Proteinograma e concentração sérica de IgG em potros, do nascimento aos trinta dias de vida, tratados com plasma / Proteinogram and serum IgG concentration in newborn foals up to thirty days of life treated with plasma

Este trabalho teve por objetivo avaliar o proteinograma e concentrações séricas de IgG (após a padronização de teste ELISA) em potros do nascimento aos trinta dias de idade, antes e depois de mamarem colostro e serem tratados com plasma por via intravenosa. Foram utilizados 20 potros e suas respectivas mães, além de quatro animais doadores de plasma. Foram colhidas amostras de sangue dos potros em cinco momentos, logo após o nascimento e antes de mamar colostro (M1), dez horas após nascimento (M2), 24 horas após nascimento e previamente administração do plasma sanguíneo (M3), 48 horas de vida e 24 horas após administração do plasma sanguíneo (M4), e 30 dias após nascimento (M5). Foram colhidos sangue e colostro das éguas progenitoras no momento do parto. A concentração de proteína total (PT) e albumina foram determinadas em analisador bioquímico, a concentração de PT também foi avaliada em refratômetro manual. O fracionamento proteico foi realizado utilizando eletroforese em gel de agarose. A densidade do colostro foi avaliada com colostrômetros de refração BRIX e de densidade específica. A concentração de IgG total de todas as amostras foi determinada por teste ELISA. Com o sistema de ELISA aqui proposto foi possível determinar concentrações de IgG em amostras de soro, plasma e colostro equino com adequada repetibilidade. A média ± desvio padrão da concentração sérica de IgG dos potros ao nascer, foi de 15±8mg/dL, com dez horas de vida foi de 2.408±608mg/dL, se manteve em níveis semelhantes até 48 horas (2.364±784mg/dL) e diminuíram significativamente aos 30 dias de vida (1.414±586mg/dL). A concentração sérica e colostral de IgG nas éguas foi de 1.746±505mg/dL e 7.714±2.619mg/dL, respectivamente. A concentração plasmática de IgG dos doadores de plasma foi de 2.026±148mg/dL. Houve correlação positiva entre as concentrações séricas de IgG e PT (r=0,69 para refratômetro e r=0,76 para bioquímico), GT (r=0,81) e gamaglobulina (r=0,85). Dez horas após o nascimento foi possível verificar a transferência de imunidade passiva, possibilitando adotar medidas profiláticas e/ou terapêuticas em haras de criação de cavalos. Considerando que a proteína total, globulinas totais e fração γ-globulina apresentam correlação com IgG, estas determinações são úteis para monitorar os potros após mamarem o colostro. Um litro de plasma administrado às 24 horas de vida não foi suficiente para aumentar as concentrações séricas de IgG, 24 horas após transfusão, em potros com adequada transferência de imunidade passiva.(AU)

The aim of this study was to evaluate serum protein and serum IgG concentrations (after a direct enzyme immunoassay test ELISA optimization) in newborns foals from birth to thirty days of life before and after colostrum consumption and intravenous treatment with plasma. Twenty foals and their respective progenitors as well as four plasma donor's horses were used. Blood samples were obtained from newborn foals at five time points, immediately after birth and before colostrum intake (M1), ten hours after birth (M2), 24 hours after birth and prior administration of blood plasma (M3), 48 hours after birth and 24 hours after plasma administration (M4), and 30 days after birth (M5). Blood and colostrum samples were collected from the progenitor mares immediately postpartum. Concentration of total protein (TP) and albumin were determined using a biochemical analyzer. The TP concentration was also measured by refractometer. Fractions of total serum protein were separated using agarose gel electrophoresis. Colostrum density was evaluated using BRIX refractometer and specific density colostrometer. Total IgG concentration was determined by an enzyme-linked immunosorbent assay. With the ELISA system proposed here it was possible to determine IgG concentrations in serum, plasma, and equine colostrum samples with adequate repeatability. Serum IgG concentration in foals at birth was 15±8mg/dL (mean ± standard deviation) raising at ten hours (2,408±608mg/dL) and remaining at similar levels up to 48 hours of life (2,364±784mg/dL), and decreasing significantly at 30 days of age (1,414±586mg/dL). Serum and colostrum IgG concentrations of mares were 1,746±505mg/dL and 7,714±2,619mg/dL, respectively. The plasma IgG concentrations from donor mares were 2,026±148mg/dL. Total protein, total globulins, and γ-globulin fraction showed correlation with IgG. Ten hours post birth was an adequate time to verify the transfer of passive immunity, allowing to adoption prophylactic and/or therapeutic measures in a horse farms. One liter of plasma administered at 24 hours of life was not sufficient to raise serum IgG concentrations in foals without passive immunity transfer failure.(AU)

Biological and biochemical studies for characterization of some Egyptian Trichomonas vaginalis isolates

Trichomoniasis has important medical, social and economical implications regarding its serious potentially associated complications, with the possibility of HIV acquisition and transmission. T. vaginalis is a very complex organism. Studying the variation in some biological and biochemical properties of the parasite can be used for characterization of the parasites. For the characterization of T. vaginalis infecting Egyptian female patients, the growth kinetics of 20 isolates, their pathogenicity, metronidazole susceptibility and electrophoretic protein patterns, were correlated with the recorded clinical manifestations associated with these isolates. Positive samples for T. vaginalis were cultured on modified Diamond`s medium. For growth pattern study, trophozoites were counted for each isolate every 24 hours for seven days. The pathogenicity assay was performed using intra-peritoneal inoculation in mice. The isolates susceptibility to different concentrations of metronidazole was recorded by determining the minimal lethal concentration [MLC] and trophozoites viability. The biochemical variability of the studied isolates was performed using 2-dimensional electrophoresis. A broad experimental variability was recorded among the 20 T. vaginalis isolates. There was a clear relationship between 3 isolates obtained from patients with severe vaginitis and the different parameters studied. These isolates had the highest number [20-25 organisms/HPF] in wet mount of vaginal discharge, and the shortest generation time [6:34-7:31 hours]; they were also highly pathogenic to mice. Only one isolate [no. 5] proved to be metronidazole resistant. The use of the first dimensional native-polyacrylamide gel electrophoresis [Native-PAGE] demonstrated the presence of some differences. The isolates were classified into two groups according to their proteins net charge. All samples in each group were considered as one isolate. However, when the 2-dimensional electrophoresis [SDS-PAGE] was applied, five different groups could be identified according to proteins molecular weights. There is a broad experimental variability among the studied Egyptian T. vaginalis isolates regarding growth kinetics, metronidazole drug susceptibility, degree of pathogenicity, as well as the electrophoretic protein patterns

Molecular characterization of hair cuticle and its extracted proteins in seven mammalian species

Hair is an of the epidermis in mammals and consists of two large groups of human hair proteins. One is hard -keratins and the other is matrix proteins. The present investigation aimed to compare the ultrastructural of the hair scale using the scanning electron microscope, and the proteins and amino acids content of the keratin in seven mammalian species. The values of the hair thickness, x/y feret and hair pattern of the species in the present study confirm the presence of species-specific characteristics and ultra structural variation. The situation in man differs from the wild mammals due to damage of hair cuticle caused by mechanical abuse, exposure to ultraviolet radiation and chemical over processing. The maximum amount of extracted proteins from hair keratin was analyzed by SDS-PAGE. The electrophoretic patterns showed an overall degree of similarity. However, differences exist between species in the intensity of stain. Quantitatively, the electrophoretic patterns scanned and analyzed using gel protein analyzer. The results showed no difference between the molecular mass of some species, but different in molecular mass distribution. Amino acid composition of keratin of mammalian hair species of the present study showed some variation, especially for methionine, isoleucine, lysine and arginine. The other amino acids studied are significantly present in most hair. One of the later amino acid is cysteine. Cysteine is a very important due to the presence of disulfate cross-links

Kinetic studies and electrophoretic separation of alkaline phosphatase isoenzyme from cancerous patients

The activity of sALP was determined by the calorimetric method King Armistrong in serum of 303 cancerous patients and 42 normal healthy controls. The results show that the ALP activity was increased in all types of cancer tissues, however ALP activity show a significant increase only in bone, liver and pancreas tissues. The increase of ALP activity could be used as tumor marker for bone, liver and pancreas tissues. The increase in sALP activity could be used as tumor marker for bone and liver cancer and to detect metastasis to the organ. No significant differences of sALP activity were found between males and females. The activity was increased with the stage of development of the disease. The kinetic studies for normal healthy human were measured at a substrate concentration 10 mM, Km 2.7 mM, temperature 370c and pH,o. The electrophoretic studies show that one band of ALP in serum of norma subjects, as compared to cancerous patients where several bands were detected in cancer patients; the intensity of band color varied with type; age and stage of disease

Identificación molecular del Gen RhD mediante PCR / Molecular identification of the RhD gene by PCR

Introducción. Con objeto de identificar al gen Rh(D) en sujetos positivo, se elaboró una estrategia basada en la reacción en cadena de la polimerasa (PCR). Material y métodos. Se estudiaron a 8 sujetos adultos, identificando el fenotipo del sistema Rh (CcDEe) con técnicas inmunohematológicas. Se extrajo el DNA genómico a partir de leucocitos de sangre periférica y se utilizaron dos iniciadores (24C y R13N) para la amplificación. Este par de iniciadores delimitan las secuencias comunes entre los exones 4 y 5 de los genes RhD y RhCE. Resultados. Inmunológicamente se identificaron cinco sujetos Rh negativo y tres Rh positivo. En la imagen electroforética, se observaron dos patrones de amplificación: uno de dos bandas (de 600 pb y de 1200 pb) y un segundo patrón de una sola banda (de 1200 pb). La banda de 1200 pb se amplificó en los sujetos Rh positivo y negativo (y corresponde al gen RhCE). La banda de 600 pb, sólo se presentó en sujetos Rh positivo (correspondiente al gen RhD). No hubo diferencias entre los resultados inmunohematológicos y el análisis molecular en los diferentes fenotipos. Conclusiones. El empleo de esta estrategia basada en la técnica de PCR, permite la tipificación del gen RhD en los sujetos Rh positivo, con diferentes fenotipos del sistema Rh

Utilidad clínica de la electrforesis de proteínas

El suero humano contiene más de 120 proteínas que pueden ser identificadas bioquímicamente,y separadas y cuantificadas electroforéticamente según sus respectivas características físico-químicas. De acuerdo con las técnicas utilizadas, en la electroforesis de proteínas en acetato de celulosa es posible identificar cinco grupos, a saber: albúmina, a1 globulinas, a2 globulinas, b globulinas y y globulinas. El análisis de la electroforesis de proteínas debe ser cuantitativo y cualitativo. La electroforesis de proteínas no aporta resultados patognomónicos y , en consecuencia, las alteraciones que en ella se encuentren deben ser confirmadas por métodos de mayor sensiblidad y especificidad. Como tal, la prueba es una excelente heramienta de tamización, muy similar al hemograma y al citoquímico de orina en la clínica. Se definen los valores de referencia, las alteraciones más significativas de cada banda electroforética y los principales patrones, en particular aquellos de mayor interés clínico. Por último, se establecen las bases para su adecuada interpretación.

Búsqueda de rotavirus en muestras fecales resuspendidas en medio de transporte utilizado para el diagnóstico bacteriológico / Rotaviruses search in rectal samples placed on transport medium for bacteriological diagnosis

Rutinariamente el diagnóstico de gastroenteritis en niños hospitalizados no incluye la búsqueda de rotavirus como agente causal. Para el examen bacteriológico se utiliza una muestra de hisopo rectal colocada en medio de transporte, la cual podría servir también, para la búsqueda de rotavirus. El objetivo del presente trabajo fue determinar la presencia de rotavirus en 105 muestras de hisopos rectales resuspendidos en medio de transporte de Stuart, provenientes de pacientes menores de 5 años de edad, hospitalizados con un cuadro gastrointestinal en el Hospital General Regional 25 del IMSS. Estas muestras previamente fueron reportadas como negativas para agentes causales de diarrea, a excepción de una, que fue positiva para Vibrio Cholerae. A las 105 Muestras se les realizó la extracción del RNA viral y se analizaron por electroforesis en geles de poliacrilamida al 10 por ciento. Se detectaron 28 positivas para rotavirus, de las cuales 26 presentaron un patrón electroforético de tipo largo y 2 de tipo corto. La distribución de los segmentos de RNA en grupos de 4, 2, 3 y 2 bandas, sugiere que en todos los casos se trata de rotavirus del grupo A. Estos resultados muestran que es posible detectar la presencia de rotavirus, utilizando la misma muestra que se toma para el diagnóstico bacteriológico y que el genoma viral presente en la muestra puede conservarse en congeladores de tipo doméstico por periodos prolongados, lo cual es muy conveniente en la realización de estudios epidemiológicos

Análise electroforética da farinha de trigo sarraceno em comazaçao com a farinha de trigo suave / Electrophoretic analysis of meal of buckwheat in comparison with common wheat flour

A composiçao das proteínas alcool (prolaminas) obtidas das farinhas de trigo suave por dois procedimentos foram analizadas por el electroforense a pH 3,1 e, após disociaçao, na presença de dodecil sulfato de sódio a pH 8,0. Os perfis obtidos da fracâo prolamina do trigo sarraceno muito diferentes tanto qualitativamente como quantitativamente daqueles da prolamina, do trigo suave. Parece, portanto, provável que ofeitos adversos associados com alimentares contendo gliadina de trigo a pacientes celíacos seriam reduzidos e possivelmente evitados se a farinha de trigo fosse substituida pela farinha de trigo sarraceno

Análisis electroforético de extractos de periplaneta australasiae y p. americana / Electrophoretic analysis of periplaneta australasiae and periplaneta americana extracts

Se prepararon extractos proteicos de todo el cuerpo de cucarachas Periplaneta americana y Periplaneta australasiae (Dictyoptera: Blattidae). Las proteínas correspondientes fueron separadas e identificadas mediante electroforesis en geles de poliacrilamida con duodecil sulfato de sodio (SDS-PAGE).Los perfiles proteicos fueron analizados, encontrándose numerosas bandas compartidas y no compartidas en los extractos de cada especie.Lo anterior podría ser la causa de la reactividad cruzada e individual a estas cucarachas mostrada por pruebas cutáneas en pacientes alérgicos a estos extractos.

La electroforesis de proteínas y su aplicación en la clínica

La experiencia clínica de muchos años ha demostrado que la electroforesis de proteínas en suero es una excelente prueba de laboratorio. Son múltiples las entidades en las que tienen gran importancia clínica. Para su adecuada utilización e interpretación se requiere tener conocimientos acerca de su composición, comportamiento y variaciones de acuerdo a cada entidad nosológica. Las publicaciones sobre este tópico son escasas y las revisiones disponibles fueron publicadas hace muchos años. En este artículo se describen las principales fracciones obtenidas en la electroforesis en acetato de celulosa y se establece una correlación con las principales patologías que la modifican. Además, se definen y esquematizan los patrones electroforéticos mas característicos y de mayor ocurrencia clínica.