ABSTRACT. Duchenne muscular dystrophy (DMD) is a human disease characterized by progressive and irreversible skeletal muscle degeneration caused by mutations in genes coding for important muscle proteins. Unfortunately, there is no efficient treatment for this disease; it causes progressive loss of motor and muscular ability until death. The canine model (golden retriever muscular dystrophy) is similar to DMD, showing similar clinical signs. Fifteen dogs were followed from birth and closely observed for clinical signs. Dogs had their disease status confirmed by polymerase chain reaction analysis and genotyping. Clinical observations of musculoskeletal, morphological, gastrointestinal, respiratory, cardiovascular, and renal features allowed us to identify three distinguishable phenotypes in dystrophic dogs: mild (grade I), moderate (grade II) and severe (grade III). These three groups showed no difference in dystrophic alterations of muscle morphology and creatine kinase levels. This information will be useful for therapeutic trials, because DMD also shows significant, inter-and intra-familiar clinical variability. Additionally, being aware of phenotypic differences in this animal model is essential for correct interpretation and understanding of results obtained in pre-clinical trials.

PURPOSE: Evaluate the bone tissue recovery following transplantation of ovine mesenchymal stem cells (MSC) from bone marrow and human immature dental-pulp stem cells (hIDPSC) in ovine model of induced osteonecrosis of femoral head (ONFH). METHODS: Eight sheep were divided in three experimental groups. First group was composed by four animals with ONFH induced by ethanol through central decompression (CD), for control group without any treatment. The second and third group were compose by two animals, six weeks after ONFH induction received transplantation of heterologous ovine MSC (CD + oMSC), and hIDPSC (CD + hIDPSC), respectively. In both experiments the cells were transplanted without application of any type of immunosupression protocol. RESULTS: Our data indicate that both cell types used in experiments were able to proliferate within injured site providing bone tissue recovery. The histological results obtained from CD+hIDPSC suggested that the bone regeneration in such animals was better than that observed in CD animals. CONCLUSION: Mesenchymal stem cell transplant in induced ovine osteonecrosis of femoral head by central decompression technique is safe, and apparently favors bone regeneration of damaged tissues.

Purpose:To evaluate different protocols to isolate stem cells from ovine umbilical cord blood and adipose tissue. Methods: There were used 5 samples of umbilical blood and 5 samples of perirenal adipose tissue from 10 female sheep. All the samples were obtained through surgery, to harvest aseptic samples. There were used 3 protocols for obtainment and culture of umbilical cord blood stem cells and 4 protocols for ovine adipose tissue stem cells. Results: It was possible to observe only one successful protocol for the obtainment of umbilical cord blood stem cells. When analyzing the techniques used to obtain adipose tissue stem cells, only one of the methods was effective as well. Through colony forming unit assay, there were obtained 58 colonies of cells after seven days in culture. Flow citometry tests revealed the cells were positive to CD44 and exhibited negative reaction to CD38, CD45, CD41/61. These cells showed a growth curve with very well defined phases LOG, LAG and PLATEAU. This phases are typically seem in mesenchymal stem cells growth curves. Conclusions: The isolation and culture of mesenchymal stem cells from ovine umbilical cord blood are complex and request more detailed assays. Stem cells from fat tissue sheep showed mesenchymal characteristics, according to their cell growth curve, ability to origin colonies of fibroblastoid cells and positive reactivity with the antibody CD44 by flow citometry. Key words: Stem Cells. Umbilical Cord. Adipose Tissue. Sheep. RESUMO Objetivo:Testar diferentes protocolos para o isolamento de células tronco a partir de sangue de cordão umbilical e tecido adiposo de ovinos. Métodos: Foram utilizadas cinco amostras de sangue de cordão umbilical e cinco amostras de tecido adiposo perirrenal de 10 fêmeas de ovelha. A coleta das amostras foi realizada através de procedimento cirúrgico para coleta do material de forma mais asséptica possível. Foram realizados três protocolos de isolamento e cultivo das células-tronco do cordão umbilical e quatro protocolos para o isolamento e cultivo das células-tronco de gordura de ovinos Resultados: Somente um dos protocolos utilizados para o isolamento das células-tronco de cordão umbilical foi efetivo. Dos quatro protocolos utilizados para isolamento das células-tronco de gordura, da mesma forma, apenas um obteve sucesso. Foi realizado o ensaio de unidades formadoras de colônias destas células, sendo contadas 58 colônias ao final de sete dias. Na citometria de fluxo essas células mostraram-se positivas para CD44 e negativas para CD38, CD45, CD41/61. Estas células apresentaram curva de crescimento com fases de LOG, LAG e PLATEAU bem definidas, características das curvas de crescimento das células-tronco de origem mesenquimal. Conclusões: O isolamento e cultivo das células-tronco mesenquimais do cordão umbilical de ovinos é de difícil realização, exigindo maiores ensaios e estudos profundos. Células tronco do tecido adiposo de ovelhas demonstraram características mesenquimais, de acordo com a curva de crescimento, habilidade de ...

This paper pretends to demonstrate the effect of the combination of transversus abdominis plane block (TAP block) and Serratus plane block (SP block) techniques in analgesia of 4 dogs undergoing total unilateral mastectomy. Dogs were premedicated with methadone (0.5mg.kg-1) intramuscularly. Anesthesia was induced with propofol (6mg.kg-1) and midazolam (0.3mg.kg-1) and maintained with isoflurane. SP and TAP block were performed unilaterally using ultrasound by the injection of bupivacaine 0.25% (0.3mL kg-1) diluted with NaCl solution 1:1. Heart rate (HR), respiratory rate (f), non-invasive arterial pressure, esophageal temperature (T), oxygen saturation (SpO2) and electrocardiogram were monitored continuously. Animals were monitored for two and four hours after extubation for pain by using the Canine Acute Pain Scale from Colorado State University. Two hours after extubation, tramadol (4mg.kg-1) and dipyrone (25mg.kg-1) was administered to all dogs. It was not observed any alteration on cardiac rhythm. HR, f, T and mean arterial pressure remained below the preincisional values for all dogs. No dog required intraoperative rescue analgesia. Recovery from anesthesia was without any complication. All animals scored 0 (0/5) at pain scale, two and four hours after extubation and none of them expressed concern over the surgical wound. Dogs were able to walk before two hours after extubation. The combination of both techniques is effective in anesthetic blocking the thoracic and abdominal walls and it is suggested both may be included in the multimodal analgesia protocols for this type of surgery.

A distrofia muscular de Duchenne (DMD) é uma alteração neuromuscular caracterizada por contínua necrose muscular e degeneração, com eventual fibrose e infiltração por tecido adiposo. O aumento progressivo da fibrose intersticial no músculo impede a migração das células miogênicas, necessárias para a formação muscular. O modelo canino constitui-se nas melhores fenocópias da doença em humanos, quando comparados com outros modelos animais com distrofia. O tratamento antifibrose de pacientes DMD, tendo como alvo os mediadores da citocina, TGF-beta, e o tratamento com antiinflamatórios, podem limitar a degeneração muscular e contribuir para a melhora do curso da doença. O presente estudo teve como objetivo observar os possíveis efeitos adversos na fisiologia renal, por meio de avaliação bioquímica sanguínea e da pressão arterial, verificando a viabilidade do uso do Losartan (um inibidor de TGF-beta) nos cães afetados pela distrofia muscular. Foram utilizados quatro cães adultos, sendo dois machos e duas fêmeas. Utilizou-se a dose de 50mg de Losartan, administrada via oral, uma vez ao dia. Os exames clínicos, bem como alterações na função renal, o nível do potássio sérico e a pressão arterial não evidenciaram reação adversa durante todo o período do experimento. O uso de Losartan, por um período de 9 semanas, mostrou-se como uma terapia segura para o tratamento antifibrótico em cães adultos, não afetando a função renal ou pressão arterial dos animais.

Objectives The aim of this study was to evaluate, using echocardiography, the effects of oral administration of a single dose of gabapentin on the physiologic variables (heart rate [HR], respiratory rate [RR] and systolic blood pressure [SBP]) and systolic and diastolic cardiac function of healthy cats. Methods This was a prospective, randomized and blinded study with 40 healthy cats aged between 6 months and 2 years. The cats’ health status was assessed on the first appointment (T1) when they underwent a physical examination, complete blood count, biochemical profile, assessment of physiologic variables and echocardiogram. The echocardiogram was used to measure the left ventricle’s (LV) internal diameter during systole and diastole, isovolumic relaxation time, transmitral flow, E-wave deceleration time and HR. The cats were randomly divided into two groups: (1) a treatment group with 20 cats that received a single oral dose of gabapentin (100 mg/cat); and (2) a control group with 20 cats that received a single oral dose of placebo. All variables of the physiologic and echocardiographic variables were re-evaluated 1–3 weeks after T1 (T2), 90 mins after medication or placebo administration. Results There was no difference in the physiologic variables evaluated in both groups. The proportion of cats in the treatment group that had their ventricular filling waves fused on T1 but did not have them fused on T2 was significantly higher (45%) compared with cats in the control group (15%; P = 0.0384). Conclusions and relevance There was no difference between the groups in regard to SBP, HR, RR and echocardiographic variables. Gabapentin improved evaluation of diastolic function on echocardiogram because it reduced the fusion of ventricular filling waves during the evaluation of the diastolic function of the LV. Gabapentin did not cause adverse effects on the cardiovascular hemodynamics of young healthy cats.

RESUMO.-A distrofia muscular de Duchenne (DMD) é uma doença de origem genética, cuja principal manifestação clínica é enfraquecimento e atrofia progressiva dos músculos. Os cães da raça Golden Retriever podem apresentar distrofia muscular, com características genotípicas e fenotípicas muito próximas à distrofia muscular humana, sendo considerado o modelo animal mais apropriado para o estudo da DMD. Foram realizadas radiografias torácicas látero-laterais e dorsoventrais de 10 cães Golden Retriever afetados pela distrofia muscular, com o objetivo de relatar as alterações radiográficas associadas a essa patologia. O exame radiográfico da cavidade torácica evidenciou: (a) padrão pulmonar intersticial e alveolar predominante, (b) um quadro de pneumonia e edema pulmonar em fase inicial, (c) a cardiomegalia como o principal achado de comprometimento circulatório na cavidade torácica, (d) O megaesôfago torácico foi observado deslocando a traquéia e silhueta cardíaca ventralmente e, (e) a cúpula diafragmática apresentou modificação morfológi-ca, mostrando protrusão para o interior da cavidade torá-cica e hérnia hiatal, com deslocamento do estômago para o espaço mediastino caudal. (DMD) is a genetic disorder with clinical signs of muscular weaknesses and progressive atrophy. Golden Retriever dogs show similar genotypic and phenotypic characteristics to Human Muscular Dystrophy and are considered a proper animal model for DMD studies. Latero-lateral and dorso-ventral thoracic radiographies were obtained from 10 Golden Retriever dogs affected by muscular dystrophy, to investigate possible radiographic alterations. Thorax radiographic examination revealed (a) interstitial and alveolar pattern, (b) initial phases of pneumonia and pulmonary edema, (c) cardiomegaly as a principal alteration in the thoracic cavity, (d) megaesophagus displacing the trachea and heart silhouette, and (e) cranial protrusion of the diaphragm lining into the thorax with development of a hiatus hernia displacing the stomach to the caudal mediastinum. Postmortem examination showed pleural effusion, pulmonary emphysema, degenerative and metaplasic processes in the diaphragm and intercostal muscles. Radiographic examination was considered essential for the diagnosis of cardiac and respiratory disease in Golden Retriever dogs affected by muscular dystrophy, and to identify the primary pulmonary process and to provide the establishment of suitable therapeutic treatment, with a reserved prognosis in advanced stage of the disease.

RESUMO A distrofia muscular de Duchenne (DMD) é um tipo de distrofia muscular em humanos caracterizada por