Comparación delmmétodo directo y precipitado, pra la determinación de los niveeles de colesterol hdl, en gallinas poeedoras / Comprison of dirrect and precipitatin methods for determining hdl ccholesterol levels in laying hens

Objetivo: Comparar un método directo y un método de precipitado en la determinación del colesterol HDL en gallinas ponedoras. Metodología: Se obtuvo suero de 40 gallinas ponedoras en ayunas de 26 semanas de edad, de la línea Hy-Line W-36. El colesterol de las lipoproteínas de alta densidad se midió por el método directo a base de un detergente específico junto con N,Nbis(4-sulfobutil)-m-toluidina, y por el método de precipitación a base de fosfotungstato + Mg2+. Resultados: Las medias ± desviación estándar para los niveles de colesterol de las lipoproteínas de alta densidad, por los métodos directo y precipitado en mg/dl fueron de: 63,1±14,3; 13,3±3,5; respectivamente. El P valor del test F es inferior a 0,05, evidenciando diferencia estadísticamente significativa, con una confianza del 95% entre métodos. Conclusiones: Es factible la utilización del método directo a base de un detergente específico y N,Nbis(4-sulfobutil)-m-toluidina para la determinación del colesterol de las lipoproteínas de alta densidad en gallinas ponedoras, debido a que es un método que no es alterado por los altos niveles de triglicéridos de estas.

Objective: To compare a direct method vs a precipitation method for determining HDL cholesterol in laying hens. Methodology: Serum of forty 26 weeks old fasting laying hens was obtained from the Hy-Line W-36. Cholesterol in high density lipoproteins was measured by direct method based on a specific detergent along with N,Nbis (4-sulfobutyl)-m-toluidine, and through the precipitation method based on phosphotungstate + Mg2+. Results: The standard deviation mean ± for high density lipoproteins though direct and precipitation methods in mg/dl were: 63.1±14.3; 13.3±3.5 respectively. The P value for the F test is < 0.05 showing significant statistical difference between the two analyzed methods with a confidence level of 95%. Conclusions: It is feasible to use the direct method using a specific detergent and N,Nbis(4-sulfobutyl)-m-toluidine to determine high density lipoprotein cholesterol in laying hens, because this method is not altered by high levels of triglyceride.

Oxygen/Glucose Deprivation and Reperfusion Cause Modifications of Postsynaptic Morphology and Activity in the CA3 Area of Organotypic Hippocampal Slice Cultures

Brain ischemia leads to overstimulation of N-methyl-D-aspartate (NMDA) receptors, referred as excitotoxicity, which mediates neuronal cell death. However, less attention has been paid to changes in synaptic activity and morphology that could have an important impact on cell function and survival following ischemic insult. In this study, we investigated the effects of reperfusion after oxygen/glucose deprivation (OGD) not only upon neuronal cell death, but also on ultrastructural and biochemical characteristics of postsynaptic density (PSD) protein, in the stratum lucidum of the CA3 area in organotypic hippocampal slice cultures. After OGD/reperfusion, neurons were found to be damaged; the organelles such as mitochondria, endoplasmic reticulum, dendrites, and synaptic terminals were swollen; and the PSD became thicker and irregular. Ethanolic phosphotungstic acid staining showed that the density of PSD was significantly decreased, and the thickness and length of the PSD were significantly increased in the OGD/reperfusion group compared to the control. The levels of PSD proteins, including PSD-95, NMDA receptor 1, NMDA receptor 2B, and calcium/calmodulin-dependent protein kinase II, were significantly decreased following OGD/reperfusion. These results suggest that OGD/reperfusion induces significant modifications to PSDs in the CA3 area of organotypic hippocampal slice cultures, both morphologically and biochemically, and this may contribute to neuronal cell death and synaptic dysfunction after OGD/reperfusion.

Sperm ultrastructure of Mytella (Bivalvia) populations from distinct habitats along the northern coast of São Paulo State, Brazil

Ultrastructural analyses of bivalve spermatozoa are relevant in studies that aim to identify taxonomic traits for the purposes of discriminating species and conducting phylogenetic studies. In the present work, spermatozoa of mussel specimens of the genus Mytella, collected from two populations living in distinct habitats, were examined by electron microscopy. The objective was to identify sperm ultrastructural taxonomic traits that could be used to differentiate Mytella species. The specimens were from populations that live in intertidal zones on the southeast coast of Brazil, either buried in muddy-sand sediment or anchored to rocky substrates. The acrosomal vesicle was conical and long, the axial rod extended from the nucleus to the acrosome, the nucleus was an oblate spheroid with a condensed chromatin, the intermediate portion contained mitochondria encircling a pair of centrioles, and there was a single flagellum. The sperm was of a primitive type. The spermatozoon ultrastructure did not distinguish the specimens buried in muddy-sand sediment from those anchored to rocky substrates. The data suggest that the specimens analyzed, despite living in distinct habitats, belong to the same species, which conchological analyses identified as M. charruana. The presence of an axial rod in their sperm cells supports the inclusion of M. charruana in the subfamily Mytilinae.

Cardiac Myocyte Cell Death in Isoproterenol-Induced Cardiac Hypertrophy in Rats

BACKGROUND: Although cardiac hypertrophy contributes to cardiac failure, the underlying mechanism has not yet been precisely determined. This study was planned in order to determine the pathogenesis of heart failure following cardiac hypertrophy induced by -adrenergic stimulation. METHODS: The extent of cardiac hypertrophy was assessed after administrating isoproterenol (ISO, 5 mg/kg) intraperitoneally for 6 hours, 1, 3, 5, 7 and 10 days. The hematoxylin-eosin, Masson's trichrome and phosphotungstic acid hematoxylin stains along with immunohistochemical stainings for proliferating cell nuclear antigen and Ki-67 were performed in the paraffin-embedded left ventricle sections. Apoptosis was assessed by DNA laddering and terminal deoxynucleotidyl transferase TdT-mediated dUTP-biotin nick end labeling (TUNEL) assay. TUNEL positive myocytes and some nonmyocytes appeared in the subepicardium at 6 hours after ISO administration. The localization of these cells was shifted to the subendocardium within 24 hours, and the TUNEL positive cells were seen throughout the myocardium on the 5th day after ISO treatment. Necrotic myocyte death occurred on the 3rd day of ISO administration in the subendocardium, and initial pericellular fibrosis was followed and increased thereafter, with replacement fibrosis accompanied by further necrotic myocyte cell death. CONCLUSIONS: Our data showed that ISO treatment induced apoptotic myocyte death and superimposed necrotic myocyte death with subsequent fibrosis. The observed cardiac myocyte death may reflect myocardial dysfunction.

Colesterol total y fracciones en mujeres embarazadas normales / Serum total cholesterol and fractions in normal pregnant women

Se reportan las variaciones del colesterol total y del HDL y no HDL colesterol, así como el Indice de Castelli, en un estudio transversal realizado en intervalos de tiempo similares en embarazadas normales. Los datos provenientes de 210 mujeres embarazadas normales en diferentes estadios, se compararon con los de una población de referencia de 30 mujeres normales no grávidas con edades similares. El colesterol HDL fue aislado con ácido fosfotúngstico/cloruro de magnesio, el colesterol se determinó enzimáticamente y el Indice de Castelli así como el no HDL colesterol fueron calculados en cada caso. Nuestro objetivo es estudiar los cambios lipídicos en esta en esta población de embarazadas con el propósito de establecer los valores de referencia e investigar si la gestación podría estar relacionada a situaciones de riesgo cardiovascular. El HDL colesterol decreció significativamente entre las semanas 24a y 28a, al tiempo que el Indice de Castelli alcanzó el máximo valor de riesgo. Desde este punto de vista se puede deducir que este proceso puede estar relacionado con el comienzo de la resistencia a la insulina y el mayor riesgo de aparición de diabetes gestacional

Estudio comparativo de los métodos de Bertrand, Lane-Eynon y Cloramina-T. Usando el Acido Fosfotungstico como sustancia precipitante / Comparative study of Bertrand, Loyne-Eynon and Chloramine-T methods using the phosphotungstic acid as precipitant substance

Se evaluaron los métodos de Bertrand, Lane-Eynon y Cloramina-T, utilizando el ácido fosfotúngstico como sustancia precipitante de las proteínas y de la grasa. Para ello se tomaron muestras de leche cruda a las cuales se les aplicó los tres métodos bajo estudio, así como a la leche cruda con adición de 1 por ciento de lactosa. Se encontró que no existen diferencias significativas entre los tres métodos cuando se utiliza el ácido fosfotúngstico como sustancia precipitante de las proteínas y de la grasa, sin embargo se pudo comprobar que el método de Bertrand ofrece mayor receptibilidad, ya que presentó el menor coeficiente de variación

Subfracionamento do colesterol HDL por precipitaçäo polianiônica e dosagem enzimática: um método simples / Subfractionation of lipoprotein HDL cholesterol by polyanionic precipitation and enzymatic determination a simple method

Propöem-se um método simples de subfracionamento do colesterol HDL onde se utiliza um só reagente precipitante tanto para VLDL e LDL quanto para a fraçäo HDL2. O método apresentou os seguintes coeficientes de variaçäo: HDL total = 5,1%, HDL2 = 6,4% e HDL3 = 3,1%. Ao comparar-se o método proposto com o método de Gidez, verificou-se que houve boa correlaçäo para HDL total (r = 0,931), para a subfraçäo HDL2 (r = 0,893) e para a subfraçäo HDL3 (r = 0,894)

Active genes visualized in pachytene and lymphocyte nuclei from normal and affected individuals: Ag-As-staining, light and electron microscopy and high-resolution autoradiography

Aglomerados de genes nucleolares transcritos foram evidenciados por microscopia óptica e eletrônica como grânulos de 0.1 a 1.0 micron m, corados com prata e ácido fosfotungstico alcoólico (PTA), em espermatócitos e linfócitos humanos. Os grânulos encontrados nos núcleos paquitênicos acham-se aglomerados numa regiäo nuclear distinta, contendo bivalentes autossômicos associados e o par sexual XY. Um grânulo corado pela prata foi encontrado estreitamente relacionado com o bivalente sexual XY. Esta específica regiäo nuclear foi designada "corpo cromatínico" (c cr), neste trabalho. Os grânulos, encontrados nos nucléolos de linfócitos, correspondem a aglomerados de sítios ativos em transcriçäo, tendo fibrilas enroladas de ribonucleoproteínas (RNP), as quais incorporam uridina [3H], em experimentos de marcaçäo de pulso. Os genes ativos foram encontrados nos componentes fibrilares do nucléolo e seus níveis de transcriçäo, estimados pela quantidade de grânulos corados, variaram em indíviduos normais e afetados e durante o tratamento com ácido fólico

A combined staining technique developed for virus particle observation in the electron microscope

Descrevemos uma tecnica de coloracao dupla para a visualizacao direta de particulas virais ao microscopio eletronico. Os virus foram contrastados com acido fosfotundstico (PIA), juntamente, com acetato de uranila (UA). Para a concentracao das particulas virais, antes da contrastacao, fizemos uso de uma tecnica descrita por Yunis et al. (1978). Esta tecnica, juntamente com o metodo de colocacao dupla, melhorou consideravelmente a visualizacao e o contraste das particulas virais encontradas nos liquidos de cultura de tecidos infectados e alantoico de ovos embrionados inoculados, tornando o exame mais confiavel, rapido e de facil execucao

Estudos sobre a contrastacao negativa de suspensoes virais. / Negative staining of viral suspension

Suspensoes virais de Rotavirus, Adenovirus Influenza, Parainfluenza, Virus Respiratorio Sincicial, Varicela, Caxumba, Rubeola Herpes Hepatite B, Astrovirus, Poliomielite e Vaccinia foram examinadas em microscopia eletronica diretamente ou atraves da imuno-microscopia eletronica, a fim de determinar a melhor substancia contrastante negativa especifica para cada um dos seus tipos morfologicos virais. Usando acido fosfotungstico (PTA) como contrastante padrao, os resultados foram aprimorados com o emprego do silico - tungstato de sodio (SST) para os Myxovirus, Adenovirus e virus da Poliomielite, do acetato de uranila (UA) para a Hepatite B e do citrato de uranila (UC) para os envoltorios do tipo morfologico da varicela e de detalhes da estrutura de outros tipos virais. Outrossim, suspensoes virais nao purificadas, ao serem contrastadas com molibdato de amonia (AM) e SST evidenciaram melhor as particulas virais integras. A fim de obter um melhor espraiamento das suspensoes virais, especialmente as obtidas de gradientes de sacarose e cloreto de cesio sobre as grades revestidas de Formvar e carbono, a poli-L-lisina mostrou-se superior as demais substancias testadas; alem de separar os agrupamentos virais das impurezas na imuno - microscopia eletronica, tambem permanencia aderido um numero maior de particulas virais isoladas

Osteonecrosis of the Femoral Head Associated with Polyvinylpyrrolidone Storage / 대한정형외과학회잡지

Eighteen cases of osteonecrosis of the femoral head associated with macromolecular polyvinylpyrrolidone (PVP) deposition were analysed on the basis of clinical, radiologic and pathologic features. The cases were observed during 8 years period from January, 1974 to December, 1981. The pathogenesis of the osteonecrosis of the femoral head due to PVP storage in reticuloendothelial system were discussed in detail. Parenteral administration of high-molecular PVP in repeated, long duration led to osteonecrosis of the femoral head. Storage of PVP in the histiocytes of the bone marrow resulted in osteonecrosis of the femoral head followed by microciculation disturbance. PVP-induced osteonecrosis were manifested as multiple foci of necrosis involving not only the femoral head, other long bones around joints, but also the visceral reticuloendo-thelial system characterized by infiltrates of histiocytes laden with PVP. The patients with PVP induced osteonecrosis complianed multiple joint pain in their early course of the disease. On roentgenogram, osteonecrosis were often noted in the hip, shoulder, knee, and ankle in order or frequency. Foamy histiocytes laden with PVP were characteristic on hematoxylin-eosin stain diagnostic on Weigert's elastica, phosphotungstic acid hematoxylin, and Congo red stains. As far as rationale of the treatment concerning a number of staging systems for Osteonecrosis, the choice of surgical procedures were similar to those given by W.F. Enneking et al. In the series, we have performed two hips in total surface replacement, 26 hips in total hip replacement mostly for 3rd generation-configuration of Charnley prosthesis. In addition, one case for free vascularized fibula graft and trans-trochanteric rotational osteotomy after Sugioka were also included for this study. The result of treatment was rather optimistic. However, complications have occured in 4 hips of 3 patients which required removal of whole prosthetic components. Therefore, we underwent revisional surgery in three out of four hips subsequently during the short post-poerative follow-up. These will be published in the future.