Efeito inibidor do soro urêmico sobre o metabolismo oxidativo dos neutrófilos de cães

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Morphological and histochemical study of the myotomal muscle in pacu (Piaractus mesopotamicus Holmberg, 1887) during the initial growth phases

Morphological and histochemical methods were used to evaluate the myotomal muscle characteristics of pacu (Piaractus mesopotamicus) from hatching to 40 days old. During the larval period, the musculature consisted predominantly of white muscle. White and red muscle mass increased at 10, 20, 30 and 40 days after hatching. The larvae had round muscle fibers with a moderate degree of maturation and central nuclei. In subsequent phases, small and immature fibers were visible near larger and more differentiated fibers. Undifferentiated cells or presumptive myoblasts located in embryogenic zones were visible in the dorsal and ventral regions, and were more evident at 30 and 40 days. The red muscle fibers located in the subdermal region, had oxidative metabolism and slow contraction, whereas the more predominant white muscle fibers had glycolytic metabolism and fast contraction. Our findings indicate that during the initial phases, myotomal muscle growth in pacu occurs by both, muscle fiber hypertrophy and hyperplasia. The analysis of frequency of red and white muscle fibers shows that hyperplastic growth is intense in this period. As the growth rate in adult fish is related to the number of muscle fibers in young fish, extrinsic factors could change the muscle fiber phenotype and influence their ultimate size.

Interleukin-15 increases Paracoccidioides brasiliensis killing by human neutrophils

Interleukin-15 is a cytokine produced by a wide range of different cell types, including macrophages, in response to lipopolysaccharide or microbial infection. This cytokine may play a crucial role in the activation of phagocytic cells against pathogens, especially during innate immune response. The effects of IL-15 on human polymorphonuclear leukocyte fungicidal activity against a highly virulent Paracoccidioides brasiliensis strain were investigated. Pretreatment of human neutrophils from healthy individuals with IL-15 for IS h increased cell fungicidal activity in a dose-dependent manner. In addition, the exposure to IL-15 induced an increase in neutrophil oxidative burst as evaluated by superoxide anion and H(2)O(2) release. Catalase inhibited fungicidal activity supporting a role for H(2)O(2) in fungus killing. In contrast, IL-8 and TNF-alpha levels were not affected by IL-15 suggesting that its effects were not mediated by these cytokines. Together, these results show that IL-15 is a potent stimulant of antifungal activities in human neutrophils, at least in part by a mechanism dependent on oxidative metabolism. (c) 2007 Elsevier Ltd. All rights reserved.

Influência do desenvolvimento etário e da suplementação com vitamina E (acetato de DL-alfa-tocoferol) no metabolismo oxidativo dos neutrófilos de bovinos da raça Holandesa (Bos taurus)

O presente trabalho teve por objetivo analisar a influência do desenvolvimento etário e da suplementação com acetato de DL-alfa-tocoferol sobre o metabolismo oxidativo de neutrófilos, em bovinos da raça holandesa, no período do nascimento até os 150 dias de idade. Foram utilizados 20 bezerros divididos em dois grupos de dez animais. Os animais do grupo Tratamento receberam 2000UI de acetato de DL-alfa-tocoferol, por via intramuscular, ao nascimento, aos 15, 30, 60, 90 e 120 dias de idade, sendo o outro o grupo Controle, que não recebeu qualquer suplementação. em ambos os grupos, o metabolismo oxidativo dos neutrófilos demonstrou pouca atividade durante os primeiros 60 dias de vida, sendo indicativo da ineficiência deste importante mecanismo bactericida. Não foi observado efeito significativo da administração do acetato de DL-alfa-tocoferol sobre o metabolismo oxidativo de neutrófilos.

Structural complexity of the nitrogen source and influence on yeast growth and fermentation

The structural complexity of the nitrogen source strongly affects both biomass and ethanol production by industrial strains of Saccharomyces cerevisiae, during fermentation in media containing glucose or maltose, and supplemented with a nitrogen source varying from a single ammonium salt (ammonium sulfate) to free amino acids (casamino acids) and peptides (peptone). Diauxie was observed at low glucose and maltose concentrations independent of nitrogen supplementation. At high sugar concentrations diauxie was not easily observed. and growth and ethanol production depended on the nature of the nitrogen source. This was different for baking and brewing ale and lager yeast strains. Sugar concentration had a strong effect on the shift from oxido-fermentative to oxidative metabolism. At low sugar concentrations, biomass production was similar under both peptone and casamino acid supplementation. Under casamino acid supplementation, the time for metabolic shift increased with the glucose concentration, together with a decrease in the biomass production. This drastic effect on glucose fermentation resulted in the extinction of the second growth phase, probably due to the loss of cell viability. Ammonium salts always induced poor yeast performance. In general, supplementation with a nitrogen source in the peptide form (peptone) was more positive for yeast metabolism, inducing higher biomass and ethanol production, and preserving yeast viability, in both glucose and maltose media, for baking and brewing ale and lager yeast strains. Determination of amino acid utilization showed that most free and peptide amino acids present, in peptone and casamino acids, were utilized by the yeast, suggesting that the results described in this work were not due to a nutritional status induced by nitrogen limitation.

Maximal lactate steady state concentration independent of pedal cadence in active individuals

The maximal lactate steady state (MLSS) is defined as the highest blood lactate concentration that can be maintained over time without a continual blood lactate accumulation. The objective of the present study was to analyze the effects of pedal cadence (50 vs. 100 rev min(-1)) on MLSS and the exercise workload at MLSS (MLSSworkload) during cycling. Nine recreationally active males (20.9 +/- 2.9 years, 73.9 +/- 6.5 kg, 1.79 +/- 0.09 m) performed an incremental maximal load test (50 and 100 rev min(-1)) to determine anaerobic threshold (AT) and peak workload (PW), and between two and four constant submaximal load tests (50 and 100 rev min(-1)) on a mechanically braked cycle ergometer to determine MLSSworkload and MLSS. MLSSworkload was defined as the highest workload at which blood lactate concentration did not increase by more than 1 mM between minutes 10 and 30 of the constant workload. The maximal lactate steady state intensity (MLSSintensity) was defined as the ratio between MLSSworkload and PW. MLSSworkload (186.1 +/- 21.2 W vs. 148.2 +/- 15.5 W) and MLSSintensity (70.5 +/- 5.7% vs. 61.4 +/- 5.1%) were significantly higher during cycling at 50 rev min(-1) than at 100 rev min(-1), respectively. However, there was no significant difference in MLSS between 50 rev min(-1) (4.8 +/- 1.6 mM) and 100 rev min(-1) (4.7 +/- 0.8 mM). We conclude that MLSSworkload and MLSSintensity are dependent on pedal cadence (50 vs. 100 rev min(-1)) in recreationally active individuals. However, this study showed that MLSS is not influenced by the different pedal cadences analyzed.

Histochemical and SEM evaluation of the neuromuscular junctions from alcoholic rats

In the present study morphological changes occurring in the neuromuscular junctions (NMJ) of the extensor digitorum longus (EDL) and soleus muscles from albino rats (Rattus norvegicus) submitted to experimental chronic alcoholism were evaluated. Seventy two male animals aged 4 months and weighing on average 400 g were divided into three groups: control, alcoholic and isocaloric. Six rats from each group were anesthetized and sacrificed after 5, 10, 15 and 18 months. The NMJ did not show detectable morphological changes in either muscle after treatment when examined by light microscopy. With respect to the dimensions, statistical analysis demonstrated a tendency to a statistically significant treatment x time interaction for the length of soleus muscle NMJ. The ultrastructural study, however, revealed that the NMJ of the soleus muscle of animals submitted to 18 months of experimental alcoholism presented important morphological alterations. Characteristically, the NMJ of these muscles is located on an elevation on the surface of the muscle fiber, presenting a regular round, oval or elliptical shape and continuous and not very deep synaptic grooves. Approximately 30% of the NMJ of alcoholic rats are irregular in shape, with the sarcolemmal elevations typical of the synapse region being flattened on at least one side, with discontinuous synaptic grooves, and deep and punctiform contacts of the synaptic buds. These data suggest that, although skeletal muscle has a greater natural resistance against the direct or indirect effects of alcohol, some submicroscopic morphological alterations are detectable in the NMJ, especially in muscles with oxidative metabolism (soleus) following long periods of ingestion of alcohol. (C) 2002 Elsevier B.V. Ltd. All rights reserved.

Aerobic fitness level typical of elite athletes is not associated with even faster VO2 kinetics during cycling exercise

The aim of this study was to address the question if the VO2 kinetics is further improved as the aerobic training status increases from trained to elite level athletes. Maximal oxygen uptake (VO(2)max), work-rate associated to VO(2)max (IVO(2)max) and VO2 kinetics of moderate (Mod) and maximal exercise (Max) were determined in fifty-five subjects. Then, they were assigned into three groups: low (LF), intermediate (IF) and high (HF) aerobic fitness level. In average, the VO(2)max of LF, IF and HF groups were, respectively, 36.0 +/- 3.1, 51.1 +/- 4.5 and 68.1 +/- 3.9 ml . kg . min(-1) (p <= 0.05 among each other). VO2 kinetics mean response time of both exercise intensities were significantly faster (p <= 0.05) in HF (Mod, 27.5 +/- 5.5 s; Max, 32.6 +/- 8.3 s) and IF (Mod, 25.0 +/- 3.1 s; Max, 42.6 +/- 10.4 s) when compared to LF (Mod, 35.7 +/- 7.9 s; Max: 57.8 +/- 17.8 s). We can conclude that VO2 kinetics is improved as the fitness level is increased from low to intermediate but not further improved as the aerobic fitness level increases from intermediate to high.

Morfologia e crescimento do músculo estriado esquelético no pirarucu Arapaima gigas Cuvier, 1817 (Teleostei, Arapaimidae)

The aim of this work was to evaluate the morphological, growth characteristics of skeletal muscle tissue III Pirarucu (Arapaima gigas) using alevins (50 days old) and juveniles (1 year old). Muscle samples were collected from dorsal, lateral cranial mid lateral caudal regions, and then frozen in liquid nitrogen. Histological frozen sections (10 [mu]m) were stained with HE and Gomori Trichrome for morphological analysis, and NADH-TR to evaluate muscle fiber oxidative metabolism. Morphometric analysis samples were obtained from dorsal and lateral cranial regions, and the smallest-diameter white fibers were measured. White dorsal muscle was thicker, and two muscle fiber compartments were identified in the lateral cranial region: red (superficial) mid white (deep) muscle. Hyperplasia muscle growth predominated in alevins mid hypertrophy in juveniles.

Histochemical study of muscle fiber types in Synbranchus marmoratus Bloch, 1795

The myotomal muscle of Synbranchus marmoratus was investigated using histochemical and immunohistochemical reactions. This musculature is composed of a superficial red compartment, uniformly distributed around the trunk circumferentially and also in the lateral line. The red compartment fibers are small in diameter and have an oxidative metabolism, a high rate of glycogen and a negative reaction to alkaline and acid myofibrillar ATPase (mATPase). The white muscle forms the bulk of the muscle mass. Its fibers are large in diameter and have a glycolytic metabolism, a negative reaction to glycogen, a strong reaction to alkaline mATPase and a negative reaction to acid mATPase. Between these two compartments there is an intermediate layer of fibers presenting a mosaic metabolism pattern with a high rate of glycogen. These fibers stained moderately for alkaline and acid m-ATPase. Several clusters of red muscles were observed inside the white muscle. Each cluster is composed of three fiber types, with a predominance of red and intermediate fibers. Reactivity to anti-MHC BA-D5 was positive only in the intermediate fibers. Reactivity to anti-MHC SC-71 was negative in all fiber types.

Metabolismo oxidativo de neutrófilos em ovelhas naturalmente infectadas por nematódeos gastrintestinais e correlação entre nível sérico de cortisol e carga parasitária

The relationships between neutrophils oxidative metabolism, cortisol serum levels and worm burden, estimated by fecal egg count (EPG), were studied in sheep naturally infected by gastrointestinal parasites at the end of pregnancy, during lactation, and after weaning. Twenty-two Suffolk sheep three to four year old, of same parity and season of parturition were used. Serum cortisol was determined by radioimmunoassay and the neutrophils oxidative metabolism by the nitroblue tetrazolium (NBT) reduction test. The highest EPG values were observed on the fifth week of lactation while the highest rates of cortisol and reduction of NBT occurred on the fourth week after weaning. A positive correlation (r = 0.52; P<0.01) was observed between the neutrophils capacity to reduce the NBT and the serum cortisol concentration in the pre-parturition period. Otherwise, the neutrophils oxidative metabolism decreased near to the parturition. A negative correlation (r = -0.39; P<0.01) between EPG and NBT reduction test was observed after weaning, which was coincident with the increase in the neutrophils capacity to reduce NBT, indicating that animals showing better immune response after weaning have neutrophils with higher oxidative metabolism and lower worm burden.

CYP-450 isoenzymes catalyze the generation of hazardous aromatic amines after reaction with the azo dye Sudan III

This work describes the mutagenic response of Sudan III, an adulterant food dye, using Salmonella typhimurium assay and the generation of hazardous aromatic amines after different oxidation methods of this azo dye. For that, we used metabolic activation by S9, catalytic oxidation by ironporphyrin and electrochemistry oxidation in order to simulate endogenous oxidation conditions. The oxidation reactions promoted discoloration from 65% to 95% of Sudan III at 1×10-4molL-1 and generation of 7.6×10-7molL-1 to 0.31×10-4molL-1 of aniline, o-anisidine, 2-methoxi-5-methylaniline, 4-aminobiphenyl, 4,4'-oxydianiline; 4,4'-diaminodiphenylmethane and 2,6-dimethylaniline. The results were confirmed by LC-MS-MS experiments. We also correlate the mutagenic effects of Sudan III using S. typhimurium with the strain TA1535 in the presence of exogenous metabolic activation (S9) with the metabolization products of this compound. Our findings clearly indicate that aromatic amines are formed due to oxidative reactions that can be promoted by hepatic cells, after the ingestion of Sudan III. Considering that, the use of azo compounds as food dyestuffs should be carefully controlled. © 2013 Elsevier Ltd.

Neutrophil dysfunction varies with the stage of canine visceral leishmaniosis

Canine visceral leishmaniosis (CVL) causes a dependent-stage alteration in neutrophil oxidative metabolism. When production of reactive oxygen species (ROS) exceeds the antioxidant capacity of neutrophils, apoptosis is triggered, impairing the viability and function of these cells, which can predispose dogs to infection. However, the uremic condition observed in late-stage CVL can also alter the viability and function of human neutrophils. To more clearly understand this relationship, the apoptosis rate and oxidative metabolism of neutrophils from control dogs (n= 20) were compared to dogs in moderate (n= 15) and very severe (n= 15) stage CVL, classified according to LeishVet Consensus. To assess neutrophil oxidative metabolism, superoxide production was measured using the nitroblue tetrazolium reduction test (NBT) in isolated neutrophils. The apoptosis rate of neutrophils was estimated using the morphological method. Moderate-stage dogs presented increased superoxide production, while dogs with very severe stage CVL presented decreased superoxide production and an increase neutrophil apoptosis rate. Leishmaniosis causes differential neutrophil dysfunction according to disease stage. In moderate stage CVL, increased superoxide production is observed with no change in neutrophil viability. However, in very severe stage CVL, decreased superoxide production and increased apoptosis occur associated with uremia. © 2013 Elsevier B.V.

Avaliação das estruturas químicas derivadas de diaminas do ferroceno no metabolismo oxidativo em cepas de Trypanosoma cruzi

Pós-graduação em Biociências e Biotecnologia Aplicadas à Farmácia - FCFAR

Efeito do exercício prévio sobre os parâmetros da cinética do VO2 durante o exercício moderado em ciclistas e indivíduos sedentários

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)