Rehabilitation therapy with insulin-like growth factor after acute skeletal muscle injury / Terapia de reabilitação com fator de crescimento semelhante à insulina após lesão muscular esquelética aguda / Terapia de rehabilitación con factor de crecimiento semejante a la insulina después de una lesión aguda del músculo esquelético

ABSTRACT Introduction Skeletal muscle injuries account for 10% to 50% of treadmill sports injuries. Insulin-like growth factor (IGF) is a family of polypeptides with both insulin-like anabolic and growth-promoting effects. Sports play a vital role in the recovery of skeletal muscle injuries. Objective The paper analyzes the ability of insulin-like growth factor 1 (IGF-1) to repair skeletal muscle injury caused by treadmill exercise. Method We injected drugs under the wound after exercise-induced injury in rats. The control group was injected with saline, and the experimental group was injected with an insulin-like growth factor. We conduct histological and electron microscopic structural analysis of rats, Results: After an injury, the experimental group formed a basal lamina protective film earlier than the control group, activated myoblasts, formed myofilaments, formed myotubes, and fused into muscle fibers earlier than the control group. The healing quality was also better. The experimental group was endogenous. The mRNA content of sex IGF-1 and IGF-2 both increased earlier than the control group. Conclusion Local injection of exogenous insulin-like growth factor-1 can stimulate the proliferation of myoblasts and accelerate the post-traumatic repair process of skeletal muscle caused by treadmill sports. Level of evidence II; Therapeutic studies - investigation of treatment results.

RESUMO Introdução As lesões do músculo esquelético representam de 10% a 50% das lesões em esteira esportiva. O fator de crescimento semelhante à insulina (IGF) é uma família de polipeptídeos com efeitos anabólicos e de promoção do crescimento semelhantes à insulina. Os esportes desempenham um papel vital na recuperação de lesões musculares esqueléticas. Objetivo o artigo analisa a capacidade do fator de crescimento semelhante à insulina 1 (IGF-1) em reparar lesões musculares esqueléticas causadas por exercícios em esteira. Método Injetamos drogas sob a ferida após lesão induzida por exercício em ratos. O grupo controle foi injetado com solução salina e o grupo experimental foi injetado com um fator de crescimento semelhante à insulina. Realizamos análises histológicas e microscópicas eletrônicas estruturais de ratos. Resultados Após a lesão, o grupo experimental formou um filme protetor da lâmina basal mais cedo do que o grupo controle, mioblastos ativados, miofilamentos formados, miotubos formados e fundidos em fibras musculares mais cedo do que o grupo controle. A qualidade da cura também foi melhor. O grupo experimental era endógeno. O conteúdo do sexo IGF-1 e IGF-2 mRNA aumentou mais cedo do que no grupo de controle. Conclusão A injeção local de fator de crescimento semelhante à insulina 1 exógeno pode estimular a proliferação de mioblastos e acelerar o processo de reparo muscular esquelético pós-traumático causado por esportes em esteira. Nível de evidência II; Estudos terapêuticos: investigação dos resultados do tratamento.

RESUMEN Introducción Las lesiones del músculo esquelético representan del 10% al 50% de las lesiones deportivas en cinta. El factor de crecimiento semejante a la insulina (IGF) es una familia de polipéptidos con efectos anabólicos y estimulantes del crecimiento semejantes a la insulina. Los deportes juegan un papel vital en la recuperación de las lesiones del músculo esquelético. Objetivo El artículo analiza la capacidad del factor de crecimiento semejante a la insulina 1 (IGF-1) para reparar la lesión del músculo esquelético causada por el ejercicio en cinta. Método inyectamos drogas debajo de la herida después de una lesión inducida por el ejercicio en ratas. Al grupo de control se le inyectó solución salina y al grupo experimental se le inyectó un factor de crecimiento semejante a la insulina. Realizamos análisis estructurales histológicos y microscópicos electrónicos de ratas, Resultados: Después de una lesión, el grupo experimental formó una película protectora de la lámina basal antes que el grupo de control, activó mioblastos, formó miofilamentos, formó miotubos y se fusionó en fibras musculares antes que el grupo de control. La calidad de curación también fue mejor. El grupo experimental fue endógeno. El contenido de ARNm de IGF-1 e IGF-2 de sexo aumentaron antes que en el grupo de control. Conclusión La inyección local de factor de crecimiento semejante a la insulina 1 exógeno puede estimular la proliferación de mioblastos y acelerar el proceso de reparación postraumático del músculo esquelético causado por los deportes en cinta. Nivel de evidencia II; Estudios terapéuticos: investigación de los resultados del tratamiento.

Effects of consumption of soft drinks on the muscular morphology of animals submitted to concurrent training / Efecto del consumo de bebidas refrescantes en la morfología muscular de animales sometidos a entrenamiento concurrente

The consumption of industrialized foods and beverages contributes to the increase in body weight and manifestation of diverse illnesses. Among these industrialized products, soft drinks can be mentioned, which when consumed in large quantities can contribute to weight gain and the emergence of chronic non-communicable diseases such as hypertension, diabetes, and obesity. In terms of performance, several training protocols have been used, among which concurrent training is presented as a tool that can aid in the control of body weight and muscle development. Thus, the identification of dietary factors, as well as the use of appropriate training methods, can be essential factors in the increase in physical performance. To study the effects of concurrent training on the skeletal muscles of animals submitted to soft drink consumption. In the present study, 32 male rats were used, of the Wistar breed, in the post-weaning biological phase (30 days). The animals performed a concurrent training protocol. Forty-eight hours after the final training session, the animals were submitted to a surgical procedure to remove samples of gastrocnemius muscle. After preparation of the histological slides, the muscle fibers of the different groups of animals were measured. The median values for the minimum diameter of muscle fibers presented the following values: Control (C): 35.78, Control Soft Drinks (CSD): 36.93, Training (T): 39.12, and Training Soft Drinks (TSD): 37.82. The CSD and TSD groups demonstrated the greatest increase in body mass in relation to the C and T groups. In addition, the groups that performed the concurrent training presented a higher mean of the smallest fiber diameter. It was concluded that the ingestion of soft drinks had a negative effect on the muscular hypertrophy of animals submitted to a concurrent training protocol.

El consumo de alimentos y bebidas industrializadas contribuyen al aumento de peso corporal y manifestación de diversas enfermedades. Entre estos alimentos, se encuentran las bebidas refrescantes, que al ser consumidas en grandes proporciones, pueden contribuir al aumento de peso y a la aparición de enfermedades crónicas, tales como la hipertensión, diabetes y obesidad. En cuanto al desempeño, diversos protocolos han sido utilizados, entre estos, el entrenamiento concurrente que puede ser una forma de ayuda en el control de peso corporal y el desarrollo muscular. Siendo así, la identificación de los factores alimentarios, como también la utilización de los métodos correctos de entrenamiento, pueden ser esenciales en el aumento del desempeño físico. El objetivo del estudio fue investigar los efectos del entrenamiento concurrente en la musculatura esquelética de animales sometidos al consumo de bebidas refrescantes. En la presente investigación fueran utilizadas 32 ratas Wistar. Los animales fueran sometidos a un protocolo de entrenamiento concurrente. Después de 48 horas de la última sesión de entrenamiento, los animales fueran sometidos a un proceso quirúrgico para la remoción de muestras de tejido muscular. Los valores de la media del diámetro de las fibras musculares fueron: Control (C): 35,78; Control y Bebida Refrescante (CBR): 36,93; Entrenamiento (ET): 39,12; y Entrenamiento y Bebida Refrescante (EBR): 37,82. Se observó que los grupos CBR y EBR presentaron mayor aumento de masa corporal en comparación con C y T. Además, los grupos que realizaran el entrenamiento concurrente obtuvieron una mayor media en el diámetro de las fibras. Es posible concluir que el consumo de bebidas refrescantes tiene una influencia negativa en la hipertrofia muscular de animales sometidos a un protocolo de entrenamiento concurrente.

Sarcomere structure: the importance of desmin protein in muscle atrophy / Estructura de sarcómera: la importancia de proteína desmina en atrofia muscular

Knowing the ultrastructure of skeletal muscle is critical to understand how it works under normal situation and the disorders caused by extreme or pathological conditions. Sarcomere is the basic structural unit of striated muscle tissue. An important element of sarcomere architecture are the intermediate filaments, including the desmin protein. Desmin protein contributes to maintenance of cell integrity, efficient transmission of force and mechanochemical signaling within the myocyte. Because of this, desmin protein has constantly been a focus of research that investigates its alterations associated to damage and muscle atrophy under different conditions. The purpose of the following literature review is to describe the basic concepts of muscle ultrastructure, emphasizing the desmin protein role under conditions of muscle disuse atrophy and aging.

Conocer la ultraestructura del músculo esquelético es crítico para entender cómo trabaja bajo situaciones normales y en desórdenes causados por condiciones extremas o patológicas. La sarcómera es la unidad de estructura básica del tejido muscular estriado. Elementos importantes en la arquitectura de la sarcómera son los filamentos intermedios, incluyendo la proteína desmina. La proteína desmina contribuye en mantener la integridad celular, la transmisión eficiente de fuerza y la señalización mecanoquímica dentro del miocito. Debido a lo anterior, la proteína desmina ha sido constante foco de investigación en trabajos que estudian sus alteraciones asociadas a daño y atrofia muscular bajo diferentes condiciones. El propósito de la siguiente revisión de la literatura es describir los conceptos básicos de la ultraestructura muscular, enfatizando en el rol de la proteína desmina bajo condiciones de atrofia muscular por desuso y envejecimiento.

Entrapment of the sural nerve on gastrocnemius muscle: histological analysis / Compresión del nervio sural en el músculo gastrocnemio: análisis histológico

Anatomical variations in the formation of the sural nerve are common, although the topographical localization of this nerve is constant. In this report, we describe the histological analysis of anomalous course of the medial sural cutaneous nerve which descended through the gastrocnemius via a tunnel formed within the muscle and fascia. This anatomical curiosity are clinically important when evaluating sensory axonal loss in distal axonal neuropathies since sural nerve mononeuropathy is less likely to occur.

Las variaciones anatómicas en la formación del nervio sural son comunes, aunque la localización topográfica de este nervio es constante. En este reporte se describe el análisis histológico del nervio cutáneo sural medial de curso anómalo, que descendió a través de los músculos gastrognemios en un túnel formado en el músculo y la fascia. Esta curiosidad anatómica es de importancia clínica cuando se evalúa la pérdida sensitiva distal en neuropatías axonales, desde mononeuropatía del nervio sural a otras de menor frecuencia.

Inserción del tendón calcáneo / Insertion of the calcaneus tendon

El tendón calcáneo (TC) en el hombre es el más voluminoso y resistente del cuerpo, constituyéndose por su inserción en un elemento vital en la mecánica articular del pie como también en las numerosas patologías que afectan las regiones talocrural y calcánea. Para el estudio se utilizaron 120 miembros inferiores (60 derechos y 60 izquierdos), de cadáveres formolizados de individuos adultos, de ambos sexos. Se disecó la cara posterior de la pierna desde la línea articular de la rodilla hasta la parte inferior del calcáneo, se retiró el tejido adiposo precalcáneo dejando libre el TC. Se observó que el TC estaba formado, exclusivamente, por la fusión de las aponeurosis de los músculos gastrocnemio y sóleo en 62 casos (51,67 por ciento). El tendón del músculo plantar contribuye a la formación parcial del TC en 38 casos (31,67 por ciento) e integralmente en 20 casos (16,66 por ciento). El ancho del TC a nivel de la terminación del vientre del músculo sóleo fue de 12,88 mm +/- 2,0 en el hombre y de 11,55 mm +/- 1,6 en la mujer y el grosor del TC al mismo nivel de 4,29 mm +/- 0,64 en el hombre y de 4,36 mm +/- 0,64 en la mujer. El ancho del TC a nivel del margen posterosuperior del calcáneo fue de 17,98 mm +/- 1,7 en el hombre y de 17,06 mm +/- 1,53 en la mujer. El grosor del TC al mismo nivel fue de 3,79 mm +/- 0,61 en el hombre y de 3,93 mm /- 0,67 en la mujer. La distancia entre el margen posterosuperior del calcáneo y el inicio de la parte insertada del TC fue de 10,99 mm +/- 2,11 en el hombre y 10,84 +/- 2,71 en la mujer. El largo de la parte insertada del TC fue de 17,78 mm +/- 2,4 en el hombre y de 17,66 mm +/- 4,75 en la mujer. El ancho del TC a nivel de su inserción en el hueso calcáneo fue de 28,77 mm +/- 2,53 en el hombre y de 27,21 mm +/- 2,93 en la mujer. La tendinopatía en la inserción del TC suele ser frecuente requiriendo en algunos casos debridamiento quirúrgico del tendón. Por lo anterior, conocer aspectos morfométricos del TC y de su inserción adq...

In man the calaneous tendon (CT) is the most voluminous and resistent tendon in the body; its insertion is a vital element of the foot joint mechanism as well as in a number of disorders that affect talocrural and calcaneal regions. For the present study 120 lower members (60 right and 60 left) of formolized cadavers, adult subjects of both sexes were used. Posterior surface of the leg was disected from the joint line of the knee to the lower part of the calcaneal removing precalcaneus adipose tissue, exposing the CT. We observed that in 62 cases (51.67 percent) the CT was formed exclusively by fusion of aponeurosis of the gastrocnemius and soleus muscles. In 38 cases (31.67 percent) tendon of the plantar muscle contributes to the partial formation of the CT, and fully integrated in 20 cases (16.66 percent). Width of the CT at the soleus muscle belly end, was 12.88 mm +/- 2.0 in men and 11.55 mm +/- 1.6 women and density at the same level was 4.29 mm +/- 0.64 in men and 4.36 mm +/- 0.64 in women. Width of the CT at the posterosuperior border of the calcaneal was 17.98 mm +/- 17 in men, and 17.06 mm +/- 1.53 in women. CT density at the same level was 3.79 mm +/- 0.61 in men and 3.93 mm +/- 0.67 in women. Distance between posterosuperior border of the calcaneal and starting point of the inserted portion of the CT was 10.99 mm +/- 2.11 in men and 10.84 +/- 2.71 in women. Length of the inserted portion of the CT was 17.78 mm +/- 2.4 in men and 17.66 mm +/- 4.75 in women. Width of the CT at the insertion level in the calcaneus bone was 28.77 mm +/- 2.53 in men and 27.21 mm +/- 2.93 in women. Insertional CT tendinopathy is a chronic condition in some cases requiring surgical debridement of the tendon. Therefore, knowledge of morphological aspects of the CT and its insertion is important at the time of surgical procedures of the talocrural region.

Action of neuwiedase, a metalloproteinase isolated from bothrops neuwiedi venom, on skeletal muscle: an ultrastructural and immunocytochemistry study

The damaging effects of neuwiedase, a non-hemorrhagic snake venom metalloproteinase from P-I class, on gastrocnemius muscle are studied herein. Following neuwiedase injection, ultrastructural alterations were detected early showing disarrangement of skeletal muscle fibers (characterized by discontinuity of Z lines), mitochondrial swelling, and disruption of plasma membrane and basal lamina. Degradation of skeletal muscle and the appearance of an amorphous substance, primarily composed of cellular debris, were noted after 24 hours. The presence of neuwiedase at the injection site (detected by immunocytochemistry) revealed highly specific labeling of myofibril components of damaged myocytes. In addition, proteolysis of muscle proteins assayed through myofibrils extracted from gastrocnemius muscle indicated that neuwiedase provoked degradation of myofibrils, especially myosin. These results suggest that skeletal muscle damage, induced by neuwiedase, is probably due to its proteolytic action on myofibrils, which are responsible for the maintenance of the cellular architecture.

Neurotoxic activity and ultrastructural changes in muscles caused by the brown widow spider Latrodectus geometricus venom / Actividad neurotóxica y cambios ultraestructurales en musculos causados por el veneno de la araña viuda marrón Latrodectus geometricus

Brown widow spider (Latrodectus geometricus) venom (BrWSV) produces few local lesions and intense systemic reactions such as cramps, harsh muscle pains, nausea, vomiting and hypertension. Approximately 16 protein bands under reducing conditions and ~ 14 bands under non-reducing conditions on a 12.5% sodium dodecyl sulfate-polyacrylamide gel electrophoresis were observed. Neurotoxic clinical manifestations were confirmed in vivo, while proteolytic activity was demonstrated on gelatine film. Severe ultrastructural damages in mice skeletal muscles were observed at 3, 6, 12 and 24 h postinjection with at total of 45 µg of venom protein. Infiltration of eosinophils and ruptures of the cellular membranes were observed in the muscles along with swelling of the nuclear cover and interruption of the collagen periodicity. Altered mitochondrias and autophage vacuoles, nuclear indentation and mitochondria without cristae, slight increment of intermyofibrillar and subsarcolemic spaces and myelinic figures formation were also observed. In the capillary, endothelial membrane unfolding into the lumen was noticed; along with myelinic figures compatible with a toxic myopathy. Swollen sarcotubular systems with lysis of membrane, intense mitochondria autophagia and areas without pinocytic vesicles were observed. Swollen mitochondria surrounded by necrotic areas, myofibrillar disorganization and big vacuolas of the sarcotubular system, degenerated mitochondrium with formation of myelinic figure was seen. Glycogenosomes with small particulate, muscle type glycogen was noticed. Autophagic vacuole (autophagolysosomes) and necrotic areas were also noticed. These damages may be due to interactive effects of the multifactorial action of venom components. However, Latrodectus geometricus venom molecules may also be utilized as neuro therapeutic tools, as they affect neuronal activities with high affinity and selectivity. To our knowledge, the present study...

El veneno de la araña viuda marrón (Latrodectus geometricus) produce pocas lesiones locales pero intensas reacciones sistémicas, tales como calambres, dolores musculares severos, nauseas, vómitos e hipertensión arterial. Se observaron ~ 16 bandas de proteina bajo condiciones reducidas y ~14 bandas bajo condiciones no reducidas en electroforesis en geles de poliacrilamida al 12.5%. Las manifestaciones neurotóxicas clínicas fueron confirmadas in vivo, mientras que la actividad proteolítica fue demostrada en una placa de gelatina. Los músculos de ratón se estudiaron durante las 3, 6, 12 y 24 horas después de ser inyectados con 45 µg de proteina de veneno. Los músculos fueron seriamente dañados por este veneno. Se demostró una infiltracción de células eosinofílicas y rupturas de membranas celulares en tejido muscular, al mismo tiempo un fuerte incremento de la membrana nuclear y una interrupción de la periodicidad del colágeno. Se observaron daños en la mitocondria y sin cristaes, vacuolas autofágicas e indentación nuclear. Se notó un aumento de la luz de los espacios intermiofibrilares y subsarcolemicos. En los capilares fue visible un desdoblamiento de la membrana endotelial hacia el lúmen vascular. Del mismo modo, fue visto un hinchamiento del sistema sarcotubular con lisis de las membranas; intensa autofagia de mitocondrias y áreas sin vesículas pinocíticas. Fue además observado, glucogenosomas con glucogeno particulado. Se observaron vacuolas autofágicas (autofagolisosomas) y áreas de necrosis. Estos daños podrían ser atribuídos a los efectos interactivos de una acción multifactorial de los componentes del veneno.

Spontaneous stage differentiation of mouse-virulent Toxoplasma gondii RH parasites in skeletal muscle cells: an ultrastructural evaluation

Although the predilection for Toxoplasma gondii to form cysts in the nervous system and skeletal and heart muscles has been described for more than fifty years, skeletal muscle cells (SkMCs) have not been explored as a host cell type to study the Toxoplasma-host cell interaction and investigate the intracellular development of the parasite. Morphological aspects of the initial events in the Toxoplasma-SkMC interaction were analysed and suggest that there are different processes of protozoan adhesion and invasion and of the subsequent fate of the parasite inside the parasitophorous vacuole (PV). Using scanning electron microscopy,Toxoplasma tachyzoites from the mouse-virulent RH strain were found to be attached to SkMCs by the anterior or posterior region of the body, with or without expansion of the SkMC membrane. This suggests that different types of parasite internalization occurred. Asynchronous multiplication and differentiation of T. gondii were observed. Importantly, intracellular parasites were seen to display high amounts of amylopectin granules in their cytoplasm, indicating that tachyzoites of the RH strain were able to differentiate spontaneously into bradyzoites in SkMCs. This stage conversion occurred in approximately 3 percent of the PVs. This is particularly intriguing as tachyzoites of virulent Toxoplasma strains are not thought to be prone to cyst formation. We discuss whether biological differences in host cells are crucial to Toxoplasma stage conversion and suggest that important questions concerning the host cell type and its relevance in Toxoplasma differentiation are still unanswered.

Apoptosis in Denervated Skeletal Muscle / Apoptosis en Músculo Estriado Desnervado

The apoptosis phenomenon happens in normal tissues and in many pathological conditions as described by several researchers. Most studied in the 70s, this phenomenon remained almost forgotten in the 90s, when, with the advent of more advanced techniques, returned to laboratories. Thus, electronic microscopy, Comet Assays and histochemistry techniques, among others, were utilized in an effort to elucidate in a thorough way, the mechanisms that lead the cell to "suicide". No one can deny the importance of the knowledge of such phenomenon, so as to control its induction or blocking in the treatment of pathologies such as cancer. In this work, the apoptosis study was aimed at denervated muscles in different periods. Furthermore, two types of skeletal striated muscles were compared: the one called red muscle (soleo) which is resistant to fatigue, having a slow contraction, and the one named white muscle (EDL) of precocious fatigue and fast contraction. Previoulsy carried out in the techniques mentioned above, this work has been accomplished using the TUNEL technique, which is most accepted in the identification of apoptosis. It was demonstrated that this phenomenon takes place in a more marked way, in the beginning of the denervation process. In addition, it was observed that the red muscle responds more intensely to denervation than the white muscle. One can infer that the removal of the main cell function, the contraction, in the case of the skeletal striated muscle, triggers the chain reactions that culminate in apoptosis. Other projects are in process using mutant animals aiming at studying degenerative muscular illnesses and correlating them with denervation.

El fenómeno apoptosis ocurre en tejidos normales y en muchas condiciones patológicas, de acuerdo a lo relatado en varios trabajos. El estudio de este fenómeno tuvo su apogeo en la década del 70, permaneciendo en el olvido hasta la década del 90, momento en el cual comienzan a surgir técnicas más avanzadas, que motivaron su retorno a laboratorios de todo el mundo. Así, técnicas de microscopía electrónica, histoquímica, ensayos cometa, etc., fueron utilizados en los esfuerzos de elucidar de forma más profunda los mecanismos que conllevan a la célula al "suicidio". No se puede negar la importancia del conocimiento de ese fenómeno o, más específicamente, del control de su inducción o bloqueo para el tratamiento de patologías como el cáncer, por ejemplo. El presente trabajo tuvo como objetivo el estudio de apoptosis en músculo inervados, por diferentes periodos. También fueron comparados dos tipos de músculo estriado, el llamado músculo rojo (m. soleo), resistente a la fatiga y de contracción lenta, un músculo blanco (m. EDL) de fatiga precoz y contracción rápida. Este trabajo hace parte de un proyecto mayor ya desarrollado por sus ejecutores, sólo que utilizando la técnica del TUNEL como herramienta principal para identificar la apoptosis. Fue demostrado que tal fenómeno ocurre de forma más acentuada en los momentos iniciales de la desnervación. Además de eso, fue observado que el músculo rojo responde de manera más intensa a la desnervación. Nos parece inferir que la retirada de la principal función celular, en el caso del músculo estriado, la contracción despierta un mecanismo cuya reacción en cadena culmina en apoptosis. Otros proyectos están siendo elaborados con el propósito de utilizar animales mutantes, para estudiar enfermedades musculares degenerativas y correlacionarlas a la desnervación.

The role of ischemic preconditioning at the gracilis muscle of rats in the early phase of reperfusion injury

OBJETIVO: Verificar o papel do precondicionamento isquêmico (PCI) na lesão de isquemia e reperfusão (I/R) do músculo grácil de ratos. MÉTODOS: Trinta ratos Wistar foram distribuídos em três grupos, sendo G-I/R e G-PCI subdivididos em relação ao tempo de isquemia. Produziu-se modelo de quase-amputação do membro posterior por meio de uma incisão, preservando o feixe vascular e o osso femoral, induzindo-se isquemia por 2h e 4h, G-I 2h/R (n=6) e G-I 4h/R (n=6), seguida por 1h de reperfusão. Os Grupos Precondicionados, G-PCI 2h (n=6) e G-PCI 4h (n=6), foram precedidos por 3 ciclos de 5min de isquemia seguidos de 5min de reperfusão, antes da isquemia sustentada. No Grupo Controle, G-C (n=6) realizou-se apenas a abordagem regional. As avaliações morfológicas foram realizadas por meio de Microscopia Óptica (MO) para avaliação da desestruturação miofibrilar, processo inflamatório e estase venosa no tecido do músculo grácil. Foram atribuídos escores semi-quantitativos às variáveis estudadas. RESULTADOS: Houve preservação miofibrilar de 76% em G-I2h/R e 0% em G-I4h/R; G-IPC 2h a preservação foi de 100% e de 67% em G-IPC 4h. A reação inflamatória foi mais acentuada em G-I 4h (100% com inflamação) do que em G-I2h (50%);G-IPC 2h apresentou 82% sem inflamação e G-IPC 4h apresentou 67% sem inflamação. A estase vascular esteve ausente em 17% do G-I 4h e em 33% do G-I 2h; em G-IPC 2h esteve ausente em33% e em G-IPC 4h em 50%. CONCLUSÃO: O PCI mostrou, em uma fase inicial, papel benéfico na lesão de I/R no músculo grácil de ratos, comprovado pela maior preservação das fibras musculares, menor reação inflamatória e menor estase venosa.

Metabolismo mitocondrial y ultraestructura de músculo esquelético de ratón con restricción calórico proteica / Mitochondrial metabolism and ultrastructure of mice's skeletal muscle with caloric proteic restriction

Para estudiar el metabolismo mitocondrial en este trabajo se utilizó el método de permeabilización del tejido muscular mediante saponina, la cual destruye el sarcolema, manteniendo las mitocondrias "in situ". Para crear un modelo de patología mitocondrial se produjo restricción calórico-proteica (RCP) en ratones adultos con un 50 por ciento de la dieta de los ratones alimentados "ad libitum" durante un período de 15 días. Se efectuaron estudios del consumo de oxígeno, relación ADP/O, así como la actividad enzimática de los Complejos I y II de la cadena respiratoria y la determinación de proteínas. Para evaluar el efecto de la saponina sobre el tejido muscular en presencia y ausencia de restricción calórica-proteica se observaron muestras del mismo al microscopio electrónico de transmisión. Los resultados revelaron en los animales una pérdida de peso, disminución en el consumo de oxígeno y de la relación ADP/O en presencia de glutamato y de succinato, por otra parte se obtuvo una disminución evidente de la actividad del Complejo I, sin modificación del Complejo II. Las observaciones obtenidas con el microscopio electrónico demuestran que la RCP en ratas tiene efecto directo sobre el aparato contráctil de las fibras musculares, produciendo desorganización de las estructuras sarcoméricas y reducción importante de los miofilamentos. La saponina, por otra parte afecta todo el sistema de citomembranas.

In this work, the method of permeabilization of muscular tissue with saponin was employed, which destroyed the sarcolemma but maintained the mitochondria "in situ". To create a model of mitochondrial pathology, a caloric-protein restriction (CPR) was produced in adult mice by reducing 50 percent the diet to fed animals for 15 days. Determinations of oxygen consumption, ADP/O ratio, total proteins, as well as enzymatic activities of respiratory chain Complexes I and II were performed. To evaluate the effect of saponin on muscular tissues under normal or CPR conditions, electron microscopy observations were done. Results of the study showed that the animals in CPR group had loss of weight, reduced total proteins, diminished oxygen consumption, lower ADP/O ratio in the presence of glutamate and succinate, and a notable decrement in the activity of Complex I but without modifications of that of Complex II. Electron microscopic observations demonstrates directs effects of CPR over the contractile apparatus of muscle fibers, such as, disorganization of sarcomeric structures and an important reduction of the myofilaments. On the other hand, saponin affected all cytomembranous systems.

Características ultraestructurales del músculo esquelético en pacientes con proteinuria glomerular / Ultrastructure of skeletal muscle in patients with glomerular proteinuria

La proteinuria produce un desbalance importante del metabolismo proteico y lipídico, que favorece la desproteinización del organismo, con grandes repercusiones sistémicas que llegan a afectar hasta el músculo esquelético. En los pacientes con falla renal, usualmente disminuye su masa muscular y se observa debilidad. Los pacientes urémicos terminales, manifiestan atrofia de las fibras musculares tipo II, además de daño miopático primario, causado probablemente por un déficit de la microcirculación. En un afán de dilucidar las posibles alteraciones ultraestructurales precoces presentes en el músculo esquelético de pacientes con glomerulopatías primarias que cursan con proteinuria, se estudiaron 4 pacientes (2 hombres y 2 mujeres) con edades entre 42 y 66 años, pertenecientes al servicio de nefrología del hospital militar ®Dr. Calos Arvelo¼ de Caracas-Venezuela. En todos ellos se realizaron mediciones de enzimas musculares (CK, LDH, AST y ALT), electromiografía de los miembros inferiores y biopsia del músculo cuadriceps femoral. En ninguno de los pacientes se observaron modificaciones en los niveles séricos de las enzimas y a excepción de uno de ellos (caso 4), los registros eletromiográficos fueron normales. A nivel ultraestructural se observó: hinchamiento de los componentes del sistema sarcotubular con presencia de tríadas prominentes, signos de atrofia muscular, núcleos hipercromáticos, glucógeno abundante y glucogenosomas. Algunos capilares intramusculares con la luz parcial o totalmente ocluida, citoplasma endotelial electrón denso, presencia de prolongaciones del citoplasma endotelial hacia la luz del capilar, vesículas pinocíticas y cavéolas. Dichos cambios nos permiten concluir el compromiso estructural del músculo esquelético en pacientes con proteinuria, el cual se hace más evidente a medida que avanza hacia la falla renal y que dicho compromiso tiene una base microvascular que condiciona cambios del metabolismo muscular.

The proteinuria produces an important unbalance of the protein and lipid metabolism that it favors thedesproteinitation of the organism, with big systemic repercussions that end up affecting the skeletal muscle. In the patients with renal failure it usually diminishes their muscular mass and weakness is observed. The terminal uremic patient, manifests atrophy of the fibers muscular type II, besides miopatic primary damage, caused probably for a deficit of the microcirculación. In a desire of elucidating the early ultraestructural alterations present in the skeletal muscle of patient with primary glomerulopaties that curse with proteinuria, four patients were studied (2 men and 2 women) with ages between 42 and 66 years, belonging to the service of nephrology of the military hospital ®Dr. Calos Arvelo¼ of Caracas-Venezuela. In all they were carried out mensurations of muscular enzymes (CK, LDH, AST and ALT), electromiografie of the inferior members and biopsy of the muscle femoral cuadriceps. In none of the patients modifications were observed in the seric levels of the enzymes and to exception of one of them (case 4); the eletromiografic registrations are normal. At ultraestructural level it was observed: swelling of the components of the system sarcotubular with presence of prominent triads, signs of muscular atrophy, hyperchromatic nuclei, abundant glucogen and glucogenosome. Some capillary ones intramuscular with the partial or completely occluded light, electron dense cytoplasm, presence of continuations of the endothelial cytoplasm toward the light of the capillary one, pinocitic vesicles and caveole. This changes allow us to conclude the structural commitment of the skeletal muscle in patient with proteinuria, which becomes more evident as it advances toward the renal failure and that said commitment has a microvascular origin that conditions changes of the muscular metabolism.

Structural and microvascular study of soleous muscle of Wistar rats after section of the sciatic nerve

Ainda não está suficientemente estabelecida a relação entre a ativação das células satélites e a microcirculação muscular, após denervação cirúrgica. Através de métodos de microscopia eletrônica de transmissão e varredura, foram estudadas comparativamente as alterações do músculo sóleo, de ratos Wistar, após sua denervação.Os resultados evidenciaram que no músculo denervado ocorre ativação, duplicação e migração das células satélites para o interior da fibra muscular, acompanhadas de aumento de densidade da rede capilar, caracterizada por maior número de anastomoses e brotamentos angiogênicos. Fica assim evidenciada a importância da microvascularização nos processos biológicos de reparação muscular.

Correlation between ultrastructural and molecular changes in denervated muscle: a paradigm based on functional eletrical stomulation

Soon after denervation, skeletal muscle undergoes ultrastructural and molecular changes that ultimately lead to muscle atrophy and cell death. Functional electrical stimulation (FES) is effective in preventing the artrophy of denervated muscles, but it is unclear whether FES can prevent the progression of ultrastructural changes in the sarcotubular system and cell death after denervation. In this work, we developed a model to study the effects of FES on the ultrastructural changes in the sarcotubular system and the cell death which follow skeletal muscle denervation. The right sciatic nerve of adult rats was sectioned and after 1 day, and 1, 2 and 3 weeks the tibialis anterioris muscle was electricall stimulated. FES was done for 1, 2, 3, 10 and 20 weeks after denervation. The stimuli (10 Hz) were given for 10s followed by a 10s rest for a total of 30 min/day. Transmission electron microscopy showed that progressive dilatation of the sarcotubular system, as well as signs of cell death, were prevented when stimulation was started one day or one week after denervation. This model could be useful for undertanding the correlation between the structural changes and the molecules involved in the dilatation of the sarcotubular system or cell death.

A comparative study of aging of the elastic fiber system of the diaphragm and the rectus abdominis muscles in rats

In the present study the age-related changes of the striated muscle elastic fiber system were investigated in the diaphragm and rectus abdominis muscles of 1-, 4-, 8- and 18-month-old rats. The activation patterns of these muscles differ in that the diaphragm is regularly mobilized tens of times every minute during the entire life of the animal whereas the rectus abdominis, although mobilized in respiration, is much less and more irregularly activated. The elastic fibers were stained by the Verhoeff technique for mature elastic fibers. Weigert stain was used to stain mature and elaunin elastic fibers, and Weigert-oxone to stain mature, elaunin and oxytalan elastic fibers. The density of mature and elaunin elastic fibers showed a progressive increase with age, whereas the amount of oxytalan elastic fibers decreased in both diaphragm and rectus abdominis muscles and their muscular fascias. These age-related quantitative and structural changes of the elastic fiber system may reduce the viscoelastic properties of the diaphragm and rectus abdominis muscles, which may compromise the transmission of tensile muscle strength to the tendons and may affect maximum total strength

Estudo morfológico no músculo gastrocnêmio de camundongos C57 BL10 submetidos à ingestäo prolongada de etanol / Study of ultrastructural alterations in gastrocnemius muscle of C57 BL10 mice after prolonged ethanol ingestion

Os efeitos do alcoolismo crônico no músculo gastrocinêmio de camundongos bem nutridos foram estudados morfologicamente com a finalidade de se avaliar a hipótese de que o etanol exerce um papel tóxico direto sobre o músculo esquelético. Foram usados 30 camundongos C57BL1O, adultos jovens, divididos em dois grupos: Grupo A (controle) consistindo de 10 camundongos que beberam água e Grupo B (alcoólico) de 2O camundongos que beberam etanol a 25 por cento. Todos os animais receberam uma raçao balanceada de laboratório e foram mantidos nesse regime ad libitum nas mesmas condiçoes ambientais durante 48 semanas e pesados uma vez por semana. O consumo diário de dieta e a ingestao calórica foram calculados tendo os animais apresentado substancial ganho de peso, nao mostrando qualquer sinal de desnutriçao. Ao final do experimento os animais foram sacrificados para estudos morfológicos. Pela microscopia óptica nao foram encontradas alteraçoes. Importantes anormalidades foram observadas pela microscopialetrônica em todos os espécimens. O retículo sarcoplasmático mostrou-se frequentemente dilatado, resultando na formaçao de grandes vesículas e envolvendo as cisternas terminais com deslocamento das tríades. Areas de estreitamente, ruptura e perda de miofibrilas ocorreram assim como zonas de completa desorganizaçao de miofibrilas com perda do padrao estriado normal. As mitocondrias estavam em linhas gerais dentro dos limites da normalidade. Agregados tubulares peculiares vistos comumente na paralisia periódica e outras condiçoes patológicas humanas, foram encontrados tanto no músculo dos camundongos controles como no dos alcoólicos. Os nervos intramusculares assim como as junçoes neuromusculares mostravam-se normais. Importantes anormalidades nos capilares musculares foram observadas, consistindo de tumefaçao das células endoteliais e de espessamente da lâmina basal. Difusa infiltraçao lipídica microvesicular foi vista no citoplasma dos hepatócitos parecendo constituir uma evidência a mais da açao tóxica do etanol sobre o organismo. Concluímos que a ingestao prolongada de etanol, representando 14,4 por cento de calorias totais, produz no músculo gastrocnêmio de camundongos C57BL1O bem nutridos um elenco de alteraçoes ultraestruturais que refletem um efeito tóxico direto sobre o músculo esquelético. As alteraçoes constatadas sao semelhantes àquelas descritas na miopatia alcoólica crônica humana.

Ultrastructure of voluntary muscle in childhood malnutrition

This paper is part of a study on the electron microscopy of protein-energy malnutrition, using a rapid autopsy protocol. Samples of voluntary muscle, obtained from eight children dying of severe oedematous malnutrition, were fised in glutaraldehyde within 75 minutes of death. Atrophy of myofibres, increased prominence of satellite cells, and segmental necrobiosis were seen by light microscopy. Electron microscopy showed variable depletion of myofibrils. In the most severe case, there was focal absence of myofibrils, also disorganized Z lines, and absent M bands. Residual atrophic myofibrils measured less than 0.1 µm in width. Other specimens showed sarcomere disorganization, mitochondrial swelling, glycogen depletion, sarcoplasmic oedema, and focal contractions of sarcomeres. Though non-specific, rigor may phosphates. These conditions may exist in severe malnutrition, complicated by terminal infection and metabolic disturbances