Estimulação elétrica neuromuscular em cães com atrofia muscular induzida / Neuromuscular electric stimulation in dogs with induced muscle atrophy

Empregou-se a estimulação elétrica neuromuscular (EENM) de baixa freqüência no músculo quadríceps femoral de cães com atrofia induzida e avaliou-se a ocorrência de ganho de massa nessa musculatura. Foram utilizados oito cães com pesos entre 15 e 30kg, distribuídos aleatoriamente em dois grupos denominados de I ou controle e II ou tratado. A articulação femorotibiopatelar esquerda foi imobilizada por 30 dias pelo método de transfixação percutânea tipo II, com retirada de aparelho de imobilização após esse período. Decorridas 48 horas da remoção, foi realizada a EENM nos cães do grupo II, cinco vezes por semana, com intervalo de 24 horas cada sessão, pelo período de 60 dias. Foram avaliadas a circunferência da coxa, a goniometria do joelho, a análise clínica da marcha, as enzimas creatina-quinase (CK) e aspartato-amino-transferase (AST) e a morfometria das fibras musculares em cortes transversais do músculo vasto lateral colhido mediante biópsia muscular. A EENM foi empregada no músculo quadríceps femoral na freqüência de 50Hz, duração de pulso de 300 milisegundos e relação de tempo on/off de 1:2. Quanto à morfometria das fibras do músculo vasto lateral, no grupo tratado houve aumento significativo (P<0,05) da área transversal aos 90 dias em relação ao dia zero. A EENM de baixa freqüência ocasiona hipertrofia do músculo vasto lateral em cães após a imobilização rígida temporária da articulação do joelho.

Low frequency neuromuscular electrical stimulation (NMES) was used on the femoral quadriceps of dogs with induced muscular atrophy and the occurrence of gain in mass in these muscles was evaluated. Eight dogs from 15 to 30kg were randomly distributed in two groups named I, or control; and II, or treated. For the induction of muscular atrophy, the left femoral-tibial-patellar joint was immobilized for 30 days by percutaneous transfixation type II. After 30 days, the immobilization device was removed. The NMES treatment began 48 hours after the removal of the immobilization device of the dogs of group II, and it was carried out five times per week with an interval of 24 hours between each session, for 60 days. The following parameters were measured: thigh circumference, goniometry of the knee, clinical gait analysis, creatine kinase (CK) and aspartate aminotransferase (AST) enzymes, and morphometry of the muscular fibers in transversal cuts of the vastus lateralis muscle collected through muscular biopsy. The NMES was applied on the femoral quadriceps at a frequency of 50 Hz, with pulse duration of 300 milliseconds, and the on/off time was at a proportion of 1:2. Regarding the morphometry of the vastus lateralis fibers, a significant increase (P<0.05) in the transversal area of the treated group at 90 days was observed when compared with that identified at the time of immobilization. Thus, it can be concluded that low frequency NMES brings about hypertrophy of the vastus lateralis muscle in dogs after temporary rigid immobilization of the knee joint.

Acetylcholinesterase activity in rats experimentally demyelinated with ethidium bromide and treated with interferon beta.

The ethidium bromide (EB) demyelinating model was associated with interferon beta (IFN-beta) to evaluate acetylcholinesterase (AChE) activity in the striatum (ST), hippocampus (HP), cerebral cortex (CC), cerebellum (CB), hypothalamus (HY), pons (PN) and synaptosomes from the CC. Rats were divided into four groups: I control (saline), II (IFN-beta), III (EB) and IV (EB and IFN-beta). After 7, 15 and 30 days rats (n = 6) were sacrificed, and the brain structures were removed for enzymatic assay. AChE activity was found to vary in all the brain structures in accordance with the day studied (7-15-30 days) (P < 0.05). In the group III, there was an inhibition of the AChE activity in the ST, CB, HY, HP and also in synaptosomes of the CC (P < 0.05). It was observed that IFN-beta per se was capable to significantly inhibit (P < 0.05) AChE activity in the ST, HP, HY and synaptosomes of the CC. Our results suggest that one of the mechanisms of action of IFN-beta is through the inhibition of AChE activity, and EB could be considered an inhibitor of AChE activity by interfering with cholinergic neurotransmission in the different brain regions.

Apyrase and 5'-nucleotidase activities in synaptosomes from the cerebral cortex of rats experimentally demyelinated with ethidium bromide and treated with interferon-beta.

Apyrase and 5'-nucleotidase activities were analyzed in an ethidium bromide (EB) demyelinating model associated with interferon-beta (IFN-beta). The animals were divided in groups: I, control (saline); II, saline and IFN-beta; III, EB and IV, EB and IFN-beta. After 7, 15 and 30 days the animals (n = 5) were sacrificed and the cerebral cortex was removed for synaptosome preparation and enzymatic assays. Apyrase activity using ATP as substrate increased in groups II, III and IV (P < 0.001) after 7 days and in groups III and IV (P < 0.001) after 15 days. Using ADP as substrate, an activation of this enzyme was observed in group III (P < 0.05) after seven and 15 days. The 5'-nucleotidase activity increased in group III (P < 0.05) after 7 days and in groups II, III and IV (P < 0.001) after 15 days. After 30 days treatment, no significant alteration was observed in enzyme activities. Results showed that apyrase and 5'-nucleotidase activities are altered in demyelination events and that IFN-beta was able to regulate the adenine nucleotide hydrolysis.

Three types of immunotherapics against pythiosis insidiosi developed and evaluated.

Pythiosis is a granulomatous disease of horses, cattle, dogs, cats and humans identified in tropical and subtropical areas and caused by Pythium insidiosum, a zoosporic fungus. Experimental models of pythiosis in naturally infected species have not yet been reported but, rabbits may be inoculated with zoospores as an experimental model for studying the disease. The present study evaluates the efficacy of three different of immunotherapics in the rabbit model. Approximately 17500 zoospores of oomycete P. insidiosum (CBS 101555 strain) were inoculated in each animal to generate the disease. Immunotherapics were produced from vortexed or sonicated cultures of the same strain. Four groups of five animals were employed: group 1, placebo; group 2, sonicated immunotherapic; group 3, mixed immunotherapic; and group 4, vortexed immunotherapic. All rabbits were inoculated with viable zoospores one month before administration of the immunotherapics. Eight doses of immunotherapic or placebo were used in each animal with a 14 day interval between injections. Rabbits receiving the vortexed immunotherapic were most effectively protected (P<0.05), showing a decrease in the area of coastal nodules due to Pythiosis insidiosum by 71.8% after 26 weeks of evaluation. Moreover, two animals in this group showed complete remission of the infection at the end of the 26 weeks. In contrast to these findings, rabbits given the sonicated immunotherapic did not show any protection and had an increase of 211.8% in the size of lesions. This failure of sonicated immunotherapic may reflect denaturation of protective antigens due to the sonication method.