767 resultados para compensatory
Resumo:
Infrared thermography (IRT) is a safe and non-invasive tool used for examining physiological functions based on skin temperature (Tsk) control. The aim of this paper was to establish the probable thermal difference between the beginning and the end of the anterior cruciate ligament (ACL) rehabilitation process after surgery. For this purpose thermograms from 25 ACL surgically operated patients (2 women, 23 men) were taken on the first and last day of a six-week rehabilitation program. A FLIR infrared camera according to the protocol established by the International Academy of Clinical Thermology (IACT). The results showed significant temperature increases in the posterior thigh area between the first and the last week of the rehabilitation process probably due to a compensatory mechanism. According to this, we can conclude that temperature of the posterior area of the injured and non-injured leg has increased from the first to the last day of the rehabilitation process.
Resumo:
La presente tesis doctoral pretende ampliar conocimientos de los sistemas de producción y tecnología asociada del cerdo Ibérico. Los trabajos de campo fueron llevados a cabo en el CIA ‘El Dehesón del Encinar’ (Oropesa, Toledo, España) con cerdos Ibéricos de la estirpe Torbiscal. En los dos primeros experimentos, utilizando dos grupos de cerdos, se estudió el efecto de la restricción de la alimentación en premontanera sobre las características de la canal y sobre la composición de la grasa al sacrificio de los cerdos posteriormente cebados en montanera. Los animales restringidos en premontanera crecieron significativamente menos que los no restringidos durante dicho periodo, y manifestaron un crecimiento compensatorio durante la montanera. Este crecimiento compensatorio no fue suficiente, y los animales restringidos llegaron con pesos significativamente inferiores al sacrificio, aunque esto no supuso diferencias para las características de la canal entre ambos grupos de animales. También al sacrificio, los animales restringidos durante la premontanera tuvieron concentraciones significativamente diferentes de ácidos grasos, sobre todo en la capa interna de la grasa subcutánea y en la fracción de lípidos polares de la grasa intramuscular; asimismo, tuvieron mayores concentraciones de tocoferoles en el músculo Longissimus dorsi. En un tercer experimento, utilizando cuatro grupos de animales, se estudió el efecto del tiempo de estancia de los cerdos en montanera exclusiva sobre las características de la canal y de la grasa al sacrificio. La duración de la montanera afectó significativamente al crecimiento, a los pesos y porcentajes de los productos del despiece y al perfil de ácidos grasos de la grasa subcutánea. Los animales cebados con pienso en confinamiento tuvieron mayor crecimiento, mejor calidad de la canal, y peor calidad de grasa que los otros tres grupos de animales que se acabaron en montanera exclusiva durante 46, 83 y 111 días. En el último experimento se evaluó si la impedancia bioeléctrica es un método adecuado para realizar un estudio tisular de los jamones de cerdos Ibéricos, aspecto especialmente relevante para la industria. La impedancia bioeléctrica resultó ser un método no invasivo adecuado, que puede ser aplicado para predecir la composición tisular de los jamones, aunque se necesitan más ensayos antes de llevar este método a la práctica cotidiana de la industria. ABSTRACT This Thesis aims to expand knowledge on production systems and technology associated to Iberian pigs. Fieldwork was conducted in the CIA ‘El Dehesón del Encinar’ (Oropesa, Toledo, Spain) with Iberian pigs Torbiscal line. In first two experiments, using two groups of pigs, the effect of feed restriction during the period previous to free-range fattening on growth and carcass characteristics in Iberian pigs finished under free-range conditions was studied. The pigs fed low feeding levels during the period previous to free-range finishing period, had significantly lower growth than those feed with a high level during that period, and showed a compensatory growth during free-range period. However, this compensatory growth was not enough to reach the same weight than the animals fed with a high level, and the slaughter weights were significantly different, although no differences were observed for the carcass characteristics between both groups of pigs. At slaughter, animals fed low feeding levels during the period previous to free-range finishing period, had significantly different fatty acid proportions, especially in the inner backfat layer and in polar lipids fraction of the intramuscular fat; also, the tocopherol concentration found in Longissimus dorsi muscle from pigs fed low feeding levels was higher than those fed high feeding levels. In a third experiment, using four groups of animals, the effect of duration under freerange exclusive conditions on carcass and fat characteristics at slaughter was studied. The growth, weights and percentages of carcass cuts, and fatty acid profile of the backfat were significantly affected by the duration under free-range exclusively conditions. The animals fed in confinement with concentrate diets during the whole fattening period had high growth, better carcass quality and worse fat quality than the other three groups of animals fed under free-range exclusive conditions during 46, 83 and 111 days. In the last experiment, it assessed whether the bioelectrical impedance is a suitable method for perform a tissue study of Iberian pig hams, particularly important aspect for industry. The bioelectrical impedance has proved to be a non-invasive suitable method, which can be applied to predict the tissues composition of the hams, but more trials are needed before carrying this method to everyday practice in the industry.
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Muchos estudios han descrito la composición y diversidad de los bosques montanos tropicales, pero los patrones espaciales y las diferentes tipos de relaciones de estos, entre especies o entre grupos funcionales ha sido poco documentada. El presente trabajo se realizó en tres parcelas completamente censadas del bosque de la Estación Biológica “Chamusquin” (Zamora Chinchipe, Ecuador), y se plantea como objetivo principal conocer la estructura espacial y dinámica del bosque montano tropical del sur del Ecuador, así como las interacciones bióticas y limitaciones abióticas que influyen en esta dinámica, para ello se planteó cuatro objetivos de estudio que son: conocer los patrones espaciales de los gremios ecológicos dentro de la zona de estudio; conocer la influencia de la dependencia negativa de la densidad sobre dos especies de helechos arborescentes (Cyatheaceae); conocer si existen especies o grupos ecológicos acumuladoras o repulsoras de diversidad en el área de estudio; y conocer cómo influyen la diversidad filogenética y la densidad del vecindario en la incidencia de herbivoría y parasitismo sobre las fases iniciales del establecimiento forestal (brinzales) así como en la supervivencia de los mismos Como paso previo para el análisis del resto de objetivos, dentro de cada parcela se marcaron todos los individuos con DAP ≥5 cm y se tomaron sus coordenadas (X, Y). Además se tomaron datos de variables superficiales y muestras del suelo. Las especies encontradas se clasificaron de acuerdo a sus características biológicas, asignándolas a cada uno de los cuatro gremios forestales usualmente distinguidos en los bosques tropicales: tolerantes a la sombra (TS), tolerantes parciales a la sombra (TPS), pioneras de vida larga (PVL) y pioneras de vida corta (PVC). Para el estudio del primer objetivo tas se emplearon funciones K de Ripley inhomogéneas, ajustando la heterogeneidad en base a la variación espacial de las variables ambientales registradas . Los resultados demostraron que tanto la frecuencia relativa como el patrón espacial de las diferentes estrategias funcionales varían a lo largo de la sucesión y que éste además está influido por la variación ambiental. En adición, tanto el patrón espacial como la respuesta a la variación ambiental de los diferentes gremios es distinta entre adultos y juveniles. Todo ello sugiere que el ensamblaje de la diversidad en los bosque montanos andinos está controlado por procesos deterministas más que por procesos neutrales. Para responder el segundo objetivo se estudiaron los efectos de la dependencia negativa de la densidad (DND) y la heterogeneidad ambiental en las poblaciones de dos especies de helechos arborescentes abundantes, Cyathea caracasana y Alsophila engelii, y cómo estos efectos cambian a través de un gradiente sucesional. Los patrones de especies albergan información sobre procesos tales como la competencia que puede ser revelado fácilmente utilizando técnicas de análisis de patrones punto. Sin embargo, su detección puede ser difícil debido a los efectos de factores de confusión heterogeneidad del hábitat. Aquí, empleamos funciones K y funciones de correlación de par homogéneas y no homogéneas para cuantificar el cambio en el patrón espacial de diferentes clases de tamaño con un diseño de casos-controles para estudiar las asociaciones entre helechos arborescentes jóvenes y adultos. Usando estimaciones espaciales de la biomasa de los cuatro tipos de gremios ecológicos (PVC, PVL, TPS, TS) como covariables, hemos ajustado modelos de Poisson heterogéneos a los patrones de puntos de de los helechos juveniles y los adultos y hemos explorado además la existencia de dependencia del hábitat en estos patrones. Nuestro estudio reveló efectos de la DND para C. caracasana y un fuerte filtrado ambiental que subyace al patrón de A. engelii. Encontramos también que las poblaciones de adultos y juveniles de ambas especies respondieron de manera diferente a la heterogeneidad del hábitat y en la mayoría de los casos esta heterogeneidad se asoció con la distribución espacial de la biomasa de los cuatro tipos de gremios. Estos resultados muestran la eficacia de controlar los efectos de la heterogeneidad ambiental para evitar su confusión con los patrones derivados de interacciones biológicas cuando se estudia la DND y demuestran la utilidad de los mapas de covariables derivados de comunidades biológicas como resumen de la heterogeneidad ambiental. Para nuestro tercer objetivo nos centramos en explorar cómo influyen las especies más abundantes en la organización espacial de la diversidad a lo largo de un gradiente sucesional en el bosque montano del sur del Ecuador. Para ello utilizamos la función ISAR (Individual Species Area Relationship). Encontramos que la frecuencia de especies neutras, repulsoras y acumuladoras de diversidad taxonómica varía dependiendo del grado de sucesión. Además se comprobó que la mayoría de los gremios forestales se comportó de forma neutral, pero la proporción de acumuladores, aumentó al avanzar la sucesión hacia estados más maduros, lo que indica el establecimiento de fuertes procesos competitivos a medida que avanza la sucesión y la mayor importancia del papel de las especies individuales en dichos estados. Finalmente, examinamos el efecto de la vecindad taxonómica y filogenética, así como la estrategia de vida, sobre la incidencia de la herbivoría y el parasitismo en las poblaciones de brinzales de tres fragmentos forestales en una secuencia sucesional del bosque montano húmedo. Evaluamos además los efectos de herbivoría, parasitismo, estrategia de vida y diferentes indicadores de la vecindad sobre la supervivencia de los brinzales. Por último contrastamos la posible existencia de una tendencia compensatoria de la comunidad (CCT) a nivel de fragmento forestal. Nuestros análisis no consiguieron detectar una CCT pero si pusieron de manifiesto la existencia de efectos locales de dependencia negativa de la densidad. Por ejemplo, la presencia de herbivoría y parasitismo sobre los brinzales se relacionó significativamente con una menor supervivencia de estos. Por otro lado, indicadores del efecto de la vecindad como la densidad de brinzales del mismo género y el área basal de árboles vecinos del mismo género incrementaron la prevalencia de la herbivoría o el parasitismo en los brinzales. El incremento de la incidencia de la herbivoría o el parasitismo no está exclusivamente ligado a tener una vecindad taxonómicamente idéntica (vecinos de la misma especie) sino que categorías taxonómicas más laxas como el "género" o simplemente relaciones de semejanza filogenética son capaces de predecir los efectos negativos de la vecindad. Los efectos detectados variaron en los diferentes grupos funcionales distinguidos. Los resultados que hemos obtenido en este trabajo parecen indicar que el funcionamiento de las comunidades de brinzales del bosque montano tropical no difiere mucho del reportado para comunidades de plántulas en otros bosques tropicales y cumple las predicciones de la hipótesis de Janzen y Connell, aunque matizadas por la mayor resistencia de los brinzales al efecto de herbivoría y parasitismo. ABSTRACT Many studies have described the composition and diversity of tropical montane forests, but the different spatial patterns and types of relationships between species or between functional groups has been poorly documented. This work was made in three completely surveyed forest plots at Biological Station "Chamusquin" (Zamora Chinchipe, Ecuador). Our main objective was to know the spatial structure and dynamics of the tropical montane forest in southern Ecuador, as well as the biotic interactions and abiotic constraints affecting this dynamic. More specifically, we aimed to understand the spatial patterns of ecological guilds; to explore the influence of negative density dependence on two species of tree ferns (Cyatheaceae); to determine whether some species or ecological groups structure spatially plant diversity in these forests; and to test the effects of biological neighborhood on the incidence of herbivory and parasitism and on the survival of saplings. We mapped within each plot all trees with DBH ≥5 cm. Besides, surface data variables and soil samples they were taken. The species found were classified according to their biological characteristics in four forest guilds: shade-tolerant (ST), partial shade tolerant (PST), long-lived pioneer (LLP) and short-lived pioneer (SLP). To analyze the spatial patterns of the ecological guilds, we employed the inhomogeneous version of Ripley's K-function and adjusted heterogeneity surfaces based on the spatial variation of the measured environmental variables. The results showed that both the relative frequency of each functional guild as well as their spatial pattern varied throughout succession and that the spatial pattern is explained by environmental variation. In addition, both spatial pattern and the response to spatial variation of each guild varied throughout ontogeny. All in all suggest that diversity assembly in the studied forests is ruled by deterministic instead of neutral processes. We also addressed the negative effects of density dependence (NDD) and environmental heterogeneity in populations of two species of abundant tree ferns, Cyathea caracasana and Alsophila engelii, and how these effects change across a successional gradient.. Here, we used homogeneous and inhomogeneous K and pair-correlation functions to quantify the change in the spatial pattern of different size classes with a case-control design to study associations between young and adult tree ferns. Using spatial estimates of the biomass of the four types of ecological guilds (SLP, LLP, PST, ST) as co-variables, we fitted heterogeneous Poisson models to juvenile and adult tree fern point patterns and explored the existence of habitat dependence. Our study revealed NDD effects for C. caracasana and strong environmental filtering underlying the pattern of A. engelii. We found that adult and juvenile populations of both species responded differently to habitat heterogeneity and in most cases this heterogeneity was associated with the spatial distribution of biomass of the four functional tree types. These findings show the effectiveness of factoring out environmental heterogeneity to avoid confounding factors when studying NDD and demonstrate the usefulness of covariate maps derived from mapped communities. For our third objective we focused on exploring how the most abundant species influence the spatial organization of tree diversity in these forests. For this, we used the individual species-area relationship function (ISAR). We found that the proportion of accumulator, repeller and neutral species, varied depending on the degree of succession. We found also that most guilds behaved neutrally but the proportion of accumulator guilds increased as succession advanced to more mature stages. This point, to the existence of strong competitive effects mediated by individual species in these mature forests. Finally, we examined the effects of life strategies and taxonomic and phylogenetic neighborhood on the incidence of herbivory and parasitism in the communities of saplings in the same forest fragments. We evaluated also the effects of life strategies, herbivory, parasitism and some indicators of neighborhood on sapling survival. Finally we tested for the existences of a compensatory community trend at plot scale. We did not found a CCT but we found proof of local NND effects. For instance, the prevalence of herbivory and parasitism were related to lower sapling survival. On the other hand the density of con-generic saplings and the basal area of neighbor con-generic trees were related to a higher prevalence of herbivory or parasitism in the saplings. We demonstrated that the increase in the prevalence of herbivory or parasitism it s not exclusive of a conspecific neighborhood but instead larger taxonomic categories such as "genus" or simple phylogenetic relationships are also able to predict NND effects. The NND effects varied among functional guilds. Our results show that the dynamic of sapling communities in Ecuadorian montane forests is similar to seedling dynamics in other tropical forest and follows the predictions of Janzen-Connell hypothesis, although softened by the strong resilience of saplings in comparison to seedlings.
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Infrared thermography (IRT) is a safe and non-invasive tool used for examining physiological functions based on skin temperature (Tsk) control. Thermograms from 25 anterior cruciate ligament (ACL) surgically operated patients (2 female, 23 male) were taken with a FLIR infrared camera according to the protocol established by the International Academy of Clinical Thermology (IACT). This work consists of 4 studies. Studies 1 and 3 were related to establish the probable thermal difference among different moments of an ACL rupture after surgery: before starting the rehabilitation (P0), at the end of rehabilitation (P1) and 18 months from the end of rehabilitation (P2). For this purpose, on the other hand, studies 2 and 4 were related to establish the skin thermal difference (Tsk) between the injured and the non-injured leg in P0, P1 and P2. Results of the first study showed significant temperature increases in the posterior thigh area between P0 and P1 probably due to a compensatory mechanism. According to this, we can conclude that temperature of the posterior area of the injured and noninjured leg has increased from the first to the last day of the rehabilitation process. In the second study we found significant temperature differences between the injured and non-injured leg in both stages of rehabilitation (p<.01). On the one hand, the temperature of the injured leg is higher in the anterior view and the temperature of the non-injured leg is higher in the posterior view. By the time the patients had recovered from the reconstruction, thermal imbalances should have not been shown between symmetrical parts, but differences seemed to be still latent.. Study 3 shows that temperatures seem to be higher after a year and a half (P2) than in P1. Study 4 shows how thermal values 18 months later seemed to be normalized between both legs. No significant differences were found between the injured leg and the noninjured leg after one year and a half of the rehabilitation process. Considering results from Study 3 and 4 we can conclude that patients seemed to have recovered from a thermal point of view. The temperature in P2 was higher but symmetrical. RESUMEN La termografía infrarroja (IRT) es una herramienta segura y no invasiva utilizada para examinar funciones fisiológicas que se basan en el control de temperatura de la piel (Tsk). Termogramas de 25 pacientes intervenidos quirúrgicamente del ligamento cruzado anterior (LCA) (2 mujeres, 23 hombres) fueron tomadas con una cámara de infrarrojos FLIR de acuerdo con el protocolo establecido por la Academia Internacional de Termología Clínica (IACT). Este trabajo consiste en 4 estudios. Los estudios 1 y 3 describen la diferencia térmica entre los diferentes momentos tras la operación del ligamento cruzado anterior: antes de comenzar la rehabilitación (P0), al final de la rehabilitación (P1) y 18 meses tras finalizar la rehabilitación (P2). Por otra parte, los estudios 2 y 4 describen la diferencia de temperatura de la piel (Tsk) entre la pierna lesionada y la pierna no lesionada en P0, P1 y P2. Los resultados del primer estudio mostraron aumentos significativos de temperatura en la zona posterior de los muslos entre P0 y P1, probablemente debido a un mecanismo de compensación. De acuerdo con esto, se puede concluir que la temperatura de la zona posterior de la pierna lesionada y no lesionada se ha incrementado desde el primero hasta el último día del proceso de rehabilitación. En el segundo estudio se encontraron diferencias significativas de temperatura entre la pierna lesionada y no lesionada en ambas etapas de la rehabilitación (p<.01). Por un lado, la temperatura de la pierna lesionada es mayor en la vista anterior. Por otro lado, la temperatura de la pierna no lesionada es mayor en la vista posterior. Una vez que los pacientes se han recuperado de todo el proceso, no deberían existir desequilibrios térmicos entre partes simétricas del cuerpo, pero las diferencias todavía estaban latentes. El tercer estudio muestra que la temperatura es más alta en P2 que en P1. El cuarto estudio muestra cómo los valores térmicos entre ambas piernas en P2 se han normalizado entre ambas piernas. No se encontraron diferencias significativas entre la pierna lesionada y la pierna no lesionada después de 18 meses tras el proceso de rehabilitación. Considerando los resultados del studio 3 y 4, podemos concluir que se ha llegado a la recuperación total desde un punto de vista térmico. La temperatura es más elevada en P2 pero simétrica.
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Este estudo teve por objetivo conhecer, discutir e analisar as motivações e aspirações dos alunos inseridos nos cursos pré-vestibulares para negros e carentes da ONG EDUCAFRO, bem como a inserção desses jovens no Ensino Superior e no mercado de trabalho, considerando os mecanismos de inclusão e exclusão dos negros no Sistema Educacional Brasileiro. Dentre muitos estudos importantes, que abordam a temática do negro no sistema educacional, gostaríamos de destacar os Movimentos Sociais, de Educação e Cidadania, a dissertação: Um Estudo sobre os Cursos Pré-Vestibulares Populares, apresentada ao Programa de Pós-graduação, como requisito parcial à obtenção do título de Mestre de Alexandre do Nascimento UERJ Universidade do Estado do Rio de Janeiro, em 1999 e a dissertação de Mestrado de Cristiane Maria Ribeiro sob o título Anti-Racismo e Educação: O Projeto Político-Pedagógico das Lideranças Negras de Uberlândia em 2000. A leitura dessas obras foi essencial para o encaminhamento dos estudos que integram este trabalho, uma vez que argumentam sobre a dívida social que o Brasil tem com os afro-descendentes no sistema educacional. Destacamos também o professor, escritor e ativista dos direitos humanos, o historiador negro nascido nos EUA, John Hope Franklin declara que as políticas compensatórias foram aplicadas desde a década de sessenta. Essas políticas pretendiam oferecer aos afro-americanos a chance de participar das mudanças sociais. De modo que as universidades foram obrigadas a implantar políticas de cotas e também implantar procedimentos que fossem favoráveis à população negra. No Brasil, essa luta está sendo organizada pela ONG EDUCAFRO que vem desenvolvendo há alguns anos mecanismos de inclusão social, justificando-os por meio da necessidade de compensar os negros pela discriminação sofrida no passado, beneficiando de alguma forma essa porcentagem da população brasileira. No decorrer da pesquisa bibliográfica, encontramos, por meio das diversas obras consultadas, uma grande preocupação dos autores com a questão das Ações Afirmativas como meio de compensar a população negra, apesar da resistência por parte daqueles que temem o progresso social dos negros, no entanto, pesquisas s indicam caminhos para reverter esse quadro negativo. Diante dessa realidade, fica o grande desafio: o que motiva e quais são as metas dos professores que ministram voluntariamente aulas nos Cursinhos Comunitários? O que almejam os alunos com o seu acesso no Ensino Superior? A pesquisa confirmou que os alunos do Núcleo estudado buscam na ONG EDUCAFRO uma forma alternativa de inserção no Ensino Superior e, que essa inserção os motiva e os inspira no vislumbre de se colocarem também no mercado de trabalho. Vale ressaltar que o presente estudo não tem a pretensão de esgotar o assunto, mas de abrir espaços
Resumo:
Este estudo teve por objetivo conhecer, discutir e analisar as motivações e aspirações dos alunos inseridos nos cursos pré-vestibulares para negros e carentes da ONG EDUCAFRO, bem como a inserção desses jovens no Ensino Superior e no mercado de trabalho, considerando os mecanismos de inclusão e exclusão dos negros no Sistema Educacional Brasileiro. Dentre muitos estudos importantes, que abordam a temática do negro no sistema educacional, gostaríamos de destacar os Movimentos Sociais, de Educação e Cidadania, a dissertação: Um Estudo sobre os Cursos Pré-Vestibulares Populares, apresentada ao Programa de Pós-graduação, como requisito parcial à obtenção do título de Mestre de Alexandre do Nascimento UERJ Universidade do Estado do Rio de Janeiro, em 1999 e a dissertação de Mestrado de Cristiane Maria Ribeiro sob o título Anti-Racismo e Educação: O Projeto Político-Pedagógico das Lideranças Negras de Uberlândia em 2000. A leitura dessas obras foi essencial para o encaminhamento dos estudos que integram este trabalho, uma vez que argumentam sobre a dívida social que o Brasil tem com os afro-descendentes no sistema educacional. Destacamos também o professor, escritor e ativista dos direitos humanos, o historiador negro nascido nos EUA, John Hope Franklin declara que as políticas compensatórias foram aplicadas desde a década de sessenta. Essas políticas pretendiam oferecer aos afro-americanos a chance de participar das mudanças sociais. De modo que as universidades foram obrigadas a implantar políticas de cotas e também implantar procedimentos que fossem favoráveis à população negra. No Brasil, essa luta está sendo organizada pela ONG EDUCAFRO que vem desenvolvendo há alguns anos mecanismos de inclusão social, justificando-os por meio da necessidade de compensar os negros pela discriminação sofrida no passado, beneficiando de alguma forma essa porcentagem da população brasileira. No decorrer da pesquisa bibliográfica, encontramos, por meio das diversas obras consultadas, uma grande preocupação dos autores com a questão das Ações Afirmativas como meio de compensar a população negra, apesar da resistência por parte daqueles que temem o progresso social dos negros, no entanto, pesquisas s indicam caminhos para reverter esse quadro negativo. Diante dessa realidade, fica o grande desafio: o que motiva e quais são as metas dos professores que ministram voluntariamente aulas nos Cursinhos Comunitários? O que almejam os alunos com o seu acesso no Ensino Superior? A pesquisa confirmou que os alunos do Núcleo estudado buscam na ONG EDUCAFRO uma forma alternativa de inserção no Ensino Superior e, que essa inserção os motiva e os inspira no vislumbre de se colocarem também no mercado de trabalho. Vale ressaltar que o presente estudo não tem a pretensão de esgotar o assunto, mas de abrir espaços
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Instability of repetitive sequences, both in intronic sequences and within coding regions, has been demonstrated to be a hallmark of genomic instability in human cancer. Understanding how these mutational events arise may provide an opportunity for prevention or early intervention in cancer development. To study the source of this instability, we have identified a region of the β-lactamase gene that is tolerant to the insertion of fragments of exogenous DNA as large as 1,614 bp with minimal loss of enzyme activity, as determined by antibiotic resistance. Fragments inserted out-of-frame render Escherichia coli sensitive to antibiotic, and compensatory frameshift mutations that restore the reading frame of β-lactamase can be selected on the basis of antibiotic resistance. We have utilized this site to insert a synthetic microsatellite sequence within the β-lactamase gene and selected for mutations yielding frameshifts. This assay provides for detection of one frameshift mutation in a background of 106 wild-type sequences. Mismatch repair deficiency increased the observed frameshift frequency ≈300-fold. Exposure of plasmid containing microsatellite sequences to hydrogen peroxide resulted in frameshift mutations that were localized exclusively to the microsatellite sequences, whereas DNA damage by UV or N-methyl-N′-nitro-N-nitrosoguanidine did not result in enhanced mutagenesis. We postulate that in tumor cells, endogenous production of oxygen free radicals may be a major factor in promoting instability of microsatellite sequences. This β-lactamase assay may provide a sensitive methodology for the detection and quantitation of mutations associated with the development of cancer.
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Sinorhizobium meliloti bacteria produce a signal molecule that enhances root respiration in alfalfa (Medicago sativa L.) and also triggers a compensatory increase in whole-plant net carbon assimilation. Nuclear magnetic resonance, mass spectrometry, and ultraviolet–visible absorption identify the enhancer as lumichrome, a common breakdown product of riboflavin. Treating alfalfa roots with 3 nM lumichrome increased root respiration 21% (P < 0.05) within 48 h. A closely linked increase in net carbon assimilation by the shoot compensated for the enhanced root respiration. For example, applying 5 nM lumichrome to young alfalfa roots increased plant growth by 8% (P < 0.05) after 12 days. Soaking alfalfa seeds in 5 nM lumichrome before germination increased growth by 18% (P < 0.01) over the same period. In both cases, significant growth enhancement (P < 0.05) was evident only in the shoot. S. meliloti requires exogenous CO2 for growth and may benefit directly from the enhanced root respiration that is triggered by lumichrome. Thus Sinorhizobium–alfalfa associations, which ultimately form symbiotic N2-reducing root nodules, may be favored at an early developmental stage by lumichrome, a previously unrecognized mutualistic signal. The rapid degradation of riboflavin to lumichrome under many physiological conditions and the prevalence of riboflavin release by rhizosphere bacteria suggest that events demonstrated here in the S. meliloti–alfalfa association may be widely important across many plant–microbe interactions.
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Long-term visual memory performance was impaired by two types of challenges: a diazepam challenge on acquisition and a sensory challenge on recognition. Using positron-emission tomography regional cerebral blood flow imaging, we studied the effect of these challenges on regional brain activation during the delayed recognition of abstract visual shapes as compared with a baseline fixation task. Both challenges induced a significant decrease in differential activation in the left fusiform gyrus, suggesting that this region is involved in the automatic or volitional comparison of incoming and stored stimuli. In contrast, thalamic differential activation increased in response to memory challenges. This increase might reflect enhanced retrieval attempts as a compensatory mechanism for restoring recognition performance.
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Muscarinic acetylcholine receptors are members of the G protein-coupled receptor superfamily expressed in neurons, cardiomyocytes, smooth muscle, and a variety of epithelia. Five subtypes of muscarinic acetylcholine receptors have been discovered by molecular cloning, but their pharmacological similarities and frequent colocalization make it difficult to assign functional roles for individual subtypes in specific neuronal responses. We have used gene targeting by homologous recombination in embryonic stem cells to produce mice lacking the m1 receptor. These mice show no obvious behavioral or histological defects, and the m2, m3, and m4 receptors continue to be expressed in brain with no evidence of compensatory induction. However, the robust suppression of the M-current potassium channel activity evoked by muscarinic agonists in sympathetic ganglion neurons is completely lost in m1 mutant mice. In addition, both homozygous and heterozygous mutant mice are highly resistant to the seizures produced by systemic administration of the muscarinic agonist pilocarpine. Thus, the m1 receptor subtype mediates M current modulation in sympathetic neurons and induction of seizure activity in the pilocarpine model of epilepsy.
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Mutation of the obese gene produces obesity, hyperinsulinemia, and compensatory “overexpression” of the defective gene. As insulin activates obese gene expression, it seemed possible that hyperinsulinemia might be responsible for overexpression of the gene. To address this question we rapidly neutralized circulating insulin by injection of an insulin antibody. Unexpectedly, insulin depletion in obese (ob/ob or db/db) mice caused massive adipose RNA degradation confirmed by histological analysis to result from adipocyte cell death by a largely necrotic mechanism. This effect was not observed in lean littermates and was completely corrected by coadministration of insulin. Comparison of multiple tissues demonstrated that the effect was restricted to adipose tissue. Insulin depletion in obese mice by administration of streptozotocin also led to cell death, but this death was less extensive and appeared to be apoptotic in mechanism. Thus insulin may promote the survival side of the physiological balance between adipocyte survival and death.
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In bacterial photosynthetic reaction centers, the protonation events associated with the different reduction states of the two quinone molecules constitute intrinsic probes of both the electrostatic interactions and the different kinetic events occurring within the protein in response to the light-generated introduction of a charge. The kinetics and stoichiometries of proton uptake on formation of the primary semiquinone QA− and the secondary acceptor QB− after the first and second flashes have been measured, at pH 7.5, in reaction centers from genetically modified strains and from the wild type. The modified strains are mutated at the L212Glu and/or at the L213Asp sites near QB; some of them carry additional mutations distant from the quinone sites (M231Arg → Leu, M43Asn → Asp, M5Asn → Asp) that compensate for the loss of L213Asp. Our data show that the mutations perturb the response of the protein system to the formation of a semiquinone, how distant compensatory mutations can restore the normal response, and the activity of a tyrosine residue (M247Ala → Tyr) in increasing and accelerating proton uptake. The data demonstrate a direct correlation between the kinetic events of proton uptake that are observed with the formation of either QA− or QB−, suggesting that the same residues respond to the generation of either semiquinone species. Therefore, the efficiency of transferring the first proton to QB is evident from examination of the pattern of H+/QA− proton uptake. This delocalized response of the protein complex to the introduction of a charge is coordinated by an interactive network that links the Q− species, polarizable residues, and numerous water molecules that are located in this region of the reaction center structure. This could be a general property of transmembrane redox proteins that couple electron transfer to proton uptake/release reactions.
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Anchorage and growth factor independence are cardinal features of the transformed phenotype. Although it is logical that the two pathways must be coregulated in normal tissues to maintain homeostasis, this has not been demonstrated directly. We showed previously that down-modulation of β1-integrin signaling reverted the malignant behavior of a human breast tumor cell line (T4–2) derived from phenotypically normal cells (HMT-3522) and led to growth arrest in a three-dimensional (3D) basement membrane assay in which the cells formed tissue-like acini (14). Here, we show that there is a bidirectional cross-modulation of β1-integrin and epidermal growth factor receptor (EGFR) signaling via the mitogen-activated protein kinase (MAPK) pathway. The reciprocal modulation does not occur in monolayer (2D) cultures. Antibody-mediated inhibition of either of these receptors in the tumor cells, or inhibition of MAPK kinase, induced a concomitant down-regulation of both receptors, followed by growth-arrest and restoration of normal breast tissue morphogenesis. Cross-modulation and tissue morphogenesis were associated with attenuation of EGF-induced transient MAPK activation. To specifically test EGFR and β1-integrin interdependency, EGFR was overexpressed in nonmalignant cells, leading to disruption of morphogenesis and a compensatory up-regulation of β1-integrin expression, again only in 3D. Our results indicate that when breast cells are spatially organized as a result of contact with basement membrane, the signaling pathways become coupled and bidirectional. They further explain why breast cells fail to differentiate in monolayer cultures in which these events are mostly uncoupled. Moreover, in a subset of tumor cells in which these pathways are misregulated but functional, the cells could be “normalized” by manipulating either pathway.
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The snowshoe hare and the Canadian lynx in the boreal forests of North America show 9- to 11-year density cycles. These are generally assumed to be linked to each other because lynx are specialist predators on hares. Based on time series data for hare and lynx, we show that the dominant dimensional structure of the hare series appears to be three whereas that of the lynx is two. The three-dimensional structure of the hare time series is hypothesized to be due to a three-trophic level model in which the hare may be seen as simultaneously regulated from below and above. The plant species in the hare diet appear compensatory to one another, and the predator species may, likewise, be seen as an internally compensatory guild. The lynx time series are, in contrast, consistent with a model of donor control in which their populations are regulated from below by prey availability. Thus our analysis suggests that the classic view of a symmetric hare–lynx interaction is too simplistic. Specifically, we argue that the classic food chain structure is inappropriate: the hare is influenced by many predators other than the lynx, and the lynx is primarily influenced by the snowshoe hare.
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Mice with a targeted mutation of the gastric inhibitory polypeptide (GIP) receptor gene (GIPR) were generated to determine the role of GIP as a mediator of signals from the gut to pancreatic β cells. GIPR−/− mice have higher blood glucose levels with impaired initial insulin response after oral glucose load. Although blood glucose levels after meal ingestion are not increased by high-fat diet in GIPR+/+ mice because of compensatory higher insulin secretion, they are significantly increased in GIPR−/− mice because of the lack of such enhancement. Accordingly, early insulin secretion mediated by GIP determines glucose tolerance after oral glucose load in vivo, and because GIP plays an important role in the compensatory enhancement of insulin secretion produced by a high insulin demand, a defect in this entero-insular axis may contribute to the pathogenesis of diabetes.