentes herramientas en línea para la búsqueda y análisis de información relacionada a las diferentes áreas de la biología. El manejo de estas tecnologías de la información facilitará el proceso de aprendizaje del estudiante. PROPÓSITO: Los conocimientos entregados en este curso pretenden capacitar al estudiante en el uso de diferentes herramientas informáticas disponibles en línea para el manejo y análisis de datos biológicos. Se revisarán bases de datos biológicas como NCBI y el Protein Data Bank, centrándose en el correcto uso de los sistemas de búsqueda de genes y proteínas. Además, se conocerán y usarán diferentes tipos de programas de alineamiento, análisis de secuencias de ADN, ARN y proteínas, y de relaciones filogenéticas. El curso proporcionará los conocimientos básicos para el diseño y análisis de secuencias de ADN, clonación molecular, mutagénesis, entre otros. Se conocerán también herramientas de visualización tridimensional de moléculas. El curso contribuye con el desarrollo de la competencia general pensamiento innovador y la competencia especifica investigación, ambas de logro 1