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Int. j. morphol ; 38(2): 505-512, abr. 2020. graf
Artigo em Inglês | LILACS | ID: biblio-1056469


Sexual dimorphism exists at all levels of the nervous system. These sex differences could underlie genderrelated differences in behavior and neuropsychological function, as well as the gender differences in the prevalence of various mental disorders such as autism, attention deficit disorders, and schizophrenia. Myelination, on the other hand, is a unique cellular process that can have a dramatic impact on the structure and physiology of an axon and its surrounding tissue. The corpus callosum (CC) is the largest of the brain commissures, which connects the cerebral cortices of the two hemispheres, and provides interhemispheric connectivity for information transfer and processing between cortical regions. Variation in the axonal properties of CC will alter the interhemispheric connectivity. The CC consists of myelinated and unmyelinated axons, glial cells and blood vessels. Several functional studies have reported that the function of CC is associated with its axons density and myelination properties. The sexual dimorphism in the axonal content of the CC has always been controversial; hence, the aim of this study was to analyze the differences in axons' diameter and myelin sheath thickness of the CC between male and female rats. For this purpose, five pairs of adult male and female rats were perfused and the CC were removed and sectioned. Four sections from different subregions of the corpus callosum that represent the genu, anterior body, posterior body, and splenium of the CC were stained and electron microscopic images were captured using stereological guidelines. Later, the axons diameter and myelin sheath thickness for each subregion were calculated and compared between males and females. Our preliminary findings of the present study indicated region specific differences in the myelinated axon thickness and diameter in the CC between male and female rats.

El dimorfismo sexual existe en todos los niveles del sistema nervioso. Estas diferencias de sexo podrían ser la base de las diferencias de comportamiento y función neuropsicológica relacionadas con el sexo, así como las diferencias en la prevalencia de diversos trastornos mentales, como el autismo, los trastornos por déficit de atención y la esquizofrenia. La mielinización, por otro lado, es un proceso celular único que puede tener un impacto dramático en la estructura y fisiología de un axón y su tejido circundante. El cuerpo calloso (CC) es la mayor comisura cerebral, que conecta las cortezas cerebrales de ambos hemisferios, y proporciona la conectividad interhemisférica para la transferencia y el procesamiento de información entre regiones corticales. La variación en las propiedades axonales de CC alterará la conectividad interhemisférica. El CC consiste en axones mielinizados y no mielinizados, células gliales y vasos sanguíneos. Varios estudios funcionales han informado que la función de CC está asociada con la densidad de axones y las propiedades de mielinización. El dimorfismo sexual en el contenido axonal del CC siempre ha sido controvertido; por lo tanto, el objetivo de este estudio fue analizar las diferencias en el diámetro de los axones y el grosor de la vaina de mielina del CC entre ratas macho y hembra. Para este propósito, se perfundieron cinco pares de ratas macho y hembra adultas y se extrajeron y seccionaron las CC. Se tiñeron cuatro secciones de diferentes subregiones del cuerpo calloso que representan el genu, el cuerpo anterior, el cuerpo posterior y el esplenio y se capturaron imágenes de microscopía electrónicas utilizando referencias estereológicas. Posteriormente se calculó el diámetro de los axones y el grosor de la vaina de mielina para cada subregión y se compararon entre machos y hembras. Nuestros hallazgos preliminares del presente estudio indicaron diferencias específicas en el grosor y diámetro del axón mielinizado en el CC entre ratas macho y hembra.

Animais , Masculino , Feminino , Ratos , Axônios/ultraestrutura , Caracteres Sexuais , Corpo Caloso/ultraestrutura , Bainha de Mielina/ultraestrutura , Microscopia Eletrônica , Corpo Caloso/citologia
Int. j. morphol ; 38(2): 305-308, abr. 2020. tab, graf
Artigo em Inglês | LILACS | ID: biblio-1056439


Fixation is one of the processes in preparing histology and pathology. The common material for fixation is buffered formalin including paraformaldehyde. However, the effect of the damaged cells, which is fixed for a long time, causes the research for other fixation materials to become necessary. In addition, paraformaldehyde is also harmful to human body and natural environment. Ethanol is one of the alternative fixation materials, which has been used for two hundred years. It has been used for many purposes, both in routine staining and immunohistochemistry. Nonetheless, no research confirms its effect on the electron microscope. The authors studied the effect of 50 % of ethanol on the cell membrane, organelles, and nucleus of Purkinje cells (Neuron purkinjense) observed on a light microscope and Transmitted Electron Microscope (TEM). Then it was compared to buffered formalin. In the light microscope, it shows that both of fixations have no different effects of the morphology of the cell membrane, cytoplasm, the nucleus of Purkinje cells and the neutrophils. We assume that our 50 % of ethanol concentration is almost the same as BF 10 % in the ability of hardening tissue and color absorption based on the previous study. In TEM, the structure of the cell membrane, organelles, and cytoplasm of Purkinje cell look broken in the cerebellum of 50 % of ethanol except for the nucleus. There was no significant difference diameter of the nucleus. It happened in general because of the shrinkage effect of ethanol. However, the authors recommend using 50 % of ethanol for routine staining.

La fijación es uno de los procesos en la preparación de muestras para histología y patología. El material más común para la fijación es la formalina tamponada. Sin embargo, el daño a las células que se mantienen en formalina durante mucho tiempo, hace necesario buscar otros materiales de fijación. Además, el paraformaldehido también es perjudicial para el cuerpo humano y el medio ambiente natural. El etanol es uno de los materiales de fijación alternativos que se ha utilizado durante muchos años, con diversos objetivos, tanto en la tinción de rutina como en la inmunohistoquímica. Sin embargo no se ha confirmdo su efecto con microscopio electrónico. Los autores estudiaron el efecto del 50 % de etanol sobre la membrana celular, los orgánulos y el núcleo de las células de Purkinje observados en un microscopio óptico y un microscopio de transmisión electrónico (TEM). Luego se comparó con la formalina tamponada. En el microscopio óptico se observó que ambas fijaciones no tienen efectos diferentes a la morfología de la membrana celular, el citoplasma, el núcleo de las células de Purkinje y los neutrófilos. Suponemos que nuestra concentración de 50 % de etanol es casi la misma que BF 10 % en la capacidad de endurecer el tejido y la absorción de color según el estudio anterior. En TEM, la estructura de la membrana celular, los orgánulos y el citoplasma de la célula de Purkinje presentaban daño en el cerebelo con un 50 % de etanol, a excepción del núcleo. No hubo diferencia significativa en el diámetro del núcleo. En general lo anterior se debió al efecto de contracción del etanol. En conclusión los autores recomiendan usar 50% de etanol para la tinción de rutina.

Animais , Masculino , Camundongos , Encéfalo/efeitos dos fármacos , Encéfalo/ultraestrutura , Fixação de Tecidos/métodos , Etanol/farmacologia , Microscopia Eletrônica , Organelas/efeitos dos fármacos , Organelas/ultraestrutura , Camundongos Endogâmicos BALB C
Int. j. morphol ; 38(2): 461-471, abr. 2020. graf
Artigo em Inglês | LILACS | ID: biblio-1056463


This experiment was designed to study the administration of normal doses of one of recent antimalarial drug and coadministration of vitamin E on the kidney tissue. A total twenty-four adult male albino rats were used and divided into four groups: the first one served as a control, the second received artemether orally for three days consecutively. The rats of the third and fourth groups received the same dose of artemether concomitantly with 50 and 100 mg/kg vitamin E orally daily for 2 weeks. After the last dose, the rats were sacrificed and the kidney tissues with blood samples obtained and processed for light, electron microscopic and biochemical analysis. Histologically, artemether treated kidneys showed atrophied glomeruli with widened urinary space and kidney tubules were degenerated with disturbed contour and some vacuoles inside it. Ultrastructurally, the glomeruli of this group showed hypertrophic endothelial cells, irregularity of its basement membrane, disrupted foot processes and filtration slits. The kidney tubule cells showed loss of basal infoldings, cytoplasmic vacuolation, polymorphic damaged swollen mitochondria a loss of its microvilli towards its capillary lumen. Artemether plus vitamin E of the rat kidney groups showed improvement of morphological changes compared to the changes seen in artemether alone. These data were confirmed by biochemical findings with marked improvement of blood urea and creatinine levels and increase of anti-oxidant enzyme activities of glutathione peroxidase and superoxide dismutase in the vitamin E treated groups. The results of this study revealed that vitamins E can improve the adverse changes of artemether of rat renal tissue.

Este proyecto fue diseñado para estudiar la administración de dosis normales de uno de los medicamentos antipalúdicos y de la administración de vitamina E en el tejido renal. Se utilizaron 24 ratas albinas machos adultas divididas en cuatro grupos: el primero sirvió como control, el segundo recibió arteméter por vía oral durante tres días consecutivos. Las ratas del tercer y cuarto grupos recibieron la misma dosis de arteméter concomitantemente con 50 y 100 mg / kg de vitamina E por vía oral diariamente durante 2 semanas. Después de la última dosis, las ratas fueron sacrificadas y se obtuvo el tejido renal de cada muestra los cuales fueron procesados para análisis con microscopías de luz y electrónica, además de exámenes bioquímicos. Histológicamente, los riñones tratados con arteméter mostraron atrofia glomerular con espacio urinario ensanchado y túbulos renales degenerados con contorno alterado y algunas vacuolas en su interior. Ultraestructuralmente, los glomérulos de este grupo mostraron células endoteliales hipertróficas, irregularidad de su membrana basal, procesos alterados del pie y hendiduras de filtración. Las células del túbulo renal mostraron pérdida de inflexiones basales, vacuolación citoplasmática, mitocondrias dañadas y pérdida de sus microvellosidades hacia la luz capilar. Arteméter más vitamina E en los grupos de riñón de rata mostraron una mejora de los cambios morfológicos, en comparación con los cambios observados en arteméter solamente. Estos datos fueron confirmados por hallazgos bioquímicos con una marcada mejoría de los niveles de urea y creatinina en sangre y un aumento de las actividades enzimáticas antioxidantes de la glutatión peroxidasa y la superóxido dismutasa en los grupos tratados con vitamina E. Los resultados de este estudio revelaron que la vitamina E puede mejorar los cambios adversos del arteméter del tejido renal de la rata.

Animais , Masculino , Ratos , Vitamina E/farmacologia , Lesão Renal Aguda/induzido quimicamente , Artemeter/toxicidade , Vitamina E/administração & dosagem , Microscopia Eletrônica , Biomarcadores/análise , Ratos Wistar , Rim/efeitos dos fármacos , Rim/patologia , Rim/ultraestrutura , Antimaláricos/toxicidade
Int. j. morphol ; 37(2): 694-700, June 2019. graf
Artigo em Espanhol | LILACS | ID: biblio-1002279


Spondylus limbatus es una especie bajo protección especial en México, de la que existe poca información biológica y nada sobre estudios histológicos o de ultraestructura del ovario. El objetivo de esta investigación fue caracterizar la morfología ultraestructural de los gametos femeninos maduros y en degeneración. La gónada femenina de S. limbatus en estado de madurez presentó ovocitos postvitelogénicos de 60-70 µm de diámetro, que presentan el aspecto característico de células metabólicamente activas y altamente sintetizadoras. La membrana citoplasmática posee especializaciones destinadas a aumentar la superficie de absorción de la célula, las microvellosidades; el citoplasma presenta numerosos sistemas membranosos relacionados con la síntesis de material de reserva y secreción; y el patrón de organización nuclear altamente lobulado, y por consiguiente con una gran superficie que asegura el intercambio núcleo-citoplasma, se incorpora de forma estructural al proceso de vitelogénesis. Finalmente, se describen los cambios ultraestructurales resultantes de la lisis de los ovocitos: colapso de las membranas nuclear y citoplásmica, y presencia de células hemocíticas macrófagas.

Spondylus limbatus is a species under special protection in Mexico, of which there is little or no information in the literature of biological, histological or ultrastructural studies of the ovary. The objective of this research was to characterize the ultrastructural morphology of mature and degenerating female gametes. The female gonad of S. limbatus in mature state presented post-vitellogenic oocytes 60-70 µm in diameter, which have characteristics of metabolically active and highly synthesizing cells. The cytoplasmic membrane has specializations designed to increase the absorption surface of the cell, the microvilli; the cytoplasm presents numerous membranous systems related to synthesis of reserve and secretion material as well as the highly lobed nuclear organization pattern; a large surface that ensures core-cytoplasm exchange, is structurally incorporated into the vitellogenesis process. Finally, ultrastructural changes resulting from the lysis of the oocytes are described: collapse of nuclear and cytoplasmic membranes, and the presence of macrophage hemocytic cells.

Animais , Feminino , Oócitos/ultraestrutura , Bivalves , Gônadas/ultraestrutura , Reprodução , Microscopia Eletrônica
Rev. odontol. Univ. Cid. São Paulo (Online) ; 31(2): 116-124, abr/jun 2019. il.
Artigo em Português | LILACS, BBO - Odontologia | ID: biblio-1021582


Atualmente o uso de instrumentos rotatórios é uma realidade, porém, estes apresentam restos de resíduos na superfície de corte. Sendo assim, o objetivo deste trabalho foi avaliar por meio de microscopia eletrônica de varredura (MEV), a presença de resíduos na superfície de corte de 20 instrumentos rotatórios de NiTi sem uso. Foram divididos em quatro grupos, a saber: Grupo A - TwistedFile® (n=5), Grupo B - BioRaCe® (n=6), Grupo C - Mtwo® (n=4) e GrupoD - EndoWave® (n=5). As pontas dos instrumentos foram microfotografadas com aumento de 190X a partir do diâmetro inicial para verificar a presença de resíduos. Em seguida, os instrumentos passaram por um rigoroso processo de limpeza e novamente microfotografados com o mesmo padrão de aumento, observando-se a ponta do instrumento e a 5mm da ponta do instrumento. Os dados obtidos foram tabulados e submetidos à análise percentual e estatística valendo-se do teste Exato de Fisher e nível de significância de 5%. No que se refere à sujidade ocorreram diferenças estatisticamente significantes entre as limas Mtwo® com os outros instrumentos (p<0,05). Concluiu-se que todos os instrumentos estavam sujos, exceto a marca Mtwo® e que após os procedimentos de limpeza a superfície de corte estava isenta de resíduos.

Currently the use of rotary instruments is a reality, however, they have leftover residue on the cut surface. Thus, the aim of this study was to evaluate by means of scanning electron microscopy (SEM), the presence of residues on the surface of cut 20 NiTi rotary instruments unused. Were divided into four groups as follows: Group A - TwistedFile® (n = 5) Group B - BioRaCe® (n = 6) Group C - Mtwo® (n = 4) Group D - EndoWave® (n = 5). The tip of the instruments was photomicrographed with 190X at magnification from the initial diameter for the presence of residues. Then, instruments passed through a rigorous cleaning and again photomicrographed with the same magnification observing the tip of the instrument and 5mm from the tip of the instrument. Data were tabulated and analyzed statistically percentage and taking advantage of the Fisher exact test and a significance level of 5%. With regard to dirt statistically significant differences between the files Mtwo® with other instruments (p <0.05). It was concluded that all instruments were dirty except Mtwo® brand and after cleaning procedures the cut surface was free of residues.

Titânio , Microscopia Eletrônica , Instrumentos Odontológicos , Níquel
Arq. Inst. Biol ; 86: e0232019, 2019. ilus
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1024605


Rust, caused by the biotrophic fungus Olivea neotectonae, stands out as an important disease in teak plantations in Brazil. Considering the current importance of this rust and the need to obtain additional basic information about its causal agent to better disease management, the present study aimed to analyze the morphology of O. neotectonae using light microscopy and demonstrate the infection process in teak leaves using scanning electron microscopy techniques (SEM). The morphological characteristics observed using light microscopy were similar to those described in the literature for this species. SEM revealed that the infection structures of the pathogen penetrated the teak leaf through the stomata complex. This study provides the first microscopic evidence of the infection process of Olivea neotectonae in leaves of teak plants.(AU)

A ferrugem, causada pelo fungo biotrófico Olivea neotectonae, destaca-se como uma doença importante nas plantações de teca no Brasil. Considerando a importância atual dessa ferrugem e a necessidade de obter informações básicas adicionais sobre seu agente causal para melhor manejo da doença, o presente estudo teve como objetivo analisar a morfologia da O. neotectonae utilizando microscopia de luz e demonstrar o processo de infecção em folhas de teca por meio de técnicas de microscopia eletrônica de varredura (MEV). As características morfológicas observadas em microscopia de luz foram semelhantes às descritas na literatura para essa espécie. A MEV revelou que as estruturas de infecção do patógeno penetraram na folha de teca através do complexo estomático. Este estudo fornece a primeira evidência microscópica do processo de infecção de Olivea neotectonae em folhas de plantas de teca.(AU)

Controle de Pragas , Floresta Úmida , Fungos , Doenças das Plantas , Microscopia Eletrônica/métodos
Int. j. morphol ; 36(4): 1316-1325, Dec. 2018. tab, graf
Artigo em Inglês | LILACS | ID: biblio-975702


The morphology of the hypophysis in Bactrian camel has not been described in the literature, despite it being the master of endocrine organs in vertebrates. In the present study, we examined the morphological features of the hypophysis in Bactrian camel by means of gross anatomy, light and electron microscope. Our findings showed that the gland was a protrusion of the bottom of the hypothalamus at the base of the brain with about 1.54 g in weight and 2 cm3 in volume. The hypophysis consists of two major parts: fully developed adenohypophysis and underdeveloped neurohypophysis, the adenohypophysis consists of pars distalis and pars intermedia. Seven type cells of the pars distalis could be distinguished with immunohistochemical techniques and electron micrographs: somatotroph, mammotroph, thyrotroph, corticotroph, gonadotroph, chromophobe and stellate cells which is in accordance with most mammals. Notably, the stellate cells could be obviously distinguished from chromophobe cells in histological oberservation. Moreover, the corpusculum neurosecretorium (Herring bodies) were rare in the external neurohypophysis, and mainly distributed in the internal neurohypophysis, this was different from most mammals. Results from this study would provide a necessary theoretical basis for ongoing investigations for Bactrian camels and their good adaptability in arid and semi-arid circumstances.

La morfología de la hipófisis en el camello bactriano no ha sido descrita en la literatura, a pesar de ser el maestro de los órganos endocrinos en los vertebrados. En el presente estudio, examinamos las características morfológicas de la hipófisis del camello bactriano por medio de anatomía general, microscocopía de luz y microscopía electrónica. Nuestros hallazgos mostraron que la hipófisis es una protuberancia ubicada en la porción inferior del hipotálamo, en la base del cerebro, con aproximadamente 1,54 g de peso y 2 cm3 de volumen. La hipófisis consta de dos partes principales: adenohipófisis, completamente desarrollada, y neurohipófisis, poco desarrollada; además, la adenohipófisis consta de una pars distalis y una pars intermedia. Con técnicas inmunohistoquímicas y micrografías electrónicas en la pars distalis se pudieron distinguir siete tipos de células: somatotrofas, mamotróficas, tirotrofas, corticotrofas, gonadotrofas, cromófobas y estrelladas, lo que es similar a la mayoría de los mamíferos. En la observación histológica las células estrelladas se pueden distinguir naturalmente de las células cromófobas. Además, es rara la presencia de corpusculum neurosecretorium (Cuerpos de Herring) en la neurohipófisis externa, hallándose distribuidos principalmente en la neurohipófisis interna, esto es diferente a lo encontrado en la mayoría de los mamíferos. Los resultados de este estudio proporcionarían una base teórica necesaria para las investigaciones en curso de los camellos bactrianos y su buena adaptabilidad en circunstancias áridas y semiáridas.

Animais , Hipófise/ultraestrutura , Camelus/anatomia & histologia , Hipófise/anatomia & histologia , Imuno-Histoquímica , Microscopia Eletrônica
Int. j. morphol ; 36(3): 792-798, Sept. 2018. tab, graf
Artigo em Espanhol | LILACS | ID: biblio-954187


RESUMEN: El cáncer de mama es la principal causa de muerte debido al cáncer en mujeres. La microscopía electrónica permite establecer características constitutivas de las células entre diferentes poblaciones celulares. Las células madre del cáncer mamario con inmunofenotipo CD44alta/CD24baja son una población de células intratumorales asociada a la quimioresistencia y metástasis, cuya ultraestructura aún no ha sido bien estudiada. El objetivo de este trabajo fue conocer características ultraestructurales de células con fenotipo de células madre del cáncer de la línea celular MDA-MB-436 de tumor mamario triple negativo comparándolas con células madre adiposas. Se utilizó microscopía electrónica de barrido y de transmisión. Previamente, mediante separación inmunomagnética positiva empleando anticuerpos anti CD44 y anti CD24 unidos a perlas magnéticas, se obtuvo una población de células con fenotipo CD44alta/CD24baja a partir de 10x106 células de la línea MDA-MB-436, la cual igual que las células madre adiposas fue cultivada en cubreobjetos para microscopía electrónica de barrido; en tanto que para microscopía electrónica de transmisión se obtuvo un pellet de células, luego se fijó con glutaraldehído al 2,5 % y post fijó con OsO4 1 %. Para microscopía óptica de alta resolución se usó azul de toluidina como tinción. Luego de obtener el fenotipo de células madre del cáncer se corroboró su pluripotencia detectando la expresión de los genes Oct4 y nanog mediante RT-PCR. Nuestros resultados muestran que las células de este fenotipo son pequeñas, redondeadas, recubiertas por microvellosidades abundantes pero cortas; el citoplasma tiene organelas de secreción celular y abundantes mitocondrias alargadas; el núcleo es excéntrico ocupando la mitad del volumen celular, el nucléolo es voluminoso y la heterocromatina está adosada a la membrana nuclear interna. Se concluye que el inmunofenotipo celular estudiado es una sub población celular dentro de la línea estudiada que difiere en tamaño y ultraestructura de las células madre adiposas.

SUMMARY: Breast cancer is the leading cause of cancer deaths in women. Electron microscopy allows establishing constitutive characteristics of cells between different cell populations. CD44 high / CD24 low mammary cancer stem cells are a population of intratumoral cells associated with chemoresistance and metastasis, whose ultrastructure has not yet been well studied. The objective of this work was to know the ultrastructural characteristics of cells with cancer stem cell phenotype, of the triple negative mammary tumor cell line MDAMB-436 436 using scanning and transmission electron microscopy, and to contrast them with adipose-derived mesenchymal stem cells. Previously, by immunomagnetic purification using anti CD44 and anti CD24 antibodies bound to magnetic beads, cells populations was obtained from 10x106 cells of the MDA-MB-436 line, which, adipose-derived mesenchymal stem whereas cultivated on coverslips for scanning electron microscopy; while for transmission electron microscopy a cell pellet was obtained, then fixed with 2.5 % glutaraldehyde and post fixed with OsO4 1 %. For high resolution optical microscopy, toluidine blue was used as staining. After obtaining the phenotype of cancer stem cells, their pluripotency was corroborated by detecting the expression of the Oct4 and nanog genes by RT-PCR. Our results show that the cells of this phenotype are small, rounded, covered by abundant but short microvilli; the cytoplasm has organelles of cellular secretion and abundant elongated mitochondria; the nucleus is eccentric occupying half of the cellular volume, the nucleolus is bulky and the heterochromatin is attached to the inner nuclear membrane. It is concluded that the cellular immunophenotype studied is a sub-cellular population within the line studied that differs in size and ultrastructure of the adipose stem cells.

Humanos , Feminino , Células-Tronco Neoplásicas/ultraestrutura , Neoplasias da Mama/patologia , Microscopia Eletrônica , Imunofenotipagem , Linhagem Celular Tumoral/ultraestrutura , Reação em Cadeia da Polimerase em Tempo Real
São Paulo; s.n; 2018. 107 p. ilus, mapas.
Tese em Português | LILACS, Sec. Est. Saúde SP, SESSP-CTDPROD, Sec. Est. Saúde SP, SESSP-ACVSES | ID: biblio-1005444


Toxoplasma gondii, protozoário causador da toxoplasmose, é transmitido dos animais para os seres humanos pela ingestão de carne contaminada ou por oocistos liberados nas fezes de felinos no ambiente. Nos seres humanos, a infecção é normalmente assintomática, mas em casos em que a infecção primária ocorre durante a gravidez ou a reativação da infecção latente ocorre em pacientes imunosuprimidos pode ser grave. Nas últimas décadas tem se estudado pequenas estruturas secretadas pelas células procarióticas e eucarióticas denominadas de vesículas extracelulares (EVs). As EVs podem transportar biomarcadores de doenças, macromoléculas biorreativas contribuindo para a patogênese e sendo uma forma de diagnóstico não invasivo. Estas pequenas estruturas podem ser isoladas por ultracentrifugação, cromatografia e observadas por microscopia eletrônica. Elas participam na comunicação entre as células, na transferência de proteínas, lipídios e ácidos nucléicos. Diante do exposto, o presente estudo teve como objetivo estabelecer um protocolo para isolar e caracterizar vesículas extracelulares produzidas e excretadas por taquizoítos da cepa RH de T. gondii. Taquizoítos provenientes de culturas de células VERO foram isolados dos sobrenadantes das culturas e centrifugados em cinco séries de lavagens. A seguir, foram incubados em meio de cultura por 24 horas para secreção das EVs. Em seguida, investigou-se o tamanho e concentração das EVs isoladas por análise de varredura de partículas (NTA) no equipamento NanoSight. Paralelamente, a morfologia e a liberação das EVs também foram investigadas por microscopia eletrônica de transmissão e de varredura. Então, as EVs foram purificadas por cromatografia em gelexclusão em alíquotas de 1 mL (24-32 frações) e imuno-selecionadas por ELISA utilizando um "pool" de soros reagente para toxoplasmose. A seguir foi investigado a presença de miRNA nas EVs; e finalmente, foi avaliado o perfil proteico das EVs de T. gondii para verificar por testes sorológicos se estas partículas poderiam ser reconhecidas pelo sistema imune hospedeiro. As análises realizadas por NTA permitiram determinar que cerca de 1 x 106 taquizoítos secretaram de 4 a 8 x 108 EVs/mL num período de 24 horas de incubação em meio de cultura celular. Adicionalmente, estas vesículas apresentaram morfologia e tamanho de 165-175 nm de diâmetro, tamanho correspondente às microvesículas. Estes resultados também foram confirmados pela avaliação das imagens fornecidas pelas microscopias eletrônicas de transmissão e varredura. Tambem foi possível determinar a presenção de small RNAs e micro RNAs através de uma corrida eletroforética micro fluídica. As análises por SDS-PAGE mostram que as proteínas que compõem as EVs apresentaram um perfil eletroforético com espectro de 15 a 70 kDa. Soros de camundongos cronicamente infectados (com 2 diferentes cepas de T. gondii) e soros humanos reconheceram distintos padrões eletroforéticos no immunoblotting. As vesículas liberadas por T. gondii podem ser um mecanismo importante pelo qual os parasitas apresentam seus antígenos ao hospedeiro e, portanto, podem ter um papel importante na patogênese da toxoplasmose

Toxoplasma gondii, a protozoan that causes toxoplasmosis, is transmitted from animals to humans through ingestion of infected meat or oocysts released by felines into the environment. In humans, infection is usually asymptomatic, but in cases where primary infection occurs during pregnancy or the reactivation of latent infection occurs in immunosuppressed patients it can be serious. In the last decades, small structures secreted by prokaryotic and eukaryotic cells called extracellular vesicles (EVs) were studied. These small structures can be isolated by ultracentrifugation, chromatography and examined by electron microscopy. EVs participate in the communication between cells, transfer of proteins, lipids and nucleic acids. In addition, them can carry disease biomarkers, bioreactive macromolecules contributing to the pathogenesis of diseases. In view of the above, the present study aimed to establish a protocol to isolate and characterize extracellular vesicles produced and excreted by tachyzoites of T. gondii RH strain. To achieve this goal, tachyzoites from VERO cell cultures were isolated from the culture supernatants and centrifuged in five sets of washes. Next, they were incubated in culture medium for 24 hours for secretion of the EVs. The size and concentration of the isolated EVs by particle scan analysis (NTA) were investigated on the NanoSight (NTA) equipment. In parallel, the morphology and the release of the EVs were also investigated by transmission and scanning electron microscopy. Then, EVs were purified by gel-exclusion chromatography in 1 mL aliquots (24-32 fractions) and immuno-selected by ELISA using a pool of reagent sera for toxoplasmosis. Next, the presence of miRNA in the EVs was investigated; and finally, the protein profile of T. gondii EVs was evaluated and verify by serological tests if these particles could be recognized by the host immune system. The results showed that the protocol for recovery of T. gondii EVs was established from 1 to 1010 tachyzoites obtained from cultured cells. Analyzes performed by NTA allowed us to determine that about 1 x 106 tachyzoites secreted 4 to 8 x 108 EVs / mL within 24 hours of incubation in culture medium. Additionally, these vesicles presented morphology and size of 165-175 nm of diameter, corresponding microvesicles size. These results were also confirmed by the evaluation of images provided by transmission and scanning electron microscopy. The miRNA purifications from the EVs and subsequent microfluidic electrophoretic run confirmed the presence of smallRNA and miRNA in the vesicles. SDS-PAGE analyzes show that the proteins carried by the EVs showed an electrophoretic profile with a spectrum of 15 to 70 kDa. Sera from chronically infected mice (with 2 different strains of T. gondii) and humam serum recognized distinct electrophoretic patterns in immunoblotting

Humanos , Masculino , Feminino , Toxoplasma , Microscopia Eletrônica , Toxoplasmose
Int. j. morphol ; 35(4): 1473-1481, Dec. 2017. tab, graf
Artigo em Inglês | LILACS | ID: biblio-893159


SUMMARY: Special features of nanoparticles have resulted in their widespread use. Small molybdenum trioxide (MoO 3) nanoparticles can translocate from the entry portals into the circulatory and lymphatic systems and ultimately to body tissues and organs depending on their composition and size. In this research, sixty Wistar rats weighting 180-250 g were divided into 6 groups (n=10) randomly: Group 1 (Control) did not receive any medicine. Group 2 (Sham) received intraperitoneal normal saline for 35 days on a daily basis. Groups 3, 4, 5 and 6 received 50, 100, 200, and 300 mg/kg MoO3, respectively, the same way in the sham group and at the same interval. At the end of the experiment, the rats were weighted again and anesthetised. Then blood samples were taken from their hearts to determine the serum levels of estrogen, progesterone, and gonadotropins. Their ovaries were removed and ovarian volume, follicular diameter, number of each follicle type, and oocyte volume were determined. Results indicated that MoO3 nanoparticles strongly reduced body and ovarian weights in the rats. Moreover, a significant decrease was observed in ovarian volume, the number of follicle types, oocyte volume and follicular diameter. The nanoparticles increased the number of atretic follicles via ovarian tissue structure. MoO3 nanoparticles decreased serum estrogen level and increased serum level of FSH that was associated with disruption in the regulation of progesterone and LH secretion. The findings showed that MoO3 nanoparticles could bear negative effects on ovarian structure and function.

RESUMEN: Las características específicas de las nanopartículas han dado lugar a su uso generalizado. Las pequeñas nanopartículas de trióxido de molibdeno (MoO3) pueden penetrar los sistemas circulatorios y linfáticos y, en última instancia, dependiendo de su composición y tamaño, también los tejidos y órganos del cuerpo. En esta investigación se dividieron 60 ratas Wistar con un peso de 180-250 g en 6 grupos (n = 10) aleatoriamente: el Grupo 1 (Control) no recibió ningún medicamento. El Grupo 2 (Sham) recibió solución salina normal intraperitoneal durante 35 días diariamente. Los grupos 3, 4, 5 y 6 recibieron 50, 100, 200 y 300 mg / kg de MoO3 respectivamente, de la misma manera en el grupo simulado, y en el mismo intervalo. Concluyendo el experimento, las ratas se pesaron nuevamente y fueron anestesiadas. Luego se tomaron muestras de sangre de los corazones para determinar los niveles séricos de estrógeno, progesterona y gonadotropinas. Se retiraron los ovarios y se determinó el volumen ovárico, el diámetro folicular, el número de cada tipo de folículo y el volumen de ovocitos. Los resultados indicaron que las nanopartículas de MoO3 redujeron significativamente los pesos corporal y ovárico en las ratas. Además, se observó una disminución importante en el volumen ovárico, el número de tipos de folículos, el volumen de ovocitos y el diámetro folicular. Las nanopartículas aumentaron el número de folículos auriculares a través de la estructura del tejido ovárico. Las nanopartículas de MoO 3 disminuyeron el nivel sérico de estrógeno y aumentaron el nivel sérico de FSH que se asoció con la interrupción en la regulación de la progesterona y la secreción de LH. Los hallazgos mostraron que las nanopartículas de MoO 3 podrían tener efectos negativos sobre la estructura y la función ovárica.

Animais , Feminino , Ratos , Molibdênio/administração & dosagem , Nanopartículas , Folículo Ovariano/efeitos dos fármacos , Oxigênio/administração & dosagem , Estrogênios/sangue , Gonadotropinas/sangue , Microscopia Eletrônica , Tamanho do Órgão/efeitos dos fármacos , Ovário/efeitos dos fármacos , Ovário/ultraestrutura , Progesterona/sangue , Ratos Wistar
Int. j. morphol ; 35(4): 1383-1390, Dec. 2017. graf
Artigo em Inglês | LILACS | ID: biblio-893146


SUMMARY: Osteoarthritis (OA) caused by ageing joints or as a secondary complication of diabetes is a common health problem. We sought to develop an animal model of OA induced by a combination of the chondrocyte glycolytic inhibitor mono-iodoacetate (MIA) and streptozotocin (STZ), the agent that induces diabetes mellitus. We then hypothesized that the extent of damages to the knee joint induced by this model can be greater than OA induced by either MIA or STZ. Rats were either injected with MIA (model 1) or STZ (model 2) or both agents (model 3). After 8 weeks, harvested tissues from the knee joint of these groups were examined using scanning and transmission electron microscopy. In addition, blood samples were assayed for tumor necrosis factor alpha (TNF-α) and interleukin -6 (IL-6) that are known to be modulated in OA and diabetes. Compared to control group, substantial damages to the articular cartilage of the knee joint were observed in the three models with the severest in model 3. In addition, rats in model 3 showed significant (P<0.0001) increase in TNF-α and IL-6 compared to model 1 and 2. Thus, we have developed a new model of knee OA in rats that mimics a type of OA that is common among elderly people who have both, "ageing" joints and diabetes.

RESUMEN: La osteoartritis (OA) es un problema generalizado de salud a causa de un envejecimiento de las articulaciones, o bien de una complicación secundaria de la diabetes. El objetivo de este estudio fue desarrollar un modelo animal de OA inducido por una combinación dos drogas, un inhibidor de los condrocitos glucolíticos, el mono-iodoacetato (MIA), y la estreptozotocina (STZ), agente que induce la diabetes mellitus. Se consideró como hipótesis que el alcance de los daños a la articulación de la rodilla inducida por este modelo puede ser mayor que la OA inducida por MIA o STZ. Las ratas fueron inyectadas con MIA (grupo 1) o STZ (grupo 2) o ambos agentes (grupo 3). Se extrajeron muestras de la articulación de la rodilla de estos grupos al término de 8 semanas, y se examinaron mediante microscopía electrónica de barrido y de transmisión. Además, se analizaron muestras de sangre para el factor de necrosis tumoral alfa (TNF-α) e interleucina-6 (IL-6), que están moduladas en OA y en la diabetes. En comparación con el grupo control, se observaron daños sustanciales en el cartílago articular de la articulación de la rodilla en los tres modelos, encontrándose los daños más severos en el grupo 3. Además, las ratas del grupo 3 mostraron un aumento significativo (P <0,0001) de los niveles de TNF-α e IL- 6, en comparación con los grupos 1 y 2. Hemos desarrollado un nuevo modelo de OA de rodilla en ratas que imita un tipo de OA el cual, además de la diabetes, es común entre las personas mayores con un nivel importante de daño en las articulaciones.

Animais , Masculino , Ratos , Estreptozocina/toxicidade , Osteoartrite do Joelho/induzido quimicamente , Ácido Iodoacético/toxicidade , Microscopia Eletrônica , Cartilagem Articular/efeitos dos fármacos , Cartilagem Articular/patologia , Cartilagem Articular/ultraestrutura , Ratos Sprague-Dawley , Osteoartrite do Joelho/patologia , Diabetes Mellitus Experimental , Modelos Animais de Doenças , Combinação de Medicamentos
An. acad. bras. ciênc ; 89(4): 3067-3082, Oct.-Dec. 2017. tab, graf
Artigo em Inglês | LILACS | ID: biblio-886832


ABSTRACT Pollen morphology characterization is important in taxonomy, conservation and plant breeding, and pollen viability studies can support breeding programs. This study investigated pollen morphology and male fertility in 18 species of Bromeliaceae with ornamental potential. For morphological characterization, pollen grains were acetolyzed and characterization of exine was done using scanning and transmission electron microscopy. Pollen viability was investigated by in vitro germination and histochemical tests. Species belonging to Aechmea and Ananas genera presented medium size pollen, except for Ae. fasciata, with large pollen. Al. nahoumii, P. sagenarius and the Vriesea species analyzed showed large pollen, except for V. carinata, with very large pollen. Pollen of Aechmea, Ananas and P. sagenarius presented bilateral symmetry, diporate, exine varying from tectate to semitectate. Al. nahoumii and Vriesea species presented pollen with bilateral symmetry, monocolpate; exine was semitectate, reticulate and heterobrochate. Germination percentage and tube growth were greater in SM and BKM media. Histochemical tests showed pollen viability above 70% for all species, except for Ananas sp. (40%). Pollen morphology is important for the identification of species, especially in this family, which contains a large number of species. High rates of viability favor fertilization and seed production, essential for efficient hybrid production and conservation.

Pólen/fisiologia , Bromeliaceae/fisiologia , Pólen/crescimento & desenvolvimento , Pólen/ultraestrutura , Imuno-Histoquímica , Microscopia Eletrônica , Bromeliaceae/classificação , Bromeliaceae/ultraestrutura
Int. j. morphol ; 35(3): 893-900, Sept. 2017. ilus
Artigo em Inglês | LILACS | ID: biblio-893070


The aim of this study was to evaluate the effect of chlorhexidine (CHX) dentin treatment on microtensile bond strength (µTBS) of adhesive systems in different storage times. Occlusal enamel was removed from ninety third-molars and flat surfaces of middle dentin were exposed. Teeth were randomly divided in 6 groups according to adhesive system (etch-and-rinse : Adper Scotchbond 1XT - ASB ; self-etch: Adper Prompt L-Pop ­ APP; and universal: Single Bond Universal - SBU) and chlorhexidine (CHX) dentin treatment (2 % CHX application for 20 s prior Primer). After resin composite build up, teeth were sectioned to obtain beam specimens and divided in 3 subgroups (n=5): 72h, 3 and 6 months storage times. After the storage times, teeth were tested in tension until failure (0.5 mm/min). SEM was performed to observe hybrid layer of adhesive systems. Data were analyzed using one-way ANOVA and Tukey tests. At 72 h, all equivalent groups (same adhesive system, different dentin treatments) maintained their µTBS when compared CHX-treatment. At 3 and 6 months, non-treated CHX groups showed less µTBS than CHX-treated ones. Six months storage time did not significantly decrease µTBS, except for G2-ASB. The effect of CHX on dentin µTBS depends on storage times and adhesive systems. While immediate µTBS was not affected by CHX treatment, CHX improved dentin µTBS after 3 and 6 months.

El objetivo de este estudio fue evaluar el efecto del tratamiento con clorhexidina (CHX) de la dentina sobre la resistencia de la unión microtensil (mTBS) de los sistemas adhesivos en diferentes tiempos de almacenamiento. Se retiró el esmalte oclusal de noventa terceros molares y se expusieron superficies planas de la dentina media. Los dientes se dividieron al azar en 6 grupos de acuerdo con el sistema adhesivo (con grabado ácido: Adper Scotchbond 1XT-ASB, auto-grabado: Adper Prompt L-Pop-APP y universal: Single Bond Universal- SBU) y el tratamiento de la dentina con clorhexidina (CHX) (aplicación de CHX al 2 % 20 s antes del Primer). Después de la aplicación de la resina compuesta, los dientes fueron seccionados para obtener muestras en forma de barras y divididos en 3 subgrupos (n = 5) con tiempos de almacenamiento de 72 h, 3 y 6 meses. Después de los tiempos de almacenamiento, los dientes se sometieron a tensión hasta la fractura (0,5 mm / min). SEM se realizó para observar la capa híbrida de sistemas adhesivos. Los datos se analizaron mediante ANOVA unidireccional y pruebas de Tukey. A las 72 h, todos los grupos equivalentes (el mismo sistema adhesivo, diferentes tratamientos de dentina) mantuvieron su mTBS cuando se comparó el tratamiento CHX. A los 3 y 6 meses, los grupos CHX no tratados mostraron menos mTBS que los tratados con CHX. Seis meses de tiempo de almacenamiento no disminuyó significativamente el mTBS, excepto para el G2-ASB. El efecto de CHX sobre la dentina mTBS depende del tiempo de almacenamiento y de los sistemas adhesivos. Mientras que el mTBS inmediato no se vio afectado por el tratamiento con CHX, CHX mejoró la mTBS a dentina después de 3 y 6 meses.

Adesivos/química , Clorexidina/farmacologia , Colagem Dentária/métodos , Adesivos Dentinários/química , Dentina , Técnicas In Vitro , Teste de Materiais , Microscopia Eletrônica , Resistência à Tração , Fatores de Tempo
Pesqui. vet. bras ; 37(9): 995-1001, Sept. 2017. ilus
Artigo em Português | LILACS, VETINDEX | ID: biblio-895521


A paca (Cuniculus paca), um dos maiores roedores da fauna brasileira, possui características inerentes à sua espécie que podem contribuir como uma nova opção de animal experimental; assim, considerando-se que há crescente busca por modelos experimentais apropriados para ortopedia e pesquisas cirúrgicas, foram analisados e descritos em detalhes a anatomia microscópica e ultraestrutural do joelho desse roedor. Os ligamentos colaterais são constituídos por feixes de fibras colágenas arranjadas paralelamente e com trajeto ondulado. Os fibroblastos formavam fileiras paralelas às fibras colágenas; quanto aos ligamentos colaterais, estes apresentaram citoplasma imperceptível à avaliação por microscopia de luz, entretanto, em análise ultraestrutural verificou-se vários prolongamentos citoplasmáticos. Microscopicamente, as estruturas presentes no joelho da paca assemelham-se às dos animais domésticos, roedores e lagomorfos.(AU)

Paca (Cuniculus paca), one of the largest rodent of the Brazilian fauna, has characteristics inherent to the species that can contribute as a new experimental animal; so, considering that there is a growing search for experimental models suitable for orthopedic and surgical research, it was analyzed and described in detail the microscopic and ultrastructural anatomy of the stifle in this rodent. The collateral ligaments are composed of bundles of collagen fibers arranged in parallel and in wavy path. Fibroblasts formed parallel rows to the collagen fibers; concerning the collateral ligaments, they presented imperceptible cytoplasm at light microscopy, but at ultrastructure analysis they presented several cytoplasmic processes. At the microscopic level, the stifle of paca resembles the domestic animals, rodents and lagomorphs.(AU)

Animais , Joelho de Quadrúpedes/anatomia & histologia , Joelho de Quadrúpedes/ultraestrutura , Cuniculidae/anatomia & histologia , Menisco/anatomia & histologia , Ligamentos/anatomia & histologia , Microscopia Eletrônica/veterinária , Modelos Animais
Mem. Inst. Oswaldo Cruz ; 112(8): 537-543, Aug. 2017. tab, graf
Artigo em Inglês | LILACS | ID: biblio-894863


BACKGROUND Silver nanoparticles (AgNPs) are increasingly being used in medical applications. Therefore, cost effective and green methods for generating AgNPs are required. OBJECTIVES This study aimed towards the biosynthesis, characterisation, and determination of antimicrobial activity of AgNPs produced using Pseudomonas aeruginosa ATCC 27853. METHODS Culture conditions (AgNO3 concentration, pH, and incubation temperature and time) were optimized to achieve maximum AgNP production. The characterisation of AgNPs and their stability were evaluated by UV-visible spectrophotometry and scanning electron microscopy. FINDINGS The characteristic UV-visible absorbance peak was observed in the 420-430 nm range. Most of the particles were spherical in shape within a size range of 33-300 nm. The biosynthesized AgNPs exhibited higher stability than that exhibited by chemically synthesized AgNPs in the presence of electrolytes. The biosynthesized AgNPs exhibited antimicrobial activity against Escherichia coli, P. aeruginosa, Salmonella typhimurium, Staphylococcus aureus, methicillin-resistant S. aureus, Acinetobacter baumannii, and Candida albicans. MAIN CONCLUSION As compared to the tested Gram-negative bacteria, Gram-positive bacteria required higher contact time to achieve 100% reduction of colony forming units when treated with biosynthesized AgNPs produced using P. aeruginosa.

Humanos , Prata/farmacologia , Contagem de Colônia Microbiana/métodos , Nanopartículas Metálicas/química , Bactérias Gram-Negativas/metabolismo , Bactérias Gram-Negativas/ultraestrutura , Bactérias Gram-Positivas/efeitos dos fármacos , Antibacterianos/biossíntese , Antibacterianos/farmacologia , Antibacterianos/química , Pseudomonas aeruginosa , Espectrofotometria , Microscopia Eletrônica/métodos
Pesqui. vet. bras ; 37(6): 613-620, jun. 2017. tab, ilus
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-895454


Canine transmissible venereal tumor (CTVT) is a naturally occurring contagious round-cell neoplasia, with poorly understood origin and transmission. This study aims to further investigate the tumor nature through immunohistochemistry, lectin histochemistry and transmission electron microscopy (TEM) analysis, and to provide support for diagnostic and differential diagnoses of CTVT. Immunohistochemistry was performed in 10 genital and six exclusively extragenital tumors, which were previously diagnosed by citology and histopathology. CTVT samples were incubated with biotinylated antibodies to specific membrane and cytoplasmic antigens (anti-lysozyme, anti-macrophage, anti-vimentin, anti-CD18, monoclonal anti-CD117, monoclonal anti-CD3, polyclonal anti-CD117, polyclonal CD3 and anti-CD79a), followed by the avidin-biotin-peroxidase complex technique. The lectins Con A, DBA, SBA, PNA, UEA-1, WGA, sWGA, GSL, JSA, PSA, PHA-L, PHA-E and RCA were additionally tested in four genital CTVTs and TEM was performed in eight genital tumors. The anti-vimentin antibody revealed strong immunoreactivity to neoplastic cells in all the assessed samples (16/16). The polyclonal anti-CD3 antibodies showed moderate to strong immunoreactivity in fourteen (14/16) and the polyclonal anti-CD117 in fifteen cases (15/16). There was no immunoreactivity to anti-lysozyme, anti-macrophage, anti-CD18, monoclonal anti-CD117, monoclonal anti-CD3 and anti-CD79a antibodies. At lectin histochemistry, it was observed strong staining of tumor cells to Con-A, PHA-L and RCA. There was no histopathological and immunoreactivity differences between genital and extragenital CTVTs. These findings do not support the hypothesis of histiocytic origin of CTVT. In contrast, the lectin histochemical results were similar to cells from lymphoid/myeloid origin.(AU)

O Tumor Venéreo Transmissível Canino (CTVT) é uma neoplasia de células células redondas, contagiosa, com origem e transmissão ainda mal compreendidas. Com a finalidade de aprofundar a investigação sobre a natureza (origem) do TVTC, bem como fornecer subsídios para o estabelecimento do diagnóstico e diagnóstico diferencial, realizaram-se avaliações imuno-histoquímica, lectino-histoquímica e ultraestrutural de TVTC(s). A avaliação imuno-histoquímica foi feita em 10 TVTCs genitais e em 6 exclusivamente extragenitais previamente diagnosticados através de citologia e da histopatologia. Os TVTCs foram testados para reagentes específicos de antígenos de membrana e citoplasmáticos (anti-lisozima, anti-macrófago, anti-vimentina, anti-CD18, anti-CD3, anti-CD79, anti-CD117) com utilização da técnica complexo avidina-biotina-peroxidase. Adicionalmente, foram utilizadas as lectinas Con A, DBA, SBA, PNA, UEA-1, WGA, sWGA, GSL, SJA, PSA, PHA-L, PHA-E e RCA em quatro TVTCs genitais. Microscopia eletrônica foi realizada em oito TVTC genitais. Em 100% dos tumores testados (16/16) com anticorpo anti-vimentina (mono e policlonal) houve forte imuno-reatividade. Não houve reatividade para os anticorpos anti-lisozima, anti-macrófago, anti-CD18, anti-CD3, anti-CD79a e anti-CD117 quando empregamos anticorpos monoclonais, entretanto, com a utilização de anticorpos policlonais verificou-se marcação dos tumores com os anticorpos anti-CD3 e anti-CD117. Na avaliação lectino-histoquímica foi verificada forte marcação das células tumorais com Con-A, PHA-L e RCA. Não houve diferença histopatológica e de imuno-reatividade entre os TVTCs genitais e extragenitais. Estes achados não corroboram com a hipótese da origem histiocítica do CTVT (ausência de reatividade dos anticorpos anti-lisozima, anti-macrófago e anti-CD18), entretanto, os resultados da avaliação lectino-histoquímica foram em parte similares aos obtidos quando células de origem linfóide/mielóide (ConA, PHA-L e RCA) foram analisadas (Gimeno et al. 1995).(AU)

Animais , Cães , Tumores Venéreos Veterinários/patologia , Tumores Venéreos Veterinários/ultraestrutura , Lectinas , Imuno-Histoquímica/veterinária , Microscopia Eletrônica/veterinária , Técnicas e Procedimentos Diagnósticos/veterinária
Int. j. morphol ; 35(1): 287-292, Mar. 2017. ilus
Artigo em Inglês | LILACS | ID: biblio-840967


The hammerhead sharks shows a head laterally expanded with eyes and nostrils on the edges, which gives the species a hammer appearance. Another strand of studies indicates that the hypothesis of head shape alterations for better adaptions in the sensorial system with the development of structures associated to binocular sight and others sensorial organs. Given the variety of characteristics, the study aim was to describe the anatomical constitution of the hammerhead shark eye bulb. The bulb and its annexes exenteration was performed, and further dissection; the morphological description of the muscle insertions and eye bulb components were based on direct observation and were further photographed and catalogued. The eye bulb fragments and its annexes were histological technical. Between the sclera and the choroid it was possible to observe, by the electronical scanning microscopy, the thin layer called supra-choroid, in this region, spaces often filled by lymphatic vessels allied to a matrix formed by loose conjunctive tissue are found. In the choroid, a layer which is rich in blood vessels, loose conjunctive tissue and collagen fibers, was observed, besides pigmentary cells full of melanin in its interior, which result in its layer's dark color. Ciliary body is a choroid's dilatation; it has the aspect of a thick ring in finger-like shape, pigmented, covering the sclera surface and containing pigmentary cells. The crystalline capsule, which shows an acellular covering that, is hyaline and homogeneous. In the electronical scanning microscopy, it was observed that the capsule is extremely thick especially in the anterior face. The capsule is very elastic, constituted mainly by thin lamellae of collagenous fibers, as illustrated by the electronically scanning microscopy. Anatomic variations related mainly to the position of the eye bulb in the skull, fibrous tunica and lens call the attention and must be related to its habitat.

El tiburón martillo tiene la cabeza lateralmente expandida con los ojos y la nariz en sus márgenes. Estudios indican que alteraciones de la forma de la cabeza de estos animales son adaptaciones que mejoran el desarrollo del sistema sensorial y estructuras asociadas a la visión binocular y otros órganos sensoriales. Dada la variedad de características, el objetivo del estudio fue describir la constitución anatómica del bulbo ocular. Se disecó el bulbo ocular junto a sus anexos. La descripción morfológica de las inserciones musculares y componentes del bulbo ocular se basaron en la observación directa, siendo éstas fotografiadas y catalogadas. Entre la esclera y la coroides se observó através de microscopía electrónica de barrido, una delgada capa supra-coroides, y vasos linfáticos junto a la matriz formada por tejidos conectivos. La coroides, era abundante en vasos sanguíneos y fibras de tejido conectivo. Se observaron escasas células pigmentarias llenas de melanina, lo que se asemeja a un anillo en forma de dedo, de manera que cubren la superficie de la esclera y células que contienen pigmentos. En la cápsula del lente, fue posible observar un recubrimiento celular, hialino y homogéneo. En la microscopía electrónica de barrido, se observó una cápsula muy gruesa, principalmente en la región frontal. La cápsula elástica,estaba constituida por láminas delgadas, principalmente, por fibras de colágeno. Se concluyó que el bulbo ocular de estos animales posee variaciones anatómicas relacionadas, principalmente, con la posición del bulbo ocular en el cráneo, túnica fibrosa y lente, lo que puede estar directamente relacionado con su hábitat.

Animais , Olho/anatomia & histologia , Tubarões/anatomia & histologia , Olho/ultraestrutura , Microscopia Eletrônica
Pesqui. vet. bras ; 37(2): 183-194, fev. 2017. ilus
Artigo em Português | LILACS, VETINDEX | ID: biblio-834017


A avaliação hematológica, de importância comprovada como um meio auxiliar de diagnóstico ao clínico de pequenos animais domésticos, vem se tornando comum em animais selvagens não apenas para a clínica, mas para a avaliação do manejo e como estudo auxiliar para a fisiologia das várias espécies. Tendo em vista o aumento da demanda para a produção de várias drogas de importância farmacêutica, a criação de serpentes peçonhentas vem se tornando comum a ponto destes animais já serem reconhecidos como sendo de produção. O conhecimento do manejo e da clínica destes animais ainda é escasso e a mortalidade é elevada nos criatórios, tornando urgente a ampliação destes. Embora alguns estudos hematológicos já tenham sido realizados em cascavéis (Crotalus durissus) os dados analisados ainda são insipientes, notadamente em relação à caracterização das células do sangue e poucos estudos em microscopia eletrônica foram realizados em serpentes. Com o objetivo de caracterizar as células sanguíneas morfologicamente, sob microscopia óptica e ultraestrutural, foram coletadas amostras de sangue de 52 de indivíduos da subespécie Crotalus durissus terrificus para a realização de esfregaços sanguíneos e avaliação ultraestrutural. Concluiu-se que a coloração hematológica de Giemsa permite a avaliação morfológica e a diferenciação das células sanguíneas em serpentes assim como a visualização de hemoparasitos. A avaliação ultraestrutural permite evidenciar as organelas celulares e a diferenciação entre as células, inclusive entre os tipos leucocitários, porém ainda são necessários outros estudos para que seja elucidada a hipótese da existência dos eosinófilos na espécie estudada assim como é necessária melhor caracterização dos grânulos dos azurófilos para que se confirme uma possível diferença entre os monócitos típicos e os azurófilos.(AU)

Hematological evaluation, important for the diagnostic by the small domestic animal clinician, has become common in wildlife clinic, and for handling and study of the physiology of various species. Given the increased demand for drug production of pharmaceutical importance, the breeding of venomous snakes has become common and is already recognized as production. Knowledge of the management and clinics of snakes is still insufficient and their mortality is high. Although some hematological studies have already been conducted in the rattlesnake (Crotalus durissus), the analyzed data are still insufficient, especially with respect to the characterization of blood cells, and few electron microscopy studies have been performed on snakes. In order to characterize morphologically blood cells with light and ultrastructural microscopy, blood samples from 52 individuals of subspecies of Crotalus durissus terrificus were collected to perform blood smears and ultrastructural evaluation. It was concluded that hematologic Giemsa staining allows morphological evaluation and differentiation of the blood cells as well as of snake hemoparasites. The ultrastructural evaluation will highlight the cell organelles and differentiation between cells, including leukocyte types; although still further studies are needed to elucidate the hypothesis of eosinophils in the species studied as also is necessary a better characterization of azurophilic beads to confirm a possible difference between the typical monocyte and the azurophilic.(AU)

Animais , Células Sanguíneas/ultraestrutura , Crotalus/sangue , Leucócitos , Testes Hematológicos/veterinária , Microscopia Eletrônica/veterinária
Mem. Inst. Oswaldo Cruz ; 112(1): 31-43, Jan. 2017. graf
Artigo em Inglês | LILACS | ID: biblio-841752


A pleiotropic response to the calpain inhibitor MDL28170 was detected in the tomato parasite Phytomonas serpens. Ultrastructural studies revealed that MDL28170 caused mitochondrial swelling, shortening of flagellum and disruption of trans Golgi network. This effect was correlated to the inhibition in processing of cruzipain-like molecules, which presented an increase in expression paralleled by decreased proteolytic activity. Concomitantly, a calcium-dependent cysteine peptidase was detected in the parasite extract, the activity of which was repressed by pre-incubation of parasites with MDL28170. Flow cytometry and Western blotting analyses revealed the differential expression of calpain-like proteins (CALPs) in response to the pre-incubation of parasites with the MDL28170, and confocal fluorescence microscopy confirmed their surface location. The interaction of promastigotes with explanted salivary glands of the insect Oncopeltus fasciatus was reduced when parasites were pre-treated with MDL28170, which was correlated to reduced levels of surface cruzipain-like and gp63-like molecules. Treatment of parasites with anti-Drosophila melanogaster (Dm) calpain antibody also decreased the adhesion process. Additionally, parasites recovered from the interaction process presented higher levels of surface cruzipain-like and gp63-like molecules, with similar levels of CALPs cross-reactive to anti-Dm-calpain antibody. The results confirm the importance of exploring the use of calpain inhibitors in studying parasites’ physiology.

Animais , Glândulas Salivares/parasitologia , Heterópteros/parasitologia , Cisteína/efeitos dos fármacos , Cisteína/metabolismo , Euglenozoários/efeitos dos fármacos , Euglenozoários/enzimologia , Euglenozoários/ultraestrutura , Interações Hospedeiro-Parasita/fisiologia , Microscopia Eletrônica , Western Blotting , Citometria de Fluxo , Dose Letal Mediana
Braz. J. Pharm. Sci. (Online) ; 53(2): e16039, 2017. tab, graf
Artigo em Inglês | LILACS | ID: biblio-839492


ABSTRACT Sustained release systems for therapeutic proteins have been widely studied targeting to improve the action of these drugs. Molecular entrapping of proteins is particularly challenging due to their conformational instability. We have developed a micro-structured poly-epsilon-caprolactone (PCL) particle system loaded with human insulin using a simple double-emulsion w/o/w method followed by solvent evaporation method. This formulation is comprised by spheric-shaped microparticles with average size of 10 micrometers. In vitro release showed a biphasic behavior such as a rapid release with about 50% of drug delivered within 2 hours and a sustained phase for up to 48 h. The subcutaneous administration of microencapsulated insulin showed a biphasic effect on glycemia in streptozotocin-induced diabetic mice, compatible with short and intermediate-acting behaviors, with first transition peak at about 2 h and the second phase exerting effect for up to 48h after s.c. administration. This study reveals that a simplified double-emulsion system results in biocompatible human-insulin-loaded PCL microparticles that might be used for further development of optimized sustained release formulations of insulin to be used in the restoration of hormonal levels.

Animais , Masculino , Feminino , Camundongos , Insulina/análise , Preparações Farmacêuticas/administração & dosagem , Microscopia Eletrônica/estatística & dados numéricos , Diabetes Mellitus/prevenção & controle , Material Particulado/farmacologia , Liberação Controlada de Fármacos/fisiologia , Hipoglicemiantes/farmacologia