Linfoma primário de células T no líquido cefalorraquidiano de cão: relato de caso

Relata-se o diagnóstico de linfoma primário no sistema nervoso central em um cão Labrador Retrievier, de 10 anos de idade, que apresentava episódios convulsivos, incoordenação nos membros posteriores, head tilt, ataxia e sensibilidade diminuída no lado esquerdo. Constataram-se alterações laterais esquerdas, como ausências de propriocepção e de posicionamento tátil, alterações posteriores nas provas de carrinho de mão e de reação ao pulo e diminuição da extensão da postura e hemilocomoção. No líquido cefalorraquidiano (LCR), houve predomínio de linfócitos atípicos, caracterizados pela presença de anisocitose e anisocariose, nucléolos evidentes e anisonucleose, basofilia e microvacuolização citoplasmáticas, mitoses atípicas e corpúsculos linfoglandulares, compatíveis com linfoma, confirmado pelo exame histológico e imunocitoquímico, o qual revelou origem T, com expressão CD3+ e CD79-. A tomografia computadorizada não foi conclusiva e evidenciou diversas áreas hipodensas e intensificação de contraste na região de sulcos e giros do parênquima encefálico. A coleta do LCR foi essencial na rapidez do diagnóstico definitivo, indicando a natureza rara desta lesão primária.

Contribuição do contraste negativo na artrografia tomográfica do joelho normal de cães

A ruptura do ligamento cruzado cranial é uma das causas mais comuns de claudicação do membro pélvico de cães. A radiografia e a ultrassonografia são métodos de diagnóstico frequentemente utilizados na rotina clínica de pequenos animais, porém a tomografia computadorizada é uma modalidade de imagem ainda pouco estudada para avaliar a articulação do joelho de cães. O presente trabalho teve por objetivo avaliar a contribuição do contraste negativo na artrografia tomográfica do joelho normal de cães, para visibilizar as estruturas intra-articulares e padronizar o uso desse contraste na articulação. Foram utilizados 24 membros pélvicos de cães de raças variadas, selecionados pela ausência de histórico de doença articular prévia do joelho e por apresentarem exames radiográficos, ultrassonográficos e macroscópicos normais. O experimento foi delineado em dois grupos, sendo o grupo I constituído de animais com peso até 20 kg e grupo II acima de 20 kg. Foram feitos cortes tomográficos com o membro flexionado e estendido. A quantidade média de ar empregada para a distensão da cápsula articular foi de 49 ml para o Grupo I e de 81 ml para o Grupo II. Utilizou-se um tubo de látex na porção distal à articulação do joelho para reduzir o escape de ar pelo tendão extensor digital profundo, que possui comunicação intra-articular. Foi possível visibilizar pela imagem tomográfica, em todas as articulações, as seguintes estruturas: ligamento cruzado cranial e caudal, meniscos medial e lateral, ligamento patelar, ligamentos colaterais e cápsula articular. Desta forma, o contraste negativo se mostrou uma alternativa eficaz para auxiliar a identificação das estruturas anatômicas do joelho na artrografia tomográfica.

Avaliação dos reflexos espinhais em bezerros

Este trabalho objetivou avaliar, quantificar e padronizar a ocorrência dos reflexos espinhais em bezerros da raça holandesa de 15 a 90 dias de idade, os quais foram submetidos a avaliação nos membros torácicos (reflexo carpo radial, reflexo bicipital, reflexo tricipital e reflexo flexor) e nos membros pélvicos (reflexo patelar, reflexo tibial cranial, reflexo gastrocnêmio, reflexo ciático e reflexo flexor). Para quantificação da resposta involuntária frente ao reflexo realizado, padronizou-se a ausência do reflexo como sendo o algarismo 0; resposta discreta do reflexo como sendo l e a presença evidente da resposta como sendo 2. Os reflexos mais evidentes e constantes foram os reflexos flexor, carpo radial, patelar e tricipital. Os reflexos menos evidentes e menos freqüentes foram os reflexos tibial cranial, bicipital, gastrocnêmio e ciático.

Avaliação dos reflexos espinhais em cordeiros da raça Suffolk

Fez-se a avaliação qualitativa e semiquantitativa dos reflexos espinhais em ovinos, utilizando-se 51 animais da raça Suffolk, machos e fêmeas, entre quatro e cinco meses de idade. Usaram-se os reflexos bilaterais dos membros torácicos, extensor carpo-radial, bicipital, tricipital e flexor, e pélvicos, isquiático, gastrocnêmio, patelar, tibial cranial e flexor, sendo zero indicativo de ausência de reflexo, 1= reflexo discreto e 2= reflexo evidente. Nos membros torácicos, as melhores respostas foram obtidas no flexor (99,0%) e no extensor carpo-radial (87,3%), seguidos de valores menos expressivos no bicipital (11,8%) e no tricipital (2,0%), com grau 2 de avaliação. Nos membros pélvicos, todos os ovinos produziram respostas em grau 2 para o reflexo flexor. Verificam-se também respostas evidentes nos reflexos patelar (98,0%) e isquiático (81,4%). Apenas 20,6% dos animais apresentaram resposta evidente ao reflexo tibial cranial, e nenhum ovino respondeu ao reflexo gastrocnêmio de forma satisfatória.

Temporal response pattern of biochemical analytes in experimental diabetes

The activities of the enzymes aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), lactate dehydrogenase (LD), creatine kinase (CK), amylase (AMS) and angiotensin converting enzyme (ACE) have been used to assess the toxic effects of xenobiotics that have hypoglycaemic action in hepatic, pancreatic, renal and muscle tissue. Using a validated experimental model of diabetes mellitus in rats, we ascertained whether this syndrome itself affected the serum activities of these enzymes over a 53-day period. Levels of hepatic enzymes AST, ALT and ALP were higher in the streptozotocin (STZ)diabetic rats (group D), but were controlled by insulin therapy (group DI). AMS was reduced in group D and unchanged in group DI rats. Proteinuria was detected 1 day after STZ administation and partially controlled by insulin (group DI); its early presence in group D rats, and the lack of any change in serum ACE in this group, indicates that proteinuria is the better marker for microangiopathy. Microscopic examination of liver, kidney, heart and skeletal muscles (soleus and extensor digitorum longus) revealed various alterations in group D rat tissues, which were less pronounced in group DI. The liver, pancreas and kidney tissue-damage was consistent with the altered serum levels of AST, ALT, ALP and AMS and proteinuria. We conclude that: (i) rigorous control is required when these serum-enzyme levels are used as indicators of tissue toxicity in experimental diabetes, and (ii) LD, CK and bilirubin serum levels, which are unaffected by diabetes, can be used when testing effects of xenobiotics on tissues.

Differential morphofunctional characteristics and gene expression in fast and slow muscle of rats with monocrotaline-induced heart failure

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

Paralyzing and myotoxic effects of a recombinant bothropstoxin-1 (BthTX-I) on mouse neuromuscular preparations

As a first step to investigate the structure-function relationship of bothropstoxin-1 (BthTX-1), a myotoxin from Bothrops jararacussu snake venom, Our group previously cloned a recombinant toxin (rBthTX-1) in Escherichia coli. The aim or this work was to characterize the biological activities of this rBthTX-1 (1.0 mu M) in both phrenic-diaphragm and extensor digitorum longus preparations in vitro, by means of myographic and morphologic techniques. Native BthTX-1 (1.0 mu M) was used as a standard. The influence of heparin (27.5 mu g/ml) upon the biological activities of both toxins was also investigated. rBthTX-1 had similar effects to the native toxin inducing blockage of both directly and indirectly evoked contractions in phrenic-diaphragm preparations, and muscle damage characterized by edema, round fibers, and cell areas devoid of myofibrils. Interestingly the paralyzing activity of rBthTX-1 was slightly more potent than the native toxin. Heparin prevented paralyzing and myotoxic effects of both the native and recombinant toxins. This work shows that rBthTX-1 was expressed in a fully active form, and presents a biological profile similar to the native toxin. (c) 2005 Elsevier GmbH All rights reserved.

Reduction of crotoxin-induced neuromuscular blockade by gamma radiation

A comparative study between crotoxin and gamma irradiated crotoxin was performed on the indirectly evoked twitches and tetani of sciatic nerve-extensor digitorum longus muscle of rats. Crotoxin (3 to 14 mu g/ml) decreased the amplitude of twitches and induced a slight tetanic fade, and irradiated crotoxin did not significantly affect either twitch amplitude or tetanic tension. Since gamma radiation reduced the neurotoxicity of crotoxin it may be useful for the production of anticrotalic serum. (C) 1998 Elsevier B.V. Ltd. All rights reserved.

Antagonism of myotoxic and paralyzing activities of bothropstoxin-I by surarnin

Polyanionic substances are known to inhibit the myotoxic effects of some crotalide snake venoms. Bothropstoxin-I (BthTX-I), a basic Lys49 phospholipase (PLA(2)) homologue from Bothrops jararacussu venom, besides inducing muscle damage, also promotes the blockade of both directly and indirectly evoked contractions in mouse neuromuscular preparation. In this work, we evaluated the ability of suramin, a polysulfonated naphtylurea derivative, to antagonize the myotoxic and the paralyzing activities of BthTX-I on mice neuromuscular junction in vitro. Myotoxicity was assessed by light and electronic microscopic analysis of extensor digitorum longus (EDL) muscles; paralyzing activity was evaluated through the recording of both directly and indirectly evoked contractions of phrenic-diaphragm (PD) preparations. BthTX-I (1 muM) alone, or pre-incubated with suramin (10 muM) at 37degreesC for 15 min was added to the preparations for 120 min. BthTX-I induced histological alterations typical of myonecrosis in 14.6 +/- 1.0% of EDL muscle fibers. In addition, BthTX-I blocked 50% of both directly and indirectly evoked contractions in PD preparations in 72.1 +/- 9.1 and 21.1 +/- 2.0 min, respectively. Pre-incubation with suramin abolished both the muscle-damaging and muscle-paralyzing activities of BthTX-I. Since suramin is a polyanionic substance, we suggested that its effects result from the formation of inactive acid-base complexes with BthTX-I. (C) 2003 Elsevier Ltd. All rights reserved.

Nandrolone Stimulates Myod Expression During Muscle Regeneration in the Condition of Myonecrosis Induced by Bothrops jararacussu Venom Poisoning

Myonecrosis with permanent loss of muscle mass is a relevant local toxic effect following envenomation with Bothrops jararacussu snake venom. Regeneration of adult skeletal muscle involves the activation of satellite cells, a process regulated by myogenic regulatory factors (MRF). MyoD is an MRF involved in both proliferation and differentiation of satellite cells. Androgens are modulators of skeletal muscle, known to increase muscle mass and strength. This study examined the hypothesis that anabolic androgens improve the muscle regeneration process in mice following envenomation by Bothrops jararacussu snake venom. Myonecrosis was induced by venom injection (30 g/50 l in physiological solution) over the extensor digitorum longus (EDL) muscles of mice. Nandrolone (ND) (6 mg/kg, sc) was administered after 12 h, 7 d, and 14 d following venom injection. The histological changes in EDL muscle at 1, 3, 7, and 21 d after muscle injury were analyzed by light microscopy. Cross-sectional areas of fibers were measured. MyoD was evaluated by immunofluorescence technique. Histological examination revealed the presence of a regeneration process in ND-treated animals, characterized by the appearance of some myotubes at 3 d, and numerous myotubes at 7 d from venom injection. Nandrolone treatment reduced the frequency of small fibers at 7 and 21 d after venom administration, and increased the frequency of large fibers at 7 d postinjury. Nandrolone also significantly augmented the expression of MyoD-positive cells at 7 and 21 d after envenomation. These results suggest that ND accelerates muscle regeneration and indicate the involvement of MyoD in this process.

Heart failure increases atrogin-1 and MuRF1 gene expression in skeletal muscle with fiber type-specific atrophy

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

Histochemical differences in the responses of predominantly fast-twitch glycolytic muscle and slow-twitch oxidative muscle to veratrine

The aim of this study was to investigate if the Na+-channel activating alkaloid veratrine is able to change the oxidative and m-ATPase activities of a fast-twitch glycolytic muscle (EDL, extensor digitorum longus) and slow-twitch oxidative muscle (SOL, soleus) in mice. Oxidative fibers and glycolytic fibers were more sensitive to veratrine than oxidative-glycolytic fibers 15, 30 and 60 min after the i.m. injection of veratrine (10 ng/kg) with both showing an increase in their metabolic activity in both muscles. In EDL, the m-ATPase reaction revealed a significant (p < 0.001) decrease (50%) in the number of type IIB fibers after 30 min while the number of type I fibers increased by 550%. Type I fibers decreased from 34% in control SOL to 17% (50% decrease) in veratrinized muscles, with a 10% decrease in type IIA fibers within 15 min. A third type of fiber appeared in SOL veratrinized muscle, which accounted for 28% of the fibers. Our work gives evidence that the changes in the percentage of the fiber types induced by veratrine may be the result, at least partially, from a direct effect of veratrine on muscle fibers and else from an interaction with the muscle type influencing distinctively the response of a same fiber type. Based on the results obtained in the present study the alterations in EDL may be related to the higher number of Na+ channels present in this muscle whereas those in SOL may involve an action of veratrine on mitochondria. Although it is unlikely that the shift of enzymes activities induced by veratrine involves genotypic expression changes an alternative explanation for the findings cannot be substantiated by the present experimental approach. (C) 2002 Elsevier B.V. Ltd. All rights reserved.

Co-60 gamma irradiation prevents Bothrops jararacussu venom neurotoxicity and myotoxicity in isolated mouse neuromuscular junction

The ability of gamma radiation from Co-60 (2000 Gy) to attenuate the toxic effects of Bothrops jararacussu venom was investigated on mouse neuromuscular preparations in vitro. A comparative study between the effects of native and irradiated venoms was performed on both phrenic-diaphragm (PD) and extensor digitorum longus (EDL) preparations by means of myographic, biochemical and morphological techniques. Native venom (10 and 20 mug/ml) induced a concentration-dependent paralysis of both directly and indirectly evoked contractions on PD preparations. At 20 mug/ml, it also caused a pronounced myotoxic effect on the EDL muscle preparation that was characterized by an increase of creatine kinase release and by several morphological changes of this preparation. By contrast, irradiated venom, even at concentrations as high as 40 mug/ml, induced neither paralyzing nor myotoxic effects. It was concluded that Co-60 gamma radiation is able to abolish both the paralyzing and the myotoxic effects of B. jararacussu venom on the mouse neuromuscular junction. These findings support the hypothesis that gamma radiation could be an important toot to improve antisera production by reducing toxicity while preserving immunogenicity. (C) 2002 Elsevier B.V. Ltd. All rights reserved.

Spermiogenesis and spermatozoa ultrastructure in five species of the Curimatidae with some considerations on spermatozoal ultrastructure in the Characiformes

A espermiogênese nas espécies Steindachnerina insculpta, Cyphocharax gillii, C. modestus, C. spilotus e Potamorhina altamazonica de Curimatidae é caracterizada pelo desenvolvimento lateral do flagelo, rotação do núcleo, formação excêntrica da fossa nuclear e cromatina compactada em fibras espessas. Estes espermatozóides exibem uma cabeça esférica contendo um núcleo com cromatina altamente condensada em fibras espessas com pequenas áreas eletronlúcidas, e sem acrossoma. A fossa nuclear é do tipo moderado e excêntrico, penetrada pelo complexo centriolar. A peça média é pequena, tem muitas vesículas alongadas e um curto canal citoplasmático. Mitocôndrias podem ser alongadas, ramificadas ou em forma de C, e são separadas do segmento inicial do axonema pelo canal citoplasmático. O flagelo contém a estrutura clássica do axonema (9+2) e tem um compartimento membranoso na região inicial; não possui expansões laterais (fins). Somente pequenas diferenças foram observadas entre as espécies e gêneros analisados de Curimatidae. A espermiogênese e os espermatozóides de Curimatidae têm muitas das características encontradas em quase todas as outras espécies de Characiformes. Por outro lado, a presença de um compartimento membranoso na região inicial do flagelo dos curimatídeos, uma estrutura comum nos espermatozóides de muitos cipriniformes, é desconhecida em outros characiformes. Discute-se sobre a espermiogênese e espermatozóides de Characiformes.

Skeletal muscles with antagonistic muscular actions: morphological, contractile and metabolic characteristics

The muscles can perform the same function in a specific segment (muscles of fast and slow contraction), and at the same time be antagonistic in relation to muscular action (flexors or extensors). The present research aimed to study the morphology, frequency and metabolism of fiber types and the contractile characteristics of extensor and flexors muscles of rabbit. We studied muscles anterior tibialis (AT), flexor digitorum supeficialis (FDS), extensor digitorum longus (EDL) and posterior tibialis (PT). The muscles were submitted to the techniques HE, NADH-TR and myofibrillar ATPase. In EDL and PT extensor muscles, the frequencies of red (SO + FOG) and white fibers (FG) were 68.77% and 31.23% versus 58.87% and 41.13%, respectively. In the AT and FDS flexor muscles, these frequencies were 75.14% and 24.86% versus 73.89% and 26.11%, respectively. In extensor muscles, the percentage of slow contraction fibers was 8.05% in EDL and 9.74% in PT, and in fast contraction, 91.95% in EDL and 90.26% in PT. In flexors, the slow contraction frequencies were 12.35% in AT and 8.17% in FDS, and in fast contraction, 87.65% and 91.83%, respectively. Skeletal muscles with antagonistic muscular actions (flexors and extensors) the morphological, contractile and metabolic characteristics are identical.