Expression profiling and sequence diversity of novel DREB genes from common bean (Phaseolus vulgaris L.) and their association with drought-related traits

Common bean is a major dietary component in several countries, but its productivity is negatively affected by abiotic stresses. Dissecting candidate genes involved in abiotic stress tolerance is a paramount step toward the improvement of common bean performance under such constraints. Thereby, this thesis presents a systematic analysis of the DEHYDRATION RESPONSIVE ELEMENT-BINDING (DREB) gene subfamily, which encompasses genes that regulate several processes during stress responses, but with limited information for common bean. First, a series of in silico analyses with sequences retrieved from the P. vulgaris genome on Phytozome supported the categorization of 54 putative PvDREB genes distributed within six phylogenetic subgroups (A-1 to A-6), along the 11 chromosomes. Second, we cloned four novel PvDREB genes and determined their inducibility-factors, including the dehydration-, salinity- and cold-inducible genes PvDREB1F and PvDREB5A, and the dehydration- and cold-inducible genes PvDREB2A and PvDREB6B. Afterwards, nucleotide polymorphisms were searched through Sanger sequencing along those genes, revealing a high number of single nucleotide polymorphisms within PvDREB6B by the comparison of Mesoamerican and Andean genotypes. The nomenclature of PvDREB6B is discussed in details. Furthermore, we used the BARCBean6K_3 SNP platform to identify and genotype the closest SNP to each one of the 54 PvDREB genes. We selected PvDREB6B for a broader study encompassing a collection of wild common bean accessions of Mesoamerican origin. The population structure of the wild beans was accessed using sequence polymorphisms of PvDREB6B. The genetic clusters were partially associated with variation in latitude, altitude, precipitation and temperature throughout the areas such beans are distributed. With an emphasis on drought stress, an adapted tube-screening method in greenhouse conditions enabled the phenotyping of several drought-related traits in the wild collection. Interestingly, our data revealed a correlation between root depth, plant height and biomass and the environmental data of the location of the accessions. Correlation was also observed between the population structure determined through PvDREB6B and the environmental data. An association study combining data from the SNP array and DREB polymorphisms enabled the detection of SNP associated with drought-related traits through a compressed mixed linear model (CMLM) analysis. This thesis highlighted important features of DREB genes in common bean, revealing candidates for further strategies aimed at improvement of abiotic stress tolerance, with emphasis on drought tolerance

Evaluación de la ingesta de líquido, pérdida de peso y tasa de sudoración en jóvenes triatletas

Introducción: El triatlón es un deporte de resistencia que comprende tres disciplinas: natación, ciclismo y carrera a pie. Es necesario establecer pautas de hidratación para prevenir deshidrataciones durante entrenamientos o competiciones y mantener un buen estado de hidratación antes, durante y después del ejercicio. El objetivo de este estudio es evaluar la ingesta de líquido, pérdida de peso y tasa de sudoración en jóvenes triatletas, durante diferentes entrenamientos. Material y Métodos: Estudio descriptivo-observacional en 14 triatletas (7 chicos y 7 chicas) durante una sesión de natación, otra de ciclismo y otra de carrera a pie. Se valoró la ingesta de líquido, pérdida de peso, % agua corporal total, % deshidratación y tasa de sudoración. Los triatletas bebieron agua en sus respectivos bidones de 750 ml y se realizó una medición de orina en containers. Resultados: Los resultados del estudio siguiendo el orden de natación, ciclismo y carrera a pie fueron: ingesta agua 2,66±1,94ml/min, 7,91±7,69ml/min y 7,08±4,13ml/min en chicos y 3,43±1,53ml/min, 6,39±5,36ml/min y 8,33±2,74ml/min en chicas; pérdida de peso 0,83±0,5kg, 0,47±0,3kg y 0,98±0,4kg en chicos y 0,79±0,3kg, 0,47±0,58kg y 0,28±0,21kg en chicas; y tasa sudoración 4,44±4,9ml/min, 11,81±6,46ml/min y 5,29±3,13ml/min en chicos y 3,89±2,4ml/min, 4,69±4,20ml/min y 7,96±5,06ml/min en chicas. Conclusiones: Se comparó el porcentaje de agua corporal y deshidratación, la pérdida de peso y la tasa de sudoración con otros estudios y se observa que nuestros resultados son inferiores a los estudios comparados, además están por debajo de la media de recomendaciones de líquido establecidas por el consenso de hidratación.

Conservação de cereja com aplicação de revestimento comestível

Dissertação apresentada à Escola Superior Agrária do Instituto Politécnico de Castelo Branco para cumprimento dos requisitos necessários à obtenção do grau de Mestre em Fruticultura Integrada.

A abordagem médica de seguimento das crianças com síndrome do intestino curto

Trabalho Final do Curso de Mestrado Integrado em Medicina, Faculdade de Medicina, Universidade de Lisboa, 2014

Conhecimento e atitudes dos pais perante a criança com febre em contexto de serviço de urgência

Enquadramento – A febre na criança constitui uma das principais manifestações que impul-siona os pais na procura dos cuidados de saúde diferenciados, resultando na utilização ina-propriada de recursos. Objetivos – Identificar as variáveis sociodemográficas que interferem no conhecimento e nas atitudes dos pais perante a criança com febre; avaliar a influência do número de filhos no conhecimento e nas atitudes; identificar as fontes de informação que influenciam o conheci-mento e as atitudes dos pais. Material e Método – Estudo quantitativo, transversal, descritivo e correlacional. Recorreu-se ao questionário que inclui a caracterização sociodemográfica, contextual da criança, variá-veis clínicas de fontes de informação e questões que avaliam o conhecimento e as atitudes dos pais perante a febre na criança. A amostragem é não probabilística por conveniência (144 pais/cuidadores). Resultados – Amostra predominantemente feminina (86.8%), os homens são mais velhos (M=34.05 anos .6.45 anos); cerca de 9 em cada 10 das crianças eram acompanhadas pela mãe. Os sintomas que mais preocupam e levaram os participantes a recorrer ao Serviço de Urgência foram: vómitos/desidratação (66.4%), mau estado geral (64.6%), dificuldade respirató-ria (52.1%); em 58.5% dos casos de febre sem foco poder-se-ia ter utilizado os cuidados de saúde primários; a idade da criança e o sexo dummy revelaram-se preditoras dos conhecimen-tos e o sexo preditor das atitudes; as mulheres e os homens com mais idade e com filhos mais jovens possuem melhores conhecimentos; as mulheres têm atitudes mais adequadas. Conclusões – As mulheres possuem conhecimentos intermédios e revelam melhor atitude perante a febre. Quanto maior a escolaridade melhor é o conhecimento e os participantes que procuram como fonte de informação um profissional de saúde, possuem atitudes mais ade-quadas. Assume grande relevância a aquisição de competências que proporcionem aos pais/cuidadores ferramentas que os ajudem a lidar com a fobia da febre. Palavras-chave: Febre; Criança; Pais; Conhecimento; Atitudes; Serviço de Urgência.

Disfagia no doente com AVC : prevalência e determinantes

Introdução: O acidente vascular cerebral (AVC) assume em Portugal elevadas taxas de morbilidade e reinternamento hospitalar. A disfagia surge como uma complicação frequente deste evento neurológico, com índices de morbilidade elevados pelo risco de desnutrição, desidratação e aspiração broncopulmonar. O diagnóstico e a sua monitorização no processo de reabilitação do doente são ações fundamentais na prevenção de aspirações alimentares, redução do internamento hospitalar e na eficácia da reabilitação do doente. Objetivo: Identificar e avaliar o grau de disfagia na pessoa com AVC e analisar a relação entre esta, e as variáveis socio-demográficas e clínicas no sentido de poder melhorar futuramente os cuidados de enfermagem de reabilitação. Métodos: Trata-se de um estudo não experimental, transversal, descritivo-correlacional de caráter quantitativo, que foi realizado numa amostra não probabilística por conveniência, constituída por 25 doentes com diagnóstico de AVC, internados na Rede Nacional Cuidados Continuados Integrados (RNCCI), em unidades de Convalescença e Reabilitação. O instrumento de colheita de dados integra uma seção de caracterização sócio-demográfica e clínica e duas escalas: Escala Gugging Swallowing Screen (GUSS) e Índice de Barthel, a fim de avaliar a disfagia e a funcionalidade, respetivamente. Resultados: A amostra apresenta uma média de idade de 76,8 anos, sendo 68% do sexo feminino e 32% do sexo masculino. Verificámos que 68% dos participantes apresenta mais de dois antecedentes clínicos e apenas 24% dos participantes não apresenta disfagia. Dos restantes, 12% apresenta disfagia grave, 36% moderada e 28% disfagia ligeira. A área de lesão parece influenciar a deglutição, demonstrando a Artéria Cerebral Média (ACM) e Artéria Cerebral Posterior (ACP) como áreas de maior sensibilidade. Denotou-se que quanto maior o grau de dependência, maior gravidade de disfagia. Conclusão: Doentes com AVC isquémico apresentam disfagia, com gravidade relacionada com a área vascular. A existência de vários antecedentes clínicos pode gerar perturbações na deglutição do doente. De igual modo, quanto maior for a dependência funcional do doente, maior é o grau de disfagia e o risco de aspiração pulmonar. Palavras-chave: AVC; Disfagia; Reabilitação.

LeProT1, a Transporter for Proline, Glycine Betaine, and γ-Amino Butyric Acid in Tomato Pollen

During maturation, pollen undergoes a period of dehydration accompanied by the accumulation of compatible solutes.Solute import across the pollen plasma membrane, which occurs via proteinaceous transporters, is required to support pollen development and also for subsequent germination and pollen tube growth. Analysis of the free amino acid composition of various tissues in tomato revealed that the proline content in flowers was 60 times higher than in any other organ analyzed. Within the floral organs, proline was confined predominantly to pollen, where it represented >70 of total free amino acids. Uptake experiments demonstrated that mature as well as germinated pollen rapidly take up proline. To identify proline transporters in tomato pollen, we isolated genes homologous to Arabidopsis proline transporters. LeProT1 was specifically expressed both in mature and germinating pollen, as demonstrated by RNA in situ hybridization. Expression in a yeast mutant demonstrated that LeProT1 transports proline and γ-amino butyric acid with low affinity and glycine betaine with high affinity. Direct uptake and competition studies demonstrate that LeProT1 constitutes a general transporter for compatible solutes.

Sediment and organic matter properties, and molecular formula from sampling locations in the Northern Black Sea

Remineralization of organic matter in reactive marine sediments releases nutrients and dissolved organic matter (DOM) into the ocean. Here we focused on the molecular-level characterization of DOM by high-resolution Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR-MS) in sediment pore waters and bottom waters from contrasting redox regimes in the northern Black Sea with particular emphasis on nitrogen-bearing compounds to derive an improved understanding of the molecular transformations involved in nitrogen release. The number of nitrogen-bearing molecules is generally higher in pore waters than in bottom waters. This suggests intensified degradation of nitrogen-bearing precursor molecules such as proteins in anoxic sediments: No significant difference was observed between sediments deposited under oxic vs anoxic conditions (average O/C ratios of 0.55) suggesting that the different organic matter quality induced by contrasting redox conditions does not impact protein diagenesis in the subseafloor. Compounds in the pore waters were on average larger, less oxygenated, and had a higher number of unsaturations. Applying a mathematical model, we could show that the assemblages of nitrogen-bearing molecular formulas are potential products of proteinaceous material that was transformed by the following reactions: (a) hydrolysis and deamination, both reducing the molecular size and nitrogen content of the products and intermediates; (b) oxidation and hydration of the intermediates; and (c) methylation and dehydration.

Lithium isotopic composition of pore fluids and sediments in the Costa Rica subduction zone

Pore fluid and sediment Li concentrations and isotopic ratios provide important insights on the hydrology, sediment contribution to the arc volcanoes and fluid-sediment reactions at the dominantly non-accretionary Costa Rica subduction zone. Ocean Drilling Program Site 1039 in the trench axis provides a reference section of 400 m of the incoming sediments, and Site 1040, situated arcward from the trench, consists of a deformed sedimentary wedge and apron sediments, the décollement, and the partially dewatered underthrust sediment section. At the reference site, pore fluids show important isotopic variations (delta6Li=-21.7 to -37.8 per mil), reflecting the interplay of in situ alteration of volcanic material and ion exchange with clay minerals. In the basal section, a reversal of Li concentration and delta6Li toward seawater values is observed, providing supporting evidence for a lateral seawater flow system in the upper oceanic basement underlying this sediment section. At Site 1040, pore fluid of the lower deformed wedge sediments and within the décollement is enriched in Li and the isotopic compositions are relatively light, suggesting infiltration of a deep-seated fluid. The delta6Li value of -22 per mil of this Li-enriched fluid (261 µM), when compared with the delta6Li value of the subducted sediment section (-11 per mil), suggests that the deep source fluid originates from mineral fluid dehydration and transformation reactions at temperatures of 100 to 150°C, consistent with the temperature range of the up-dip seismogenic zone and of transformation of smectite to illite. The distribution of Li and its isotopes in the underthrust section are similar to those at the reference site, indicating near complete subduction of the incoming sediments and that early dewatering of the underthrust sediments occurs predominantly by lateral flow into the ocean. The hemipelagic clay-rich sediment section of the subducting plate carries most of the Li into this subduction zone, and the pelagic diatomaceous and nannofossil calcareous oozes contain little Li. The Li isotopes of both the clay-rich hemipelagic sediments and of the pelagic oozes are, however, similar, with delta6Li values of -9 to -12 per mil. The observations that (1) the delta6Li values of the underthrust sediments are distinctly lower than that of the mantle, and (2) the lavas of the Costa Rican volcanoes are enriched in Li and 7Li, provide an approximation of the contribution of the subducted sediments to the arc volcanoes. A first order mass balance calculation suggests that approximately half of the Li flux delivered by subducted sediments and altered oceanic crust into the Middle American Trench is recycled to the Costa Rican arc and at most a quarter of sedimentary Li is returned into the ocean through thrust faults, primarily the décollement thrust.

National training course, emergency medical technician - paramedic. Instructor's lesson plans. Module III: shock and fluid therapy.

Mode of access: Internet.

Using a non-invasive stable isotope tracer to measure the absorption of water in humans

The development of solutions that prevent dehydration or promote adequate re-hydration play a vital role in preventing fatigue during exercise, however, the methods commonly used to assess the hydration ability of such solutions are invasive and often assess the components of absorption separately. This paper describes using a non-invasive deuterium tracer technique that assesses gastric emptying and intestinal absorption simultaneously to evaluate the uptake of water during rest and exercise. The kinetics of absorption are further examined by mathematical modelling of the data generated. For the rest group, 0.05 g/kg of body weight of deuterium, contained in gelatine capsules, was ingested with ordinary tap water and saliva samples were collected every 5 min for one hour while the subject remained seated. The deuterium was administered as above for the exercise group but sample collection was during one hour of exercise on a treadmill at 55% of the subject's maximum heart rate. The enrichment data for each subject were mathematically modelled and the parameters obtained were compared across groups using an independent samples t-test. Compared with the rest condition, the exercise group showed delayed absorption of water as indicated by significant differences for the modelling parameters t(2), t(1/2), maximum absorption rate and solution absorption amount at t(1). Labelling with a deuterium tracer is a good measure of the relative rate ingested fluids are absorbed by the body. Mathematical modelling of the data generates rates of maximum absorption and allows calculation of the percentage of the solution that is absorbed at any given time during the testing period. Copyright (C) 2004 John Wiley Sons, Ltd.

Acute bovine laminitis: A new induction model using alimentary oligofructose overload

Twelve dairy heifers were used to examine the clinical response of an alimentary oligofructose overload. Six animals were divided into 3 subgroups, and each was given a bolus dose of 13, 17, or 21 g/kg of oligofructose orally. The control group (n = 6) was sham-treated with tap water. Signs of lameness, cardiovascular function, and gastrointestinal function were monitored every 6 h during development of rumen acidosis. The heifers were euthanized 48 and 72 h after administration of oligofructose. All animals given oligofructose developed depression, anorexia, and diarrhea 9 to 39 h after receiving oligofructose. By 33 to 45 h after treatment, the feces returned to normal consistency and the heifers began eating again. Animals given oligofructose developed transient fever, severe metabolic acidosis, and moderate dehydration, which were alleviated by supportive therapy. Four of 6 animals given oligofructose displayed clinical signs of laminitis starting 39 to 45 h after receiving oligofructose and lasting until euthanasia. The lameness was obvious, but could easily be overlooked by the untrained eye, because the heifers continued to stand and walk, and did not interrupt their eating behavior. No positive pain reactions or lameness were seen in control animals. Based on these results, we conclude that an alimentary oligofructose overload is able to induce signs of acute laminitis in cattle. This model offers a new method, which can be used in further investigation of the pathogenesis and pathophysiology of bovine laminitis.

Physique traits of lightweight rowers and their relationship to competitive success

Objectives: Physique traits and their relationship to competitive success were assessed amongst lightweight rowers competing at the 2003 Australian Rowing Championships. Methods: Full anthropometric profiles were collected from 107 lightweight rowers (n = 65 males, n = 45 females) competing in the Under 23 and Open age categories. Performance assessments were obtained for 66 of these rowers based on results in the single sculls events. The relationship between physique traits and competitive success was then determined. Results: Lower body fat ( heat time estimate -8.4 s kg(-1), p< 0.01), greater total body mass ( heat time estimate -4.4 s kg(-1), p = 0.03), and muscle mass ( heat time estimate -10.2 s kg(-1), p< 0.01) were associated with faster 2000 m heat times. Conclusions: The more successful lightweight rowers were those who had lower body fat and greater total muscle mass.

Post-crystallization treatments for improving diffraction quality of protein crystals

X-ray crystallography is the most powerful method for determining the three-dimensional structure of biological macromolecules. One of the major obstacles in the process is the production of high-quality crystals for structure determination. All too often, crystals are produced that are of poor quality and are unsuitable for diffraction studies. This review provides a compilation of post-crystallization methods that can convert poorly diffracting crystals into data-quality crystals. Protocols for annealing, dehydration, soaking and cross-linking are outlined and examples of some spectacular changes in crystal quality are provided. The protocols are easily incorporated into the structure-determination pipeline and a practical guide is provided that shows how and when to use the different post-crystallization treatments for improving crystal quality.

Seasonal patterns of growth, dehydrins and water-soluble carbohydrates in genotypes of Dactylis glomerata varying in summer dormancy

Background and Aims Summer dormancy in perennial grasses has been studied inadequately, despite its potential to enhance plant survival and persistence in Mediterranean areas. The aim of the present work was to characterize summer dormancy and dehydration tolerance in two cultivars of Dactylis glomerata (dormant 'Kasbah', non-dormant 'Oasis') and their hybrid using physiological indicators associated with these traits. Methods Dehydration tolerance was assessed in a glasshouse experiment, while seasonal metabolic changes which produce putative protectants for drought, such as carbohydrates and dehydrins that might be associated with summer dormancy, were analysed in the field. Key Results The genotypes differed in their ability to survive increasing soil water deficit: lethal soil water potential (ψ(s)) was -3(.)4 MPa for 'Kasbah' (although non-dormant), -1(.)3 MPa for 'Oasis', and -1(.)6 MPa for their hybrid. In contrast, lethal water content of apices was similar for all genotypes (approx. 0(.)45 g H2O g d. wt(-1)), and hence the greater survival of 'Kasbah' can be ascribed to better drought avoidance rather than dehydration tolerance. In autumn-sown plants, 'Kasbah' had greatest dormancy, the hybrid was intermediate and 'Oasis' had none. The more dormant the genotype, the lower the metabolic activity during summer, and the earlier the activity declined in spring. Decreased monosaccharide content was an early indicator of dormancy induction. Accumulation of dehydrins did not correlate with stress tolerance, but dehydrin content was a function of the water status of the tissues, irrespective of the soil moisture. A protein of approx. 55 kDa occurred in leaf bases of the most dormant cultivar even in winter. Conclusions Drought avoidance and summer dormancy are correlated but can be independently expressed. These traits are heritable, allowing selection in breeding programmes.