Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2015-12-04T19:34:33Z No. of bitstreams: 1 RaniereFagundesDeMeloSilveira_TESE.pdf: 3512000 bytes, checksum: 19fe99baef9b0082648ee82a63fbf577 (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2015-12-09T23:17:35Z (GMT) No. of bitstreams: 1 RaniereFagundesDeMeloSilveira_TESE.pdf: 3512000 bytes, checksum: 19fe99baef9b0082648ee82a63fbf577 (MD5) / Made available in DSpace on 2015-12-09T23:17:35Z (GMT). No. of bitstreams: 1 RaniereFagundesDeMeloSilveira_TESE.pdf: 3512000 bytes, checksum: 19fe99baef9b0082648ee82a63fbf577 (MD5) Previous issue date: 2014-10-20 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico - CNPq / A prospec??o de polissacar?deos farmacologicamente ativos provenientes de subprodutos agr?colas ainda ? uma pr?tica pouco explorada no meio cient?fico. Dessa forma, este trabalho pretendeu ampliar o conhecimento relacionado ?s atividades farmacol?gicas de polissacar?deos extra?dos do sabugo de milho. A partir da farinha de sabugo de milho, um extrato polissacar?dico foi obtido ao combinar ondas de ultrassom em meio alcalino, e ao final do processo o produto foi denominado de EPSM. Esse extrato foi caracterizado fisico-quimicamente e, atrav?s de ensaios in vitro, foi avaliado como agente antioxidante, citot?xico, anticoagulante e imunomodulator. Os resultados indicaram que o EPSM apresentou significativa a??o quelante de metal, al?m de n?o apresentar efeito t?xico para c?lulas de linhagem normal, mas evidenciar efeito citot?xico contra as c?lulas tumorais HeLa, quando causou a morte celular por apoptose. Em adi??o, outros efeitos farmacol?gicos foram observados, o EPSM diminuiu a produ??o de ?xido n?trico (NO) por macr?fagos ativados, e estendeu o tempo de coagula??o sangu?nea quando avaliado pelo ensaio de APTT. Posteriormente, fra??es metan?licas, etan?licas e cet?nicas foram obtidas a partir do fracionamento dos polissacar?deos presentes no EPSM. Foram obtidas cinco fra??es metan?licas, seis fra??es etan?licas e duas cet?nicas; e todas avaliadas quanto ?s atividades antioxidante, citot?xica, anticoagulante e imunomodulat?ria. Dentre as fra??es, E1.4 exibiu significativo efeito qualante de metal, a??o t?xica em c?lulas HeLa por indu??o da apoptose, reduziu a produ??o de NO por macr?fagos ativados, e ampliou o tempo de coagula??o sangu?nea. Esses resultados sinalizaram que os polissacar?deos farmacologicamente ativos do EPSM estariam presentes na fra??o E1.4. A partir do fracionamento da E1.4 foram obtidas seis subfra??es polissacar?dicas com tamanhos distintos: -3; 3-10; 10-30; 30-50; 50-100 e +100 KDa. Cerca de 80% dos polissacar?deos de E1.4 apresentou tamanho inferior a 10 KDa, e todas as subfra??es apresentaram mais de 61% de a??car em suas composi??es qu?micas. Essas subfra??es exibiram diferentes composi??es monossacar?dicas, mas todas apresentaram xilose. As subfra??es evidenciaram distintos efeitos nos ensaios farmacol?gicos in vitro. As subfra??es de menor tamanho (< 30 KDa) demonstraram maior atividade quelante de metal e maior atividade citot?xica contra c?lulas tumorais. As intermedi?rias (entre 30 e 100 KDa) diminu?ram mais a produ??o de NO por macr?fagos ativados, mas a de maior tamanho (+100 KDa) modulou um maior n?mero de citocinas inflamat?rias, e demonstrou um maior efeito anticoagulante. Portanto, a an?lise conjunta dos resultados indica que os polissacar?deos farmacologicamente ativos do ESPM s?o heteroxilanas e foram concentrados nas subfra??es de E1.4, al?m disso, os efeitos farmacol?gicos dessas heteroxilanas dependem do seu tamanho molecular. / Prospecting pharmacological active polysaccharides from agricultural byproducts, such as corncobs, is an underexplored practice in the scientific community. Thus, this work aims to expand knowledge about pharmacological activities of polysaccharides extracted from corncobs. From corn cob flour a extract was obtained by ultrasound waves in an alkaline medium, and the end of the process the product was termed PECC (polysaccharidic extract from corncobs). This extract was physicochemical characterized and evaluated by in vitro assays as an antioxidant, cytotoxic, anticoagulant and imunomodulator agent. Results indicated significant activity metal chelating by PECC, and the use of PECC in cell culture cells showed no toxic effects to normal cell lines, but toxic action against HeLa tumor cells due promoting cell death by apoptosis. In addition, other pharmacological effects were observed, the PECC decreased nitric oxide (NO) production by activated macrophages, and prolonged blood clotting time through APTT assay. Then methanolic, ethanolic and ketone fractions were obtained from fractionation of PECC polysaccharides. Five methanolic fractions, six ethanolic fractions and two ketones were obtained; and all fractions were evaluated for antioxidant, cytotoxic, anticoagulant, immunomodulatory activities. E1.4 fraction exhibited significant metal chelating effect, a toxic action to induce apoptosis in HeLa cells, decreased NO production by activated macrophages, and extended blood clotting time. These results showed that the PECC pharmacological active polysaccharides would be present in the fraction E1.4. From fractionation of E1.4 polysaccharide six subfractions with different sizes were obtained: <3; 3-10; 10-30; 30-50; 50-100 and >100 KDa. About 80% of E1.4 polysaccharides had lower size to 10 KDa, and all the subfractions showed over 61% sugar in their chemical compositions. These subfractions exhibited different monosaccharide compositions, but xylose was presented in all of them. The subfractions exhibited distinct pharmacological effects in in vitro assays. Smaller subfractions (<30 KDa) had highest metal chelating activity and greater toxic action in tumor cells. The intermediate fractions (between 30-100 KDa) decreased more NO production of activated macrophages, for other side, the larger size (>100 KDa) modulated a greater number of inflammatory cytokines, and the had greatest anticoagulant effect. Therefore, when analyzing all the results together it is evident that the PECC pharmacological polysaccharides are heteroxylans, and were concentrated in E1.4 fraction, and heteroxilanas pharmacological effects depends on their molecular size. Thus, corncobs could be used as source from molecules with biotechnology potential

Submitted by Celso Magalhaes (celsomagalhaes@ufrrj.br) on 2018-09-12T18:08:44Z No. of bitstreams: 1 2017 - Julia Dias Villas Boas.pdf: 1378660 bytes, checksum: 2a1537e1b92056b0327c8e50b73feb34 (MD5) / Made available in DSpace on 2018-09-12T18:08:44Z (GMT). No. of bitstreams: 1 2017 - Julia Dias Villas Boas.pdf: 1378660 bytes, checksum: 2a1537e1b92056b0327c8e50b73feb34 (MD5) Previous issue date: 2017-04-28 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico - CNPq / The horse has a natural predisposition for the sport, however, its use in competitions can result in stress related problems that impair its sporting performance and especially its health. In this way it is fundamental not only to understand how the different risk and resilience factors to different stressors influence the response to stress, but also to develop strategies that can prevent or minimize the deleterious effects of stress. In this sense, acupuncture is an ancient technique of Traditional Chinese Medicine that has been used in the treatment and prevention of stress-related diseases. The present study proposed the use of two models of stress: one physical (physical exercise) and another psychological (startle model) to verify the reactivity to the stress of athletes horses. In addition, it was also evaluated if horses of different sporting modalities present different psychological stress responses and if acupuncture can alter the responses to physical stress. In the experiment 1, 16 Thoroughbred race horses were submitted to a exercise in the field of high intensity and short duration (12 m / s, 4 min). The RR intervals for analysis of the Heart Rate Variability were acquired through the Polar Equine ? heart rate monitor and blood samples were collected before and immediately after 2h, 4h, 6h, and 24h after exercise. The exercise promoted autonomic alterations in the sympatho-vagal balance since it significantly increased the low frequency component (LF), the heart rate and the LF / HF ratio, and decreased the high frequency component (HF) (p <0.01). There was an increase in hematocrit, plasma proteins, glucose and plasma lactate immediately after exercise (p <0.001). There was an increase (p <0.01) in serum cortisol values after 30 minutes, returning to baseline values after 60 min. However, no significant difference was observed in plasma cytokines IL-1? and IL-6 between moments after exercise and baseline. In experiment 2, horses of the experimental group 1 after exercise were randomly divided into two groups: CTL (C2): Control (without manipulation) and ACUP (C2)): animals submitted to weekly sessions of acupuncture at points VG1, C7, VG20 and B52 for 10 weeks. After the treatment period the animals repeated the same exercise and the same parameters were analyzed. Acupuncture reduced the LF / HF ratio, promoting a faster recovery of the animals, showing no influence on the other parameters analyzed. In the experiment 3, 24 equines were used, from three equestrian modes: Polo (PSI) (n = 9), Dressage (Brazilian Horse Riding) (n = 6) and Endurance (n=6) were subjected to the experimental model of startling through the abrupt opening of an umbrella. The results showed that endurance horses presented a significantly less intense startle-induced autonomic response than Polo and Dressage horses (lower LF / HF ratio at the time of the jump), paradoxically Enduro horses have cortisol levels in response in response to the startle than Polo horses. However, there was no difference between the modalities in the behavioral response after the startle, and Polo horses had significantly higher baseline levels of cortisol than the other modalities and did not change their cortisol levels in response to stress. Thus, we can conclude that 1) the exercise in the field of high intensity and short duration promoted adaptive changes characteristic of stress, being able to be used in studies of reactivity to stress in race horses; 2) acupuncture has a modulating effect on the stress-induced autonomic response in athletic horses, and 3) the equestrian modalities of Dressage, Polo and Endurance present different stress reactivity / O cavalo tem uma predisposi??o natural para o esporte, no entanto, o seu uso em competi??es pode resultar em problemas relacionados ao estresse que prejudicam seu desempenho esportivo e principalmente a sua sa?de. Desta forma ? fundamental n?o apenas entender como os diferentes fatores de risco e de resili?ncia a diferentes estressores influenciam a resposta ao estresse, como tamb?m desenvolver estrat?gias que possam prevenir ou minimizar os efeitos delet?rios do estresse. Neste sentido a acupuntura ? uma t?cnica milenar da Medicina Tradicional Chinesa que tem sido utilizada no tratamento e preven??es de doen?as relacionadas ao estresse. O presente estudo prop?s o uso de dois modelos de estresse: um f?sico (exerc?cio f?sico) e outro psicol?gico (modelo de sobressalto) para verificar a reatividade ao estresse de cavalos atletas. Al?m disso, tamb?m foi avaliado se cavalos de diferentes modalidades esportivas apresentam respostas ao estresse de psicol?gico distintas e se acupuntura pode alterar as respostas ao estresse f?sico. No experimento 1, 16 equinos de corrida da ra?a Puro Sangue Ingl?s foram submetidos ao exerc?cio a campo de alta intensidade e curta dura??o (12 m/s, 4min). Os intervalos RR para an?lise da Variabilidade da Frequencia Card?aca foram adquiridos atrav?s do frequenc?metro card?aco Polar Equine? e as amostras de sangue foram coletadas antes e, imediatamente, 2h, 4h, 6h, e 24h ap?s o exerc?cio. O exerc?cio promoveu altera??es auton?micas no balan?o simpato-vagal uma vez que aumentou significativamente o componente de baixa frequ?ncia (LF), a frequ?ncia card?aca e a raz?o LF/HF e diminuiu o componente de alta frequ?ncia (HF) (p < 0.01). Houve aumento do hemat?crito, das prote?nas plasm?ticas, glicose e lactato plasm?tico imediatamente ap?s o exerc?cio (p < 0.001). Houve aumento (p<0.01) nos valores s?ricos de cortisol ap?s 30 minutos, retornando aos valores basais ap?s 60min. No entanto, n?o foi observado diferen?a significativa nas citocinas plasm?ticas IL-1? e IL-6 entre os momentos ap?s exerc?cio e o momento basal. No experimento 2: os equinos do experimento 1 ap?s o exerc?cio foram aleatoriamente divididos em dois grupos: CTL (C2): Controle (sem manipula??o) e ACUP (C2)ACUP (C2): animais submetidos a sess?es semanais de acupuntura nos pontos VG1, C7, VG20 e B52 durante 10 semanas. Ap?s o per?odo de tratamento os animais repetiram o mesmo exerc?cio e foram analisados os mesmos par?metros. A acupuntura reduziu a raz?o LF/HF, promovendo uma recupera??o mais r?pida dos animais n?o apresentando influ?ncia nos demais par?metros analisados. No experimento 3, foram utilizados 24 equinos, pertencentes a tr?s modalidades equetres: P?lo (ra?a PSI) (n=9), Adestramento (ra?a Brasileiro de Hipismo) (n=6) e Enduro (Puro Sangue ?rabe) (n=9) submetidos ao modelo experimental de sobressalto atrav?s da abertura abrupta de um guarda-chuva. Os resultados mostraram que cavalos de enduro apresentaram resposta auton?mica induzida pelo sobressalto significativamente menos intensa que cavalos de Polo e Adestramento (menor raz?o LF/HF no momento do sobressalto), paradoxalmente cavalos de Enduro possuem n?veis de cortisol em resposta ao sobressalto mais altos que cavalos de Polo. N?o houve diferen?a entre as modalidades na resposta comportamental ap?s o sobressalto, no entanto cavalos de P?lo apresentam n?veis basais de cortisol significativamente mais altos que as demais modalidades e n?o variaram seus n?veis de cortisol em resposta ao estresse. Dessa forma, podemos concluir que 1) o exerc?cio a campo de alta intensidade e curta dura??o promoveu altera??es adaptativas caracter?stica de estresse, podendo ser utilizado em estudos de reatividade ao estresse em cavalos de corrida; 2) a acupuntura tem um efeito modulador da resposta auton?mica induzida pelo estresse em cavalos atletas e 3) as modalidades equestres de Adestramento, Polo e enduro apresentam reatividade ao estresse distintas