Impairment of nitric oxide (NO) - cyclic GMP signaling pathway is likely to contribute to human begnin prostate hyperplasia (BPH). In the present study we have used a model of chronic NO synthesis inhibition to evaluate the functional alterations of prostate smooth muscle (PSM) machinery, and involvement of Rho-kinase pathway. Wistar rats were treated with the NO inhibitor N(ω)-nitro-l-arginine methyl ester (L-NAME, 20mg/kg/day; 4 weeks), after which contractile responses to phenylephrine (α1-adrenoceptor agonist; 1nM to 100µM), carbachol (muscarinic agonist; 1nM to 1mM) and α,β-methylene ATP (P2X receptor agonist; 1-10µM), as well as to electrical-field stimulation (EFS; 1-32Hz) were evaluated. PSM relaxations to isoproterenol (non-selective β-adrenoceptor agonist, 0.1nM to 10µM) and sodium nitroprusside (NO donor, 1nM to 10mM) were also evaluated. The ratio prostate weight/body weight was 22% greater (P<0.05) in L-NAME compared with control group. The PSM contractions to phenylephrine, carbachol and α,β-methylene ATP were higher in L-NAME (Emax: 3.85±0.25, 3.52±0.35 and 2.03±0.2mN, respectively) compared with control group (Emax: 3.08±0.17, 2.37±0.18 and 1.57±0.18mN, respectively). The PSM contractions induced by EFS were also significantly greater in L-NAME group. Prior incubation with the Rho-kinase inhibitor Y27632 (1µM) fully reversed the enhanced contractions to phenylephrine and carbachol. Isoproterenol-induced PSM relaxations were 34% lower in L-NAME group, which was associated with reduced levels of cAMP in prostate tissue. The relaxations to sodium nitroprusside remained unaltered in L-NAME group. In summary, chronic NO deficiency leads to increased Rho-kinase-mediated PSM contractile responses accompanied by impairment of β-adrenergic-cAMP-signaling pathway.
RESUMO -Objetivou-se avaliar o consumo e as digestibilidades aparentes de matéria seca (MS), proteína bruta (PB), extrato etéreo (EE), carboidratos totais (CT) e fibra em detergente neutro (FDN), o ganho de peso e a conversão alimentar em bovinos Nelore, recebendo rações contendo feno de capim-tifton 85 com 35, 42 e 56 dias de rebrota. Foram utilizados 18 novilhos, não-castrados, com peso vivo médio inicial de 345 kg, distribuídos em um delineamento em blocos casualizados, com seis repetições. Todas as rações continham 60% de volumoso e 40% de mistura concentrada constituída por fubá de milho e minerais, na base da matéria seca. O experimento teve duração de 100 dias, 16 dias de adaptação e três períodos de 28 dias para a avaliação do desempenho. O avanço da idade de rebrota do capimtifton 85 produziu comportamento linear decrescente para consumos de MS, PB e EE, enquanto os consumos de CT e FDN não foram afetados pela idade de rebrota. O ganho médio diário de peso (1,23 kg) e a conversão alimentar (6,35) não foram influenciados pela idade de rebrota. Os coeficientes de digestibilidade de MS (59,6), PB (48,0), EE (53,2), CT (63,6) e FDN (41,5) também não foram influenciados pela idade de rebrota do feno de capim-tifton 85. Embora as dietas utilizadas sejam eqüivalentes nutricionalmente, recomenda-se a idade de corte de 42 dias de rebrota.Palavras-chave: conversão alimentar, ganho de peso Intake, Digestibility and Performance of Steers Fed Diets Containing Tifton-85Bermudagrass Hays at Different Regrowth Ages ABSTRACT -Intake and apparent digestibilities of dry matter (DM), crude protein (CP), ether extract (EE), total carbohydrates (TC) and neutral detergent fiber (NDF), average daily weight gain and feed: gain of diets containing Tifton 85 bermudagrass hays with 35, 42 and 56 days of regrowth were evaluated with Nellore steers. Eighteen animals averaging 345 kg were allotted in a randomized complete blocks design with six replicates. All diets contained 60% forage and 40% of a concentrate mix based on ground corn and minerals, in dry matter basis. The experiment lasted 100 days, in which 16 were used for the animal adaptation and three periods of 28 days for the animal performance evaluation. As the hay age increased, there was a linear decrease in DM, CP and EE intake, while the TC and NDF intakes were not affected by the regrowth age. The average daily weight gain (1.23kg) and the feed: gain (6.35) was not influenced by the regrowth age. The coefficients of digestibility of DM (59.6), CP (48.0), EE (53.2), TC (63.6) and NDF (41.5) were not affected by the regrowth age of the Tifton 85 bermudagrass hay. Although the diets were nutritionally similar, the recommendation for the regrowth age was 42 days.Key Words: feed:gain ratio, weight gain Introdução A bovinocultura de corte constitui-se no maior segmento da economia rural brasileira, tendo, neste contexto, relevante papel social, tanto por gerar empregos, como por se constituir na principal fonte de proteína animal da dieta da população do país (NOGUEIRA NETT...
Immunohistochemistry (IHC) technique is an alternative toxicological analysis to detect drugs in insects of forensic importance, but it requires thorough histological procedures. In this study, we tested different fixatives--phosphate-buffered paraformaldehyde 4% (PP), Carnoy's fluid (CF), Kahle's solution (KS), ethanol in different concentrations, and ethanol associated to PP and CF, time of fixation and histological processes for dipteran larvae's tissue, aiming to develop a sample preparation protocol for IHC application. A suitable fixation was achieved using PP for 12 and 24 h, CF for 3 h, 70% ethanol for 19 days, and 70% ethanol/CF for 2 h/3 h. Postfixation using negative pressure, two immersions in xylene for 30 min each, and one in xylene plus paraffin for 45 min increased tissue preservation. An immunohistochemical test for cocaine detection was performed using monoclonal benzoylecgonine antibody from mouse, peroxidase-conjugated anti-mouse IgG and visualized by 3,3'-diaminobenzidine method showed these histological procedures didn't compromise antigenicity.
Dedico este trabalho à meus pais, Marcos José Alves e Solange de Oliveira Alves por permitirem que toda essa trajetória tenha sido possível investindo, acreditando e incentivando sempre, além do amor e carinho incondicionais.viii ix AGRADECIMENTOSAo Prof. Dr. André Almeida Schenka, pela oportunidade oferecida ao acreditar em mim para a realização desse trabalho ciente de todas as dificuldades que esta ação traria. Agradeço também por toda orientação neste projeto, sempre contribuindo com seu conhecimento científico e com sua amizade.Aos meus pais, à minha irmã Juliana e aos meus avós (vivos ou não), cuja convivência me fazem desenvolver meu caráter e princípios, além do apoio, torcida, carinho e incentivo constantes.Aos amigos André, Cristielle, Marina, Philipi e Valéria por toda "assessoria técnico-científica", amizade, carinho e momentos de descontração.
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