Thesis (Ph. D.)--University of Missouri--Columbia, 2002. / "December 2002." Typescript. Vita. Includes bibliographical references (leaves 202-211). Also available on the Internet.

Thesis (Ph. D.)--Florida State University, 2006. / Advisor: Emily Haymes, Florida State University, College of Human Sciences, Dept. of Food, Nutrition, and Exercise Sciences. Title and description from dissertation home page (viewed Sept. 19, 2006). Document formatted into pages; contains xi, 123 pages. Includes bibliographical references.

It has been hypothesized that an increase in the ratio of plasma tryptophan (TRP) to branched-chain amino acid (BCAA) concentrations may mediate an increase in cerebral serotonin synthesis, through an increased cerebral tryptophan uptake. It is postulated that the increased brain serotonin content may induce central fatigue during prolonged exercise. Until present, this postulate had not been subject to rigorous scientific testing during prolonged exercise. Therefore the aim of this study was to investigate whether ingesting a BCAA supplement during prolonged exercise improves physical performance and central fatigue. The use of such a supplement during prolonged exercise could then be expected to have a large effect on performance. Eight trained cyclists (VO₂ max= 61.9 ± 4.3 ml 02/kg/min) ingested, in random order, a drink containing either 10% carbohydrate (CHO), 10% CHO and 0.16% branched-chain amino acid (BCAA) or 0.16% BCAA. Every hour, for the duration of the exercise (4 hours, 55% VO₂ max) blood samples were analysed for amino acids, ammonia, free fatty acids, glycerol, glucose and insulin concentrations. Urine was analysed for urea and creatinine concentrations. Heart rate, oxygen consumption (VO₂), respiratory exchange ratio (RER) and rating of perceived exertion were also analysed. Thereafter, subject's 40km time trial performance and RPE was assessed on a Velodyne windtrainer. Central fatigue following the time trial was quantified using the Sternberg reaction-time paradigm. The serum concentration of the BCAA's declined as a result of the exercise, in the BCAA only trial. Tryptophan concentration, however, did not change during the exercise. The serum TRP:BCAA ratio increased (0.16 ± 0.06 to 0.20 ± 0.10; p≤0.05) in the CHO trial only. The BCAA trial differed from the two trials in which CHO was ingested because plasma ammonia and glucose concentrations did not increase, while free fatty acids (FF A's) and glycerol concentrations increased significantly (p≤0.05). The lower RER in the BCAA trials suggests a higher proportion of fat was oxidised in these trials, compared to the other two trials. Cycling performance, over a 40km time trial, (CHO= 68.59 ± 6.02; CHO+ BCAA = 68.00 ± 3.01; BCAA = 69.43 ± 5.35 min/sec), ratings of perceived exertion, submaximal or maximal heart rates, and mental performance were not different between trials. Data from this study appears to refute the thesis hypothesis that an increase in serum TRP:BCAA decreases physical performance and central fatigue, during prolonged exercise.

Thesis (Ph. D.)--University of Missouri--Columbia, 2002. / Typescript. Vita. Includes bibliographical references (leaves 202-211). Also available on the Internet.

Orientador: Jose Barreto Campello Carvalheira / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-25T02:56:20Z (GMT). No. of bitstreams: 1 Mendes_MariaCarolinaSantos_D.pdf: 2658779 bytes, checksum: 153ed5344815e7e59a41c04c4a965670 (MD5) Previous issue date: 2014 / Resumo: A proteína mTOR é um proteína reguladora chave de vários processos celulares, dentre eles proliferação, crescimento e sobrevivência celular. Fatores de crescimento, oxigênio, status energético e a presença de aminoácidos são fundamentais para que todos esses processos ocorram normalmente. Descobertas realizadas nas últimas décadas mostraram que a via da mTOR encontra-se ativada em vários processos celulares, incluindo formação tumoral e angiogênese. A leucina é um aminoácido de cadeia ramificada que tem o maior potencial em ativar a via da mTOR. Devido sua capacidade de promover a síntese proteica e ganho de massa muscular, seu uso é constantemente estimulado em pacientes com câncer. No entanto, seus efeitos no crescimento tumoral não está claro. Dessa forma, realizamos um estudo cujo objetivo principal foi investigar os efeitos da dieta suplementada com leucina na modulação do crescimento tumoral em diferentes linhagens de células tumorais que se diferenciem em relação à ativação constitutiva da via IRS1/Akt/mTOR. Estudos in vivo e in vitro realizados demonstraram que as células que se diferenciam em relação à ativação da via IRS1/AKT/mTOR respondem de maneira distinta à suplementação com leucina. Linhagens de células tumorais que possuem a via da mTOR constitutivamente ativada, PC-3 e MCF-7, quando suplementadas com doses elevadas de leucina in vitro reduziram a proliferação celular e causaram retenção das células na fase G1 do ciclo celular. Já o xenoenxerto tumoral da PC-3 reduziu sua proliferação e aumentou a morte celular quando os animais foram suplementados com leucina na dieta. Nós também observamos aumento da atividade da mTOR e da p70S6K em todas as linhagens celulares quando suplementadas com leucina. O aumento da atividade da proteína mTOR foi acompanhado de redução na fosforilação de AKTser473 nas células que possuíam a via da PI3K hiperativada (PC-3 e MCF-7). Esse fato pode estar ocorrendo devido a ativação das alças de contraregulação ocasionadas pela estimulação excessiva provocada pela suplementação com leucina, naquelas linhagens celulares que já possuem a via hiperativada. Fato este comprovado pelo aumento da fosforilação em serina 307 da proteína IRS1. Dessa forma, nossos resultados sugerem que a ativação da via da mTOR é central para determinar a sensibilidade de tumores à dieta suplementada com leucina, podendo modular o desenvolvimento tumoral naquelas células que já possuem a via IRS1/AKT/mTOR constitutivamente ativada. O mecanismo pelo qual a leucina pode retardar o desenvolvimento tumoral em células que possuem a via da mTOR hiperativada parece estar relacionado com o eixo de regulação negativa p70S6K-PI3K, com consequente redução da fosforilação de AKT e liberação das vias apoptóticas nos tecidos tumorais / Abstract: mTOR is a key regulatory protein in various cellular processes including proliferation, cell growth and survival. Growth factors, oxygen, energy status and amino acids are all essential to these processes. New findings in the last few decades have shown that the mTOR pathway is activated in many cellular processes, including tumorigenesis and angiogenesis. The branched chain amino acid leucine has the greatest potential to activate the mTOR pathway. Due to its ability to promote protein synthesis and muscle mass gain, use of leucine is frequently utilized in patients with cancer. However, the effect of leucine on tumor growth is not clear. The aim of this study is therefore to investigate the effect of diet-supplemented leucine on the modulation of tumor growth in several tumor cell lines that differ in the constitutive activation status of the insulin receptor substrate 1 (IRS1)/AKT/mTOR pathway. Both in vitro and in vivo experiments demonstrated different cell proliferation responses when cells were exposed to high doses of leucine. Tumor cell lines PC-3 and MCF-7, which have a constitutively activated mTOR signaling, displayed reduced cell proliferation and G1 phase cell cycle arrest when supplemented with high doses of leucine in vitro. Likewise, leucine-supplemented PC-3 cell tumor xenografts displayed reduced proliferation and increased cell death. We also observed increased activity of mTOR and its downstream substrate p70S6K in all cell lines supplemented with leucine. Increased mTOR activity was accompanied by a reduction in AKT serine 473 (ser473) phosphorylation in cell lines with a hyperactivated PI3K pathway (PC-3 and MCF-7). This most likely occurred because leucine supplementation further increased mTOR and p70S6K activity, triggering the inhibitory p70S6K/IRS1 axis. In fact, we found increased IRS1 ser307 phosphorylation in hyperactivated cell lines (PC-3 and MCF-7) supplemented with high doses of leucine. Therefore, our results suggest that mTOR pathway activation is central to determining the sensitivity of tumors to leucine supplementation. Furthermore, this could affect the response to leucine-supplemented therapies of those tumors in which the PI3K pathway is constitutively activated. The mechanism for this appears to be related to the negative p70S6K/IRS1 regulation axis, with consequent reduction of AKT phosphorylation and the release of apoptotic pathways in tumor tissues / Doutorado / Fisiopatologia Médica / Doutora em Ciências