Made available in DSpace on 2016-12-06T17:07:25Z (GMT). No. of bitstreams: 1 Lilian.pdf: 1551232 bytes, checksum: c1c7318479a8b0ae5bf784930ff5f056 (MD5) Previous issue date: 2009-10-30 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / This study was to investigate the effects of taurine supplementation on biomarkers of oxidative stress after eccentric exercise. Twenty four male rats Wistar were divided into the following groups (n=6): control (C); eccentric exercise (EE); eccentric exercise plus taurine (EET); eccentric exercise plus saline (EES). Taurine was administered with solution of 1ml water 300mg/Kg body weight (BW)/day by oral lavage, for two weeks before and 2, 12, 24 and 48 hours after the exercise session. The animals were submitted to one downhill run session with duration of 90 minutes and velocity constant of 16,6 m.min-1. Forty-eight hours after the exercise session, the animals were killed and the quadriceps muscle were surgically removed. Production of superoxide anion, lipoperoxidation, carbonylation, total thiol content and antioxidant enzyme were analyze. The results show the EET group had a significant reduction (p <0.05) in the production of superoxide anion (0,09 + 0.2 nmol / min / mg protein) when compared with the EE group (1.62 0.3 nmol / min / mg protein) and the EES group (1,55 0,3 nmol/ min/ mg protein). Lipid peroxidation was diminished as EET showed 0,13 0,001nmol/ mg protein of tiobarbituric acid reactive species (TBARS) significantly lower (p < 0,05) than other groups (EES - 0,18 0,03 nmol/mg protein and EE - 0,18 0,05 nmol/mg protein). The total thiol content were higher (p < 0,05) in EET (18,8 0,3 nmol TNB/ mg protein) when compared with EES (13,8 1 nmol TNB/ mg protein) and EE (12,2 1 nmol TNB/ mg protein). The EET group showed a lower value (p < 0.05) of proteins carbonyl (0,20 0,2 nmol/ mg protein) compared to other groups (EES - 0,30 0,03 nmol/ mg protein and EE - 0,30 0,025 nmol/ mg protein). The activity of antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT) had no significant difference (p <0.05) with supplementation of taurine. The values of SOD activity in the groups were: EE 1,22 0,25 U of SOD/ mg protein; EET 1,18 0,15 U of SOD / mg protein; EES 1,22 0,12 U of SOD / mg protein; and the values of CAT were: EE - 6,12 0,19 U of CAT/ mg protein; EET- 5,85 0,62 U of CAT/ mg protein; EES 5,61 0,45 U of CAT/ mg protein. In conclusion, the present study demonstrates that taurine supplementation decreased damage oxidative but did not affect antioxidant enzyme activity after eccentric exercise. / O objetivo deste estudo foi investigar os efeitos da suplementação de taurina sobre biomarcadores do estresse oxidativo após o exercício excêntrico. Vinte e quatro ratos da raça Wistar machos foram divididos nos seguintes grupos (n = 6): controle (C); exercício excêntrico (EE); exercício excêntrico com a suplementação de taurina (EET); exercício excêntrico com a suplementação de salina (EES). A quantidade de taurina administrada foi de 300mg/Kg de peso corporal / dia em solução de 1 ml de água deionizada por gavagem oral, por duas semanas antes e 2, 12, 24 e 48 horas após a sessão de exercício. Os animais foram submetidos a uma sessão única de corrida em declive com duração de 90 minutos e velocidade constante de 16,6 m.min-1. Quarenta e oito horas após a sessão de exercício, os animais foram sacrificados e os músculos do quadríceps removidos cirurgicamente. Produção do ânion superóxido, lipoperoxidação, carbonilação das proteínas, conteúdo total de tióis e atividade de enzimas superóxido dismutase (SOD) e catalase (CAT) foram analisados. Os resultados mostram que o grupo EET apresentou uma redução significativa (p<0,05) na produção do ânion Superóxido (0,9 0,2 nmol/min/mg proteína) quando comparado com o grupo EE (1,62 0,2 nmol/min/mg proteína) e o grupo EES (1,55 0,3 nmol/min/mg proteína). A lipoperoxidação foi diminuída já que EET apresentou quantidade de espécies reativas ao ácido tiobarbitúrico (TBARS) de 0,13 0,001nmol/mg proteína, significativamente menor (p<0,05) aos outros grupos (EES - 0,18 0,03 nmol/mg proteína e EE - 0,18 0,05 nmol/mg proteína). O conteúdo total de tióis apresentou-se aumentado (p<0,05) no grupo EET (18,8 0,3 nmol TNB/mg proteína) em comparação com EES (13,8 1 nmol TNB/mg proteína) e EE (12,2 1 nmol TNB/mg proteína). O grupo EET demonstrou um valor menor (p<0,05) na carbonilação de proteínas (0,20 0,2 nmol/mg proteína) em relação aos demais grupos (EES - 0,30 0,03 nmol/mg proteína; e EE - 0,30 0,025 nmol/mg proteína). Entretanto, a atividade das enzimas antioxidantes SOD e CAT não teve diferença significativa (p<0,05) com a suplementação da taurina. Os valores da atividade da SOD nos grupos foi de: EE 1,22 0,25 U de SOD/mg proteína; EET 1,18 0,15 U de SOD /mg proteína; EES 1,22 0,12 U de SOD /mg proteína; e da CAT foi: EE - 6,12 0,19 U de CAT/mg proteína; EET- 5,85 0,62 U de CAT/mg proteína; EES 5,61 0,45 U de CAT/mg proteína. Em conclusão, o estudo demonstra que a suplementação de taurina diminuiu o dano oxidativo, mas não afetou a atividade das enzimas antioxidantes após exercício excêntrico.

Orientador: Everardo Magalhães Carneiro / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-13T21:03:55Z (GMT). No. of bitstreams: 1 Batista_ThiagoMartins_M.pdf: 1168177 bytes, checksum: 6c327ab4c396fab9844364b47f69ad79 (MD5) Previous issue date: 2009 / Resumo: A desnutrição ainda é um problema de saúde pública que afeta principalmente países em desenvolvimento e sua prevalência chega a ser crescente em algumas áreas. Vários estudos obtiveram êxito em correlacionar a má nutrição em estágios iniciais de vida com o desenvolvimento de doenças cardiovasculares e diabetes tipo 2 na vida adulta. No modelo de desnutrição pós desmame verifica-se menor secreção de insulina estimulada por glicose e outros agentes insulinotrópicos bem como menor expressão de várias proteínas envolvidas com a funcionalidade da célula b. Estudos realizados por nosso grupo e outros laboratórios mostram que a suplementação de camundongos com taurina aumenta a secreção de insulina além de regular o influxo de íons Ca2+ para as células b, etapa crucial para o processo secretório. Para avaliar os efeitos da taurina sobre animais desnutridos, utilizamos ratos wistar, machos com 21 dias de vida. Os animais receberam dieta contendo 17% de proteína (normoprotéica) (C) ou 6% de proteína (hipoprotéica) (D). Animais C e D receberam suplementação com taurina a 2,5% na água de beber por 30 dias (CT30 e DT30) ou 90 dias (CT90 e DT90). Em seguida avaliamos parâmetros biométricos e bioquímicos, tolerância à glicose, secreção de insulina estimulada por glicose e pelo agonista colinérgico carbacol, expressão de proteínas envolvidas no controle da secreção de insulina e, finalmente, registramos os movimentos citoplasmáticos de íons Ca2+ após estímulo com glicose e carbacol. Verificamos que a restrição protéica retardou o crescimento dos animais além de reduzir a concentração plasmática de proteínas totais (C = 6,81±0,04; CT30 = 7,15±0,54; CT90 = 6,87±0,19; D = 5,35±0,24; DT30 = 5,37±0,28; DT90 = 5,70±0,09 g/dl; n = 3-5) e albumina (C = 3,20±0,11; CT30 = 3,41±0,02; CT90 = 3,18±0,05; D = 2,74±0,07; DT30 = 2,49±0,09; DT90 = 2,67±0,04 g/dl; n = 5-9) sem efeito da suplementação com taurina. Os animais D se mostraram mais tolerantes à glicose e a suplementação com taurina por 90 dias restaurou parcialmente a tolerância desses animais (C = 30249±2682; CT30 = 37255±6691; CT90 = 29365±2257; D = 16916±1609; DT30 = 18791±2859; DT90 = 23425±3856 AAC; n = 5-9). Nesse trabalho mostramos que a suplementação com taurina corrige a hipoinsulinemia verificada em animais desnutridos alimentados (C = 4,97±0,34; CT90 = 3,56±0,52; D = 1,39±0,10; DT90 = 3,31±0,70 ng/ml; n = 5-8) bem como a responsividade de ilhotas isoladas a concentrações crescentes de glicose. Verificamos também que a taurina normaliza a secreção de insulina potencializada pelo carbacol (C = 9,4+0,8; CT90 = 12,4+0,7; D = 6,4+0,5; DT90 = 9+0,7 ng/ml; n = 12). As respostas secretórias foram observadas em conjunto com a regulação da expressão das proteínas SERCA3 (C = 100+21; CT90 = 174+17; D =96+90; DT90 = 149+11 % do C; n = 6), receptor muscarínico M3 (C = 100+24; CT90 = 155+80; D = 51+10; DT90 = 108+14 % do C; n = 5) e sintaxina 1 (C = 100+30; CT90 = 92+40; D = 50+12; DT90 = 77+11 % do C; n = 5) que participam do controle de diferentes etapas do processo de secreção de insulina. Por fim, verificamos que a suplementação com taurina melhorou o padrão de oscilação de íons Ca2+ após estímulo com glicose. Concluímos então que a suplementação com taurina por 90 dias restaura a sensibilidade das ilhotas à glicose e ao carbacol possivelmente pela regulação do fluxo de cálcio para as células b bem como pela modulação da expressão de proteínas que controlam o processo de secreção de insulina. / Abstract: Malnutrition still is a public health issue, especially in developing countries. Many studies correlate malnourishment during early life and the development of cardiovascular disease and type 2 Diabetes Mellitus on latter stages. Animal models of malnutrition reveal impaired insulin secretion stimulated by glucose and other insulinotropic agents as well as lower expression of key proteins for b cell function. The literature shows that taurine supplementation increases insulin secretion and regulates calcium dynamics on b cells. Male, 21 days old, wistar rats received diet containing 17% (C) or 6% (D) of protein. Both groups received taurine supplementation on the drinking water for 30 (CT 30 and DT 30) and 90 (CT90 and DT 30) days. Next we assessed biometric and biochemical parameters, glucose tolerance, glucose and carbacholstimulated insulin secretion, protein expression of muscarinic M3 receptor, Phospholipase C b2, SERCA3, Syntaxin 1 and, finally, we registered cytoplasmic Ca2+ after stimulus with glucose and carbachol. Protein restricted rats showed lower body weight, plasma proteins (C = 6,81±0,04; CT30 = 7,15±0,54; CT90 = 6,87±0,19; D = 5,35±0,24; DT30 = 5,37±0,28; DT90 = 5,70±0,09 g/dl; n = 3-5), albumin (C = 3,20±0,11; CT30 = 3,41±0,02; CT90 = 3,18±0,05; D = 2,74±0,07; DT30 = 2,49±0,09; DT90 = 2,67±0,04 g/dl; n = 5-9) and increased glucose tolerance (C = 30249±2682; CT30 = 37255±6691; CT90 = 29365±2257; D = 16916±1609; DT30 = 18791±2859; DT90 = 23425±3856 AUC; n = 5-9). Taurine supplementation had no effect upon nutritional status parameters and partially restored glucose tolerance and insulinemia to C levels. Taurine increased secretory response to glucose and carbachol (C = 9,4+0,8; CT90 = 12,4+0,7; D = 6,4+0,5; DT90 = 9+0,7 ng/ml; n = 12). It also increased protein expression of M3 receptor (C = 100+24; CT90 = 155+80; D = 51+10; DT90 = 108+14 % of C; n = 5), SERCA 3 (C = 100+21; CT90 = 174+17; D =96+90; DT90 = 149+11 % of C; n = 6) and syntaxin 1 (C = 100+30; CT90 = 92+40; D = 50+12; DT90 = 77+11 % of C; n = 5). Finally, taurine supplementation for 90 days improved Ca2+ dynamics when the islets were stimulated with glucose. In conclusion, these data show that taurine supplementation restores secretory responsiveness to glucose and carbachol possibly through Ca2+ dynamics modulation and increased expression of key proteins for insulin secretion. / Mestrado / Fisiologia / Mestre em Biologia Funcional e Molecular

(1) Introduction: Sulfonates, which can be diet- or host-derived, are a class of compounds detected in the gut, are involved in host–microbiome interactions and have several health effects. Our aim was to develop a method to quantify five of the sulfonates in the intestine and apply it in a simplified human microbiome model. These were taurine, its metabolic precursor cysteate and one of its degradation products isethionate, as well as sulfoquinovose and one of its most relevant degradation products 2,3-dihydroxy-1-propanesulfonate. (2) Methods: An extraction and sample preparation method was developed, without the need for derivatization. To detect and quantify the extracted sulfonates, a multiplexed LC-MS/MS-MRM method was established. (3) Results: The accuracy and precision of the method were within GLP-accepted parameters. To apply this method in a pilot study, we spiked either taurine or sulfoquinovose into an in vitro simplified human microbiota model with and without Bilophila wadsworthia, a known sulfonate utilizer. The results revealed that only the culture with B. wadsworthia was able to degrade taurine, with isethionate as an intermediate. After spiking the communities with sulfoquinovose, the results revealed that the simplified human microbiome model was able to degrade sulfoquinovose to 2,3-dihydroxypropane-1-sulfonate, which was probably catalyzed by Escherichia coli. In the community with B. wadsworthia, the 2,3-dihydroxypropane-1-sulfonate produced was further degraded by B. wadsworthia to sulfide. (4) Conclusions: We successfully developed a method for sulfonate quantification and applied it in a first pilot study.

Caffeine is the most widely consumed socially acceptable drug in the world and is commonly used for its ergogenic properties with demonstrated performance enhancing effects in endurance, power and strength based activities. Despite a wealth of evidence concluding a caffeine induced performance benefit, the direct effects of the drug on peripheral physiological processes have not been fully examined. Early works showed high dose caffeine has direct force potentiating effect on skeletal muscle, a notion that has only recently been confirmed by James et al. (2005) to also occur at a maximal human physiological concentration (70µM). The present research, using mouse muscle as a model for mammalian muscle in general, provides an in-depth assessment of the direct effect of physiological concentrations of caffeine on isolated skeletal muscle performance. This research uniquely: quantifies the dose response relationship; assesses the effects of caffeine on maximal and sub maximal muscle power output and fatigue; looks at the relationship between muscles with different fiber type compositions. As high concentrations of caffeine and taurine are a constituent of many energy drinks, the suggested interaction of these ingredients to further potentiate muscle mechanical performance was also assessed. The study further examines how mammalian muscle mechanical properties change over an age range of development to aged, and how this differs between muscles with predominantly different anatomical locations and functions. In light of this the age related direct effect of physiological concentrations of caffeine was assessed in order to examine whether the ergogenic benefit changed with age. The present results demonstrate a direct muscle performance enhancing effect of physiological concentrations of caffeine that is likely to promote greater benefit on long duration endurance based activities. Furthermore, the present study demonstrates that there is no direct effect of physiological concentrations of taurine and no further performance enhancing benefit when combined with caffeine. Finally this research uniquely highlights the muscle specific age related changes in mechanical performance and further indicates that the direct effect of caffeine changes with age.

Made available in DSpace on 2017-07-10T14:17:06Z (GMT). No. of bitstreams: 1 Luiz Caetano.pdf: 1961056 bytes, checksum: 05aefff53c5671351bad5763039bd60b (MD5) Previous issue date: 2013-11-10 / Among the several organic alterations arising from obesity, chronic inflammation is associated with the balance of cytokines TNF-&#945;, IL-1&#946;, IL-6, IL-2, IFNg, IL-4 and IL-10, and there is evidence the amino acid taurine (Tau) has anti-inflammatory effect. Therefore, this study investigated the inflammatory profile in plasma and retroperitoneal adipose tissue of MSG-obese rats, supplemented or not, with the TAU. Male Wistar rats received subcutaneous injections of MSG (4mg/kg body weight/day) or hyperosmotic saline during the first 5 days of life, composing the control (CON) and MSG groups. After 21 days, half of each group received TAU 2.5% in drinking water, and separated into 04 groups: CON, CON with TAU (CTAU), MSG and MSG with TAU (MTAU). At 120 days of age, the animals were euthanized. The MSG rats showed an increase in Lee Index, retroperitoneal and perigonadal fat pads deposition, insulin and triglycerides plasmatic concentrations and HOMA-IR, when compared to CON animals, showing that the treatment with MSG led to obesity. The TAU supplementation attenuated retroperitoneal fat deposition, as well as TG concentration. The MSG treatment did not alter the expression of JNK and I&#954;B&#945;. However, the supplementation with TAU increased 61% the expression of IkB&#945; in CTAU group compared to the CON and 107% in the MTAU animals compared to the MSG. The expression of TNF-&#945;, IL-1&#946; and IL-6 in the retroperitoneal adipose tissue were similar in the four groups of animals, as well as plasma concentrations of TNF-&#945;, IL-1&#946;, IL-6, IL-2, IFN&#947;, IL-4 and IL-10. It is possible to conclude that neonatal treatment with MSG does not influence the inflammatory profile of the animals. We also conclude that the TAU increased 61% of IkB&#945; protein expression in the control group and 107% in the MSG-obese animals, without affecting the inflammatory cytokines. Thus we suggest that TAU can exert their anti-inflammatory effects in adipose tissue, via NF-kB / Dentre as várias alterações orgânicas decorrentes da obesidade, está o processo inflamatório crônico associado ao balanço das citocinas TNF-&#945;, IL-1&#946;, IL-6, IL-2, IFN&#947;, IL-4 e IL-10, e, há evidências de que o aminoácido taurina (TAU) possui efeito anti-inflamatório. Assim, neste trabalho investigamos o perfil inflamatório plasmático e do tecido adiposo retroperitoneal de ratos obesos-MSG, suplementados ou não, com o aminoácido TAU. Ratos Wistar receberam injeções subcutâneas de MSG (4mg/kg de peso corporal/dia) ou salina hiperosmótica, durante os primeiros 5 dias de vida e foram distribuídos nos grupos MSG e CON, respectivamente. Após os 21 dias de vida, metade de cada grupo recebeu 2,5% de TAU na água de beber, sendo separados nos grupos CON, CON + TAU (CTAU), MSG e MSG + TAU (MTAU). Aos 120 dias de vida os animais foram eutanasiados. Ratos MSG apresentaram obesidade acompanhada de hipertrigliceridemia e resistência à insulina (RI). Todavia, não afetou a expressão de I&#954;B&#945; e JNK. A suplementação com TAU aumentou 61% a expressão do IkB&#945; no grupo CTAU em relação ao grupo CON e 107% nos animais MTAU em comparação com os obesos-MSG. As expressões de TNF-&#945;, IL-1&#946; e IL-6 no tecido adiposo retroperitoneal foram semelhantes nos 4 grupos de animais estudados, assim como as concentrações plasmáticas do TNF-&#945;, IL-1&#946;, IL-6, IL-2, IFN&#947;, IL-4 e IL-10. É possível concluir que o tratamento neonatal com MSG não influencia o perfil inflamatório dos animais. Concluímos também que a TAU aumentou a expressão proteica do IkB&#945; nos animais controle e MSG, sem afetar as citocinas inflamatórias. Desta forma sugerimos que a TAU possa exercer seus efeitos anti-inflamatórios no tecido adiposo, via NF-&#954;B

All living cell functions require an ongoing supply of energy derived from carbohydrates, lipids and proteins with their own pathways of breakdown. All of them end up in the oxidation of reduced coenzymes, yielding chemically-bound energy in the form of adenosine triphosphate (ATP). One broad definition of energy would be the capability to do work and, therefore, the more work that has to be done, the more energy is needed, which may under extreme conditions put the cell into a state of energy metabolic stress. This complex of problems has been examined in the present thesis, where individuals representing different degrees of training status, have been subjected to various types of stressful work-loads as regards intensity and duration. Meanwhile, the energy turnover has been monitored on different levels as whole body (organism)-, single organ/tissue-, cellular and molecular levels. Combined methodologies have been developed and utilized to examine carefully and in some detail energy expenditure and biochemical variables with study subjects under long-term, (outfield) physically and mentally stressful conditions. When the individuals were in a well-controlled energy balance, a diet rich in saturated fatty acids did not elicit any major metabolic stress signs concerning serum lipoproteins and/or insulin/glucose homeostasis during the test period including high volume and low intensity energy turn over. Only a slight decrease in the Apo-B / Apo-A1 ratio was observed, despite a period of totally sedentary life style among the participants. Mental stress combined with a varying energy balance during off-shore sailing races was shown to cause such an energy metabolic stress situation that development of abdominal obesity and signs of a metabolic syndrome in embryo affected the participants who were young, non-obese men and despite their fairly healthy lifestyle concerning the diet they were on and their physical activity habits. Even well-trained young individuals of both sexes, subjected to exhaustive endurance (high intensity exercise session), developed signs of insulin resistance with a deteriorated intracellular glucose availability leading to a supposed ion pump failure and a disturbed osmoregulation on a cellular level. Hence, they presented themselves as having acquired an energy metabolic stress like condition. In conclusion, an energy metabolic stress syndrome has been described, basically due to impaired fuelling of ion pumps with a cluster of signs and symptoms on single organ/tissue-, cellular and molecular levels manifested by muscular intracellular swelling, tendency towards erythrocyte shrinkage as a consequence of a relative insulin resistance concomitant with ion distribution disturbances (Gardos effect), oxidative stress and osmoregulatory taurine leakage.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The aim of this study was to evaluate the viability of using diets composed of ingredients of plant and animal origin in the feeding of post-larvae jundiá (Rhamdia quelen) and its influence on the development of animals. Experiments were carried out where it is tested in the first (E1) the total replacement (30%) and partial (15%) of the liver poultry for fish meal (FJ) and / or protein soy concentrate (CPS) on the standard diet containing 30% liver poultry. In the second (E2) substitution levels (5; 10; 15; 20 and 25%) of the liver by FJ and third (E3) of the liver levels for CPS replacement (15, 20, 25 and 30%) supplemented with taurine (CPST). Performance parameters were analyzed (weight, total length, condition factor, specific growth rate, daily weight gain, survival and product weight versus survival), muscle development (fiber diameter, number of fibers / mm² and total number of fibers ) and enzymatic activity (trypsin, chymotrypsin, lipase, amylase and maltase). The best performance of jundiá post-larvae was from 15FJ diets (E1 and E2) and 15CPST (E3). In the muscle development is found larger diameter and total number of fibers with the above mentioned diets. The development of the digestive system was not affected by the diets provided to post-larvae in this study. The enzymes assessed were already present and active at the first feeding. The enzyme activity varied during all experimental periods, with reduced activity of trypsin and chymotrypsin for diets with higher percentages of CPST over those with a lower percentage. According to the results we conclude that the combination of the sources of animal and plant improved the diet, helping improve the development of post-larvae of jundiá. / O objetivo deste trabalho foi avaliar a viabilidade da utilização de dietas compostas por ingredientes de origem animal e vegetal na alimentação de pós-larvas de jundiás (Rhamdia quelen) e sua influência no desenvolvimento dos animais. Foram realizados três experimentos onde testou-se no primeiro (E1) a substituição total (30%) e parcial (15%) do fígado de aves por farinha de peixe (FJ) e/ou concentrado proteico de soja (CPS) na dieta padrão contendo 30% de fígado de aves. No segundo (E2) níveis de substituição (5; 10; 15; 20 e 25%) do fígado por FJ e no terceiro (E3) níveis de substituição do fígado por CPS (15; 20; 25 e 30%) suplementado com taurina (CPST). Foram analisados parâmetros de desempenho (peso, comprimento total, fator de condição, taxa de crescimento específico, ganho em peso diário, sobrevivência e produto peso versus sobrevivência), desenvolvimento muscular (diâmetro da fibra, número de fibras/mm² e número total de fibras) e atividade enzimática (tripsina, quimotripsina, lipase, amilase e maltase). O melhor desempenho das pós-larvas de jundiá foi a partir das dietas 15FJ (E1 e E2) e 15CPST (E3). No desenvolvimento muscular encontrou-se maiores diâmetros e número total de fibras com as dietas citadas acima. O desenvolvimento do sistema digestório não foi prejudicado pelas dietas fornecidas às pós-larvas neste estudo. As enzimas analisadas já estavam presentes e ativas no momento da primeira alimentação. A atividade das enzimas oscilou durante todos os períodos experimentais, apresentando redução da atividade da tripsina e quimotripsina para as dietas com maiores percentuais de CPST em relação aquelas com menor percentual. De acordo com os resultados concluimos que a combinação das fontes de origem animal e vegetal aprimorou a dieta, contribuindo para melhorar o desenvolvimento das pós-larvas de jundiá.

Chapter 1: Amphiphilic self-assembled systems as nanocarriers Nanocarriers are the nanometric size molecular assemblies that are used for the transport of small molecules into their non-solvating environments. These systems find major applications as drug delivery systems (DDS) in pharmacological research. These drug delivery systems improves solubility and stability of the drug molecules through encapsulation and also offer additional advantages like target specificity and stimuli responsive release of the drug molecules. Several types of DDS are reported in the literature, which can be prepared by a variety of processing techniques. Of these, molecular self- Chart 1: Developments in the design of amphiphilic nanocarriers assembly has attained considerable attention due to its greater tunability and control in the preparation of nanocarriers. In this chapter we discussed about the amphiphilic nanocarriers which are prepared through self-assembly of amphiphiles through hydrophobic interactions. Several developments in the area of amphiphilic nanocarriers such as di-block polymeric systems, dendritic systems and core-shell architectures are also mentioned. We also highlighted some recent developments in the design of amphiphilic nanocarriers through supramolecular interactions and advantages of such systems. Chapter 2: Bile acid derived dendrons and their application as nanocarriers Host-guest chemistry is well known for dendritic systems. To understand the influence of steric crowding, dendritic effect and importance of number of hydroxyl groups on the bile acid backbone in the host-guest chemistry of bile acid dendrons, we designed and synthesized a new series of C3 symmetric systems and studied the above-mentioned objectives through extraction of polar dyes into nonpolar media. Dye extraction experiments performed using trimeric molecules suggested that only the cholate derivatives (3 and 4) showed considerable extraction of the polar dyes into chloroform; deoxycholate derivatives did not show any extraction, thus emphasizing the importance of the number of hydroxyl groups for dye extraction in these molecular architectures. The effect of steric crowding at the core of these trimeric molecules was shown by efficient extraction of the dyes with the triethylbenzene core (4) compared to the benzene core (3). Greater influence of the aggregates in the case of triethylbenzene core on the extracted dye was also manifested in the Chart 2: Structures of the designed molecules 1-6 value of the induced circular dichroism signal. Surprisingly, a higher analogue in these molecular architectures showed lesser efficiency in dye extraction (on a per bile acid residue basis) compared to the trimers, suggesting a more compact structure for the higher analogue. This was supported by molecular modeling studies. Generality of these systems as nanocarriers for hydrophilic dyes was investigated by screening several other dyes and polar molecules, which are diverse in their structure and functionalities. All these experiments suggested a dependency of the extraction profile on the size of the dye molecule. This was also examined by dynamic light scattering studies, which showed larger size and wider distribution in the size of the aggregates in the case of larger dyes. We also demonstrated selective extraction of a single dye molecule from a blended food color (apple green) using one of the trimer (4) and demonstrated solvent dependent morphological changes in these compounds using electron microscopy. The self-assembly of these amphilic molecules at the air-water interface was studied through Langmuir monolayer studies. Chart 3: Structure of polar guest molecules (Cresol red (7). Erioglaucine (8), Eriochrome black T (9),) phenyl β-D-glucopyranoside (10) and Eosin B (11) Chapter 3: Design and synthesis of bile acid derived surfactants: Study of their aggregation and potential applications Bile acids are facially amphiphilic systems and their amphiphilicity can be improved by attaching polar groups on the bile acid back bone or by synthesizing oligomeric systems which show better self-assembly compared to their monomeric units. To study and improve the amphiphilicity of bile acids, we designed and synthesized a new tripodal surfactant system, with a phosphine oxide based central core to which the bile acids were attached through the C-3 position using click chemistry. Our molecular design also offers added advantage of studying the influence of the stereochemistry at the C-3 position on the aggregation of these molecular architectures. We synthesized trimeric systems with both cholic and deoxycholic acids attached to the central phosphine oxide core with α and β stereochemistry at the C-3 position. Aggregation of these molecules was studied by surface tension measurements, dye extraction studies and NMR. All these compounds showed aggregation at micromolar concentrations. NMR studies suggested changes in the structure of the aggregates at higher temperature and these changes were studied by DLS, which suggested thermodynamically stable monodispersed aggregates for cholic acid derivatives (13 and 15) at higher temperature. These aggregates are stable even after cooling to room temperature and with time. The aggregates of these derivatives were also characterized by atomic force microscopy. Gelation was observed in the case of α derivatives (12 and 13) in phosphate buffer (0.1 M) at pH 7.5 for both deoxy and cholic derivatives, which emphasized the influence of stereochemistry at C-3 position in these architectures. These gels were characterized by rheology experiments. Finally, the possible utility of these micellar systems as model systems to study photophysical processes was demonstrated through lanthanide sensitization experiments in these micellar solutions. Chart 4: Structure of the designed molecules Chapter 4: Synthesis of oligomeric bile acid-taurine conjugates: Study of their aggregation and efficiency in cholesterol solubilization Bile acids are bio-surfactants that are used for the emulsification of fats, vitamins etc. in our body. Bile salts also solubilize the excess cholesterol in our body through mixed micelle formation in the bile and when the bile gets saturated with cholesterol, it leads to cholesterol gallstone formation, which needs to be treated. Ursodeoxycholic acid (UDCA) is used as drug in some cases for the solubilization of (small) cholesterol gallstones, even though the efficiency to solubilize cholesterol is less for UDCA compared to the other bile acids (UDCA is less toxic than the others). So there is a need to develop new cholesterol solubilizing agents. Since oligomeric systems can aggregate better, we designed and synthesized two tetramer taurine conjugates, which differ in the spacer between the bile acid units. Since these conjugates are not soluble in water, their solubility and aggregation was studied in 10% MeOH/Water using pyrene fluorescence experiments. Aggregation studies suggested better aggregation for these molecules compared to their monomeric analogues. These aggregates were also characterized byDLS and electron microscopy. These systems were subsequently studied as nanocarriers for liphophilic dye molecules into aqueous media. Finally, the influence of oligomeric effect in cholesterol solubilization was investigated by cholesterol solubilization studied using these two tetramer taurine compounds and a control, sodium taurocholate. These studies suggested efficient solubilization of cholesterol by oligomers compared to monomeric analogues.(For structural formula pl see the abstract file)

Orientador: Alexandre Leite Rodrigues Oliveira / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-18T14:50:45Z (GMT). No. of bitstreams: 1 Inacio_RodrigoFabrizzio_M.pdf: 6359187 bytes, checksum: 67cbbe4ec81cf9bbd0b48c1dd3b13843 (MD5) Previous issue date: 2011 / Resumo: Foi demonstrado que a regulação positiva do MHC I por tratamento exógeno com interferon beta (IFN beta) influencia no processo de eliminação das sinapses. Também, o aminoácido taurina mostrou ter influencia positiva na sobrevivência e plasticidade neuronal. No entanto, o estabelecimento de um modelo in vitro para estudo do processo de formação/eliminação sináptica e sua relação com a expressão de MHC I ainda não foi proposto. Portanto, o presente estudo tem como objetivo investigar os efeitos do tratamento com IFN e taurina, sozinhos ou diluídos em meio glial (derivado do glioma NG97), na expressão de MHC classe I e na formação de sinapses em células PC12. Células PC12 foram tratadas com NGF para indução do fenótipo semelhante a neurônio e as culturas estabelecidas foram submetidas ao tratamento com IFN beta (500 e 1000 IU) e taurina (0.025 and 0.050mg/mL) por 15 dias em meio normal e por 10 dias em meio condicionado. Finalizado o período de cultivo, as células foram fixadas e processadas para imunocitoquímica com anticorpos anti-MHC I (OX18) e anti-sinaptofisina. A imunomarcação foi mensurada com o software Image J. Nesse contexto, quatro campos representativos foram usados, a partir de cada poço de cultivo. Os resultados mostraram que o IFNbeta (500UI) e a taurina (0.025 mg) modulam a expressão de MHC em células PC12, especialmente após 10 dias de tratamento. IFN e taurina apresentaram efeitos opostos, sendo que o IFN induz o aumento do MHC I, enquanto a taurina causa sua diminuição. Em ambos os casos, o aumento das doses causa degeneração da cultura. Interessantemente, a regulação diferencial do MHC I ocorreu paralelamente a um aumento ou diminuição da plasticidade sináptica, respectivamente. O uso do meio condicionado de NG97, juntamente com IFNbeta ou taurina, leva a uma diminuição da estabilidade sináptica. De uma maneira geral, os presentes dados indicam que as células PC12 podem ser usadas como modelo in vitro para estudos de modulação de MHC I e plasticidade sináptica. Também, reforçam o papel do IFNbeta na eliminação sináptica e indicam que a taurina é capaz de aumentar a formação da rede sináptica / Abstract: It has been demonstrated that MHC I up regulation by exogenous treatment with interferon beta (IFN beta) influences the glial reaction and the synaptic elimination process. Also, the amino acid taurine has been shown to positively influence neuronal survival and plasticity. Nevertheless, the establishment of an in vitro model for studying the synaptic formation/elimination process and its relationship with MHC I expression has not yet been proposed. Therefore, the present study aimed to investigate the effects of the IFN beta and taurine treatments, alone or diluted in glial medium (derived from the NG97 gliome), on the expression of MHC I and synaptic formation in PC12 cells. Established cultures were subjected to the IFN beta (500 and 1000 IU) and taurine treatments (0.025 and 0.050mg/ml) for 5 and 10 days. Finally the cells were fixed and processed for immuno-cytochemistry with antisera against MHC I (OX18) and synaptophysin. The results were compared with control cultures only treated with basal or conditioned medium. The results showed that IFNbeta (500 IU) and taurine (0.025 mg) modulated the MHC I expression in PC12 cells, especially after 10 days of treatment. IFN and taurine displayed opposite effects, such that IFN induced MHC I up regulation, while taurine induced down regulation. In both cases, the highest doses caused culture degeneration. Interestingly, the differential regulation of MHC I was paralleled by enhancement or a decrease in synaptic plasticity, respectively. The use of the NG97 conditioned medium together with IFNbeta or taurine led to a decrease in synaptic stability. Altogether, the present data indicate that PC12 cells may be used as an in vitro model for studying MHC I modulation and synaptic plasticity. It also reinforced the role of IFNbeta on synaptic elimination and indicated that taurine was able to increase the synaptic network formation / Mestrado / Anatomia / Mestre em Biologia Celular e Estrutural

[pt] Câncer é o nome dado a um conjunto de mais de 100 doenças e entre as possibilidades de tratamento está a quimioterapia. Após a descoberta das propriedades antitumorais do complexo de coordenação comumente chamado cisplatina, um dos compostos mais utilizados em neoplasias malignas, o estudo dos complexos metálicos teve um grande impulso e alguns compostos promissores de cobre(II) já foram desenvolvidos. Por outro lado, bases de Schiff derivadas de aminas e aldeídos aromáticos têm apresentado uma ampla aplicação em muitas áreas de pesquisa, sendo que algumas são farmacologicamente utilizadas na terapia anti-hipertensiva, hipnótica e antineoplásica. Neste contexto, no presente trabalho, foi sintetizado e caracterizado um ligante imínico binucleante sulfonado derivado da taurina, já conhecido na literatura: (LIT) e, a partir deste, seus complexos inéditos de Zn(II), Cu(II) e Ni(II), que foram caracterizados pelas seguintes técnicas: espectroscopia vibracional e eletrônica, análise elementar de CHNS, análise termogravimétrica, espectroscopia de ressonância paramagnética eletrônica (EPR) e modelagem molecular computacional. Os novos compostos obtidos neste trabalho são, a saber: composto (1), composto (2) e composto (3), em que LIT representa uma forma parcialmente hidrolisada de LIT. Os complexos 1 e 2 são os primeiros compostos binucleares do ligante LIT descritos. Neles, os centros metálicos são tetracoordenados e apresentam uma ponte exógena acetato coordenada nas formas bidentada, para o composto 1, e monodentada, para 2. Esta diferença na coordenação da ponte se dá, provavelmente, devido aos distintos arranjos geométricos em torno dos metais: enquanto o zinco apresenta um arranjo tetraédrico, o cobre mostra um do tipo quadrático. O complexo 3 é binuclear, composto por um dímero altamente simétrico envolvendo, como dito acima, uma forma parcialmente hidrolisada de LIT. Os centros metálicos são hexacoordenados, ligados por pontes endógenas fenólicas. Tanto 2 quanto 3 são silenciosos ao EPR. Foi realizado também um ensaio de toxicidade aguda em Artemia salina para as espécies hidrossolúveis LIT e complexo 1. Este ensaio mostra boa correlação com a atividade citotóxica para alguns tumores sólidos humanos. / [en] Cancer is a name given to a set of more than 100 diseases and among the possibilities of treatment is chemotherapy. After the discovery of the antitumor properties of the coordination complex commonly called cisplatin, it is one of the compounds most used in malignancies. The study of metal complexes had a big boost and some promising copper(II) compounds have been developed. Furthermore, the Schiff bases derived from aromatic aldehydes and amines present a wide range of applications in many areas of research, some of which are pharmacologically used in antihypertensive, hypnotic, and antineoplastic therapy. In this context, in the present study, we synthesized and characterized a binucleating imine ligand, derivative of taurine, already known in the literature: (LIT). New dinuclear Zn(II), Cu(II) and Ni(II) complexes of this ligand were synthesized, and were characterized by the following techniques: vibrational and electronic spectroscopies, CHNS elemental analysis, thermogravimetric analysis, electron paramagnetic resonance (EPR) and computational molecular modeling. The new compounds obtained in this work are: composite (1), composite (2) composite (3), where LIT represents a LIT partially hydrolyzed form. Complexes 1 and 2 are the first dinuclear compounds of the LIT ligand described. In these, metal centers are tetracoordinated, with the presence of an exogenous acetate bridge, which shows a bidentate coordination mode for compound 1 and a monodentate coordination pattern for 2. This difference occurs probably due to different geometrical arrangements around the metal centers: while zinc has a tetrahedral coordination geometry, copper shows one of the square planar type. On the other hand, compound 3 a dinuclear complex, which is composed of a highly symmetric dimer involving, as mentioned above, a partially hydrolyzed form of LIT. The hexacoordinated metal centers are connected by two endogenous phenolic bridges. Both 2 and 3 are EPR silent. An acute toxicity test on Artemia salina shrimp was also carried out for the hydro-soluble species LIT and 1. This assay shows good correlation with cytotoxic activity for some human solid tumors.