Motor physical therapy affects muscle collagen type I and decreases gait speed in dystrophin-deficient dogs.

Golden Retriever Muscular Dystrophy (GRMD) is a dystrophin-deficient canine model genetically homologous to Duchenne Muscular Dystrophy (DMD) in humans. Muscular fibrosis secondary to cycles of degeneration/regeneration of dystrophic muscle tissue and muscular weakness leads to biomechanical adaptation that impairs the quality of gait. Physical therapy (PT) is one of the supportive therapies available for DMD, however, motor PT approaches have controversial recommendations and there is no consensus regarding the type and intensity of physical therapy. In this study we investigated the effect of physical therapy on gait biomechanics and muscular collagen deposition types I and III in dystrophin-deficient dogs. Two dystrophic dogs (treated dogs-TD) underwent a PT protocol of active walking exercise, 3×/week, 40 minutes/day, 12 weeks. Two dystrophic control dogs (CD) maintained their routine of activities of daily living. At t0 (pre) and t1 (post-physical therapy), collagen type I and III were assessed by immunohistochemistry and gait biomechanics were analyzed. Angular displacement of shoulder, elbow, carpal, hip, stifle and tarsal joint and vertical (Fy), mediolateral (Fz) and craniocaudal (Fx) ground reaction forces (GRF) were assessed. Wilcoxon test was used to verify the difference of biomechanical variables between t0 and t1, considering p<.05. Type I collagen of endomysium suffered the influence of PT, as well as gait speed that had decreased from t0 to t1 (p<.000). The PT protocol employed accelerates morphological alterations on dystrophic muscle and promotes a slower velocity of gait. Control dogs which maintained their routine of activities of daily living seem to have found a better balance between movement and preservation of motor function.

Opportunities for bead-based multiplex assays in veterinary diagnostic laboratories.

Bead-based multiplex assays (BBMAs) are applicable for high throughput, simultaneous detection of multiple analytes in solution (from several to 50-500 analytes within a single, small sample volume). Currently, few assays are commercially available for veterinary applications, but they are available to identify and measure various cytokines, growth factors and their receptors, inflammatory proteins, kinases and inhibitors, neurobiology proteins, and pathogens and antibodies in human beings, nonhuman primates, and rodent species. In veterinary medicine, various nucleic acid and protein-coupled beads can be used in, or for the development of, antigen and antibody BBMAs, with the advantage that more data can be collected using approximately the same amount of labor as used for other antigen and antibody assays. Veterinary-related BBMAs could be used for detection of pathogens, genotyping, measurement of hormone levels, and in disease surveillance and vaccine assessment. It will be important to evaluate whether BBMAs are "fit for purpose," how costs and efficiencies compare between assays, which assays are published or commercially available for specific veterinary applications, and what procedures are involved in the development of the assays. It is expected that many veterinary-related BBMAs will be published and/or become commercially available in the next few years. The current review summarizes the BBMA technology and some of the currently available BBMAs developed for veterinary settings. Some of the human diagnostic BBMAs are also described, providing an example of possible templates for future development of new veterinary-related BBMAs.

Bortezomib (PS-341) treatment decreases inflammation and partially rescues the expression of the dystrophin-glycoprotein complex in GRMD dogs.

Golden retriever muscular dystrophy (GRMD) is a genetic myopathy corresponding to Duchenne muscular dystrophy (DMD) in humans. Muscle atrophy is known to be associated with degradation of the dystrophin-glycoprotein complex (DGC) via the ubiquitin-proteasome pathway. In the present study, we investigated the effect of bortezomib treatment on the muscle fibers of GRMD dogs. Five GRMD dogs were examined; two were treated (TD- Treated dogs) with the proteasome inhibitor bortezomib, and three were control dogs (CD). Dogs were treated with bortezomib using the same treatment regimen used for multiple myeloma. Pharmacodynamics were evaluated by measuring the inhibition of 20S proteasome activity in whole blood after treatment and comparing it to that in CD. We performed immunohistochemical studies on muscle biopsy specimens to evaluate the rescue of dystrophin and dystrophin-associated proteins in the muscles of GRMD dogs treated with bortezomib. Skeletal tissue from TD had lower levels of connective tissue deposition and inflammatory cell infiltration than CD as determined by histology, collagen morphometry and ultrastructural analysis. The CD showed higher expression of phospho-NFκB and TGF-ß1, suggesting a more pronounced activation of anti-apoptotic factors and inflammatory molecules and greater connective tissue deposition, respectively. Immunohistochemical analysis demonstrated that dystrophin was not present in the sarcoplasmic membrane of either group. However, bortezomib-TD showed higher expression of α- and ß-dystroglycan, indicating an improved disease histopathology phenotype. Significant inhibition of 20S proteasome activity was observed 1 hour after bortezomib administration in the last cycle when the dose was higher. Proteasome inhibitors may thus improve the appearance of GRMD muscle fibers, lessen connective tissue deposition and reduce the infiltration of inflammatory cells. In addition, proteasome inhibitors may rescue some dystrophin-associated proteins in the muscle fiber membrane.

Estudo morfofuncional dos rins de cães da raça Golden Retriever afetados pela distrofia muscular (GRMD) / Morphofunctional study in kidneys of Golden Retriever dogs affected by muscular dystrophy (GRMD)

A Distrofia Muscular de Duchenne (DMD) é uma miopatia severa de caráter recessivo ligada ao cromossomo X e o modelo animal de estudo mais relevante é o Golden Retriever Muscular Dystrophy (GRMD). Além das severas alterações que ocorrem na musculatura estriada, muitos estudos mostram que outras estruturas, inclusive viscerais, podem se mostrar alteradas nesta patologia. Desta forma, este trabalho objetivou análisar e comparar possíveis alterações estruturais e funcionais do rim em cães GRMD. Neste modelo de estudo, foi possível observar a presença das faces convexa e côncava, do hilo renal e dos pólos craniais e caudais dos rins. O órgão mostrou-se envolto por uma cápsula fibrosa. Em um corte sagital do órgão, notou-se a presença das regiões cortical e medular e da pelve renal. Na análise microscópica foi possível identificar a zona medular e cortical com suas estruturas: os corpúsculos renais formados pelo glomérulo e pela cápsula de Bowman, os túbulos contorcidos proximais e distais, os ductos coletores, vasos sanguíneos e os segmentos das Alças de Henle. As dosagens séricas de creatinina e uréia encontram-se dentro dos limites de normalidade. Desta forma, de acordo com os nossos resultados, podemos concluir que os animais afetados estudados, não apresentaram alterações estruturais ou funcionais dos rins, o que nos permitir sugerir que apesar da ingestão hídrica comprometida, a estrutura renal, mantem- se preservada nos animais GRMD.

Duchenne muscular dystrophy (DMD) is a severe myopathy of recessive X-linked character and the most relevant animal study model is the Golden Retriever muscular dystrophy (GRMD). In addition to the severe changes occurring in the striated musculature, several studies show that other structures, including viscera, may prove to be altered in this pathology. Thus, this study aimed to analyze and compare possible structural and functional alterations of the kidney in GRMD dogs. In this study model, it was possible to observe the presence of convex and concave faces, the renal hilum, and the cranial and caudal poles of the kidneys. The organ was surrounded by a fibrous capsule. In a sagittal section of the organ, the presence of the cortical and medullary regions and the renal pelvis were noticed. On microscopic examination, it was possible to identify the medullary and cortical zones and their structures: the renal corpuscles formed by the glomerulus and Bowman's capsule, the proximal and distal convoluted tubules, the collecting ducts, the blood vessels, and the segments of the loops of Henle. The serum creatinine and urea were within normal limits. Thus, according to our results, we may conclude that the affected animals under study showed no structural or functional changes in the kidneys, something which allows us to suggest that, despite the impaired water intake, renal structure remains preserved in GRMD animals.

Study of the cardiac left atrioventricular valvar complex in water buffaloes (Bubalus bubalis) of the Jafarabadi breed / Estudo do complexo valvar atrioventricular cardíaco esquerdo em búfalos (Bubalus bubalis) da raça Jafarabadi

Atrioventricular valve complex of 30 Jafarabadi water buffaloes, adult males were studied in this research with no heart diseases. The animals were obtained from a slaughterhouse in Brazilian State of Parana. The hearts were opened at the third portion affording access to the valve complex. The complexes had its area, number and type of tendinous cords submitted to analysis. The results showed that the complex is composed by two cusps and four accessory cusps, two or three papillary muscles in which 10-25 tendinous cords fix on the cusps that face the ventricle wall. The total area of the complex was on average 38.56cm², with a minimum of 24.96cm² and a maximum of 55.54cm². Statistically, no relation between the number of cords and the cusps' area where they are inserted or with the number of papillary muscle where they originated from was observed.

Foram estudados os complexos valvares atrioventricular esquerdo de 30 búfalos da raça Jafarabadi, machos e adultos, sem alterações cardíacas, provenientes de abatedouros do Estado do Paraná. Os corações foram examinados em seu terço médio para acesso ao complexo valvar, que foi submetido a estudos de área, número e tipificação de cordas tendíneas. Os resultados demonstram que este complexo é formado por duas cúspides principais e quatro cúspides acessórias, apresentam em sua formação de 2-3 músculos papilares, nos quais se inserem de 10-25 cordas tendíneas, que se fixam em cúspides voltadas para a parede do ventrículo. A área total deste complexo apresenta uma média de 38,56cm² com um mínimo de 24,96cm² e um máximo de 55,54cm². Estatisticamente não há relação entre número de cordas e a área da cúspide onde estas estão inseridas, nem com o número de músculos papilares dos quais elas provem.

Morfofisiologia da inervação do diafragma de ovinos / Morphophysiology of diaphragm innervation in sheep

Thirty diaphragms of sheep of Santa Inês breed were studied regarding their origin, division and arrangement of the right and left phrenic nerves (Fde), and the participation of other nerves in the innervation of the diaphragm. By fixing and dissecting pieces, it was found that phrenic nerves (F) frequently come from the ventral branches of the 5th (C5) and 6th (C6) cervical spinal nerves (Ec), at right (46.67 percent) and at left (43.33 percent). The F often form a lumbocostal trunk, sternal branches at right (40.00 percent) and lumbar, costal and esternal branches at left (36.68 percent). The lumbar branches of F innervate frequently at left (96.67 percent) the homolateral pillar of the diaphragma, and at right (50.00 percent) they give fillets to Vena cava caudalis. The costal branches of the F innervate at left (90.00 percent) and at right (76.66 percent) the dorsal and ventral regions of the pars costalis. The sternal branches of the F innervate at right (100.00 percent) and at left (83.33 percent) the pars sternalis and the ventral region of the pars costalis at the same side. The intercostal nerves (VII to XII pairs, 63.33 percent) contribute to innervate the diaphragm of Santa Inês sheep.

Foram estudados em 30 diafragmas de ovinos da raça Santa Inês, a origem, a divisão e a distribuição dos nervos frênicos direito e esquerdo (Fde) e a participação de outros nervos na inervação do diafragma. Mediante fixação e dissecação das peças foi observado que os nervos frênicos (F) originam-se a partir dos ramos ventrais do 5º (C5) e 6º (C6) nervos espinhais cervicais (Ec) tanto à direita (46,67 por cento) como à esquerda (43,33 por cento). Os F finalizam em tronco lombocostal e ramo esternal à direita (40,00 por cento) e em ramo lombar, costal e esternal à esquerda (36,68 por cento). Os ramos lombares dos F inervam à esquerda (96,67 por cento) o pilar homolateral do diafragma e, à direita (50,00 por cento) fornecem filetes à veia caudal. Os ramos costais dos F ramificam à esquerda (90,00 por cento) e à direita (76,67 por cento) as regiões dorsal e ventral da pars costalis. Os ramos esternais dos F inervam à direita (100,00 por cento) e à esquerda (83,33 por cento) a pars sternalis e a região ventral da pars costalis do mesmo lado. Os nervos intercostais (VIII ao XII pares, 63,33 por cento) contribuem na inervação do diafragma de ovinos da raça Santa Inês.

Comportamento dos nervos glossofaríngeo e vago, na região retrofaríngea de ovinos: origem aparente no crânio, trajeto, ramificação e distribuição / Behavior of the glossopharyngeal and vagus nerves in the retropharyngeal region of sheep: apparent origin in cranium, course, branching and distribution

In 60 hemiheads of sheep of the Santa Ines breed the apparent origin in the skull of itinerary, ramification and distribution of the glossopharingeal nerve (Gf) and the vagus nerve (Vg) in the retropharyngeal region (Rr) were studied. By fixation and dissection of the specimens it was seen that the glossopharyngeal nerve and the vagus nerve arise from the jugular foramen in 100 percent of the cases. The right and the left glossopharingeal nerve (Glde) are frequently (86.6 percent) located more medially to the tympanic bulla, pass caudally to the stylohyoid bone, bypass the margin of the caudal stylopharyngeal muscle, the tonsilla, of the pharyngeal and the lingual mucous membrane. These branches are distributed, respectively, in the carotid sinus, pharyngeal musculature, soft palate, stylopharyngeal muscle, palatine tonsil, pharyngeal mucosa and the caudal third of the tongue (100 percent). The right and the left vagus nerve run caudally in association with the accessory nerves (Ac) up to the atlas (70 percent) and go then medially to the occipital artery, dorsally to the common carotid and the sympathetic trunk in the cervical region (80 percent). The vague nerves have pharyngeal (86.6 percent) and cranial laryngeal (100 percent) branches.

Em 60 hemicabeças de ovinos da raça Santa Inês foram estudadas a origem aparente no crânio, trajeto, ramificação e distribuição do nervo glossofaríngeo (Gf) e do nervo vago (Vg), na região retrofaríngea (Rr). Mediante fixação e dissecação das peças, foi observado que os nervos glossofaríngeos e vagos originam-se no forame jugular em 100 por cento dos casos. Os nervos glossofaríngeos direito e esquerdo (Gfde) são localizados, com maior freqüência (86,6 por cento), medialmente à bula timpânica, passam caudalmente ao osso estiloióide, contornam a margem caudal do músculo estilofaríngeo caudal, tonsilar, da mucosa faríngea e lingual. Estes ramos distribuemse, respectivamente, no seio carotídeo, musculatura faríngea, palato mole, músculo estilofaríngeo caudal, tonsila palatina, mucosa faríngea e terço caudal da língua (100 por cento). Os nervos vagos direito e esquerdo (Vgde) correm caudalmente em associação com os nervos acessórios (Ac) até o atlas (70 por cento), após o que passam medialmente à artéria occipital, dorsalmente à carótida comum e ao tronco simpático, na região cervical (80 por cento). Os ramos dos nervos vagos são os faríngeos (86,66 por cento) e os laríngeos craniais (100 por cento).