Made available in DSpace on 2016-04-28T14:59:00Z (GMT). No. of bitstreams: 1 Cleiton Mateus Sousa.pdf: 6410770 bytes, checksum: 26a93d90d24ec7420524c2e0478aec38 (MD5) Previous issue date: 2008-07-25 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / The determination of plant hormones can be an excellent tool for to study the plant development. Today, the complexity of methods, equipment and reagents of high cost limit the use with practice of routine. Some advantages of methods immunoassay can exceed those limitations, showing a potential of practical use in determination of plant hormones. The test Enzyme-Linked Immunosorbent Assay (ELISA) is utilized in detection of molecules with low molecular weight. However, the ELISA requires specific antibodies against molecules of interest, in the case, plant hormones. Today in the market there is not available antibodies for the determination of hormonal molecules. Faced with that, proposed be produced and characterize antibodies against molecules of AIA, 2ip and zeatin and subsequently, for use the detection those molecules. For that, the hormonal molecules were conjugated with protein (BSA) for subsequent immunization of chicken or rabbits. From the serum of rabbits or yolks eggs the antibodies were purified and characterized through the test of precipitation of double diffusion, SDS-PAGE and test ELISA. The antibodies that presented better results were utilized in detection of plant hormones molecules in samples of plantlets in vitro of gerbera. The test of precipitation of double diffusion revealed that the antibodies productized were capable of will detect the molecule in study. Through in the SDS-PAGE was verified that the antibodies obtained in eggs yolks of chicken presented superior purity than them obtained in serum of rabbits. From the results of the test ELISA, observed itself that the antibodies against AIA did not present potential of practical use. The detection of plant hormones in samples of tissue stayed restricted to 2ip and zeatin. The detection of those two molecules in crude extract, obtained from plantlets in vitro of gerbera, revealed that the level endogenous of zeatin was higher of level 2ip. Plantlets with six weeks after multiplication presented higher level of zeatin than plantlets with one week after multiplication. For the 2ip, that difference was not evident. The use of antibodies obtained in eggs yolks of chicken permitted the determination of zeatin and 2ip in samples plant utilizing the test ELISA. / A determina??o do n?vel hormonal end?geno pode ser uma excelente ferramenta para estudar o desenvolvimento vegetal. Atualmente, fica limitada devido a complexidade das metodologias adotadas, uma vez que demandam equipamentos e reagentes de alto custo e ainda apresentam baixo rendimento. Por outro lado, os ensaios imunoenzim?ticos possuem algumas vantagens que superam essas limita??es, demonstrando um potencial de uso pr?tico na dosagem de horm?nios vegetais. Entre os ensaios imunoenzim?ticos, destaca-se o teste Enzyme-Linked Immunosorbent Assay (ELISA), o qual vem sendo utilizado na detec??o de mol?culas com baixo peso molecular. No entanto, o teste ELISA exige anticorpos espec?ficos e que sejam capazes de reconhecer as mol?culas de interesse, no caso os horm?nios vegetais. Hoje no mercado n?o h? anticorpos dispon?veis para a determina??o das mol?culas hormonais. Diante disso, prop?s-se produzir e caracterizar anticorpos contra mol?culas de AIA, 2ip e zeatina e posteriormente, us?-los na detec??o dessas mol?culas. Para isso, as mol?culas hormonais foram conjugadas com prote?na (BSA) para posterior imuniza??o de galinhas poedeiras ou coelhos. A partir do soro de coelhos ou de gemas de ovos de galinhas os anticorpos foram purificados e caracterizados atrav?s do teste de imunodifus?o, SDSPAGE e teste ELISA. Os anticorpos que apresentaram melhores resultados foram utilizados na detec??o de mol?culas hormonais em amostras de tecidos de pl?ntulas mantidas in vitro. O teste de imunodifus?o revelou que os anticorpos obtidos foram capazes de detectarem a mol?cula em estudo. Atrav?s do SDS-PAGE verificou-se que os anticorpos obtidos em gemas de ovos de galinhas apresentaram maior pureza que os obtidos em coelhos, sugerindo que os mesmos possuem maior potencial de uso pr?tico. A partir dos resultados do teste ELISA, observou-se que os anticorpos contra AIA n?o apresentaram potencial de uso pr?tico na determina??o dessa mol?cula em amostras vegetais. Sendo assim, a detec??o de mol?culas hormonais em amostras de tecidos vegetais ficou restrita a 2ip e a zeatina. A detec??o dessas duas mol?culas em extrato bruto, obtidos a partir de pl?ntulas de g?rbera mantidas in vitro, revelou que o n?vel end?geno de zeatina foi superior ao n?vel de 2ip. Pl?ntulas com seis semanas ap?s a repicagem apresentaram maior n?vel de zeatina do que pl?ntulas rec?m repicadas, enquanto para o 2ip, essa diferen?a n?o foi evidente. O uso de anticorpos obtidos em gemas de ovos de galinhas permitiu a detec??o e quantifica??o de zeatina e 2ip em amostras vegetais utilizando o teste ELISA.

Submitted by Celso Magalhaes (celsomagalhaes@ufrrj.br) on 2017-05-03T14:02:59Z No. of bitstreams: 1 2014 - Natalia Soares Carvalho de Souza.pdf: 1833535 bytes, checksum: 4574c94ab4a477d9f8d0ddf31d6b4069 (MD5) / Made available in DSpace on 2017-05-03T14:03:00Z (GMT). No. of bitstreams: 1 2014 - Natalia Soares Carvalho de Souza.pdf: 1833535 bytes, checksum: 4574c94ab4a477d9f8d0ddf31d6b4069 (MD5) Previous issue date: 2014-08-26 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Premenopausal women are less prone to develop cardiovascular diseases than men and this advantage do not persist in postmenopausal women. Thus herein we aimed to investigate the gender difference and the estrogen influence in cardiac function, fluid balance and thyroid status, in Wistar rats subjected to experimental model of myocardial infarction (MI). In the first step, adult male (n = 18) and female (n = 21) rats underwent experimental MI (MIm and MIf,) or sham-operation (ShamM and ShamF) respectively. One and four weeks post-MI rats were placed in metabolic cages, subjected to echocardiography (ECHO), electrocardiography and then euthanized for blood sample collection and tissue collection (heart, lung and liver). In the second step female rats were ovariectomized (n = 24) or continued intact (n = 21), two weeks later they were subjected to MI (MIOVX and MIINT, respectively) or sham operation (ShamOVX and ShamINT). Four weeks post-MI, they were subjected to the same evaluations of the first step, not only the electrocardiography. In the third step, female rats were subjected to ovariectomy and treated with estrogen (E2) (n = 13) or vehicle (n = 22). Two weeks later they underwent experimental MI (MIOVX+E2 and MIOVX+Veh). Four weeks post-MI they were subjected to the same evaluations of the second step. Male infarcted rats developed cardiac dysfunction (shortening fraction, SF, reduction, ~ 70%) and fluid homeostasis changes (sodium intake increasing, ~ 146% and urinary volume reduction, ~55%) earlier than female, in the first week post-MI while female presented these changes (SF reduction, ~28% and sodium intake increasing, ~143%) only in the fourth week and attenuated compared to male. MIM showed reduction in LF/HF ratio (~70%), one week post-MI. And only male rats presented hypothyroidism after MI (T4 ~52% and T3 ~38%, reduction). We also verified reduction in SF (~55%), increasing in LA/Ao ratio (~75%) and changes in fluid balance (sodium intake reduction, ~67% and urinary volume reduction, ~40%) more pronounced in MIOVX than in MIINT. MIOVX group reduced thyroid hormone levels after MI (T3 ~35%). MIOVX+Veh showed more pronounced reduction in SF (~55%) and increasing in LA/Ao ratio (~75%) than the MIOVX+E2 group. The sodium intake reduced in MIOVX+Veh (~67%) and in the urinary volume we verified significant reduction in ShamOVX+Veh and MIOVX+Veh groups compared to ShamOVX+E2 and MIOVX+E2 groups (P < 0.05). Serum T3 reduced significantly (~35%) only in MIOVX+Veh group. The pathophysiological development of heart failure post-MI was attenuated in female compared to male. And female rats subjected to MI presented fluid balance more favorable and related to the less pronounced development of heart failure. Estrogen seems to influence positively the cardiac function and attenuate the dysfunction that occur post-MI. The euthyroid status in female intact do not seems to be determinant to the less pronounced development of heart failure / Sabe-se que mulheres na pr?-menopausa apresentam menor preval?ncia de doen?as cardiovasculares do que homens e, esta diferen?a desaparece ap?s a menopausa. Sendo assim, o presente estudo buscou avaliar o dimorfismo sexual e a influ?ncia do estr?geno nas altera??es da fun??o card?aca, do equil?brio hidroeletrol?tico e do status tireoidiano de ratos Wistar submetidos ao infarto do mioc?rdio (IM) experimental. Na primeira etapa ratos wistar machos (n = 18) e f?meas (n = 21) foram submetidos ao infarto experimental (INF.M e INF.F) ou ? falsa cirurgia (Sham.M e Sham.F). Na primeira e quarta semana p?s-IM foram colocados em gaiolas metab?licas e submetidos ? ecodopplercardiografia (ECO) e eletrocardiografia (an?lise espectral), seguido de eutan?sia para coleta de sangue (dosagem s?rica de horm?nios tireoidianos) e de tecidos (cora??o, pulm?o e f?gado, para biometria). Na segunda etapa, f?meas foram ovariectomizadas (n = 24) ou mantidas intactas (n = 21) e ap?s duas semanas submetidas ao infarto do mioc?rdio (INFOVX e INFINT) ou ? falsa cirurgia (ShamOVX e ShamINT). Quatro semanas ap?s p?s-IM, as mesmas avalia??es da etapa anterior foram realizadas exceto a eletrocardiogr?fica. Na terceira etapa, foi realizada ovariectomia e reposi??o com estr?geno (E2) ( n = 13) ou ve?culo (n = 22). Ap?s duas semanas foi realizada a cirurgia de indu??o ao infarto (INFOVX+E2 e INFOVX+Veic) e a falsa cirurgia (ShamOVX+E2 e ShamOVX+Veic). Decorridas quatro semanas foram feitas as mesmas avalia??es da segunda etapa. O grupo INFM desenvolveu disfun??o card?aca (fra??o de encurtamento, FEnc%, ~70% de redu??o) e altera??es na regula??o hidroeletrol?tica (aumento do apetite por s?dio, ~146% e redu??o do volume urin?rio, ~55%), uma semana p?s-IM e, portanto, mais cedo que as f?meas, que apresentaram altera??es na fun??o card?aca (FEnc%, ~28% de redu??o) e regula??o hidroeletrol?tica (aumento do apetite por s?dio, ~143%) na quarta semana p?s-IM. O grupo INFM apresentou redu??o na rela??o LF/HF (~70%), uma semana p?s-IM. E, apenas os ratos machos desenvolveram hipotireoidismo ap?s o infarto. Tamb?m foi observada redu??o da FEnc% (~55%), aumento da rela??o ?trio esquerdo/ aorta (AE/Ao, ~75%) e altera??es na regula??o hidroeletrol?tica (redu??o do apetite por s?dio, ~67% e do volume urin?rio, ~40%) mais pronunciadas em INFOVX do que em INFINT. O grupo INFOVX tamb?m apresentou redu??o dos n?veis s?ricos de T3 (~35%) p?s-IM. O grupo INFOVX+Vei mostrou redu??o da FEnc% (~55%) e aumento na rela??o AE/Ao (~75%) mais pronunciadas que o grupo INFOVX+E2. INFOVX+Vei reduziu o apetite por s?dio (~67%) e, no volume urin?rio, foi observada redu??o significativa nos grupos ShamOVX+Vei e INFOVX+Vei em rela??o aos grupos ShamOVX+E2 e INFOVX+E2 (P<0,05). O T3 s?rico reduziu significativamente (~35%) apenas no grupo INFOVX+Vei. Houve diferen?a no desenvolvimento fisiopatol?gico da insufici?ncia card?aca (IC) p?s- IM entre machos e f?meas, sendo mais brando nas f?meas. E f?meas infartadas apresentaram uma regula??o hidroeletrol?tica mais favor?vel e compat?vel com o desenvolvimento menos acentuado da IC. O estr?geno influenciou positivamente a regula??o hidroeletrolitica de f?meas infartadas, o que favoreceu a fun??o card?aca e atenuou desta forma, a disfun??o que ocorre ap?s o infarto. A manuten??o do status eutire?ideo n?o pareceu ser determinante para o desenvolvimento menos pronunciado da IC

Submitted by Jos? Henrique Henrique (jose.neves@ufvjm.edu.br) on 2017-03-02T20:23:00Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) ana_cristina_resende_camargos.pdf: 3572613 bytes, checksum: ed282931c95236a0305016db8c75f026 (MD5) / Approved for entry into archive by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2017-03-06T11:37:07Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) ana_cristina_resende_camargos.pdf: 3572613 bytes, checksum: ed282931c95236a0305016db8c75f026 (MD5) / Made available in DSpace on 2017-03-06T11:37:08Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) ana_cristina_resende_camargos.pdf: 3572613 bytes, checksum: ed282931c95236a0305016db8c75f026 (MD5) Previous issue date: 2016 / A obesidade infantil ? considerada um dos maiores problemas de sa?de p?blica do mundo. Estudos demonstram altera??o do padr?o de secre??o de adipocinas, cortisol e do fator neurotr?fico derivado do c?rebro (BDNF), bem como inflama??o cl?nica cr?nica sublimiar e desequil?brio redox em crian?as com sobrepeso e obesidade na idade escolar. Al?m disso, o excesso de peso tamb?m est? associado a reduzido desenvolvimento cognitivo e motor. Apesar da literatura indicar que os primeiros 24 meses de vida representam um per?odo importante para o desenvolvimento da obesidade infantil, n?o existem evid?ncias se crian?as com sobrepeso e obesidade nesta faixa et?ria j? apresentam altera??es em par?metros neuroend?crino-inflamat?rios, com poss?vel impacto no desenvolvimento infantil. Dessa forma, os objetivos deste estudo foram: 1) analisar as concentra??es plasm?ticas de adipocinas e BDNF, as concentra??es s?ricas de cortisol e o estado redox de crian?as com sobrepeso/obesidade entre seis e 24 meses de idade; 2) avaliar o desenvolvimento cognitivo e motor de crian?as com sobrepeso/obesidade entre seis e 24 meses de idade e; 3) verificar associa??es entre os biomarcadores avaliados com o desenvolvimento cognitivo e motor nessa faixa et?ria. Foi realizado um estudo transversal com crian?as com sobrepeso/obesidade e eutr?ficas entre seis e 24 meses de idade, cadastradas nas Estrat?gias de Sa?de da Fam?lia. As concentra??es plasm?ticas de leptina, adiponectina, resistina, receptores sol?veis do fator de necrose tumoral 1 e 2 (sTNFR1 e sTNFR1), BDNF e as concentra??es s?ricas de cortisol foram mensuradas pelo m?todo ELISA. As quimiocinas foram mensuradas pela t?cnica cytometric bead arrays e o estado redox foi determinado por meio da detec??o das concentra??es de subst?ncias reativas ao ?cido tiobarbit?rico (TBARS), da atividade das enzimas catalase (CAT) e super?xido dismutase (SOD) e da habilidade de redu??o do ferro (FRAP). O desenvolvimento infantil foi avaliado por meio do instrumento Bayley Scales of Infant and Toddler Development, 3? edi??o (Bayley-III). Foi utilizado teste t para amostras independentes para comparar os grupos e correla??o de Pearson ou Spearman para verificar a associa??o entre as vari?veis. Al?m disso, foi utilizado um modelo de regress?o linear m?ltipla stepwise para verificar a associa??o entre os biomarcardores selecionados com o desenvolvimento cognitivo e motor. As concentra??es plasm?ticas de leptina (p=0,0001), adiponectina (p=0,0007), BDNF (p=0,003) e as concentra??es s?ricas de cortisol (p=0,048) foram significativamente superiores no grupo de crian?as com sobrepeso/obesidade. Em contrapartida, as crian?as deste grupo apresentaram menores concentra??es de TBARS (p=0,004) e menor atividade das enzimas antioxidantes CAT (p=0,045) e SOD (p=0,02). N?o foram encontradas diferen?as significativas nas concentra??es de quimiocinas, sTNFR1, sTNFR2, resistina e FRAP entre os grupos (p>0,05). Al?m disso, as crian?as do grupo sobrepeso/obesidade apresentaram menores escores de desenvolvimento cognitivo (p=0,03) e motor (p=0,04). Foi ainda encontrada associa??o significativa entre as concentra??es plasm?ticas de leptina e sTNFR1 com o escore composto cognitivo (p=0,001) e das concentra??es plasm?ticas de sTNFR1 com o escore composto motor (p=0,003). Todos esses resultados apontaram a presen?a de altera??es neuroend?crino-inflamat?rias em crian?as com sobrepeso/obesidade entre seis e 24 meses de idade. Al?m disso, embora a maior parte das crian?as com sobrepeso/obesidade apresentem desenvolvimento infantil dentro dos limites de normalidade, evidencia-se pior desempenho cognitivo e motor. Por fim, foi demonstrado que maiores concentra??es de sTNFR1 e menores concentra??es de leptina foram associadas com melhores desfechos de desenvolvimento infantil nessa faixa et?ria. / Tese (Doutorado) ? Programa Multic?ntrico de P?s-Gradua??o em Ci?ncias Fisiol?gicas, Universidade Federal dos Vales do Jequitinhonha e Mucuri, 2016. / Childhood obesity is one of the world?s most serious public health issues. Studies have demonstrated dysregulated secretion pattern of adipokines, cortisol, brain-derived neurotrophic factor (BDNF), as well as chronic low-grade inflammation and redox imbalance in school-age overweight or obese children. Overweight/obese infants also had lower cognitive and motor development scores. Although the literature points out that the first 24 months of life represent an important period for the development of childhood obesity, it is not known whether overweight/obese infants in this age interval present alterations in neuroendocrine inflammatory parameters, with possible impact on child development. Then, the objectives of this study were: 1) to analyze the plasmatic levels of adipokines and BDNF, serum cortisol and redox status in overweight/obese infants between 6 and 24 months of age; 2) to evaluate the cognitive and motor development in overweight/obese infants between 6 and 24 months of age and; 3) to verify the association of the biomarkers evaluated with cognitive and motor development in this age interval. A cross-sectional study was conducted with infants with overweight/obesity and normal-weight between 6 and 24 months enrolled in Family Health Strategies. Plasma leptin, adiponectin, resistin, BDNF, soluble tumour necrosis factor receptors 1 and 2 (sTNFR1, sTNFR2), and serum cortisol levels were measured using conventional ELISA kits. Plasma chemokines were measured using the cytometric bead arrays kit, and oxidative stress was assessed by thiobarbituric acid reactive substances (TBARS) concentration, enzymes catalase (CAT) and dismutase superoxide (SOD) activity, as well by ferric reducing antioxidant power (FRAP). Infant development was performed using the Bayley Scales of Infant and Toddler Development, 3rd edition (Bayley-III). A t test for independent samples was performed to compare the groups and Pearson and Spearman correlation was used to verify the association between parameters. Multiple linear stepwise regression models were utilized to verify the association between the biomarkers selected and cognitive and motor composite scores. Plasma levels of leptin (p=0.0001), adiponectin (p=0.0007), BDNF (p=0.003) and serum cortisol (p=0.048) were significantly higher in overweight/obese infants. In contrast, concentration of TBARS (p=0.004), CAT (p=0.045) and SOD activity (p=0.02) were lower in overweight/obese infants. There were no differences in the levels of chemokines, sTNFR1, sTNFR2, resistin and FRAP between groups (p>0.05). Moreover, overweight/obese infants had lower cognitive (p=0.03) and motor (p=0.04) development scores than normal-weight infants. A significant association of plasma leptin and sTNFR1 levels with cognitive composite scores (p=0.001) were found and plasma sTNFR1 levels were associated with motor composite scores (p=0.003). All these results point out neuroendocrine inflammatory response changes in overweight/obese infants between 6 and 24 months. Moreover, although most of overweight/obese infants have presented cognitive and motor development within normal limits, there is evidence of worse cognitive and motor performance. Finally, high sTNFR1 and low leptin levels were associated with increase developmental outcomes in infants in this age interval.

Made available in DSpace on 2014-12-17T15:36:52Z (GMT). No. of bitstreams: 1 MarcioVR.pdf: 229432 bytes, checksum: 1ee93b8bd1e9e53f934d759c8f562466 (MD5) Previous issue date: 2008-03-31 / Many behavioral and biological variables of animals are expressed in the form of biological rhytms, down by the Circadian Timing System, that synchronize them with the environment from external stimuli such as light. One of them is the secretion profile of most circulating hormones regulated by the hypothalamuspirtuitary axis, which controls functions essential for the survival and reproduction of organisms. The sag?i, Callithrix jacchus, one of the most studied species about their endocrine physiology, is an appropriate subject for evaluating the profile of plasma prolactin and cortisol of adult males and females born in captivity throughout the year. Three male and two adult femelas were housed individually and subjected to natural environmental conditions over two years. Blood samples were used to measure the circulating levels of both hormones by methods radioimmunoassay (RIA) and immunoassay (ELISA), respectively. The analysis during the year of the plasmatic values of both hormones test was performed by ANOVA for repeated measures, the correlation of Spearman, and the test of Friedman and Student's t-test. The levels of prolactin in plasma were higher during the months in which there is a greater incidence of births of baby in the colony, possibly serving for modulating the expression of the behavior of parental care in both sexes. The plasma cortisol showed a lift in anticipation of the station with the highest birth rate and may be associated with the preparation of individual participation in caring for the baby, and also with the establishment of emotional bond between reproductive partners. Thus, this study shows that, despite the variations observed in the environment in which the animals live, plasma levels of prolactin and cortisol vary little throughout the year. / Muitas vari?veis biol?gicas e comportamentais dos animais se expressam sob a forma de ritmos biol?gicos, determinadas pelo Sistema de Temporiza??o Circadiana, que as sincronizam com o ambiente a partir de est?mulos externos, com a luz. Uma delas ? o perfil de secre??o da maioria dos horm?nios circulantes regulado pelo eixo Hipot?lamo-Hipofis?rio, que control fun??es essenciais para a sobreviv?ncia e a reprodu??o dos organismos. O sag?i, Callithrix jacchus, uma das esp?cies mais estudadas quanto ? sua fisiologia end?crina, ? um sujeito adequado para a avalia??o do perfil plasm?tico de prolactina e cortisol de machos e f?meas adultas nascidos em cativeiro ao lonto do ano. Tr?s machos e duas f?meas adultas foram alojados individualmente e submetidos ?s condi??es ambientais naturais durante dois anos. Amostras de sangue foram usadas para dosar os n?veis circulantes de ambos os horm?nios pelos m?todos Radioimunoensaio (RIA) e Imunoenzim?tico (ELISA), respectivamente. A an?lise ao longo do ano dos valores plasm?ticos de ambos os horm?nios foi realizada pelo teste de ANOVA para Medidas Repetidas, pela correla??o de Spearman, e pelos testes de Friedman e t de Student. Os n?veis de prolactina no plasma se mostraram mais elevados durante os meses nos quais h? uma maior ocorr?ncia de nascimentos de filhotes na col?nia, servindo possivelmente para modular a express?o do comportamento de cuidado parental em ambos os sexos. O cortisol plasm?tico mostrou uma eleva??o em antecipa??o ? esta??o com maior natalidade, podendo estar associada com a prepara??o do indiv?duo para participa??o no cuidado aos filhotes, e tamb?m com o estabelecimento de la?o afetivo entre os parceiros reprodutivos. Assim, o presente estudo mostra que, apesar das varia??es observadas no ambiente no qual os animais vivem, os n?veis plasm?ticos de prolactina e cortisol oscilam pouco ao longo do ano.