O presente trabalho teve o objetivo de verificar a influência da inoculação de ingredientes intra ovo aos 18 dias de incubação de ovos oriundos de matrizes jovens e de pesos distintos. Ovos oriundos de matrizes com 30 semanas de idade foram separados em ovos leves e pesados, sendo incubados na mesma máquina incubadora. Aos 18 dias de incubação, no momento da transferência para o nascedouro, os ovos foram inoculados com soluções de Maltose, Polivitamínico, Zinco-Glicina, Glutamina, Mistura de todos os produtos descritos anteriormente e Cloreto de Sódio (controle). Como via de inoculação, as soluções foram utilizadas como diluentes da vacina de Marek efetuada intra-ovo aos 18 dias de incubação. Após o nascimento, 2460 pintinhos machos foram enviados para o aviário experimental onde foram divididos em 60 boxes em um delineamento inteiramente casualizado em esquema fatorial 2x6 (2 pesos e 6 soluções) totalizando 12 tratamentos com 5 repetições de 40 aves. Foram sacrificadas uma ave por repetição aos 00, 07 e 21 dias de idade para pesagem do saco da gema, intestino e fígado. Amostras de duodeno, jejuno e íleo foram enviadas para histologia para determinação de profundidade de criptas e altura de vilosidades. Amostras de sangue foram coletadas e enviadas para laboratório para determinação de nível de anticorpos contra reovírus e bronquite aviária. Os animais e a ração fornecida foram pesados semanalmente (07, 14, 21, 28, 35 e 42 dias) para determinação do peso, consumo e conversão alimentar. Aos 43 dias de idade 3 aves por repetição foram pesadas e sacrificadas para determinação do peso e rendimento de carcaça, peito com osso e pele e perna com osso e pele. Animais oriundos de ovos mais pesados obtiveram uma maior eclosão, peso ao nascimento e peso de fígado e intestino aos 00 dias. O peso aos 42 dias foi superior em aves oriundas de ovos pesados, produzindo uma carcaça e peito mais pesado, porém sem diferença de rendimento. Não houve diferença de conversão aos 42 dias de idade. Viabilidade de animais oriundos de ovos pesados foi superior de 00 a 07, 14 a 21 e 21 a 28 dias de idade, mas não afetou a viabilidade final. Peso do ovo não interferiu com o nível de anticorpos. As inoculações de soluções aos 18 dias de incubação obtiveram resultados variáveis dependendo do produto utilizado, tendo maior influência sobre altura de vilosidades e profundidade de criptas e sobre a produção de anticorpos. Não afetaram, entretanto, os parâmetros zootécnicos (ganho de peso, consumo de ração, conversão alimentar e viabilidade). A inoculação de produtos intra-ovo já é uma técnica possível de ser utilizada na avicultura industrial, entretanto, novos estudos devem ser ainda desenvolvidos no intuito de definir o melhor produto ou composto de produtos a ser utilizado / This trial had the objective to verify the influence of the in-ovo inoculation of ingredients at 18th day of incubation of eggs from different weights. Eggs from a broiler breeder flock with 30 weeks of age were separated in light and heavy eggs and were incubated in the same machine. At the 18th day of incubation, when the eggs were been transferred, they were inoculated with solutions of Maltose, Vitamins, Zinc-Glicine, Glutamine, mixture of all the ingredients and sodium chloride (control). The solutions were inoculated as Marek\'s vaccine diluter. After the eclosion, 2460 male chicks were send to the experimental house were they were divided on 60 boxes at a completely random design and a factorial 2x6 (two egg weigths and six solutions) design, summing 12 treatments with 5 repetitions of 40 chicks. One chick per repetition was sacrificed at 00, 07 and 21 days of age to weigth the yolk sac, intestine and liver. Samples of duodenum, jejunum and ileum were sent to histology to determinate villus high and cripts deep. Blood sample of the same birds were collected and sent to the laboratory to determinate anti body levels against reovirus and avian bronquitis. Animals and feed were weighted every week to determine the animal weight gain, feed consumption and feed conversion. At 43 days of age, 3 birds per repetition were weighted and sacrificed to determinate the carcass, breast and leg weight and yield. Animals from heavy eggs had a higher born weight, eclosion and liver and intestine weight at 00 days. At 42 days of age, birds from heavier eggs had a higher weight producing a heavier carcass and breast, but without yield variation. There was no difference on feed conversion at 42 days. Liveability of birds from heavier eggs were higher form 00 to 07, 14 to 21 and 21 to 28 days of age, but it didn\'t interfere the total livibility. Egg weight didn\'t interfere on the anti body level. The solutions inoculated at 18th day of incubation had variable results depending on the product utilized, influencing the villus height and cripts deep and anti body production. However, the solutions inoculation doesn\'t interfere on zoothecnical parameters as weight gain, feed consumption, feed conversion and livability. The in-ovo inoculation is a technique possible to be used on broiler production, however, new studies have to be done searching from the best product or ingredient to be used

Neste estudo focamos no papel de cinco transportadores da família ABC (ATP-binding cassete) envolvidos com a captação ativa de oligopeptídeos, poliaminas, fosfato inorgânico, glutamato e glutamina, em duas espécies bacterianas: Streptococcus mutans, que causa a cárie, e Escherichia coli enterohemorrágica (EHEC), responsável por diarreias e síndrome hemolítica urêmica em humanos. Com relação a inativação do sistema de transporte de oligopeptídeos de S. mutans não observamos alteração do crescimento bacteriano ou da aderência à superfícies abióticas. A deleção do sistema de captação de poliaminas não interferiu com o crescimento em meio rico, porém aumentou a resistência à ambiente ácidos. Inativação do sistema de transporte de fosfato inorgânico reduziu a aderência de S. mutans. Sobre os sistemas de transporte de glutamato e glutamina, mutantes de S. mutans apresentaram alterações nas taxas de crescimento e adesão à superfícies. Inativação da proteína OppA de EHEC não afetou a produção da toxina Stx bem como a patogenicidade in vitro e in vivo de EHEC. Em suma, o presente estudo demonstra que o papel dos transportadores ABC na fisiologia e patogenicidade de bactérias pode variar de acordo com a espécies bem como com o substrato transportado. / This study focuses on the role of five ABC (ATP-binding cassette) transport systems related to active uptake of oligopeptides, polyamines, inorganic phosphate, glutamate and glutamine. In this work we study two bacterial species: Streptococcus mutans, which causes caries, and enterohaemorrhagic Escherichia coli (EHEC), responsible for diarrhea and hemolytic uremic syndrome in humans. The inactivation of the S. mutans oligopeptide transport system did not change the bacterial growth and adherence to abiotic surfaces. Inactivation of the polyamine uptake system did not interfere with growth in rich medium, but increased the survival of bacteria in acid environments. Inactivation of the inorganic phosphate transport system reduced the adherence of S. mutans. Regarding the glutamate and glutamine transport systems, the S. mutans mutants showed changes in growth rates and adhesion to abiotic surfaces. Inactivation of the EHEC OppA protein, did not affect the production of the Stx toxin neither affected the in vitro and in vivo pathogenicity of the strain. Collectively, the present study shows that the roles of ABC transporters in the physiology and pathogenicity of bacteria may vary according to the species involved as well as the substrate transported.

A desnutrição proteico-energética (DPE) altera a hemopoese e, portanto, a geração de células imunológicas, bem como compromete o sistema imune. Desta forma, indivíduos desnutridos apresentam maior susceptibilidade a infecções. As células tronco mesenquimais (CTMs) possuem propriedades imunomodulatórias e são importantes na formação do estroma medular que sustenta a hemopoese. Visto que a L-glutamina (GLUT) é o aminoácido condicionalmente essencial mais consumido por CTMs, e que também apresenta capacidade imunomoduladora, investigou-se, neste trabalho, se a GLUT exerceria efeito sobre aspectos imunomodulatórios das CTMs em um modelo experimental de DPE. Para tanto, utilizou-se camundongos da linhagem BALB/c, os quais receberam rações normoproteica ou hipoproteica isocalóricas contendo, respectivamente, 12% e 2% de proteína por um período de 5 semanas. Após o isolamento e a caracterização de CTMs provenientes dos grupos controle (CTMct) e desnutrido (CTMdesn), cultivou-se essas células em 0, 0,6, 2 e 10mM GLUT, a fim de determinar a influência deste aminoácido sobre a expressão de fatores de transcrição e produção de citocinas por CTMct e CTMdesn. Adicionalmente, avaliou-se o efeito dos sobrenadantes das culturas de CTMct e CTMdesn sobre a proliferação e produção de citocinas por macrófagos e linfócitos esplênicos. Os animais desnutridos apresentaram anemia, leucopenia, hipoplasia medular e diminuição na concentração de proteínas séricas, albumina e préa-lbumina. A DPE não modificou a morfologia e o fenótipo das CTMs, bem como não alterou a expressão de proteínas reguladoras do ciclo celular. Por outro lado, a expressão de NFkB e STAT-3 e a produção de IL-1β, IL-6, IL-10 e TGF-β por CTMs foram alteradas pela DPE e variaram de acordo com as concentrações de GLUT testadas. O aumento na concentração de GLUT diminuiu a expressão de NFkB e induziu a expressão de STAT-3 por CTMs obtidas de ambos os grupos. Quanto a produção de citocinas por essas células, observou-se uma diminuição nos níveis de IL-β e IL-6 e uma elevação nos níveis de IL-10 e TGF-β com o aumento na concentração de GLUT. Variações na concentração desse aminoácido não alteraram a produção de IL-17 ou IFN-γ por CTMct e CTMdesn. Ademais, a concentração de GLUT alterou, de forma diretamente proporcional, a taxa de proliferação das CTMs. Os meios condicionados de CTMct e CTMdesn diminuíram a proliferação de macrófagos e linfócitos esplênicos estimulados com LPS, induziram aumento na produção da citocina antiinflamatória IL-10 por ambos os tipos celulares e diminuíram a produção das citocinas pró-inflamatórias IL-12 e TNF-α por macrófagos e IL-17 por linfócitos. Portanto, conclui-se que a GLUT possui efeito sobre a proliferação das CTMs, bem como a capacidade de imunomodular estas células. / Protein-energy malnutrition (PEM) alters hemopoiesis and, therefore, the generation of immune cells, and compromises the immune system. In this way, malnourished individuals are more susceptible to infections. Mesenchymal stem cells (MSCs) have immunomodulatory properties and are important in the formation of bone marrow stroma that supports hemopoiesis. Since L-glutamine (GLUT) is a conditionally essential amino acid, which is most consumed by MSCs, and present immunomodulatory capacity, this work investigated whether GLUT would have an effect on immunomodulatory aspects of MSCs in a PEM experimental model. For this purpose, BALB/c mice were used, which received isocaloric normoproteic or hypoproteic diets, containing respectively, 12% and 2% of protein for a period of 5 weeks. After isolation and characterization of MSCs from control (MSCct) and malnourished (MSCmaln) groups, these cells were cultured with 0, 0.6, 2 and GLUT 10mM in order to determine the influence of this amino acid on the expression of transcription factors and cytokine production by MSCct and MSCmaln. Besides that, the effect of MSCct and MSCmaln culture supernatants on proliferation and cytokine production by macrophages and splenic lymphocytes was evaluated. Malnourished animals presented anemia, leucopenia, marrow hypoplasia and decreased concentration of serum proteins, albumin and prealbumin. PEM did not change morphology and phenotype of MSCs or altered the expression of cell cycle regulatory proteins. On the other hand, the expression of NFkB and STAT-3 and the production of IL-1β, IL-6, IL-10 and TGF-β by MSCs were modified by PEM and varied according to the tested GLUT concentrations. An increase in GLUT concentration decreased NFkB expression and induced STAT-3 expression by MSCs obtained from both groups. Regarding the production of cytokines by these cells, an increase in GLUT concentration resulted in decreased IL-1β and IL-6 levels and increased IL- 10 and TGF-β levels. Changes in the concentration of this aminoacid did not alter IL- 17 or IFN-γ production by MSCct and MSCmaln. Furthermore, the concentration of GLUT changed, in direct proportion, the proliferation of MSCs. The conditioned media MSCct and MSCmaln decreased the proliferation of macrophages and splenic lymphocytes stimulated with LPS, induced an increase in the production of the antiinflammatory cytokine IL-10 by both cell types, and decreased the production of proinflammatory cytokines IL-12 and TNF-α by macrophages and IL-17 by lymphocytes. Therefore, it can be concluded that GLUT has an effect on the proliferation of MSCs and it has the capacity to immunomodulate these cells.

Este trabalho foi realizado com a finalidade de avaliar atividade da redutase do nitrato (RN), sintetase da glutamina (GS) e conteúdo de proteína total solúvel (PTS) em folhas de cafeeiro durante o desenvolvimento dos frutos em condições de campo. O experimento foi instalado em blocos ao acaso em esquema fatorial 3x6, constituído pela combinação de 3 doses de N (0, 150 e 350 kg ha-1) e avaliado em seis diferentes fases fenológicas (janeiro, fevereiro, março, abril, maio e junho), com sete repetições. No experimento foi utilizada a cultivar Catuaí Vermelho IAC 44 com seis anos implantada num Nitossolo Vermelho Eutroférrico, no município de Piracicaba-SP. Com relação à atividade da RN, GS e PTS em função das doses de N verificou incremento nos valores da atividade destas enzimas e do teor de proteína total solúvel em função da dose de N. As maiores atividades da RN foram obtidas no mês de janeiro (0,834, 1,370 e 1,781 µmol NO2 - g-1 MF h-1), bem como para a GS (82,47, 95,31 e 136,72 µmol γGH h-1 mg-1 PTS). O teor de proteína solúvel total (PTS) apresentou a mesma tendência (0,62, 5,66 e 6,52 mg mL-1). Estes valores foram obtidos em frutos na fase chumbinho. Durante o desenvolvimento dos frutos do cafeeiro a atividade das enzimas estudadas e o conteúdo de PTS foliar diminuíram, ao longo do tempo de janeiro a junho (maturação frutos). / The aim of this work was to evaluate the nitrate reductase (NR) and glutamine synthetase (GS) activity, total soluble protein content (PTS) in coffee leaves during the fruits development in field conditions. The study was carried out in ESALQ/USP (University of São Paulo), Piracicaba, São Paulo, Brazil in a Red Nitosol eutrofic. The experimental design was a complete randomized, in a factorial outline 3 x 6, constituted by combination of 3 levels (0, 150 and 350 kg ha-1) of nitrogen in six different periods (January, February, March, April, May and June) in plants (Cultivar Catuaí Vermelho IAC 44). The RN, GS activity and leaves PTS content was linear increased in relation of levels of N applied. In accordance with the present study, the enzymes had presented greater activity during the January than other months (0,834, 1,370 and 1,781 µmol NO2 - g-1 DF h-1) of RN; (82,47, 95,31 and 136,72 µmol γGH h-1 mg-1 PTS) of GS and (0,62, 5,66 and 6,52 mg mL-1) of total soluble protein (PTS), in the treatments T0, T1 and T2, respectively. In January, the fruits were young grains and this period the coffee plant has high enzymatic activity. However, the capacity of nitrate assimilation during the annual cycle of coffee plants can be change through the environment and climatic variations and this phase (January) was verified a great precipitation. During the coffee fruits development, the RN and GS activity and the foliar PTS content had decreased, this occur due to the process of senescence of leaves, promote it the remobilization to other plant organs, as seed filling.

Trypanosoma cruzi e Trypanosoma brucei são os agentes etiológicos da doença de Chagas e da doença do sono, respectivamente. Ambos são tripanossomatídeos, apresentam um ciclo de vida que alterna entre os hospedeiros mamíferos e os hospedeiros invertebrados e apresentam o metabolismo baseado no consumo de aminoácidos e/ou glicose, dependendo da disponibilidade de nutrientes no ambiente. Neste trabalho foi demonstrado a importância da glutamina (Gln) em diferentes aspectos da biologia do T. cruzi e a relevância da Gln e da enzima glutamina sintetase (GS) para formas sanguícolas de T. brucei. A Gln é transportada pelo T. cruzi e pelo T. brucei a partir do meio externo. Em T. cruzi foi demonstrado que esse transporte é realizado por um único sistema, saturável, específico, dependente de ATP e do gradiente de H+ na menbrana do parasita. Também foi demonstrado que a Gln é importante para replicação das formas amastigotas e epimastigotas, além de promover o processo de metaciclogênese. Tratamento com análogos estruturais da Gln dimuiu a proliferação do estágio epimastigota e também a diferenciação para tripomastigota metacíclico. Além do mais células infectadas e tratadas com os análogos apresentaram redução do número de tripomastigotas que eclodiram das células, demonstrando que a Gln também é importante para os estágios intracelulares. Em formas sanguícolas de T. brucei, a enzima GS é ativa, mas é incapaz de suprir a necessidade de Gln do parasita, fazendo com que seja completamente dependente do transporte a partir do meio externo. A Gln é importante para a proliferação formas sanguícolas e correta progressão do ciclo celular. Em meio sem Gln os parasitas são incapazes de manter a proliferação normalmente, sendo que este processo é dependente da concentração de Gln no meio externo. Também foi demonstrado que a Gln participa do processo de modificação pós-traducional de glutamilação da tubulina. Conclui-se portanto que a Gln é um aminoácido fundamental para sobrevivência do T. cruzi e do T. brucei. / Trypanosoma cruzi and Trypanosoma brucei are the etiologic agent of Chagas disease and sleeping sickness, respectively. Both parasites are trypanosomatids that have a complex life cycle, which alternates between a mammalian host and insect vector. T. cruzi and T. brucei are able to use carbohydrates and amino acids as energy source, depending on availability of nutrients in the different environments that parasites go through in the life cycle. In this work we demonstrate that glutamine (Gln) is an important metabolite that participates in many biological processes in T. cruzi, and the relevance of the enzyme glutamine synthetase and Gln in bloodstream forms of T. brucei. T. cruzi and T. brucei are able to uptake Gln from the medium. T. cruzi incorporate Gln through a single and saturable transport system. Gln uptake system is dependent on ATP intracellular levels and H+ gradient and is a highly specific system. Also was demonstrated that Gln is important to replicative stages amastigote and epimastigote, and promotes the metacyclogenesis process. The treatment with Gln analogs impared the epimastigote replication and the differentiation from epimastigote to trypomastigote metacyclic. Moreover, analogs treatment in the infected cells decrease the number of trypomastigotes released from the cells, suggesting that Gln is important to intracellular development of T. cruzi. This work also demonstrates that the enzyme glutamine synthetase is active in bloodstream forms from T. brucei, but is not enough to produce the amount of Gln required by the parasite. T. brucei, bloodstream forms are completely dependent of Gln uptake from the medium. The proper proliferation rate and correct cell cycle progress are dependent of Gln concentration in the medium. Moreover Gln participates in the tubulin glutamylation process in bloodstream forms; this is a post translational modification that is important to microtubules dynamics and cytokinesis process. We concluded that Gln is a fundamental amino acid to maintenance of T. cruzi and T. brucei.

O processo de criopreservação promove danos celulares que podem comprometer a qualidade espermática em termos de motilidade e dos índices de fertilidade, principalmente, devido ao estresse oxidativo. Assim, o objetivo deste estudo foi avaliar o efeito de diferentes concentrações de cisteína e glutamina na motilidade, morfologia, integridade da membrana, danos no DNA e índices de estresse oxidativo no sêmen de jundiá (Rhamdia quelen) pós-descongelamento. O sêmen de 5 machos (369,6 ± 71,75 g), com motilidade espermática superior a 80%, foi criopreservado em solução crioprotetora (frutose 50 g / L, leite em pó 50 g / L e metanol 100 mL / L) contendo diferentes concentrações de cisteína (0, 2,5, 5, 10 e 20 mM) e/ou glutamina (2,5 e 5,0 mM). O pool de sêmen foi diluído na proporção de 1:3, armazenado em palhetas de 0,25 mL, congelado em vapor de nitrogênio e mergulhado em nitrogênio líquido. Após o descongelamento (25°C por 10s) foram avaliados: motilidade (motilidade total, 0-100%), morfologia (Rosa de Bengala), fertilização, integridade da membrana (Eosina-Nigrosina), dano ao DNA (teste cometa), peroxidação lipídica (TBARS), atividade das enzimas SOD, CAT, GST e GPx, e a concentração de grupos carbonilas e sulfidrilas. Em relação aos parâmetros de motilidade, fertilização e morfologia espermática, nenhum tratamento apresentou diferença significativa em relação ao controle Na avaliação da integridade de membrana não foi observada diferença entre os tratamentos (P=0,7323). No ensaio do cometa e peroxidação lipídica os tratamentos que apresentaram os piores resultados foram os com maiores concentrações de cisteína e glutamina combinadas (P<0,0001) em relação ao controle. Observou-se uma maior atividade da SOD nos tratamentos 20C, 2,5G e 5G menor atividade no controle (P<0,0001). A atividade da CAT, GST e GPx foi maior no tratamento com as maiores concentrações dos antioxidantes (20C+5G; P<0,0001) e menor no controle. A concentração de grupos carbonilas foi maior no tratamento 20C+5G e menor controle (P<0,0001). Já a concentração de grupos sulfidrilas foi maior no controle e no tratamento 5C+5G (P<0,0001). Os achados deste estudo mostram que cisteína e glutamina, nas concentrações testadas, não apresentaram resultados satisfatórios e sim efeitos prejudiciais à qualidade espermática nos parâmetros de motilidade, morfologia, fertilização, peroxidação lipídica, índice de danos ao DNA e oxidação de proteínas. Portanto, as concentrações testadas não são recomendadas para a suplementação da solução crioprotetora para congelamento de sêmen de Rhamdia quelen. / The cryopreservation process promotes cellular damage that could compromise sperm quality in terms of motility and fertility rates, mainly due to oxidative stress. Thus, aim of this study was to assess the effects of different concentrations of cysteine and glutamine on post-thaw sperm motility, morphology, membrane integrity, fertility, DNA damage and indices of oxidative stress in the South American silver catfish (Rhamdia quelen). Sperm collected from five males (369.6 ± 71.75g), with sperm motility higher than 80%, was cryopreserved in cryoprotectant solution (fructose 50 g/L, powdered milk 50 g/L and methanol 100mL/L) containing different cysteine concentrations (0, 2.5, 5, 10 and 20 mM) and/or glutamine (2.5 and 5.0 mM). The semen pool was diluted 1:3, filled in 0.25 mL straws, frozen in nitrogen vapor, and plunged into liquid nitrogen. After thawing (25°C for 10 s) were measured: motility (total motile, 0-100%), morphology (Bengal Rose Staining), fertilization, membrane integrity (Eosin-Nigrosine), DNA damage (cometa assay), lipid peroxidation (TBARS), the activity of SOD, CAT, GST and GPx enzymes, and the concentration of Carbonyl and Sulfhydril groups. In relation to parameters of motility, fertilization and sperm morphology, no treatment presented a significant difference in relation to the control In the evaluation of membrane integrity, no difference was observed between treatments (P = 0.7323). In the comet and lipid peroxidation assay the treatments with the worst results were those with the highest concentrations of cysteine and glutamine combined (P <0.0001) in relation to the control. A higher activity of SOD was observed in treatments 20C, 2.5G and 5G lower activity in the control (P <0.0001). The activity of CAT, GST and GPx was higher in the treatment with the highest concentrations of antioxidants (20C + 5G; P <0.0001) and lower in the control. The concentration of carbonyl groups was higher in the 20C + 5G treatment and lower control (P <0.0001). The concentration of sulfhydryl groups was higher in control and 5C + 5G treatment (P <0.0001).The findings of this study show that cysteine and glutamine, at the concentrations tested, did not present satisfactory results, but rather, damaging effects on sperm quality in the parameters of motility, morphology, fertilization, lipid peroxidation, DNA damage and protein oxidation. Therefore, the concentrations tested are not recommended for the supplementation of the cryoprotectant solution for freezing semen of Rhamdia quelen.

During brain activity neurons release the major excitatory transmitter glutamate, which is taken up by astrocytes and converted to glutamine. Glutamine returns to neurons for re-conversion to glutamate. This glutamate-glutamine cycle is energy demanding. Glutamate turnover in injured brain was studied using an animal iron-induced posttraumatic epilepsy model and using neurointensive care data from 33 patients with spontaneous subarachnoid hemorrhage (SAH). Immunoblotting revealed that the functional form of the major astrocytic glutamate uptake protein GLT-1 was decreased 1-5 days following a cortical epileptogenic iron-injection, presumably due to oxidation-induced aggregation. Using microdialysis it was shown that the GLT-1 decrease was associated with increased interstitial glutamate levels and decreased interstitial glutamine levels. The results indicate a possible posttraumatic and post-stroke epileptogenic mechanism. Analysing 3600 microdialysis hours from patients it was found that the interstitial lactate/pyruvate (L/P) ratio correlate with the glutamine/glutamate ratio (r =-0.66). This correlation was as strong as the correlation between L/P and glutamate (r=0.68) and between lactate and glutamate (r=0.65). Pyruvate and glutamine correlated linearly (r=0.52). Energy failure periods, defined as L/P&gt;40, were associated with high interstitial glutamate levels. Glutamine increased or decreased during energy failure periods depending on pyruvate. Energy failure periods were clinically associated with delayed ischemic neurological deficits (DIND) or development of radiologically verified infarcts, confirming that L/P&gt;40 is a pathological microdialysis pattern that can predict ischemic deterioration after SAH. DIND-associated microdialysis patterns were L/P elevations and surges in interstitial glutamine. Glutamine and pyruvate correlated with the cerebral perfusion pressure (r=0.25, r=0.24). Glutamine and the glutamine/glutamate ratio correlated with the intracranial pressure (r=-0.29, r=0.40). Glutamine surges appeared upon substantial lowering of the intracranial pressure by increased cerebrospinal fluid drainage. Increased interstitial glutamine and pyruvate levels may reflect augmented astrocytic glycolysis in recovering brain tissue with increased energy demand due to a high glutamate-glutamine turnover.

Calf scours are caused by a variety of infectious agents. Oral rehydration therapy solutions are formulated with the objective of correcting dehydration and acidosis. Currently, oral rehydraton therapy does not promote gut healing in diarrheic calves. However, investigators are examining the role of nutrition in promoting gut healing. Previous work has shown that the amino acid glutamine is important in nitrogen transport between tissues and is an indispensable nutrient for rapidly dividing cells such as lymphocytes, fibroblasts and enterocytes. Small intestinal epithelial cells depend mainly on glutamine, glucose, and ketone bodies for their energy under normal physiological conditions. The oxidative substrate preferred by large intestinal epithelial cells appears to be butyrate, followed by acetate, glutamine, and glucose. Research shows that glutamine supplementation increases intestinal protein synthesis. This may be one of the mechanisms by which glutamine exerts its protective effect on gut integrity and mucosal barrier function during critical illness. However, questions concerning the optimum dose and route by which glutamine is to be administered have yet to be addressed. A surgical model was developed to chronically study the nutrient concentration differences across the portal-drained viscera of preruminant calves. The experimental design consisted of a series of infusions conducted on four different study days in each calf. On the study days, 4 separate 1h infusions of acetate, glucose, glutamine, saline (control) were administered intravenously via the jugular vein at 200 mmol/L/h in a different order. Venous and arterial blood were collected over the last 15 min of each 1h infusion. Blood flow was also measured. Intestinal uptake in ìmol/kg^0.75/min was 0.3 ± 1.1 for glutamine and 1.9 ± 3.1 for glucose during saline infusion. During acetate, glucose, and saline infusions, glucose was a greater source of energy than glutamine for the intestine. However, during glutamine infusion, intestinal glutamine uptake (29.9 ± 11.2 ìmol/kg^0.75/min) increased significantly which was associated with a rise in ammonia production (7.0 ± 0.5 ìmol/kg^0.75min). A second experiment was designed to determine if glutamine uptake could be further stimulated either by longer term intravenous infusion or by chronic oral supplementation in neonatal calves. Intestinal metabolism was investigated by measuring nutrient uptake during three intravenous infusions of glutamine over a 5 h period after an overnight fast. Prior to the first infusion, calves diet consisted of milk only. Diet was supplemented with oral glutamine for the second and third infusions. Glutamine was administered via the jugular vein at a rate of 200 mmol/L/h. Venous and arterial blood was collected in duplicate every hour for 5 h. Blood flow was also measured. During glutamine infusion, there was an absolute increase in PDV uptake of glutamine associated with a significant production of ammonia. Feeding glutamine orally did not alter the PDV glutamine uptake. Glutamine infusion did not increase the intestinal uptake of essential amino acids. Neither chronic oral supplementation with glutamine, or infusion for periods longer than an hour, further increased intestinal glutamine uptake. Arterial leucine concentration and intestinal uptake declined during glutamine infusion suggesting that its supply became limiting. Thus glutamine supplementation may require the provision of a mixture of amino acids to be effective.

Fourteen Ball State University softball players participated in this study to examine the effects of an amino acid supplement, Rebuild II TM, on strength and performance. Rebuild II TM contains high levels of BCAAs and glutamine. Two groups strength trained for ten weeks with the treatment group consuming Rebuild II TM, and the control group consuming a placebo. Subjects were pre and post-tested on the bench press, squat and machine shoulder press for strength, and performed a 90-foot sprint and a vertical jump for performance measures. LBW was calculated from the subjects body weight and percent fat. Pre and post measures were analyzed using a two way Anova variance with repeated measures test with a significant level set at p< 0.05. Both groups showed significant increases in strength and vertical jump performance, but only the Rebuild II TM group had a significant increase in LBW and decrease in percent fat. It was concluded that with a controlled weight training program, softball players will increase strength and vertical jump performance, and by adding an amino acid supplement to an athletes diet, there may be larger increases in LBW and decreases in percent fat while strength training. / School of Physical Education

Thirteen Ball State University, male basketball players participated in this study to examine the effects of an amino acid supplement, Rebuild IITM and glutamine, on strength and vertical jump performance. Rebuild IITM contains high levels of BCAA and glutamine.The thirteen subjects were divided into two groups, Rebuild IIT"'' (n=7) and placebo (n=6). Both groups participated in a ten week strength and conditioning training program. Subjects were pre and post-tested on the hang clean, bench press, squat, and the vertical jump. Changes in percent body fat were also compared. Pre and post measures were analyzed using a 2x2 ANOVA variance with a significant level set at p<0.05. Both groups showed significant increases between pre and post-test measurements in all of the variables measured, but could not be significantly proven to be due to treatment effect. It was concluded that with a controlled strength and conditioning program, basketball players would increase strength and vertical jump performance and decrease percent body fat. Yet in order to determine whether an amino acid supplement may further increase these measurements, a dietary recall must be obtained. / School of Physical Education