955 resultados para serum retinol
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Fluoroacetate is a highly toxic species naturally found in plants and in commercial products (compound 1080) for population control of several undesirable animal species. However, it is non-selective and toxic to many other animals including humans, and thus its detection is very important for forensic purposes. This paper presents a sensitive and fast method for the determination of fluoroacetate in blood serum using capillary electrophoresis with capacitively coupled contactless conductivity detection. Serum blood samples were treated with ethanol to remove proteins. The samples were analyzed in BGE containing 15 mmol/L histidine and 30 mmol/L gluconic acid (pH 3.85). The calibration curve was linear up to 75 mu mol/L (R(2) = 0.9995 for N = 12). The detection limit in the blood serum was 0.15 mg/kg, which is smaller than the lethal dose for humans and other animals. Fluoride, a metabolite of the fluoroacetate defluorination, could also be detected for levels greater than 20 mu mol/L, when polybrene was used for reversion of the EOF. CTAB and didecyldimethylammonium bromide are not useful for this task because of the severe reduction of the fluoride level. However, no interference was observed for fluoroacetate.
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CD and EPR were used to characterize interactions of oxindole-Schiff base copper(II) complexes with human serum albumin (HSA). These imine ligands form very stable complexes with copper, and can efficiently compete for this metal ion towards the specific N-terminal binding site of the protein, consisting of the amino acid sequence Asp-Ala-His. Relative stability constants for the corresponding complexes were estimated from CD data, using the protein as competitive ligand, with values of log K(CuL) in the range 15.7-18.1, very close to that of [Cu(HSA)] itself, with log K(CuHSA) 16.2. Some of the complexes are also able to interfere in the a-helix structure of the protein, while others seem not to affect it. EPR spectra corroborate those results, indicating at least two different metal species in solution, depending on the imine ligand. Oxidative damage to the protein after incubation with these copper(II) complexes, particularly in the presence of hydrogen peroxide, was monitored by carbonyl groups formation, and was observed to be more severe when conformational features of the protein were modified. Complementary EPR spin-trapping data indicated significant formation of hydroxyl and carbon centered radicals, consistent with an oxidative mechanism. Theoretical calculations at density functional theory (DFT) level were employed to evaluate Cu(II)-L binding energies, L -> Cu(II) donation, and Cu(II) -> L back-donation, by considering the Schiff bases and the N-terminal site of HSA as ligands. These results complement previous studies on cytotoxicity, nuclease and pro-apoptotic properties of this kind of copper(II) complexes, providing additional information about their possibilities of transport and disposition in blood plasma. (C) 2009 Elsevier Inc. All rights reserved.
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The interaction of bovine serum albumin (BSA) with the ionic surfactants sodium dodecylsulfate (SDS, anionic), cetyltrimethylammonium chloride (CTAC, cationic) and N-hexadecyl-N,N-dimethyl-3-ammonio-1-propanesulfonate (HPS, zwitterionic) was studied by electron paramagnetic resonance (EPR) spectroscopy of spin label covalently bound to the single free thiol group of the protein. EPR spectra simulation allows to monitor the protein dynamics at the labeling site and to estimate the changes in standard Gibbs free energy, enthalpy and entropy for transferring the nitroxide side chain from the more motionally restricted to the less restricted component. Whereas SDS and CTAC showed similar increases in the dynamics of the protein backbone for all measured concentrations. HPS presented a smaller effect at concentrations above 1.5 mM. At 10 mM of surfactants and 0.15 mM BSA, the standard Gibbs free energy change was consistent with protein backbone conformations more expanded and exposed to the solvent as compared to the native protein, but with a less pronounced effect for HPS. In the presence of the surfactants, the enthalpy change, related to the energy required to dissociate the nitroxide side chain from the protein, was greater, suggesting a lower water activity. The nitroxide side chain also detected a higher viscosity environment in the vicinity of the paramagnetic probe induced by the addition of the surfactants. The results suggest that the surfactant-BSA interaction, at higher surfactant concentration, is affected by the affinities of the surfactant to its own micelles and micelle-like aggregates. Complementary DLS data suggests that the temperature induced changes monitored by the nitroxide probe reflects local changes in the vicinity of the single thiol group of Cys-34 BSA residue. (C) 2011 Elsevier B.V. All rights reserved.
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EPR spectra of 5- and 16-doxyl stearic acid nitroxide probes (5-DSA and 16-DSA, respectively) bound to bovine serum albumin (BSA) revealed that in the presence of ionic surfactants, at least, two label populations coexist in equilibrium. The rotational correlation times (tau) indicated that component I displays a more restricted mobility state, associated to the spin labels bound to the protein; the less immobilized component 2 is due to label localization in the surfactant aggregates. For both probes, the increase of surfactant concentration leads to higher motional levels of component 1 followed by a simultaneous decrease of this fraction of nitroxides and its conversion into component 2. For 10 mM cethyltrimethylammonium chloride (CTAC), the nitroxides are 100% bound to the protein, whereas at 10mM N-hexadecyl-N,N-dimethyl-3-ammonio-1-propanesulfonate (HPS) and sodium dodecyl sulfate (SDS) the fractions of bound nitroxides are reduced to 18% and 86%, respectively. No significant polarity changes were observed in the whole surfactant concentration range for component 1. Moreover, at higher surfactant concentration, component 2 exhibited a similar polarity as in the pure surfactant micelles. For 16-DSA the surfactant effect is different: at 10mM of HPS and CTAC the fractions of bound nitroxides are 76% and 49%, respectively, while at 10 mM SDS they are present exclusively in a micellar environment, consistent with 100% of component 2. Overall, both SDS and HPS are able to effectively displace the nitroxide probes from the protein binding sites. while CTAC seems to affect the nitroxide binding to a significantly smaller extent. (C) 2008 Elsevier B.V. All rights reserved.
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A modulação fenotípica da célula estrelada hepática (CEH) é um dos eventos primários do processo de fibrose hepática. Durante este evento, a CEH modifica seu fenótipo lipocítico (quiescente) para miofibroblástico (ativado), fenômeno chamado de ativação. Estudos recentes sugerem que fatores de transcrição adipogênicos atuam no processo de ativação das CEH, e conseqüentemente teriam um papel chave na evolução do processo fibrótico. Neste estudo analisamos a expressão gênica de vários fatores de transcrição adipogênicos, como os receptores ativados por proliferadores de peroxissomos (RAPP), receptor X do fígadoα (RXFα), proteína ligante de CCAAT/enhancerα (PLCEα) e proteína ligante do elemento regulatório de esteróis-1 (PLERE-1), em uma linhagem representativa de CEH, a GRX. A linhagem GRX, é capaz de ser induzida in vitro a expressar o fenótipo quiescente através do tratamento com indometacina ou retinol. Após 24 horas ou 7 dias de tratamento, a expressão gênica foi analisada através de PCR em tempo real. Tratamento com indometacina aumentou a expressão de todos os genes avaliados, exceto PLCEα e RAPPβ. O mesmo resultado obtido com desmetil indometacina (LM 4511) sobre RAPPα e γ sugere que a ação da indometacina sobre a diferenciação fenotípica da GRX é independente de sua ação inibitória sobre ciclooxigenase, já que a LM 4511 não possui esta atividade. Tratamento com retinol levou à modulação de um número menor de genes, diminuindo a expressão de PLCEα e aumentando a de RAPPγ, e RAPPβ. Adipsina, gene característico de adipócitos, teve sua expressão induzida apenas nas células tratadas com indometacina. Pex16 e catalase, genes modulados pelos RAPPs, foram diferencialmente expressos, dependendo do tratamento. Os resultados apresentados sugerem que a linhagem representativa de CEH, GRX, tem sua indução fenotípica modulada por fatores de transcrição adipogênicos, que são diferencialmente expressos dependendo do tratamento de conversão.
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The caprine milk is a product of high biological value and high digestibility. Due to these characteristics it is quite used by newly born children that are not breastfed or that are intolerant to the bovine milk. The vitamin deficiency is a public health problem in underdeveloped areas as the Northeast of Brazil and where areas the caprine ones adapt very well. The present study was led to analyze the influence of the feeding in the vitamin levels in the caprine milk. The animals used were the races Saanen and Murciana, divided in three groups. The first group with 38 animals of race Saanen and the second with 30 animals of race Murciana were, fed with concentrated and voluminous. A third group with 20 animals of the race Saanen was fed exclusively with voluminous. The four group was added with 10. 000 UI of retinol palmitato, administered directly, like capsule, in the mouth of animal. Parallel it was verified the level of retinol of milk in the beginning and final of the sucked, in the goats of the second group (race Murciana n =30). The retinol of caprine milk was determined through the system of liquid cromatografia of high efficiency (HPLC). The retinol levels in the studied groups were respectively: first (38. 5 ± 12. 7 μg/100ml), second (40. 5 ± 9. 7 μg/100ml); third, with 20 animals of race Saanen fed exclusively with voluminous (23. 1 ± 6. 7 μg/100ml) and in the group a, suplementation with 10. 000 UI of retinol palmitato (43,7 ± 18,8 μg/100ml) before, and (61,9 ± 26,9 μg/100ml) after the supplementation. It was not found significant difference between the averages from animals of the first and second group, that were fed with the same concentrate diet and voluminous, showing that the retinol levels in the milk of these two races are equivalent. Already in the animals of the first and third group that they were fed with different diets, in those which diet was just voluminous, a drastic reduction was verified in the retinol levels. In relation to the retinol of the milk in different moments from the same sucked, it was observed in the beginning of the breast-feeding (22. 6 ± 9. 8 μg/100ml) and at the end of the sucked (49. 6 ± 14. 7μg/100ml), being the difference between the averages, statistically significant (p < 0,0001). Already in the animals that were supplemented, a significant increase was observed in the retinol concentration, being obtained a medium response of 41,85%
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The intake of adequate quantities of food, including those rich in vitamins, is necessary for a healthy life. The lack of vitamin A has been characterized as a public health problem in developing countries, however, a high intake of vitamin A can result in toxic and teratogenics effects. High concentrations of vitamin A have been observed in the livers of animals. The objective of this study was to assess the levels of retinol in chicken livers and verify the effect of frozen storage on these levels. 64 livers from two chicken strains, Cobb and Ross, were used, came from four different farms. We examined 32 livers from each strain, 8 samples from each farm. Liver sample were homogenized individually, then 4 aliquots were taken from each sample. One of aliquots was analyzed immediately after slaughter (T0), the others were analyzed after 30, 60 and 90 days of storage at -18oC (T30, T60 and T90, respectively). Retinol dosage in the liver was determined by High Performance Liquid Chromatography (HPLC). The levels of retinol varied significantly according to the strain. The mean retinol value in the fresh samples was 6678.0 ± 1337.7 and 8324.1 ± 1158.5 µg/100g in the Cobb and Ross strain, respectively. Values of 4258 ± 918.7 ± 1391.7 and 4650.5 ± 1391.7 μg/100g were found after 90 days of storage for Cobb and Ross strain, respectively. The liver freezing caused a significant reduction in their levels of retinol, causing a loss of up to 44% with respect to fresh livers. The reduction in retinol levels occurred from 30 days of storage. Even with the losses from the frozen, the ingestion of a typical portion of 100 g of liver, regardless the chicken strain analyzed, surpass all recommendations of consumption and the maximum tolerable intake of vitamin A (3000 μg/day) for adults
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Vitamin A deficiency is a serious public health problem in developing countries, and it causes death and blindness among children in the developing countries. The fortification of food could be an important source of vitamins to control deficiency. 60 Coturnix coturnix japonica quails were used in a randomized design with duration of seven weeks. The birds were assigned into five treatments with four repetitions. The objective was to evaluate the influence of the supplementation with different levels of retinyl palmitate (2,000 IU, 4,000 IU, 8,000IU and 16,000 IU) in quails under the levels of retinyl in egg yolks. The method used to dose retinyl in yolks of quail eggs was High Performance Liquid Chromatography and the enzymatic method to quantify the cholesterol concentration. The weight and production of eggs was significantly modified by the supplementation with retinyl in the birds. The results showed a gradual increase in the incorporation of retinyl in the egg yolk as a response to the supplementation, reaching values 384% higher than the control values. By the end of the supplementations a significant reduction in the concentrations of retinyl in the eggs yolk was observed. The most lasting supplementations were with 8,000 IU and 16,000 IU which lasted for three weeks. The cholesterol content in eggs was not significantly modified. The consumption of one egg enriched with 16000UI of retinol palmitate in the present study, by day, would probably reach 10 and 7,3% of the daily recommendations of this micronutrient for children of 1 to 3 years of age, and for 4 to 8 years, respectively. The nutritional value of eggs, related to the vitamin A, can be improved by supplementation of quails
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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
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Foram examinados 10 bezerros da raça Holandesa com duas a quatro semanas de idade, distribuídos aleatoriamente em dois grupos, controle e infectado. Os bezerros do grupo-controle foram inoculados por via intrabronquial com 5ml de solução salina fosfato-tamponada Dulbecco (DPBSS). Os do grupo infectado receberam um inóculo contendo 5x10(9) unidades formadoras de colônia-UFC de Mannheimia (Pasteurella) haemolytica, suspensas em 5ml de DPBSS. As amostras de sangue foram colhidas 15 minutos antes e uma, duas, quatro e seis horas após a inoculação. O proteinograma sérico foi obtido por eletroforese em gel de acrilamida. As concentrações séricas das proteínas de pesos moleculares 125.000 D (ceruloplasmina), 60.000 D (a1-antitripsina), 45.000 D (haptoglobina) e 40.000 D (glicoproteína ácida) foram significativamente maiores em bezerros infectados do que nos do grupo-controle. Os resultados indicam que as concentrações das proteínas de fase aguda se elevaram mais rapidamente que o previamente imaginado. O proteinograma sérico pode ser útil no monitoramento da evolução da pneumonia de bezerros causada por M. haemolytica.
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Foram examinados 100 bezerros da raça Nelore com 6 a 12 meses de idade, distribuídos em: grupo controle (G1; 50 bezerros sadios) e grupo fotossensibilização (G2; n= 50). As amostras de sangue foram coletadas 12 a 24 horas após o início da dermatite (M1) e 15 a 30 dias após (M2), época da cura das lesões cutâneas. O proteinograma sérico foi obtido por eletroforese em gel de acrilamida. em todos os bezerros foram identificadas 18 proteínas com pesos moleculares (PM) entre 16.000 a 189.000 dáltons (Da). em M1 e M2, as concentrações séricas das proteínas de PM 115.000Da (ceruloplasmina), 61.000Da (1-antitripsina), 45.000Da (haptoglobina) e 40.000Da (glicoproteína ácida) foram significativamente maiores em bezerros com fotossensibilização hepatógena em comparação com aquelas dos animais do grupo-controle. A determinação dos teores séricos de proteínas de fase aguda pode ser útil no monitoramento da progressão da fotossensibilização hepatógena em bovinos, inclusive orientando possíveis alterações em procedimentos terapêuticos.
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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Cisplatin is an antineoplastic drug that is used to treat carcinomas and osteosarcomas in dogs. However, it can cause kidney damage which in turn leads to the loss of electrolytes. Concentration of magnesium was evaluated in the blood serum of eight dogs that had undergone therapy with cisplatin. The dogs were divided into two groups. Group 1 was supply with cisplatin (70 mg/m(2), iv) and to prevent nephrotoxcity, saline solution at 0.9% was administered (25 ml/kg/hour, iv in three hours). The animals in Group 2 were not supplied with cisplatin. There was no significant difference (P < 0.05) between the two groups and the results obtained indicated that the protocol used for Group 1 did not cause changes in the serum concentrations of magnesium. Therefore, it can be concluded that the administration of cisplatin does not cause variations if it is combined with the fluid therapy.
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)