RESUMEN
In order to explore the nature of glucose-6-phosphate dehydrogenase (G6PD) deficiency in south-east Sicily, we have analysed the G6PD gene in 25 unrelated males with abnormal G6PD activity and/or electrophoretic mobility, by using the analysis of the appropriate PCR-amplified fragment of DNA and subsequent digestion by appropriate restriction-enzymes, looking for the presence of certain known G6PD mutations. We amplified the entire G6PD coding sequence into eight fragments, followed by single-strand conformation polymorphism (SSCP) analysis and sequencing of those individual fragments that were found to be abnormal by SSCP. Through these methods we found a total of twelve G6PD Mediterranean variants with the association of a silent mutation 1311 (also known as polymorphic site Bcl I), one G6PD Mediterranean without this association, four G6PD A-Val 68 and two G6PD Santamaria and five G6PD Chatham. In a subject with normal activity a mutation was found in exon 5, designated as G6PD Sao Borja. This is the first report on the molecular analysis of G6PD mutations in Sicily and we have obtained evidence for four distinct classes of variants.
Asunto(s)
Heterogeneidad Genética , Deficiencia de Glucosafosfato Deshidrogenasa/genética , Femenino , Deficiencia de Glucosafosfato Deshidrogenasa/epidemiología , Humanos , Masculino , Mutación Puntual , Reacción en Cadena de la Polimerasa , Análisis de Secuencia de ADN , Sicilia/epidemiologíaRESUMEN
In the present study we have assayed the effect of divicine in G6PD-deficient red blood cells in the presence of deferoxamine (iron-chelating drug) and NaN3 (inhibitor of catalase). The effect of divicine has been compared to oxidative stress by H2O2; haemolysis is regarded as an index of cellular toxicity. In addition, we have tested antioxidant enzymatic systems. No significant change in antioxidant enzymatic systems was found in RBCs from subjects with G6PD deficiency when compared to the control group, either in oxidative haemolysis by divicine or by H2O2; a significant decrease in oxidative haemolysis by H2O2 was observed in the presence of deferoxamine, whereas no change was found in oxidative haemolysis by divicine. The replacement of incubation medium by homologous plasma or the supplementation with bovine serum albumin resulted in a marked decrease of percentage of haemolysis by divicine.
Asunto(s)
Deferoxamina/farmacología , Eritrocitos/enzimología , Deficiencia de Glucosafosfato Deshidrogenasa/sangre , Adulto , Catalasa/sangre , Eritrocitos/efectos de los fármacos , Glutatión Peroxidasa/sangre , Glutatión Reductasa/sangre , Hemólisis/efectos de los fármacos , Humanos , Peróxido de Hidrógeno/farmacología , Técnicas In Vitro , Masculino , Oxidación-Reducción , Pirimidinonas/farmacologíaRESUMEN
Intensive desferrioxamine treatment may negatively affect zinc stores in thalassemic patients. Two groups of young thalassemic patients, with good and poor compliance to desferrioxamine treatment, and a control group were investigated. Leukocyte alkaline phosphatase (LAP), zinc content in neutrophils, plasma zinc levels, and urinary zinc excretion were determined. LAP score, and neutrophil zinc and plasma zinc levels of the good compliance patients were statistically lower than those seen in the controls and in the poor compliance patients. Urinary zinc excretion was statistically higher in good compliance patients than in controls and poor compliance patients only during high-dose intravenous desferrioxamine. On the contrary, poor compliance patients had LAP scores, neutrophil zinc, plasma zinc, and urinary zinc excretion values within the normal range. We conclude that high-dose desferrioxamine produces a reduction of zinc stores in thalassemic patients. LAP scores determinations, which are a reliable and inexpensive marker of body zinc, could become a routine test for monitoring thalassemic patients undergoing hypertransfusion programs.