Efeito da fisioterapia na amplitude de movimento articular e deposição de colágeno muscular no modelo golden retriever muscular dystrophy (GRMD)

OBJETIVO: Elucidar o efeito da fisioterapia na Amplitude de Movimento Articular (ADM) e na fibrose muscular em animais GRMD. MÉTODOS: Estudo não randomizado, com grupo controle, cego, seis meses de intervenção, avaliação antes e depois da intervenção. Seis animais da raça Golden Retriever, distróficos, machos, média de idade 10,16±3,46 meses e peso de 17,75±6,01 kg foram separados em grupo tratado (n=3) e não tratado. Esses grupos de animais foram nomeados: G1=grupo tratado antes do tratamento; G2=grupo tratado após tratamento; G3=grupo não tratado antes do tratamento; G4=grupo não tratado após tratamento. O G1 participou do programa de fisioterapia que consistiu em um circuito de 300 metros com obstáculos. As ADMs do joelho, tarso, cotovelo e carpo foram avaliadas com goniômetro antes e após o tratamento. A área de colágeno do músculo vastus lateralis foi mensurada por histomorfometria, e a localização dos tipos de colágeno I, III e IV foi estudada por Imuno-histoquímica (IHC). RESULTADOS: Os valores da ADM do tarso do G2 apresentaram uma tendência a aumentar. A área de colágeno muscular foi diferente entre os grupos após o tratamento, e uma tendência ao aumento desses valores no G4 foi observada. Os colágenos dos tipos I e III foram os mais observados, constituindo feixes largos no perimísio nos dois grupos (G2 e G4). O colágeno do tipo I foi mais observado no endomísio do que o colágeno do tipo III. O colágeno do tipo IV foi observado apenas na lâmina basal. CONCLUSÃO: A Fisioterapia parece aumentar a ADM do tarso dos animais do grupo tratado sem aumentar a fibrose muscularOBJECTIVE: To elucidate the effect of physical therapy on joint range of motion (ROM) and muscle fibrosis in GRMD animals.METHODS: This was a nonrandomized blinded study with a control group, with six months of intervention evaluated beforehand and afterwards. Six dystrophic male Golden Retrievers of mean age 10.16±3.46 months and weight 17.75±6.01 kg were divided into a treated group (n=3) and an untreated group. These groups of dogs were named: G1=treated group before treatment; G2=treated group after treatment; G3=untreated group before treatment; and G4=untreated group after treatment. G1 underwent a physical therapy program that consisted of a 300-meter circuit with obstacles. Stifle, tarsal, elbow and carpal ROM were assessed using a goniometer before and after treatment. The area of collagen in the vastus lateralis muscle was measured using histomorphometry. The locations of collagen types I, III and IV were studied usingmmunohistochemistry. RESULTS: The tarsal ROM values in G2 presented an increasing trend. The area of muscle collagen differed between the groups after treatment and an increasing trend in these values was observed in G4. Collagen types I and III were the ones most frequently observed, forming broad bands in the perimysium of both G2 and G4. Type I collagen was observed in the endomysium more than type III collagen. Type IV collagen was observed only in the basal layer. CONCLUSION: Physical Therapy seemed to improve tarsal ROM in the treated group without increasing muscular fibrosi

The subplacenta of the red-rumped agouti (Dasyprocta leporina L)

BACKGROUND: Hystricognath rodents have a lobed placenta, comprising labyrinthine exchange areas and interlobular trophoblast. These correspond to the labyrinthine and spongy zones of other rodent placentae. Beneath them, however, is a structure unique to hystricognath rodents called the subplacenta. We here describe the subplacenta of the red-rumped agouti and examine the possible functional correlates of this structure. METHODS: Placentae were collected from early in midgestation to near term of pregnancy and examined by standard histological techniques, immunohistochemistry and transmission electron microscopy. In addition, to study the microvasculature of the subplacenta, vessel casts were inspected by scanning electron microscopy RESULTS: In the subplacenta, lamellae of connective tissue support a layer of mononuclear cytotrophoblast cells. Beneath this is found syncytiotrophoblast. Clusters of multinuclear giant cells occur in the transition zone between the subplacenta and decidua. There are prominent intercellular spaces between the cytotrophoblast cells. The basal membrane of these cells is often close to fetal blood vessels. The syncytiotrophoblast surrounds an extensive system of lacunae. Microvilli project into these lacunae from the plasma membrane of the syncytiotrophoblast. The syncytial cytoplasm contains electron-dense granules. This is probably the amylase-resistant PAS-positive material identified by histochemistry. The subplacenta is supplied entirely from the fetal circulation. Within it the vessels pursue a tortuous course with sinusoidal dilatations and constrictions. CONCLUSION: The functions that have been attributed to the subplacenta include hormone production. Our findings are consistent with this interpretation, but suggest that hormone secretion is directed towards the fetal circulation rather than the maternal tissues

Quantification of endothelin-1 level in placentae and human fetal membranes

A endotelina é um peptídeo vasoconstrictor potente que possui funções angiogênicas, mitogênicas e neurotróficas. Neste estudo procurou-se quantificar os níveis de Endotelina -1 (ET-1) em vilos coriônicos placentários e no âmnio de placentas de fetos gemelares, cujas mães submeteram-se à reprodução assistida e fecundação espontânea. Foram colhidas oito amostras de placentas, que após o delivramento placentário, foram colocadas em nitrogênio líquido e, posteriormente, congeladas e mantidas à -80ºC até o processamento por ensaio imunoenzimático (ELISA). Os níveis de concentração de ET-1 em amostras de vilos coriônicos de placentas de fetos gemelares variaram de 0,52 a 0,70fmol/ml, enquanto que de fetos únicos variou de 0,47 a 0,86fmol/ml. A mesma determinação em amostras de âmnio de placentas de gemelares variou de 0,61 a 1,16 fmol/ml, enquanto que de fetos únicos variou de 0,65 a 1,04fmol/ml. Estes achados indicam que em vilos coriônicos, os níveis de concentração de ET-1 não variaram entre gemelares e fetos únicos. No âmnio, os níveis de ET-1 são mais elevados em gemelares que em fetos únicos, dados que sugerem que o âmnio é a principal fonte de ET-1 presente no líquido amniótico.The endothelin is a potent vasoconstricor that possess angiogenic, mitogenic and neurotrophical functions. In this study we aimed to quantify the Endothelin-1 (ET-1) levels in chorionic placental villi and amnion of placentae of twins childbirths whose women were submitted to an assisted pregnancy and spontaneal fecundation. Were collected 08 placental samples. After the placental delivery the samples had been placed in liquid nitrogen and later in a -80ºC until the Enzyme-Linked Immunosorbent Assay (ELISA) technique. The concentration levels of ET-1 in samples of chorionic villi in twins placentae varied from 0,52 to 0,70 fmol/ml, while in chorionic villi single gestation varied from 0,47 to 0,86 fmol/ml. The same determination in samples of amnion of twins placentae varied from 0,65 to 1,16 fmol/ml, while in single gestation varied from 0,65 to 1,04 fmol/ml. These findings suggests that in the chorionic villi the levels of ET-1 didn't vary between twins and single gestation. In the amnion the levels of ET-1 are higher in twins than in single gestation, suggesting that the amnion is the source of ET-1 in the amniotic fluid

Quantification of endothelin-1 level in placentae and human fetal membranes

The endothelin is a potent vasoconstricor that possess angiogenic, mitogenic and neurotrophical functions. In this study we aimed to quantify the Endothelin-1 (ET-1) levels in chorionic placental villi and amnion of placentae of twins childbirths whose women were submitted to an assisted pregnancy and spontaneal fecundation. Were collected 08 placental samples. After the placental delivery the samples had been placed in liquid nitrogen and later in a -80ºC until the Enzyme-Linked Immunosorbent Assay (ELISA) technique. The concentration levels of ET-1 in samples of chorionic villi in twins placentae varied from 0,52 to 0,70 fmol/ml, while in chorionic villi single gestation varied from 0,47 to 0,86 fmol/ml. The same determination in samples of amnion of twins placentae varied from 0,65 to 1,16 fmol/ml, while in single gestation varied from 0,65 to 1,04 fmol/ml. These findings suggests that in the chorionic villi the levels of ET-1 didn't vary between twins and single gestation. In the amnion the levels of ET-1 are higher in twins than in single gestation, suggesting that the amnion is the source of ET-1 in the amniotic fluid.A endotelina é um peptídeo vasoconstrictor potente que possui funções angiogênicas, mitogênicas e neurotróficas. Neste estudo procurou-se quantificar os níveis de Endotelina -1 (ET-1) em vilos coriônicos placentários e no âmnio de placentas de fetos gemelares, cujas mães submeteram-se à reprodução assistida e fecundação espontânea. Foram colhidas oito amostras de placentas, que após o delivramento placentário, foram colocadas em nitrogênio líquido e, posteriormente, congeladas e mantidas à -80ºC até o processamento por ensaio imunoenzimático (ELISA). Os níveis de concentração de ET-1 em amostras de vilos coriônicos de placentas de fetos gemelares variaram de 0,52 a 0,70fmol/ml, enquanto que de fetos únicos variou de 0,47 a 0,86fmol/ml. A mesma determinação em amostras de âmnio de placentas de gemelares variou de 0,61 a 1,16 fmol/ml, enquanto que de fetos únicos variou de 0,65 a 1,04fmol/ml. Estes achados indicam que em vilos coriônicos, os níveis de concentração de ET-1 não variaram entre gemelares e fetos únicos. No âmnio, os níveis de ET-1 são mais elevados em gemelares que em fetos únicos, dados que sugerem que o âmnio é a principal fonte de ET-1 presente no líquido amniótico

Effectiveness of sequential viscosupplementation in temporomandibular joint internal derangements and symptomatology : a case series

Viscosupplementation is a minimally invasive technique that replaces synovial fluid by intra-articular injection of hyaluronic acid (HA). Although effective in some joints, there is not conclusive evidence regarding temporomandibular disorders. +is case series described the efficacy of a viscosupplementation protocol in intra-articular temporomandibular disorders. Ten patients with a diagnosis of disc displacement and/or osteoarthritis by Research Diagnostic Criteria for Temporomandibular Disorders (RDC/TMD) were submitted to four monthly injections of low or medium molecular weight HA. Pain, mandibular function, image analysis by tomography and magnetic resonance, and quality of life were assessed at baseline and follow-ups (1 and 6 months). Pain, jaw range-of-motion, mandibular function, and quality of life improved at follow-up evaluations. Osteoarthritis changes decreased, and 20% of patients improved mandibular head excursion after treatment. Resolution of effusion and improvement in disc morphology were observed for most patients. +is viscosupplementation protocol reduced pain and symptoms associated with internal derangement of temporomandibular joint, improved quality of life, and showed benefits from both low and medium molecular weight HA in alternate cycles

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, 36/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 functionFAPESP, 06/61561-

Morphological aspects of yolk sac from rodents of hystricomorpha subordem: paca (Agouti paca) and agouti (Dasyprocta aguti)

Este trabalho visou caracterizar macro e microscopicamente o saco vitelino em pacas (Agouti paca) e cutias (Dasyprocta aguti) no início de gestação. Três embriões/fetos de pacas e três de cutias foram utilizados para a análise do saco vitelino, durante as fases iniciais de gestação. Fragmentos do saco vitelino foram removidos do embrião/feto e rotineiramente processados para inclusão em parafina (técnica histológica rotineira) e em resina Spurr (análise ultra-estrutural). Macroscopicamente, a placenta vitelínica em ambas as espécies inseria-se na superfície da placenta principal, com suas margens projetando-se completamente sobre o embrião/fetos. Na microscopia de luz, a placenta vitelínica apresentava-se constituída pelo epitélio endodérmico e um mesenquima com inúmeros vasos vitelínicos. Ultraestruturalmente, a placenta vitelínica visceral da paca era formada por células endodérmicas com núcleos na região mediana e da cutia por núcleos dispostos apicalmente; outra característica foi o grande número de mitocôndrias, vesículas de conteúdo eletrodenso e com microvilosidades. Com base nos resultados concluímos, que (1) a placenta vitelínica das duas espécies apresenta inserção na superfície da placenta principal; (2) a placenta vitelínica de paca se apóia na membrana de Reichert, diferentemente da cutia, que não possui tal membrana; (3) o cório e alantóide apresentam-se fusionados, formando a placenta corioalantoídea; e (4) o saco vitelino em ambas as espécies é invertido e vascularizadoThe study aimed to characterize gross and microscopic features of the yolk sac in paca (Agouti paca) and agouti (Dasyprocta aguti) in early gestation. Fragments of the yolk sac of 3 paca and 3 agouti fetuses at early gestation were taken and processed for histological and ultrastructural analyses. Gross features of the vitelline placenta in both species showed its insertion over the main placenta surface and projections to the embryos/fetuses. Microscopically, the vitelline placenta was constituted by endoderm epithelium and mesenchyme, in which vitelline vessels are abundant. The ultrastructure of the samples showed that the visceral yolk sac of the paca was formed by endodermic cells with nuclei in the median region, and that the visceral yolk sac of the agouti was formed by nuclei arranged apically; other characteristic was the large number of mitochondrias, eletrodense vesicles with microvilosities We conclude that (1) the vitelline placenta of the two species presents insertion in the surface of the main placenta; (2) the vitelline placenta of paca rests on the Reichert's membrane, whereas the agouti vitelline placenta does not have this membrane; (3) the chorion and allantoic are fusioned; and (4) the chorioallantoic placenta and the yolk sac in both species are reversed and vascularize

Morphology and functionality of the type II pneumocytes and their variation in relation to bovine gestational age

Este estudo objetivou caracterizar a presença de pneumócitos tipo II e o início da produção de lipoproteína surfactante em bovinos, correlacionando a idade gestacional com a síntese de surfactante durante o desenvolvimento fetal. Pulmões de fetos com quatro meses de idade gestacional estavam na fase canalicular de desenvolvimento, sem a presença de pneumócitos tipo II ou bandas eletroforéticas compatíveis com a presença de proteínas surfactante. No 5&deg; mês gestacional, os pulmões dos fetos encontravam-se em fase de saculação terminal, com a presença de alvéolos por epitélio cúbico, com áreas formadas por pneumócitos I e II. Nesse período ainda não foi possível identificar proteína surfactante nos pulmões. Esses órgãos em fetos com seis meses de idade gestacional estavam em fase de saco terminal, com presença de pneumócitos tipo I e II. Nessa fase a análise para determinação protéica do surfactante de feto bovino (SDS - PAGE) demonstrou presença de bandas entre 26 e 36kDa, confirmando produção de SP - A, proteína surfactante encontrada em maior quantidade. A partir do 7&deg; mês gestacional, a fase de saco terminal é mais evidente e complexa, com desenvolvimento de intensa vascularização. O pneumócito tipo I apresentava aspecto mais pavimentoso, e o tipo II apresentava aspecto mais globoso. Na análise SDS - PAGE do lavado bronco - alveolar, bandas de proteína surfactante com aspecto similar ao de animais recém-nascidos foram encontradas. Em recém-nascidos, pulmões na fase alveolar foram observados com pneumócitos tipo I e II característicos. O perfil das bandas do lavado bronco-alveolar dos recém-nascidos foi igual ao de animais adultos. Esses achados sugerem que um animal nascido precocemente, a partir dos sete meses de gestação, teria sua sobrevivência garantida devido a uma possível funcionalidade do sistema respiratório do feto, pois o pulmão possuiria as características necessárias para a síntese de proteínas surfactantes. Entretanto, mais estudos clínicos sobre a funcionalidade do sistema respiratório abrem novas fronteiras de experimentos sobre fisiologia respiratória em recém-nascidos bovinos.The aim of this study was to characterize the presence of type II pneumocyte and the beginning of the surfactant protein production in bovines to the correlated gestational age. Lungs of fetuses at 4 months gestational age were found to be in the canalicular development phase, without presence of type II pneumocytes or appearance of electrophoretic bands compatible with the presence of the surfactant protein. In fetuses at 5 months of gestational age, the lungs were at the terminal sac phase, with the presence of primitive alveolus, formed by cubical epithelium and areas formed by pneumocytes I and pneumocytes II. No electrophoretic bands compatible to the surfactants proteins were identified. In contrast, lungs of fetuses at 6 months of gestational age showed, development of terminal sac phase, with the presence of type I and type II pneumocytes. In this phase, proteic determination for the SDS - PAGE analysis showed the presence of bands between 26 to 36kDa, demonstrating SP - A production, which is the surfactant protein found in highest amount. From the 7th gestational month on, the phase of terminal sac is more evident and complex, with intense sprouting vascularization. The type I pneumocyte had a more squamous aspect and type II pneumocyte were more globular. In the SDS - PAGE analysis of the bronchial - alveolar wash, surfactant protein bands had been observed with a profile similar to that of the newborn animals. In newborn animals, lungs in alveolar phase showed the development of type I and II pneumocytes. The profile of the bronchial - alveolar wash from the newborn was similar to an adult animal, presenting the same bands in triplets. These results suggest that early born fetuses, from 7 months of pregnancy could have a guarantee of surviving due to possible functionality of respiratory system because the lung presented all necessary features for surphactant protein syntheses. However, new clinical studies related to respiratory system functionality open new venues for newborn bovine's experiments respiratory physiology

Characterization of the fetal membranes in buffalo cows in the first three months of gestation

Este estudo visou à caracterização das membranas fetais em búfalas (Bubalus bubalis, Linnaeus 1758) no terço inicial da gestação. As membranas fetais foram analisadas macroscópica e microscopicamente (luz e microscopia eletrônica de transmissão). O córion possui uma camada simples de células circulares, com núcleos de forma esférica, denominadas trofobláticas; há outro tipo celular, as células trofoblásticas gigantes, com dois ou mais núcleos. Ambas possuem uma grande quantidade de vesículas no citoplasma e retículo endoplasmático à microscopia de transmissão. O alantóide possui vasos preenchidos com eritrócitos, e contêm células alongadas, que formam um epitélio estratificado simples. O âmnion é uma membrana transparente, ou esbranquiçada; constituído por epitélio estratificado simples. A diferença principal entre o alantóide e o âmnion é que o último é avascular. O saco vitelínico é uma membrana opaca que desaparece durante a gestação; é a única membrana que não está em contato com as outras e apresenta três tipos diferentes de células que dão forma a três camadas distintas (endoderma, mesotélio, mesênquima)The aim of the study was to characterize the fetal membranes in buffalo cows (Bubalus bubalis, Linnaeus 1758) in the first 3 months of the gestation. The fetal membranes were analyzed gross and microscopically (light and ultrastructural analysis). The corion is a simple layer of circle cells, with spherical nucleus, named trofoblastics; there is another cellular type, named trophoblastic giant cells, with two or more nucleus. Both of them have many cytoplasmic and reticule endoplasmic vesicles on transmission electron microscopy. The allantoic has many vessels filled with eritrocytes, elongated cells which form a stratified simple epithelium. The amnion is transparent or sometimes whitish, consisting of a stratified simple epithelium. The main difference between the allantoic and amnion is that the latter is avascular. The yolk sac is an opaque membrane that disappears during gestation, and is the only membrane that is not in contact with the others; it presents three different types of cells which form three distinct layers (endoderm, mesothelium and mesenquimal