Resazurin reduction-based assays revisited: guidelines for accurate reporting of relative differences on metabolic status

© 2023 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).Cell viability and metabolic activity are ubiquitous parameters used in biochemistry, molecular biology, and biotechnological studies. Virtually all toxicology and pharmacological projects include at some point the evaluation of cell viability and/or metabolic activity. Among the methods used to address cell metabolic activity, resazurin reduction is probably the most common. At variance with resazurin, resorufin is intrinsically fluorescent, which simplifies its detection. Resazurin conversion to resorufin in the presence of cells is used as a reporter of metabolic activity of cells and can be detected by a simple fluorometric assay. UV–Vis absorbance is an alternative technique but is not as sensitive. In contrast to its wide empirical “black box” use, the chemical and cell biology fundamentals of the resazurin assay are underexplored. Resorufin is further converted to other species, which jeopardizes the linearity of the assays, and the interference of extracellular processes has to be accounted for when quantitative bioassays are aimed at. In this work, we revisit the fundamentals of metabolic activity assays based on the reduction of resazurin. Deviation to linearity both in calibration and kinetics, as well as the existence of competing reactions for resazurin and resorufin and their impact on the outcome of the assay, are addressed. In brief, fluorometric ratio assays using low resazurin concentrations obtained from data collected at short time intervals are proposed to ensure reliable conclusions.The project leading to these results has received funding from “la Caixa” Foundation and FCT, I.P. under the project code [LCF/PR/HR21/00605], BREAST-BRAIN-N-BBB.info:eu-repo/semantics/publishedVersio

Avaliação preliminar de populações de cenoura para reação à mistura populacional de Meloidogyne incognita raça 1 e Meloidogyne javanica.

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Rendimento industrial de cenourete® em função do tamanho da matéria-prima e do tempo de processamento.

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Induction of systemic resistance in tomato by the autochthonous phylloplane resident Bacillus cereus.

The objective of this work was to verify if the induced resistance mechanism is responsible for the capacity of a phylloplane resident bacteria (Bacillus cereus), isolated from healthy tomato plants, to control several diseases of this crop. A strain of Pseudomonas syringae pv. tomato was used as the challenging pathogen. The absence of direct antibiosis of the antagonist against the pathogen, the significant increase in peroxidases activity in tomato plants exposed to the antagonist and then inoculated with the challenging pathogen, as well as the character of the protection, are evidences wich suggest that biocontrol efficiency presented by the antagonist in previous works might be due to induced systemic resistance (ISR)

Herdabilidade e resposta à seleção para peso de raízes e tolerância a queima-das-folhas em populações de cenoura.

Objetivou-se neste trabalho, estimar os parâmetros genéticos e a resposta à seleção em três populações de cenoura (CNPH-561, CNPH-586 e CNPH-587) com diferentes propourções de germoplasma de clima temperado e tropical.bitstream/item/72167/1/bpd-811.pd