Comparative biochemical studies of myotoxic phospholipase A(2) from Bothrops venom

Venoms from Bothrops jararacussu, Bothrops asper, Bothrops atrox, Bothrops pirajai, Bothrops moojeni, Bothrops alternatus and Bothrops (Bothriopsis) bilineata were fractionated using a simplified procedure based on ion-exchange chromatography on CM-Sepharose at pH 8.0 or reverse phase HPLC. The resulting elution profiles showed important differences in the myotoxin content of these venoms. The venoms from B. alternatus, B. atrox and Bothriopsis bilineata did not contain the major myotoxin found in the other venoms. The amino acid sequence of the first 50 residues of the N-terminal region of the PLA(2)-like myotoxins showed a homology of 90-96% with other bothropic myotoxins. All of the myotoxins isolated induced rat paw edema, increased the level of plasma creatine kinase and produced myonecrosis together with polymorphonuclear cell infiltration.

The use of the oxidative stress responses as biomarkers in Nile tilapia (Oreochromis niloticus) exposed to in vivo cadmium contamination

Water contaminants have a high potential risk for the health of populations. Protection from toxic effects of environmental water pollutants primarily involves considering the mechanism of low level toxicity and likely biological effects in organisms who live in these polluted waters. The biomarkers assessment of oxidative stress and metabolic alterations to cadmium exposure were evaluated in Nile tilapia, Oreochromis niloticus. The fish were exposed to 0.35, 0.75, 1.5, and 3.0 mg/l concentrations of Cd2+ (CdCl2) in water for 60 days. Fish that survived cadmium exposure showed a metabolic shift and a compensatory development for maintenance of the body weight gain. We observed a decreased glycogen content and decreased glucose uptake in white muscle. Lactate dehydrogenase (LDH) and creatine phosphokinase (CK) activities were also decreased, indicating that the glycolytic capacity was decreased in this tissue. No alterations were observed in total protein content in white muscle due to cadmium exposure suggesting a metabolic shift of carbohydrate metabolism to maintenance of the muscle protein reserve. There was an increase in glucose uptake, CK increased activity, and a clear increase of LDH activity in red muscle of fish with cadmium exposure. Since no alterations were observed in lipoperoxide concentration, while antioxidant enzymes glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) were changed in the liver and the red and white muscle of fish with cadmium exposure, we can conclude that oxygen free radicals are produced as a mediator of cadmium toxicity. Resistance development is related with increased activities of antioxidant enzymes, which were important in the protection against cadmium damage, inhibiting lipoperoxide formation. (C) 2002 Elsevier B.V. Ltd. All rights reserved.

Myotoxic effects of mastoparan from Polybia paulista (Hymenoptera, Epiponini) wasp venom in mice skeletal muscle

In a previous study, we showed that the Polybia paulista wasp venom causes strong myonecrosis. This study was undertaken to characterize the myotoxic potency of mastoparan (Polybia-MPII) isolated from venom (0.25 mu g/mu l) and injected in the tibial anterior (TA) muscle (i.m.) of Balb/c mice. The time course of the changes was followed at muscle degenerative (3 and 24 h) and regenerative (3, 7, and 21 days) periods (n = 6) after injection and compared to matched controls by calculation of the percentage of cross-sectional area affected and determination of creatine kinase (CK) activity (n = 10). The results showed that although NIP was strongly myotoxic, its capacity for regeneration was maintained high. Since the extent of tissue damage was not correlated with the CK serum levels, which remained very low, we raised the hypothesis that the enzyme underwent denaturation by the peptide. Evidence suggested that MP induced the death of TA fibers by necrosis and apoptosis and had the sarcolemma as its primordial target. Given its amphiphilic polycationic nature and based on the vast spectrum of functions attributed to the peptide, we suggest that MP interaction with cell membrane impaired the phosphorylation of dystrophin essential for sarcolemma mechanical stability, and disturbed Ca2+ mobilization with obvious implications on sarcoplasmic reticulum and mitochondrial functioning. (c) 2007 Elsevier Ltd. All rights reserved.

Efeito protetor após sessões de exercício excêntrico: comparação entre membros superiores e inferiores

O objetivo foi comparar a magnitude do efeito protetor (EP) contra o dano muscular (DM) induzido por uma sessão de exercícios excêntricos (EEM) entre os extensores do joelho e os flexores do cotovelo. Doze sujeitos do gênero masculino foram divididos em 2 grupos, braços (GB) e pernas (GP), e realizaram 2 sessões de EEM. Foram coletados 3 marcadores de DM, sendo eles, pico de torque isométrico (PTI), creatina quinase (CK) e percepção subjetiva de dor (PSD), antes, imediatamente após (com exceção da CK) e 48 horas após cada sessão de EEM. Foi encontrada queda significante de PTI e aumento significante de CK e PSD tanto imediatamente e 48 horas após a primeira sessão de EEM para o GB. No GP houve aumento significante de CK 48 horas após os EEM e da PSD imediatamente após os EEM, decorrentes da primeira sessão. No GB, a segunda sessão apenas provocou queda de PTI imediatamente após os EEM, enquanto no GP houve aumento significante apenas na PSD imediatamente após a segunda sessão de EEM. Apenas a CK apresentou EP para ambos os grupos. Pudemos concluir que o EP foi maior para o GB em comparação com o GP. Esse fenômeno pode ter ocorrido em detrimento da existência de um EP prévio para o GP, uma vez que este membro realiza contrações excêntricas intensas com maior freqüência no dia-a-dia, quando comparados com os GB.

Padronização de um protocolo experimental de treinamento periodizado em natação utilizando ratos Wistar

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Psychological, biochemical and physiological responses of Brazilian soccer players during a training program

Aims of the study. - The aim of this study is to investigate the behavior of the physiological, biochemical and psychological parameters in Brazilian soccer players during a training program.Materials. - Fifteen athletes were evaluated at the beginning (T1), in the middle (T2) and at the end (T3) of the training program. on the first day, at 7:30 am, before the blood collecting at rest for the determination of serum creatine kinase (CK), serum creatinine and serum urea, the athletes had their psychological parameters assessed by the profile of mood state questionnaire (POMS). After 90 min, they performed a 250-m sprint. on the second day at 8:30 am, the athletes had their alactic anaerobic performance measured and, after 40 min, they completed the aerobic test. Friedman test was used to verify the behavior of overtraining markers.Results. - There was a decrease in vigor score in T3 (p=0.01) compared with T1 and T2. In T3 (p=0.01), the athletes also showed an increase in serum creatinine levels compared to T1. Furthermore, in the same period, we verified a diminishing in the team performance.Conclusion. - The training program developed between T2 and T3 led to the fall of the vigor score, the increase in serum creatinine concentrations and the diminishing in team performance. (c) 2007 Elsevier Masson SAS. All rights reserved.

Heavy-intensity aerobic exercise affects the isokinetic torque and functional but not conventional hamstrings:quadriceps ratios

Running exercises are frequently related to muscular injuries, which may be a result of muscular imbalance. The present study aimed to verify the effects of heavy-intensity continuous running exercise on the functional and conventional hamstrings: quadriceps ratios, and also in the knee flexors and extensors EMG activity in active non-athletic individuals. Sixteen active males performed maximal isokinetic concentric and eccentric knee flexions and extensions at 60 degrees s(-1) and 180 degrees s(-1). In another session, the same procedure was conducted after a continuous running exercise at 95% onset of blood lactate accumulation. Torque and electromyographic ratios were calculated from peak torque and integrated electromyographic activity (knee flexor and extensors). Creatine kinase was measured before and 24 h after running exercise. Eccentric torque (knee flexion and extension) decreased significantly after running only at 180 degrees s(-1) (p < 0.05). No differences were found for the conventional torque ratios (p > 0.05), however, the functional torque ratios at 180 degrees s(-1) decreased significantly after running (p < 0.05). No effects on the electromyographic activity and electronnyographic ratios were found (p > 0.05). Creatine kinase increased slightly 24 h after running (p < 0.05). Heavy-intensity continuous running exercise decreased knee flexor and extensor eccentric torque, and functional torque ratios under fast velocities (180 degrees s(-1)), probably as result of peripheral fatigue. (C) 2008 Elsevier Ltd. All rights reserved.

Muscle damage slows oxygen uptake kinetics during moderate-intensity exercise performed at high pedal rate

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Toxicological evaluation of long-term intravenous administration of amitraz in horses

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Ação do ácido trissódio-cálcio-dietileno-triaminopentaacético (CaNa3DTPA) nas lesões de isquemia-reperfusão em membro posterior de rato

OBJETIVO: Ação do ácido trissódio-cálcio-dietileno-triaminopentaacético (CaNa3DTPA), quelante de ferro com ação ainda anti-viral, antiinflamatória e imunológica, na atenuação de lesões de reperfusão em músculos esqueléticos de ratos. MÉTODOS: 52 ratos Wistar, pesando 188±22g, foram anestesiados e submetidos a semi-amputação de membro posterior direito (MPD), poupando-se o fêmur, artéria e veia femorais. Foram então randomizados e distribuídos: G1-CTAN - controle anestesia, sem cirurgia e sem isquemia; G2-CTCIR - controle cirurgia, sem isquemia; G3-IRCT e G3-IRDTPA - com isquemia (4 hora) e reperfusão ( 2 horas). O G3-IRCT foi tratado, ao final da isquemia, com cloreto de sódio 0,9% e G3-IRDTPA com (CaNa3DTPA).Parâmetros: Circunferência do pé direito e peso do rato, dosagem sérica de CPK, dosagem de malonaldeído e microscopia óptica de músculos soleus bilateral. RESULTADOS: Aumento da circunferência nos G3-IRCT e G3-IRDTPA (significante no G3-IRCT quando comparado ao G1-CTAN); CPK elevado nos G3-IRCT e G3-IRDTPA comparados aos controles; MDA mais alto no membro contralateral do G3-IRDTPA, comparado ao MPD do G3-IRDTPA e ao G1-CTAN; maior edema intersticial em G3-IRCT, maior infiltrado inflamatório em G3-IRDTPA e recuperação dos níveis de glicogênio semelhantes em G3-IRCT e G3-IRDTPA. CONCLUSÃO: Apesar do menor edema no G3-IRDTPA comparado ao G3-IRCT, o CaNa3DTPA não alterou CPK sérico, MDA muscular e morfologia muscular dos animais.

Tempo de jejum na granja sobre o perfil hormonal e os parâmetros fisiológicos em suínos de abate pesados

O objetivo deste estudo foi avaliar o tempo de jejum na granja e a posição dos animais na carroceria do caminhão durante o transporte ao abatedouro sobre o status hormonal e fisiológico de suínos de abate pesados visando obter melhorias no manejo pré-abate e reduzir perdas na qualidade de carne. Foram utilizadas 64 fêmeas com peso médio de 133+11kg, oriundas de duas granjas de terminação. Os tempos de jejum avaliados foram nove, 12, 15 e 18h, enquanto que as posições consideradas na carroceria foram box (frente, meio e atrás), piso (inferior e superior) e lado (lateral direita e esquerda). Ao abate, foram medidos os níveis de glicose, lactato e CPK no sangue. A concentração de cortisol na saliva (CCS) foi medida nas granjas (24 horas antes e após embarque) e no abatedouro (logo após o descarregamento e antes do abate). A freqüência cardíaca foi monitorada durante todo o manejo pré-abate. Foi observado o efeito da interação entre TJG e o local de avaliação sobre a CCS e a freqüência cardíaca. A CCS e a freqüência cardíaca aumentaram significativamente da granja ao desembarque no abatedouro em relação ao descanso pré-abate no abatedouro foi observada uma redução (P<0,05) nos valores. A CCS variou em função do TJG e o local de avaliação da seguinte maneira: suínos com 18 horas de jejum apresentaram menor (P<0,05) variação na CCS durante o transcorrer das diferentes etapas do manejo pré-abate do que suínos com TJG menores e, entre estes, os animais com TJG de nove horas apresentaram a maior (P<0,05) variação. Antes do abate, os suínos com TJG de nove horas apresentaram o maior valor (P<0,05) de CCS quando comparados aos outros TJG. Conclui-se que o TJG promove mudanças (P<0,05) nos valores do cortisol na saliva e na freqüência cardíaca no manejo pré-abate, mas não afetam (P>0,05) os níveis de glicose, lactato e CPK no abate dos suínos.

Efeito do manejo pré-abate sobre alguns parâmetros fisiológicos em fêmeas suínas pesadas

O objetivo neste trabalho foi avaliar o efeito do período de descanso (3, 5, 7 e 9 horas) dos suínos no frigorífico (PDF) e da localização dos suínos na carroceria do caminhão (PBO), quando transportados, no inverno ou verão, sobre alguns parâmetros fisiológicos avaliados em 64 fêmeas, com peso médio de 130kg para abate, durante o manejo pré-abate. Para a análise estatística, foram considerados, no modelo de análise da variância, os efeitos de bloco, PDF, PBO e da interação (bloco x PDF), entre outros. O PDF influenciou, significativamente, as concentrações de lactato no sangue e cortisol na saliva. Suínos que descansaram 5 e 7 horas apresentaram maior concentração de lactato em relação aos animais que descansaram 3 e 9 horas. No transporte, a freqüência cardíaca foi muito maior em relação aos demais locais avaliados. Concluiu-se que o incremento do PDF não promove mudanças na freqüência cardíaca, nas concentrações de glicose e CPK no sangue e cortisol na saliva, mas interfere na concentração de lactato no sangue dos suínos.

Parâmetros bioquímicos de equinos submetidos à simulação de prova de enduro recebendo dietas com adição de óleo de soja

Com o objetivo de verificar o efeito da adição de níveis crescentes de óleo no concentrado sobre a atividade plasmática das enzimas creatina quinase (CK), aspartato aminotransferase (AST) e lactato desidrogenase (LDH) como indicativo de metabolismo energético, foram fornecidas dietas experimentais compostas de cinco níveis de óleo (controle, 6, 12, 18 e 24% do concentrado). Utilizaram-se 20 equinos da raça Árabe, peso médio de 400 kg, submetidos a prova de enduro de 80 km em esteira rolante. O enduro foi dividido em quatro anéis de 20 km, com duração média de 1 hora e dez minutos. A adição de óleo e a distância percorrida tiveram efeitos sobre as variáveis AST, CK e LDH, que apresentaram as respectivas expressões: AST (7,045-0,2292x+0,007991x2+0,008517z- 0,0003282xz), CK (8,06-,07020x+0,05546x2-0,001262x3+0,01204z+0,0006207xz) e LDH (6,624-0,3522x+0,03448x2-0,0008382x3+0,02401z-0,0007489xz) . O óleo é uma importante e bem aproveitada fonte de energia para equinos em exercício, pois sua adição na dieta de animais submetidos a prova de enduro promoveu alteração metabólica que favorece a produção de energia. O metabolismo animal poupou suas reservas energéticas oriundas da glicose, favorecendo a utilização do óleo. A menor atividade plasmática das enzimas AST, CK e LDH com a adição de óleo nas dietas indica direcionamento do metabolismo energético para a β-oxidação. Como apresentam várias isoenzimas, as enzimas estudadas atuam amplamente no metabolismo energético, favorecendo a constante reposição de ATP ao longo do exercício.

Dietary macronutrients, endocrine functioning and intermediary metabolism in broiler chickens - Pair wise substitutions between protein, fat and carbohydrate

Three pens of male broiler chicks were raised under standard conditions and fed from 7 to 42 days of age three isocaloric diets each with 15.8; 19.6 and 19.5% of CP; and 51, 51, and 44% of CHO; and 6.5; 3.0 and 7.7% of fat, and designated as the low protein (LowCP), low lipid (LowL) and low carbohydrate (LowCHO) diets, respectively. Body weights and feed intake were monitored weekly and blood samples were collected at the same time for posterior analysis of hormone and metabolite content. Chickens fed the LowCP diet were characterized by a reduced body weight gain and feed intake and poorer feed conversion efficiency compared to those fed the LowL and LowCHO diets, which were very similar in this respect. Plasma corticosterone and glucose levels and creatine kinase activity were not significantly changed by diet composition. LowCP chickens were characterised by the lowest plasma T-4 and uric acid levels (indicative for reduced protein breakdown and lower protein ingestion) but highest plasma triglyceride levels (congruent with their higher fat deposition) compared to the LowL and LowCHO chickens. LowL chickens had on average higher plasma T-3 and free fatty acid levels compared to the LowCP and LowCHO chickens.In conclusion, a limited substitution of carbohydrate for fat in iso-nitrogenous, iso-energetic diets has no pronounced effects on plasma hormone and metabolite levels, except for the elevation in T-3 (may enhance glucose uptake) and free fatty acid levels in the plasma of the chickens fed the LowL diet. The protein content of the diet has a greater impact on zootechnical performance, and underlying endocrine regulation of the intermediary metabolism compared to the dietary lipid and CHO fraction. (C) 2003 Elsevier B.V. All rights reserved.

ANTAGONISM OF THE MYOTOXIC EFFECTS OF BOTHROPS-JARARACUSSU VENOM AND BOTHROPSTOXIN BY POLYANIONS

The effects of heparin and other polyanions on the myotoxicity of Bothrops jararacussu venom and purified bothropstoxin (BthTX) were investigated. The release rate of creatine kinase (CK) from isolated extensor digitorum longus muscle and the plasma CK activity of mice were used to quantify the results. The myotoxic effects of B. jararacussu venom or BthTX were inhibited by preincubation of these agents with one of the following: a heterogeneous heparin preparation (designated 'heparin'), low mol. wt heparin (H-4500) or dextran sulfates (DS-8000 and DS-500,000). Non-sulfated dextran (D-40,000) and two chondroitin sulfates were ineffective. The antimyotoxic effects of the polyanions are ascribed to their forming inactive acid-base complexes with the basic myotoxins of Bothrops venoms. Gel-filtration experiments in Sephadex provided direct evidence for complex formation between heparin and BthTX. Intravenous (i.v.) administration of H-4500 or DS-8000 opposed the increase in plasma CK activity induced by a subsequent i.m. injection of venom or BthTX. In contrast, pretreatment with i.v. heparin or DS-500,000 enhanced the venom-induced increase in plasma CK activity. This effect was not observed (1) when the animals were treated with a polyvalent antivenom, which inhibits the coagulation and local stasis induced by Bothrops venoms, and (2) when BthTX, which has no thrombotic or hemorrhagic properties, was the myotoxic agent. The potentiation of the venom-induced increase in plasma CK activity by heparin and DS-500,000 is ascribed to improved washout of the CK released from damaged fibers, because of the anticoagulant properties of the drugs.