ABSTRACT
Se estudió la actividad enzimática (AE) de la enzima glucosa-6-fosfato deshidrogenasa eritrocitaria (G6PD) y la movilidad electroforética (ME) en una población de hombres y mujeres de la ciudad de Rosario (provincia de Santa Fe), Argentina y zona de influencia. Para la determinación de AE se utilizó la técnica cinética de Glock y McLean y para la electroforesis de la enzima, la técnica de M.C. Rattazzi y L.C. Bernini en acetato de celulosa. Los valores normales de actividad enzimática (AE) para hombres y mujeres adultos fueron de 8,1 ± 1,4 UI G6PD/g Hb. Se demostró que los valores de AE son independientes de la edad, sexo y concentración de hemoglobina. En todos los grupos etarios estudiados no se observaron diferencias significativas de AE con respecto a los adultos normales a excepción de los neonatos que presentaron un significativo aumento de la misma, lo cual está directamente relacionado con las características fisiológicas de los eritrocitos del recién nacido. Entre los 686 individuos estudiados se detectaron 2 pacientes deficientes de G6PD, lo que dio una prevalencia de 0,3% y el patrón electroforético correspondiente a esta población fue 98% (n: 672) para G6PD B y 2% (n: 14) para G6PD con movilidad rápida tipo A.
Enzymatic activity (EA) of erythrocyte glucose-6-phosphate dehydrogenase (G6PD) and electrophoretic mobility (EM) have been studied in a population of males and females in the city of Rosario and its area of influence. To determine EA, the Glock and McLean kinetic technique was used. Electrophoretic mobility assay was performed by M.C. Rattazzi and L.C. Bernini technique in cellulose acetate gel. Results demonstrated that the EA values in normal individual are independent of age, sex and hemoglobin values. The normal values of EA were: 8.1±1.4 IU of G6PD/g Hb. There were no significant differences in different age groups studied regarding healthy adults, except for neonatal group that yielded a significant EA increase which is directly related to the physiological characteristics of newborn erythrocytes. Two patients out of 686 individuals bearing G6PD deficiency were detected, corresponding to 0.3% prevalence. The electrophorectic mobility pattern was 98% (n: 672) for G6PD B, and 2% (n: 14) for G6PD A fast mobility variant.
Foi estudada a atividade enzimática (AE) da enzima glicose-6-fosfato desidrogenase eritrocitária (G6PD) e a mobilidade eletroforética (ME) numa população de homens e mulheres da cidade de Rosario, província de Santa Fe, Argentina e zona de influência. Para a determinação da AE foi utilizada a técnica cinética de GlocK e Mc Lean e para a eletroforese da enzima a técnica de M.C. Rattazzi e L.C. Bernini em acetato de celulose. Os valores normais de atividade enzimática (AE) para homens e mulheres adultos foram de 8,1 ± 1,4 UI G6PD/g Hb. Foi demonstrado que os valores da AE são independentes da idade, sexo e concentração de hemoglobina. Em nenhum dos grupos etários estudados foram observadas diferenças significativas de AE no que diz respeito aos adultos normais, com exceção dos neonatos que apresentaram um significativo aumento da mesma, o qual está diretamente relacionado com as características fisiológicas dos eritrócitos do recém-nascido. Entre os 686 indivíduos estudados foram detectados 2 pacientes deficientes de G6PD, o que deu uma prevalência de 0,3% e o padrão eletroforético correspondente a esta população foi de 98% (n: 672) para a G6PD B e 2% (n: 14) para G6PD com mobilidade rápida tipo A.
Subject(s)
Humans , Male , Female , Glucosephosphate Dehydrogenase/blood , Glucosephosphate Dehydrogenase/metabolism , Glucosephosphate Dehydrogenase/physiology , Argentina , Electrophoretic Mobility Shift Assay , Glucosephosphate Dehydrogenase DeficiencyABSTRACT
Hemoglobine (Hb) Q-India is an innocuous αglobin variant: α64 Asp → His. DNA sequencing studies have shown that the Hb Q India mutation is GAC → CAC in codon 64 of the α1 gene. Hb Q-India is a well-known hemoglobin variant in South-East Asia but only isolated case reports exist in literature to describe this rare entity in the rest of de world. The variant has been found with various forms of αand ß thalassemia. This hemoglobin has the same electrophoretic mobility as Hb S. We report, for the first time, the identification of Hb Q-India in an Argentinian woman (her parents came from Gibraltar), referred to our laboratory bearing a mild microcytic hypocromic anemia; a co-inherited α+ thalassemia (-α3.7 th) was also found.
La hemoglobina (Hb) Q India es una hemoglobina anormal e inocua que afecta la cadena α de esta. Los análisis de secuencia han demostrado que la mutación se encuentra en el codon 64 GAC → CAC del gen α1. Si bien es una variante muy conocida en el sudeste asiático, solo se han reportado pocos casos en el resto del mundo. Esta hemoglobina anormal se ha encontrado asociada con diversas formas de α y ß talasemia y su posición electroforética es idéntica a la de la Hb S. Reportamos, por primera vez, la identificación de la Hb Q India en una mujer Argentina (cuyos padres procedían del Peñón de Gibraltar), enviada a nuestro laboratorio por padecer de anemia microcítica hipocrómica, en la que se encontró también la coexistencia de α+ talasemia (-α3,7 th).
ABSTRACT
Los mecanismos que determinan la senescencia de los eritrocitos han sido extensamente estudiados, sin embargo, no se han logrado conclusiones definitivas debido a la ausencia de una técnica que permita el aislamiento de grupos etáreos bien definidos. Los métodos más comúnmente empleados se basan en el aumento de densidad de los eritrocitos durante el envejecimiento. En este trabajo desarrollamos una técnica para la separación de glóbulos rojos de distintas edades empleando gradientes preformados de Percoll, un polímero sintético con propiedades fisicoquímicas adecuadas para trabajar con células vivas. En las suspensiones eritrocitarias obtenidas se realizaron determinaciones hematológicas, actividades de enzimas antioxidantes y el ensayo de eritrofagocitosis. Los valores de los parámetros hematológicos evaluados fueron significativamente mayores en las suspensiones de glóbulos rojos jóvenes. Las actividades enzimáticas mostraron una disminución de la capacidad antioxidante en las poblaciones de eritrocitos senescentes. Este proceso favorecería la interacción de los hematíes envejecidos con las células fagocíticas, demostrada mediante el ensayo de eritrofagocitosis. Los resultados obtenidos indican que el método de gradientes de Percoll permite una adecuada separación de las suspensiones eritrocitarias de distintas edades, con una eficiencia comparable a la observada en la técnica de centrifugación diferencial considerada de referencia.
The mechanisms that determine the senescence of the erythrocytes have been extensively studied; however. definitive conclusions have not been achieved mainly because of the lack of a technique that allows the isolation of well-defined etarian groups. The methods most commonly used for separating erythrocytes from different ages are based on the increase in density that these cells present during their aging. In the present work we have developed a technique for obtaining red blood cells from different ages using Percoll preformed gradients, a synthetic polymer with adequate physic-chemic properties to work with lives cells. In the erythrocytes suspensions we have made hematological determinations. activities of antioxidants enzymes and the essay of erythrophagocytosis. The values of the hematological parameters were significantly higher in the suspensions of young red blood cells. In the measurements of the enzymatic activity we observed a decrease of the antioxidant capacity in the populations of senescent erythrocytes. This process would promote the interaction between the old erythrocytes and the phagocyte cells, demonstrated by the erythrophagocytosis essay. The results obtained indicate that the method Percoll density gradients allows an appropriate separation of the erythrocytes suspensions of different ages with a comparable efficiency to that observed in the technique differential centrifugation, considered as reference.
Subject(s)
Erythrocyte Aging/physiology , Erythrocyte Aging/immunology , Povidone , Centrifugation, Density Gradient/methods , Blood Physiological Phenomena , Chemistry Techniques, Analytical/methods , Ultracentrifugation/methodsABSTRACT
Hematological parameters in newborn umbilical cord blood samples (n=476), collected at the Hospital Provincial del Centenario, Rosario, were studied. They were divided into 3 groups: (I) full term newborns with weight according to gestational age; II) low weight and normal gestational age; (III) preterm newborns. The results were as follows: group (I) Hb: 15.5 + 1.1 g/dl; RBC: 4.66 + 0.33 x 1012/I; PCV:49 + 4.3 percent, MCV 105.1 + 5.3 fl; MHC: 33.2 + 1.2 pg. Decreased Hb concentration (p<0.05) and increased MCV (p<0.01) were observed in preterm newborns in comparison with normal ones, and a slight PCV increase and RBC values in low weight newborns compared to the control group (p<0.05). Erytrocyte morphology was normal as well as reticulocyte values in these samples. The electrophoretic pattern was (FA) with the following Hb F values 66.3 + 6.8 percent, and Hb A2 0.45 + 0.3 percent in group (I), with a significant increase of Hb F in 30-35 weeks preterm newborns. Group(I) values are considered as normal hematological parameters in newborns in our country, whereas MCV< 94.7 fl is considered as a neonatal microcytosis marker, consequently an alert to investigate alpha-thalassemia. There was no influence on Hb concentration due to maternal smoking habit. The present work could be of relevance for our region since up to the present time there are no similar records.