Escherichia coli, un petit gran organisme

Els processos essencials que tenen lloc dins les cèl·lules, tant fisiològics com genètics o bioquímics, estan altament conservats entre tots els éssers vius. Els bacteris són microorganismes d'estructura cel·lular senzilla en comparació de les cèl·lules dels organismes superiors. A més, molts bacteris es divideixen molt ràpidament, són fàcilment manipulables i es poden fer créixer en una àmplia varietat de medis de cultiu. Tot això ha fet que els bacteris s'hagin fet servir molt extensament com a model biològic per respondre qüestions sobre processos bàsics de la biologia que són comuns a tots els éssers vius. Per exemple, l'estat actual del coneixement biològic, el desenvolupament de la biologia molecular i moltes de les aplicacions biotecnològiques derivades són resultat, en gran mesura, de l'estudi en bacteriologia. Entre tots els bacteris utilitzats en laboratoris de recerca Escherichia coli ha estat, sens dubte, el més utilitzat, i ha donat lloc a molts dels coneixements de què disposem avui dia sobre el funcionament, l'estructura i l'evolució dels éssers vius. En aquest capítol fem una visió sobre l'ús passat, present i futur com a model biològic dels bacteris en general i d'E. coli en particular.Escherichia coli, a little big organism. The basic physiological, biochemical and genetic processes that take place inside a cell are highly conserved among the different organisms. When compared with high organisms, bacteria are microorganisms that have a simple cell structure. Moreover, many bacteria can divide very quickly, are easy to handle and can grow in a broad variety of culture media. All these properties promoted that bacteria were used as model organisms to answer relevant biological questions common in all living beings. Present knowledge in biology, development of molecular biology and many biotechnological applications are mainly due to bacteriological studies. Among all, there is one bacterium that has been very extensively used in biological studies, Escherichia coli. Most of knowledge we have nowadays on the structure, physiology and evolution of the living creatures is based in data from E. coli based studies. In this chapter, an overview on the pass, present and future use of bacteria, and more specifically of E. coli, as a model organism is done

Pulsed light inactivation of mushroom polyphenol oxidase: a fluorometric and spectrophotometric study

Polyphenol oxidase (PPO) is one of the most important food enzymes, it is responsible for the browning of many foods. Pulsed light (PL) is a non-thermal method of food preservation that is able to inactivate PPO. The aim of this work was to gain insight into the mechanism of PPO inactivation by PL. To this, the kinetics of PPO inactivation by PL was measured, together with associated changes in tryptophan fluorescence, KI fluorescence quenching and turbidity; and results were analysed by parameter A and phase diagram methods. Enzyme inactivation followed the Weibull model. Tryptophan fluorescence decreased during PL treatment, as well as the parameter A, while Stern-Volmer constants increased and turbidity was constant. The phase diagram showed only two populated states. There was a high correlation between the loss of activity and parameter A. Results indicate that under the experimental conditions, the inactivation of PPO by PL is an all-or-none process where the enzyme progressively unfolds with no evidence of aggregation.Fundación Universitaria San Antonio de CartagenaCiencias de la Alimentació

QTL mapping in population originated from a bi-parental commercial cross of sugarcane

Orientadores: Anete Pereira de Souza, Monalisa Sampaio CarneiroTese (doutorado) - Universidade Estadual de Campinas, Instituto de BiologiaResumo: A cana-de-açúcar é uma fonte renovável de energia e com potencial para expansão. A complexidade genética da cana-de-açúcar decorrente de seu alto nível de ploidia e aneuploidia, aliada à natureza quantitativa da maioria dos caracteres agronômicos tem dificultado, atualmente, a obtenção de elevados índices de ganho genético através do melhoramento convencional. O desenvolvimento de marcadores moleculares e a construção de mapas genéticos podem auxiliar na elaboração de estratégias a serem introduzidas nos programas de melhoramento de forma a aumentar a eficiência dos processos de seleção e acelerar o desenvolvimento de novas cultivares. Desta forma, a proposta desta tese foi a construção de um mapa genético em cana-de-açúcar visando a identificação de regiões genômicas que controlam características de interesse através do mapeamento de QTL (Quantitative Trait Loci). Uma população de cana-de-açúcar com 153 indivíduos oriundos do cruzamento comercial bi-parental entre as cultivares SP80-3280 e RB835486 foi utilizada para alcançar os objetivos citados. O experimento de campo foi instalado em duas localidades, Araras-SP e Ipaussu-SP, usando o delineamento de blocos aumentados incompletos com 3 repetições. As avaliações fenotípicas foram realizadas ao longo de três anos (2011, 2012 e 2013). Empregou-se a abordagem de modelos mistos para análise das características fenotípicas relacionadas com componentes de produção e resistência à ferrugem marrom. Os dados de severidade à ferrugem marrom foram analisados, como uma primeira abordagem, via modelo misto linear generalizado. As estimativas de herdabilidade das características fenotípicas foram altas, variando de 0.78 (altura de colmos) a 0.92 (diâmetro de colmos), e a análise de severidade à ferrugem marrom mostrou que 66% dos clones possuem, no mínimo, 90% de probabilidade de serem resistentes à doença. Para construção do mapa genético integrado foram utilizados marcadores moleculares do tipo microssatélites (Simple Sequence Repeats, SSR), TRAP (Target Target Region Amplification Polymorphism), além de SNPs (Single Nucleotide Polymorphims) e indels (inserções e deleções) oriundos da técnica de GBS (Genotyping-by-Sequencing). Para descoberta de marcadores baseados em GBS foram utilizadas quatro pseudo-referências: genoma de sorgo (Sorghum bicolor), genoma metil-filtrado da cana-de-açúcar, transcriptoma da cana-de-açúcar (RNAseq) e sequências do projeto SUCEST. A ploidia e dosagem de cada loco bi-alélico foi estimada através do software SUPERMASSA. Utilizando o software Onemap (v. 2.0-4) e empregando-se LOD > 9.0 e fração de recombinação 9.0 and recombination fraction <0.10, a total of 993 markers in single dose were mapped. These markers were distributed throughout 223 linkage groups, which were clustered in 18 homo(eo)logous groups. The total length of the map was 3,682.04 cM with an average marker density of 3.70 cM. Using composite interval mapping (Composite Interval Mapping, CIM) were mapped seven QTLs considering four of the 11 phenotypic traits evaluated. The results suggest the presence of a stable QTL across locations to soluble solid content (BRIX) and sucrose content of the cane (POL%C). Furthermore, QTLs to BRIX and fiber content (FIB) traits had associated markers with candidate genes, which had great potential for validation and future use for molecular breeding in sugarcane. This study is the first to report the use of GBS for large-scale variant discovery and genotyping of a population in sugarcane with posterior analysis to composite interval mappingDoutoradoGenetica Vegetal e MelhoramentoDoutor em Genetica e Biologia Molecular2010/50091-4 , 2008/52197-4FAPES

Use of machine-learning and load–velocity profiling to estimate 1-Repetition maximums for two variations of the bench-press exercise

The purpose of the current study was to compare the ability of five different methods to estimate eccentric–concentric and concentric-only bench-press 1RM from load–velocity profile data. Smith machine bench-press tests were performed in an eccentric–concentric (n= 192) and a concentric-only manner (n= 176) while mean concentric velocity was registered using a linear position transducer. Load–velocity profiles were derived from incremental submaximal load (40–80% 1RM) tests. Five different methods were used to calculate 1RM using the slope, intercept, and velocity at 1RM (minimum velocity threshold—MVT) from the load–velocity profiles: calculation with individual MVT, calculation with group average MVT, multilinear regression without MVT, regularized regression without MVT, and an artificial neural network without MVT. Mean average errors for all methods ranged from 2.7 to 3.3 kg. Calculations with individual or group MVT resulted in significant overprediction of eccentric–concentric 1RM (individual MVT: difference = 0.76 kg, p= 0.020, d= 0.17; group MVT: difference = 0.72 kg, p= 0.023, d= 0.17). The multilinear and regularized regression both resulted in the lowest errors and highest correlations. The results demonstrated that bench-press 1RM can be accurately estimated from load–velocity data derived from submaximal loads and without MVT. In addition, results showed that multilinear regression can be used to estimate bench-press 1RM. Collectively, the findings and resulting equations should be helpful for strength and conditioning coaches as they would help estimating 1RM without MVT data