Experimental poisoning by Brachiaria decumbens in rabbits / Intoxicação experimental por Brachiaria decumbens em coelhos

Brachiaria spp. are important sources of forage for ruminants in Brazil, due to the easy cultivation, good resistance to drought, good adaptation to different soils and low maintenance cost. However, the ingestion of this grass has been related to photosensitization outbreaks in cattle and sheep with significant economic losses. The hepatotoxic effects related to the ingestion of grass are the formation of crystals and foamy macrophages due to the accumulation of toxic metabolites. The use of cattle and sheep in experiments involving the plant presents several obstacles in the ethical, economic and animal management. The objective of this study was to evaluate the sensitivity of rabbits as an experimental model for B. decumbens poisoning. Two experiments were carried out. In Experiment 1 four rabbits received the fresh plant in daily doses of 10, 20, 40 and 80g/kg body weight for 120 days. In Experiment 2 three rabbits received the fresh plant in amounts of 500g daily with duration of 210 days. The animals of Experiment 1 showed no clinical signs and no macroscopic and microscopic changes characteristic of B. decumbens poisoning. In Experiment 2 the animals also showed no clinical signs or significant macroscopic alterations. Histological analysis showed isolated foamy macrophages or present in random groups of cells in the liver and mesenteric lymph nodes. Samples of liver and mesenteric lymph nodes of the rabbits of Experiment 2 were submitted to the lectin-histochemistry technique. The WGA, sWGA and RCA lectins showed reactivity in foamy macrophages in both organs. This is the first study of our knowledge that demonstrates histopathological lesions caused expetimentally by Brachiaria spp. in rabbits, demonstrating its potential as an animal model.(AU)

Brachiaria ssp. são importantes fontes de forragem para ruminantes no Brasil, devido ao fácil cultivo, boa resistência a seca, boa adaptação a diferentes solos e baixo custo de manutenção. Entretanto, a ingestão desta gramínea está relacionada a surtos de fotossensibilização, em bovinos e ovinos, principalmente, ocasionando prejuízos econômicos significativos. Os efeitos hepatotóxicos relacionados à ingestão da gramínea são a formação de cristais e macrófagos espumosos causados pelo acúmulo de metabólitos tóxicos. A utilização de bovinos e ovinos em experimentos envolvendo a planta apresenta vários empecilhos, tanto no âmbito ético, econômico e no manejo dos animais. O objetivo do presente trabalho foi avaliar a sensibilidade de coelhos como modelo experimental para intoxicação por B. decumbens. No presente estudo foram realizados dois experimentos. O Experimento 1 utilizou quatro coelhos que receberam a planta fresca em doses diárias de 10, 20, 40 e 80 g/Kg de peso vivo durante 120 dias. O Experimento 2 utilizou três coelhos recebendo a planta fresca em quantidades de 500g diárias por animal com duração de 210 dias. No Experimento 1, os animais não apresentaram sinais clínicos e nem alterações macroscópicas e microscópicas características de intoxicação por B. decumbens. No Experimento 2 os animais também não apresentaram sinais clínicos e alterações macroscópicas significativas. Na análise histológica observou-se presença de macrófagos espumosos isolados ou em grupos aleatórios de células no fígado e nos linfonodos mesentéricos. Amostras de fígado e linfonodos mesentéricos dos animais do Experimento 2 foram submetidos à técnica de lectino-histoquímica. As lectinas WGA, sWGA e RCA apresentaram reatividade em macrófagos espumosos nos dois órgãos. Este é o primeiro trabalho de nosso conhecimento que demonstra lesões histopatológicas por Brachiaria spp conduzido de forma experimental em coelhos, demonstrando seu potencial como modelo animal nesse campo de estudo.(AU)

Immunohistochemical, lectin histochemical and ultrastructural studies of canine transmissible venereal tumor in Brazil / Estudos imuno-histoquímico, lectino-histoquímico e ultraestrutural do Tumor Venéreo Transmissível Canino no Brasil

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)

Avaliação lectino-histoquímica de fígado e linfonodo mesentérico de búfalos mantidos em pastagens de Brachiaria spp / Lectin histochemistry evaluation of liver and mesenteric lymph node of buffaloes kept on Brachiaria spp. pastures

Animais que se alimentam em pastos de Brachiaria spp. comumente apresentam macrófagos espumosos isolados ou agrupados no fígado, além de cristais no interior de ductos biliares. A patogênese da formação e a natureza do material armazenado nestas células, contudo, ainda não são completamente conhecidas. Através da avaliação lectino-histoquímica, saponinas esteroidais (metabólitos glicosilados secundários) têm sido identificadas nos cristais e no citoplasma das células espumosas, e provavelmente são responsáveis por danificar o fígado e levar ao acúmulo de filoeritrina. Por meio deste trabalho, objetivou-se padronizar e caracterizar a utilização da lectino-histoquímica na detecção de metabólitos glicosilados nos tecidos de búfalos mantidos em diferentes pastos de Brachiaria spp. no Brasil. Fragmentos de fígado e linfonodo mesentérico de 40 animais foram analisados: 10 búfalos mantidos em pastagem predominante de B. decumbens por aproximadamente 12 meses; 10 búfalos mantidos em pastagem predominante de B. brizantha por aproximadamente 18 meses; 10 búfalos mantidos em pastagem de B. brizantha por aproximadamente quatro anos; e, como controle negativo, 10 búfalos mantidos em pastagem livre de Brachiaria spp. desde o nascimento. Quatorze lectinas foram testadas (Con-A, SBA, WGA, DBA, UEA, RCA, PNA, GSL-I, PSA, LCA, PHA-E, PHA-L, SJA e SWGA), em um total de 1120 fragmentos avaliados. Estudos anteriores demonstraram que a lectina PNA possui marcada reatividade para macrófagos espumosos de bovinos e ovinos. No presente estudo, a lectina SWGA apresentou acentuada reatividade e alta especificidade para macrófagos espumosos; WGA, GSL, PHA-E e PHA-L mostraram moderada a acentuada reatividade, mas baixa especificidade aos macrófagos espumosos; as outras lectinas não apresentaram reatividade ou especificidade relevantes. Além disso, não houve diferença relevante de marcação entre os fragmentos coletados de animais que se alimentaram de B. decumbens por 12 meses e B. brizantha por 18 meses. Porém, a diminuição da presença e marcação lectino-histoquímica dos macrófagos espumosos nos tecidos dos búfalos que ingeriram Brachiaria brizantha durante mais tempo indica que os animais podem passar por um processo de adaptação de acordo com o tempo de ingestão da planta. A avaliação lectino-histoquímica pode ser utilizada para caracterizar o material armazenado em macrófagos espumosos presentes no fígado e linfonodo mesentérico de búfalos que se alimentam em pastagens de Brachiaria spp. e ajuda na compreensão da patogênese de formação destas células.(AU)

Animals grazing Brachiaria spp. commonly present foamy macrophages isolated or grouped in the liver, and crystals within biliary ducts. The pathogenesis of formation and the nature of the material stored in these cells however are not completely known. Through lectin histochemistry evaluation, steroidal saponins (secondary glycosylated metabolites) have been identified in the crystals and within the cytoplasm of the foam cells, which are probably liable for damaging the liver, leading to accumulation of phylloerythrin. This study aims to standardize and characterize the use of lectin histochemistry to detect glycosylated metabolites in tissues of buffaloes kept on different Brachiaria spp. pastures in Brazil. Fragments of liver and mesenteric lymph node from 40 buffaloes were analyzed: 10 buffaloes that were kept in predominant pasture of B. decumbens for 12 months; 10 buffaloes that were kept in pasture with a predominance of B. brizantha for 18 months; 10 buffaloes that were kept on pasture of B. brizantha for about four years; and as a negative control, 10 buffaloes that were maintained on native pasture without Brachiaria spp. since birth. Fourteen lectins were tested (Con-A, SBA, WGA, DBA, UEA, RCA, PNA, GSL-I, PSA, LCA, PHA-E, PHA-L, SJA and SWGA), in a total of 1120 evaluated samples. Previous studies demonstrated that PNA showed great binding reactivity for foamy macrophages in cattle and sheep. In the present study, SWGA showed high specificity and marked binding reactivity for foamy macrophages; WGA, GSL, PHA-E and PHA-L showed moderate to marked reactivity, but low specificity for foamy macrophages. The other lectins had not relevant reactivity or specificity. Moreover there was no relevant reactivity difference between the collected samplesd from buffaloes that grazed B. decumbens for 12 months and Brachiaria brizantha for 18 months. However the decreased presence of foamy macrophages and its lectin histochemical binding in animals that fed on B. brizantha for a longer time, indicates that the buffaloes can pass through an adaptation process according to the plant intake time. Lectin histochemistry analysis can be used to characterize the material stored in foamy macrophages present in liver and mesenteric lymph node of buffaloes that graze on Brachiaria spp. pastures and helps to clarify the pathogenesis of these cells.(AU)

Avaliação lectino-histoquímica de fígado e rim de ovinos com fotossensibilização causada por Brachiaria decumbens / Lectin histochemistry evaluation of liver and kidney of sheep with photosensitization caused by Brachiaria decumbens

Embora sejam as forrageiras mais importantes para a pecuária de corte (bovinocultura de corte) no Brasil, em certas épocas ou condições, Brachiaria spp. podem ser tóxicas e causar surtos de fotossensibilização hepatógena que determinam significativas perdas econômicas. Animais que se alimentam em pastos de Brachiaria spp. comumente apresentam macrófagos espumosos isolados ou agrupados, além de cristais no interior de ductos biliares. Saponinas esteroidais têm sido identificadas nestes cristais e são responsabilizadas por lesar o fígado levando ao acúmulo de filoeritrina. Por vezes, imagens negativas desses cristais podem estar presentes no citoplasma de macrófagos espumosos. A patogênese de formação e o tipo de material armazenado nas células espumosas ainda são desconhecidos. A técnica de lectino-histoquímica visa auxiliar na detecção desses macrófagos e, consequentemente, no diagnóstico, além de identificar quais os resíduos de açúcares específicos que estão presentes no citoplasma das células espumosas. Assim, este trabalho teve por objetivo identificar quais lectinas são mais indicadas na detecção de saponinas esteroidais no fígado e rim de ovinos com fotossensibilização causada por Brachiaria decumbens. Fragmentos de fígado e rim de quinze ovinos, de ambos os sexos e idade variável, oriundos de Conceição do Mato Dentro, Minas Gerais, e um ovino mantido em pastagem livre de Brachiaria spp. foram avaliados pela técnica de lectino-histoquímica. Quatorze lectinas foram utilizadas (Con-A, DBA, SBA, PNA, SJA, RCA-I, UEA-I, WGA, SWGA, GSL, PSA, PHA-L, PHA-E e LCA). Verificou-se que, no fígado de ovinos com fotossensibilização provocada pela ingestão de Brachiaria decumbens, a lectina PNA apresentou especificidade e acentuada reatividade aos macrófagos espumosos, bem como especificidade e leve reatividade aos hepatócitos; a lectina WGA teve especificidade e moderada reatividade aos macrófagos espumosos do fígado e especificidade e leve reatividade aos...

Inspite that Brachiaria spp. are most important forage grasses for beef cattle raising in Brazil, they can under certain conditions be toxic, cause outbreaks of hepatogenic photosensibilization and determine severe economic losses. Animals grazing toxic Brachiaria spp. commonly show in their liver foamy macrophages, isolated or grouped together, and crystals inside the biliary ducts. Steroidal saponins have been identified in these crystals and are liable for damage the liverleading to accumulation of phylloerytrin. Occasionally negative images of the crystals may be seen in the cytoplasm of foamy macrophages. The pathogenesis of formation and the type of material stored in the foamy cells is still unknown. The lectin histochemistry aims to assist in the detection of foamy macrophages and consequently in the diagnosis of the poisoning. Lectin histochemistry detects what kind of specific sugar residues exists in the cytoplasm of the foamy cells. The purpose of the present study was to identify the most suitable lectins the detection of steroidal saponins in the liver and kidney of sheep with photosensitization caused by Brachiara decumbens. Fragments of liver and kidney of fifteen sheep of both sexes and variable age, from Conceição do Mato Dentro, Minas Gerais, southeastern Brazil, and one sheep kept free grazing Brachiaria spp. were evaluated by lectin histochemistry. Fourteen lectins were used (Con-A, DBA, SBA, PNA, SJA, RCA-I, UEA-I, WGA, SWGA, GSL, PSA, PHA-L, PHA-E and LCA). It was found that the PNA lectin detected in the liver of sheep with photosensitization, caused by ingestion of Brachiaria decumbens, showed specificity and great binding reactivity for foamy macrophages, and also specificity and slight binding reactivity for hepatocytes. The WGA lectin showed specificity and moderate binding reactivity for foamy macrophages of the liver, and also specificity and slight binding reactivity for hepatocytes. The SBA, GSL and LCA lectins showed...

CONA and UEA-I ligands during tooth development reveals differences between human and rodents models and shows the beginning of mineral extracelular matrix secretion / Ligandos CONA y UEA-I revelan diferencias entre humanos y en modelos roedores durante el desarrollo dental y muestran el comienzo de la secreción de la matriz mineral extracelular

Many of Glycobyological aspects of human development remain still unclear, mainly in oral science, and this could be observed in a lack of literature with few and old papers about this subject. During tooth histo-morphogenesis changes occur in basement membrane composition, expression of signaling molecules and in localization of cell surface components, where glyco components could be involved. In this sense, this work aimed to analyze the ConA ligands commonly founded in glycoproteins cores and UEA-I ligands since fucolization is a Key event in many physiological and pathological processes. Therefore 15 jaws of human fetuses were ethically obtained, histologicaly processed and then submitted to lectin histochemistry with appropriated controls. The results showed that lectins staining increase their intensity during dental development; ConA only starts to recognize glucose/mannose residues on ectomesechymal cells in the crow phase revealing its ligands when the enamel matrix starts to be secreted. Interestingly, Con A ligands were not founded in the basement membrane of the stratum intermedium of the enamel differing from rodents models. The staining pattern of UEA-I was different, starting to be positive in the ectomesenchyma since the bud stage and shown variable expression in cell type and staining intensity, which appeared be directly proportional to the progress of odontogenesis. Thus, this work shows that Con A and UEA-I exhibit a growing staining directly proporcional to ameloblasts and odontoblasts cytodiferenciation and revels some glycan differences between human odontogenesis and rodents models...

Los aspectos glicobiológicos del desarrollo humano siguen siendo poco investigados, sobre todo en odontología, y esto puede ser observado en la literatura por los escasos y antiguos artículos sobre el tema. Durante la histomorfogénesis del diente se producen cambios en la composición de la membrana basal, en la expresión de moléculas de señalización y en la localización de la superficie celular de los componentes, donde los glico componentes podrían estar involucrados. Este trabajo tuvo como objetivo analizar los ligandos de la ConA ya que glucosa/manosa son comúnmente encontrados en núcleos de glicoproteínas y ligandos de la UEA-I debido a fucolización es un evento clave en muchos procesos fisiológicos y patológicos. Fueron obtenidas 15 mandíbulas de fetos humanos, procesadas y tratadas mediante histoquímica de lectinas con controles apropiados. Los resultados mostraron que la intensidad de tinción de las lectinas aumenta durante el desarrollo del diente. ConA sólo comienza a reconocer residuos de glucosa/manosa en células ectomesénquimales en la fase de corona revelando cuando la matriz de esmalte empieza a ser secretada. Curiosamente, ligandos de la ConA no se encontraron en la membrana basal de la capa intermedia del esmalte, difiriendo de los modelos de roedores. El patrón de tinción de la UEA-I fue diferente, empieza a ser positivo en el ectomesenquima desde la etapa de brotación y muestra variable expresión en el tipo de célula y la intensidad de la tinción, que parecía ser directamente proporcional al progreso de la odontogénesis. Por lo tanto, este trabajo demuestra que la Con A y la UEA-I presentan una coloración que crece directamente proporcional a citodiferenciación de los ameloblastos y odontoblastos, y revela algunas diferencias entre el estándar glicano de odontogénesis humanos y los modelos roedores...

Pathological findings in fetuses of goats and cattle poisoned by Sida carpinifolia (Malvaceae) / Achados patológicos em fetos de caprinos e bovinos intoxicados por Sida carpinifolia (Malvaceae)

In Brazil, the consumption of Sida carpinifolia by livestock has been associated with neurological diseases linked to lysosomal storage disorders. This paper describes the pathological findings in two caprine fetuses from dams that were experimentally poisoned with S. carpinifolia. The goats were orally dosed with 10 and 13g/kg of a paste of green chopped S. carpinifolia for 30 days and were observed for an additional 15 days period after the last dosage with the plant; thereafter they were euthanized and necropsied. The dams showed only slight clinical signs. The study also includes the findings in one bovine fetus from a naturally S. carpinifolia poisoned cow which showed mild incoordination, generalized tremors, staggering, and frequent falls. The cow was euthanized and necropsied. While there were no significant histopathological changes in the goats, in the cow vacuolation of Purkinje neurons of the cerebellum, pancreatic acinar cells, and thyroid follicular cells were observed. The main microscopic changes observed in the caprine and bovine fetuses were vacuolation in the epithelium of renal tubules, thyroid follicular cells, and Purkinje neurons of the cerebellum. Transmission electron microscopy of sections from CNS of the cow and its fetus revealed vacuoles containing fine granular material surrounded by membrane. Lectin-histochemistry of CNS sections from goat fetuses marked lightly to sWGA lectins, WGA, and Con-A.

No Brasil, o consumo de Sida carpinifolia por animais de produção tem sido associada a doenças neurológicas relacionadas com doença de depósito lisossômico. Este trabalho descreve os achados patológicos observados em dois fetos caprinos de mães que foram experimentalmente intoxicadas por S. carpinifolia. As cabras foram intoxicadas experimentalmente com S. carpinifolia nas doses de 10 e 13g/kg durante 30 dias e foram acompanhadas durante 15 dias após o consumo da planta. Após este período foram eutanasiadas e necropsiadas. O estudo também inclui os achados patológicos encontrados em um feto de uma fêmea bovina intoxicada naturalmente pela planta, que mostrou leve incoordenação, tremores generalizados, andar desequilibrado e quedas frequentes. A vaca foi eutanasiada e necropsiada. Embora não houvesse alterações histológicas significativas nas cabras, vacuolização dos neurônios de Purkinje do cerebelo, das células acinares do pâncreas e nas células foliculares da tireoide foram observadas na vaca. As principais alterações histológicas observadas nos fetos caprinos e no feto bovino foram vacuolização no epitélio dos túbulos renais, nas células foliculares da tireoide e nos neurônios de Purkinje do cerebelo. Na microscopia eletrônica de transmissão do sistema nervoso central da vaca e de seu feto revelaram-se vacúolos contendo material finamente granulado e delimitado por membrana. Na técnica de lectina-histoquímica dos fetos caprinos houve marcação leve no SNC para as lectinas sWGA, WGA e para Con-A.

Histochemical study of lectin-binding patterns in the rat vomeronasal organ during postnatal development / 대한수의학회지

Histochemical patterns of lectin binding during development of the rat vomeronasal organ (VNO) were studied to determine whether glycoconjugates are differently expressed after birth. Three types of lectins, Dolichos biflorus agglutinin (DBA), wheat germ agglutinin (WGA), and Ulex europaeus agglutinin I (UEA-I), were studied histochemically in the rat VNO at various stages post-birth: postnatal days 1 and 7, the preweaning period (4 weeks after birth), and at sexual maturity (8 weeks after birth). The free border of the vomeronasal sensory epithelium was positive for both WGA and UEA-I in rats of all ages; whereas, VNO receptor cells and supporting cells were positive only for both WGA and UEA-I from 4 weeks after birth. DBA reactivity was detected in the free border but less so in receptor cells and supporting cells. WGA and UEA-I, but not DBA, showed similar patterns in various ages. In the Jacobson's gland, WGA, UEA-I and DBA were detected in some acini from 4 weeks after birth but not at postnatal days 1 or 7. Collectively, reactivity for three lectins, WGA, UEA-I and DBA, increased in receptor cells and gland acini during postnatal development, possibly contributing to the enhanced chemoreception in rats.

Histochemical Localization of Carbohydrates in Morphological Stages of Developing Human Minor Salivary Glands: A Comparative Study with Cytoskeletal Markers / Localización Histoquímica de Carbohidratos en el Desarrollo de Glándulas Salivales Menores Humanas: un Estudio Comparativo con los Marcadores del Citoesqueleto

Carbohydrates play a critical role in many cellular processes like disease, growth and development. In this work lectins, proteins that recognizes carbohydrate free or conjugated, were used as histochemical probes for carbohydrates localization in developing human minor salivary gland. Immunohistochemistry for traditional cytoskeleton markers (Cks 7, 8, 13, 14, 19, SMA and Vimentin) was performed and then compared whit lectin histochemistry for PNA, WGA, ConA and UEA-I, specifics for D-galactose, N-acetyl-glucosamine, glucose/mannose and L-fucose respectively. For this, specimens were obtained from tongues and lips of 15 human foetuses at 10-28 weeks of gestation. None of immune cytoskeleton markers were identified in the first stage of development differing from carbohydrate markers. UEA-I, WGA and PNA recognized their specific carbohydrate residues in all stages analyzed varying the staining intensity and cell types. Ck8 and N-acetyl-glucosamine were expressed in canalicular, branching and cytodifferentiation stages while SMA and glucose/mannose were observed in the cytodifferentiation stage one. ConA only recognized myoepithelial cells on cytodifferentiation stages because of this specificity ConA could be used as biomarker of myoepithelial cells on cytodifferentiation. Lectin histochemistry suggests that L-fucose, D-galactose e N-acetyl-glucosamine are intensily and previously expressed than traditional cytoskeletal markers in human minor salivary gland during development.

Los hidratos de carbono tienen un papel crítico en muchos procesos celulares, como la enfermedad, el crecimiento y el desarrollo. Fueron utilizadasas lectinas, proteínas que reconocen los hidratos de carbono libres o conjugados, como sondas de localización histoquímica de los carbohidratos en el desarrollo humano de la glándula salival menor. Se realizó inmunohistoquímica de los marcadores tradicionales del citoesqueleto (CKs 7, 8, 13, 14, 19, SMA y vimentina) y posterior comparación con la histoquímica de lectinas para PNA, WGA, ConA y la UEA-I, específicas para D-galactosa, N-acetil-glucosamina, glucosa/manosa y L-fucosa, respectivamente. Para ello, se obtuvieron muestras de la lengua y de los labios de 15 fetos humanos entre 10-28 semanas de gestación. Ninguno de los marcadores inmunológicos del citoesqueleto se identificaron en la primera etapa del desarrollo, diferente de los marcadores de hidratos de carbono. UEA-I, WGA y PNA reconocen sus residuos específicos de hidratos de carbono en todas las etapas analizadas variando la intensidad de la tinción y los tipos de células. CK8 y N-acetil-glucosamina se expresaron en etapas de canalización, ramificación y citodiferenciación mientras que SMA y la glucosa/manosa se observaron solamente en la etapa de citodiferenciación. ConA sólo se reconoció en las células mioepiteliales en etapas de citodiferenciación. Así, debido a esta especificidad, ConA podría utilizarse como marcador biológico de las células mioepiteliales en la citodiferenciación. La histoquímica de lectinas sugiere que L-fucosa, D-galactosa y N-acetil-glucosamina son intensamente expresadas durante el desarrollo como los marcadores tradicionales del citoesqueleto humanos en las glándulas salivales menores .

Evaluation of the antischistosomal activity of sulfated α-D-glucan from the lichen Ramalina celastri free and encapsulated into liposomes

The antischistosomal activity of the sulfated polysaccharide α-D-glucan (Glu.SO4) extracted from Ramalina celastri was evaluated after encapsulation into liposomes (Glu.SO4-LIPO) in Schistosoma mansoni-infected mice. The effect of treatment with Glu.SO4 and Glu.SO4-LIPO (10 mg/kg) on egg elimination, worm burden and hepatic granuloma formation was assessed using female albino Swiss mice, 35-40 days of age, weighing 25 ± 2 g, infected with 150 cercariae/animal (Biomphalaria glabrata, BH strain). Four groups (N = 10) were studied, two controls (empty liposomes and NaCl) and two treated groups (Glu.SO4-LIPO and Glu.SO4) using a single dose. Parasitological analysis revealed that Glu.SO4-LIPO was as efficient as Glu.SO4 in reducing egg elimination and worm burden. Treatment with free Glu.SO4 and Glu.SO4-LIPO induced a statistically significant reduction in the number of granulomas (62 and 63 percent, respectively). Lectin histochemistry showed that wheat germ agglutinin intensely stained the egg-granuloma system in all treated groups. On the other hand, peanut agglutinin stained cells in the control groups, but not in the treated groups. The present results suggest a correlation between the decreasing number of hepatic egg-granulomas and the glycosylation profile of the egg-granuloma system in animals treated with free Glu.SO4 or Glu.SO4-LIPO.

Expression patterns of influenza virus receptors in the respiratory tracts of four species of poultry

The primary determinant of influenza virus infectivity is the type of linkage between sialic acid and oligosaccharides on the host cells. Hemagglutinin of avian influenza viruses preferentially binds to sialic acids linked to galactose by an alpha-2,3 linkage whereas hemagglutinin of human influenza viruses binds to sialic acids with an alpha-2,6 linkage. The distribution patterns of influenza receptors in the avian respiratory tracts are of particular interest because these are important for initial viral attachment, replication, and transmission to other species. In this study, we examined the distribution patterns of influenza receptors in the respiratory tract of chickens, ducks, pheasants, and quails because these species have been known to act as intermediate hosts in interspecies transmission. Lectin histochemistry was performed to detect receptor-bearing cells. Cell-specific distribution of the receptors was determined and expression densities were compared. We observed species-, site-, and cell-specific variations in receptor expression. In general, receptor expression was the highest in quails and lowest in ducks. Pheasants and quails had abundant expression of both types of receptors throughout the respiratory tract. These results indicate that pheasants and quails may play important roles as intermediate hosts for the generation of influenza viruses with pandemic potential.

Avian and human influenza A virus receptors in trachea and lung of animals.

Background: Influenza A viruses are capable of crossing the specific barrier between human beings and animals resulting in interspecies transmission. The important factor of potential infectivity of influenza A viruses is the suitability of the receptor binding site of the host and viruses. The affinities of avian and human influenza virus to bind with the receptors and the distributions of receptors in animals are different. Objective: This study aims to investigate the anatomical distribution of avian and human influenza virus receptors using the double staining lectin histochemistry method. Methods: Double staining of lectin histochemistry was performed to identify both SA α2,3 Gal and SA α2,6 Gal receptors in trachea and lung tissue of dogs, cats, tigers, ferret, pigs, ducks and chickens. Results: We have demonstrated that avian and human influenza virus receptors were abundantly present in trachea, bronchus and bronchiole, but in alveoli of dogs, cats and tigers showed SA α2,6 Gal only. Furthermore, endothelial cells in lung tissues showed presence of SA α2,3 Gal. Conclusion: The positive sites of both receptors in respiratory tract, especially in the trachea, suggest that all mammalian species studied can be infected with avian influenza virus. These findings suggested that dogs and cats in close contact with humans should be of greater concern as an intermediate host for avian influenza A in which there is the potential for viral adaptation and reassortment.

Doença do armazenamento lisossomal induzida pelo consumo de Sida carpinifolia em bovinos do Rio Grande do Sul / Lysosomal storage disease caused by Sida carpinifolia in cattle in Rio Grande do Sul

This paper reports the natural poisoning by Sida carpinifolia (guanxuma, chá-da-índia) in cattle in Rio Grande do Sul, Brazil. Five cattle were affected in the period 2001-2008. Clinical signs included weight loss, incoordination, walking difficulty, generalized tremors, frequent falls, and death. Microscopically, the main changes were vacuolation of Purkinje neurons in the cerebellum, pancreatic acinar cells, and thyroid follicular cells. Transmission electron microscopy revealed vacuoles bordered by membrane containing finely granular material. Lectin histochemistry showed positive staining in neurons with the lectins Concanavalia ensiformis (Con-A), Triticum vulgaris (WGA), and Succinyl Triticum vulgaris (sWGA).

Relata-se a intoxicação natural por Sida carpinifolia (guanxuma, chá-da-índia) em bovinos no Rio Grande do Sul. Foram afetados cinco bovinos no período 2001-2008. O quadro clínico foi caracterizado por emagrecimento, incoordenação, dificuldade de locomoção, tremores generalizados, quedas frequentes e morte. Microscopicamente, as principais alterações foram vacuolização dos neurônios de Purkinje do cerebelo, das células acinares do pâncreas e das células foliculares da tireoide. A microscopia eletrônica evidenciou vacúolos com conteúdo finamente granulado e delimitado por membrana. Na lectina-histoquímica, observou-se marcação em neurônios com as lectinas Concanavalia ensiformis (Con-A), Triticum vulgaris (WGA) e Succinyl Triticum vulgaris (sWGA).

Formaldehyde Gas Induced Morphological Changes in the Rat Olfactory Epithelium / 대한해부학회지

Experimental animals were inhalated 50 ppm formaldehyde gas for 3 times with one hour exposure and one hour rest. The olfactory mucosa were taken from the animals on 4, 7, 9, 11 days and 2-6 weeks after the inhalation. The characteristics of the various glycoproteins and the mitotic activity of the olfactory epithelial cells were investigated using lectins and 5'-bromodeoxyuridine (BrdU, 50 mg/kg) which injected one hour before sacrifice of the animals. The results were as follows; 1. In experimental animals, the degenerative changes of the olfactory epithelium such as atrophy and squamous metaplasia were observed until 2 weeks after formaldehyde gas inhalation. 2. In control animals, positive reactions appeared in the supporting cell to PNA, SBA, WGA, ECL, PHA-L and in the olfactory cells to PNA, SBA, WGA, UEA and in the proper basal cell to GS-I, SBA, WGA, PHA-L. In experimental groups, the positive reaction was increased in the supporting cells to SBA, ECL, PHA-L and in Bowman's gland to used lectins except ECL, GS-I. 3. The number of BrdU labelled cells in the olfactory epithelium was 14.8+/-1.2/mm in the control animals. The mitotic activities were decreased to 4.8+/-0.8 mm in 2 weeks and recovered in 3 weeks after the gas inhalation. 4. To find the stem cells of olfactory receptor cells, double labelling method was performed with lectins which were specific for proper basal cells (GS-I or PHA-L) and BrdU immunohistochemistry. The ratio of globose/proper basal cells in BrdU labelled cells with GS-I lectin positive reaction was 82%/18% in control group, 39.3%/60.7% in lesion group and 55.5%/44.5% in recovery group.