Miositis osificante progresiva: ultraestructura, bioquímica e histoquímica de músculo macroscópicamente sano

Se estudió un caso de miositis osificante progresiva en una niña de 13 años, a la cual se le tomó una muestra de músculo gastronecmio lateral, aparentemente no afectado, en el curso de una intervención quirúrgica ortopédica. La muestra se procesó mediante métodos histológicos, histoquímicos, bioquímicos, inmunocitoquímicos y ultraestructurales. Se encontró un predominio de fibras musculares tipo I (83%) con alta capacidad oxidativa y baja capacidad glicolítica. Las fibras del tipo II eran pequeñas (área promedio 2.084 Um2 ) y mostraron otros signos de atrofia al examen ultraestructural. La densidad capilar fue relativamente alta, (573) siendo normal el índice capilar/fibra (1,76). Sin embargo, algunos capilares se mostraron engrosados y con la luz ocluida, con la tinción de amilasa-PAS, lo cual fue corroborado con la microscopía electrónica, donde se vio la membrana basal engrosada, e inclusive algunos capilares totalmente degenerados. No se encontró reacción de inmunofluorescencia con las globulinas anti-IgG ni anti-IgM en los cortes de músculo. El espacio intersticial se encontró agrandado. Se concluye que no hay evidencias de la participación de un mecanismo autoinmune en la miositis osificante progresiva, que existe un daño capilar y alteración de las fibras musculares, aún en el músculo que no manifiesta a simple vista proceso de osificación.During an orthopedic operation a sample of the apparently normal lateral gastronecmius muscle was taken from a 13 year old female patient affected by myositis ossificans progressiva. The muscle sample was analyzed by light, electron and fluorescence microscopy, and some enzymes were assayed. Muscle fibers were classified by the adenosintriphosphatase reaction. The percentage of type I fiber was high (83%). Atrophy was found in type II fibers as shown by small mean area (2.084 Um2 ) and some ultrastructural features as infoldings of the sarcolemma. Capillary density was high (573 capillaries/mm2 ), and capillaries per fiber index was normal (1.76), as were oxidative enzymes. However many capillaries were occluded, with thick basal membrane and abnormal endothelial cells and pericytes. No immunofluorescence was found with anti IgG or anti IgM in the muscle fibers. Intersticial spaces in the cross section of the muscle were enlarged. In conclusion, no evidence of autoimmune involvement was found in myositis ossificans progressiva, but alteracions of capillaries and muscle fibers were found in a muscle apparently not affected yet by the ossification process

Trypanosoma vivax Adhesion to Red Blood Cells in Experimentally Infected Sheep

Trypanosomosis, a globally occurring parasitic disease, poses as a major obstacle to livestock production in tropical and subtropical regions resulting in tangible economic losses. In Latin America including Venezuela, trypanosomosis of ruminants is mainly caused by Trypanosoma vivax. Biologically active substances produced from trypanosomes, as well as host-trypanosome cellular interactions, contribute to the pathogenesis of anemia in an infection. The aim of this study was to examine with a scanning electron microscope the cellular interactions and alterations in ovine red blood cells (RBC) experimentally infected with T. vivax. Ovine infection resulted in changes of RBC shape as well as the formation of surface holes or vesicles. A frequent observation was the adhesion to the ovine RBC by the trypanosome's free flagellum, cell body, or attached flagellum in a process mediated by the filopodia emission from the trypanosome surface. The observed RBC alterations are caused by mechanical and biochemical damage from host-parasite interactions occurring in the bloodstream. The altered erythrocytes are prone to mononuclear phagocytic removal contributing to the hematocrit decrease during infection.This research was supported by Project G-98003462-Fondo Nacional de Ciencia, Tecnología e Innovación (FONACIT), Caracas, Venezuela, and the Instituto de Estudios Científicos y Tecnológicos from Universidad Nacional Experimental Simón Rodríguez. The authors thank Beatriz Cajade for critical reading of this paper.S

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 is the first ultrastructural report in the literature of muscle injuries and neurological and proteolytic activities caused by BrWSV.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

PATOLOGÍA ULTRAESTRUCTURAL DEL INFILTRADO INFLAMATORIO EN LA DISPLASIA EPITELIAL Y CARCINOMA ESPINOCELULAR BUCAL

Se ha observado que muchas neoplasias se desarrollan en asociación con la inflamación, infección e irritación crónica, siendo éste el caso del Carcinoma Espinocelular bucal y los estados previos de Displasia Epitelial. El propósito del presente trabajo fue estudiar ultraestructuralmente el infiltrado inflamatorio en lesiones epiteliales con Displasias y Carcinomas Espinocelulares bucales, mediante técnicas convencionales de la microscopia electrónica de transmisión. La presencia de un infiltrado inflamatorio y las frecuentes interacciones observadas entre sus componentes con las células epiteliales neoplásicas, sugieren su participación activa en el desarrollo y progresión del Carcinoma Espinocelular bucal dada la etiopatogenia de estas lesiones y la estrecha relación entre el microambiente generado por la inflamación crónica y las células epiteliales tumorales.ABSTRACT It has been observed that many neoplasms are developed in association with inflammation, infection and chronic irritation, which is the case of oral Spinocellular Carcinoma and previous states of Epithelial Dysplasias. The purpose of the present work was to study ultrastructurally : inflammatory infiltrate in epithelial lesions related to Dysplasia and oral Spinocellular Carcinomas by routine techniques for transmission electron microscopy. Observed presence of inflammatory infiltrate and the frequent interactions between cells of infiltrate and with neoplastic cells suggests its active participation in the development and progression of oral Spinocellular Carcinoma given the etiopathogenia of these lesions and the intimate relation among microenvironment generated by chronic inflammation and tumor epithelial cells

Construcción de un modelo de regresión multivariante predictivo de cáncer de cuello uterino

The microscopic characteristics predictive of cancer have been synthesized in the presence of coagulative necrosis of tumor cells, which is a strong predictor of malignancy, other features are the unconventional forms of tumor cells and degenerate hyperchromatic pleomorphic nuclei.This research tried to identify ultrastructural variables in premalignant and malignant cells which could have greater predictive strength. Methods: 52 biopsies were taken from patients with premalignant cervical cancer and 13 with invasive cancer, samples were treated with conventional techniques for transmission electron microscopy, and data was processed with the program SPSS18®. Results and conclusions: in tissues with premalignant cells were found lymphocytes (I), altered smooth muscle fibers (X), plasma membrane discontinuous cells (L), degenerate capillaries (H), vacuolated cytoplasm (C), fibroblasts with abundant rough endoplasmic reticulum (O), mitochondria degenerate (K), nuclei with irregular contours (D), absence of desmosomes (Y). These elements were also found in invasive cancer samples, based on these results, the following equation was constructed: Cancer ̂=0,674+1,241*I+0,645*X+0,439*L+0,426*H+0,275*C+0,242*O+0,227*K+0,223*D-0,397*YLas características microscópicas predictivas de cáncer se han sintetizado en la presencia de necrosis coagulativa de células tumorales, que es un sólido predictor de malignidad, otras son formas no convencionales de las células tumorales y núcleos degenerados, hipercromáticos y pleomórficos. Objetivo de la investigación: identificar las variables ultraestructurales de las células malignas y premalignas de mayor fortaleza predictiva de cáncer. Metodología: se tomaron 52 biopsias con diagnóstico de lesiones premalignas de cuello uterino y 13 con cáncer invasor. Las muestras fueron tratadas con técnicas convencionales para estudio de microscopía electrónica de transmisión, la data fue procesada con programa SPSS18. Resultados y conclusiones: En células premalignas se encontraron linfocitos(I), fibras musculares lisas alteradas(X), membrana plasmática discontinua (L), vasos capilares degenerados(H), citoplasma vacuolizado(C), fibroblastos con abundantes retículo endoplásmico rugoso(O), mitocondrias degeneradas(K), núcleos de contornos irregulares(D), ausencia de desmosomas(Y). Estos elementos fueron igualmente encontrados en muestras de cáncer invasivo,estos resultados permitieron el diseño de la ecuación: Cancer = 0,674 + 1,241 * I + 0,645 * X + 0,439 * L + 0,426 * H + 0,275 * C + 0,242 * O + 0,227 *K + 0,223 * D - 0,397 *