Isolement par chromatographie d'affinite de fractions membranaires specialisees, a partir de microsomes d'hepatocytes de chat

AbstractMicrosomal glucokinase is solubilized by incubation in the presence of several metabolites. After solubilization of the enzymes, the membranes present free sites for specific binding of glucokinase, therefore, they can purified by affinity chromatography on Sepharose-ATP-glucokinase. This method yields membranous vesicles which contain, in addition to glucokinase, uridylyl-transferase, phosphoglucomutase, sialyl-transferase and adenylate cyclase. Galactosyl-transferase, glucose-6-phosphatase and NADPH cytochrome c reductase are absent. It appears that functionally related enzyme from UDP-glucose biosynthesis are aggregated onto specific patches of the membrane, most likely from Golgi apparatus

Pediatric reference values for urine particle quantification by using automated flow cytometer: Results of a multicenter study of Italian urinalysis group.

OBJECTIVES: The purpose of this Italian multicenter study was to define pediatric upper reference values for urine particle quantification by using automated flow cytometry. DESIGN AND METHODS: Four hospital-based clinical laboratories participated in this multicenter investigation, which included a total study population of 161 Italian children aged from 1 to 12years. Two laboratories used Sysmex UF-100 and analyzed 86 children, whereas the other two used Sysmex UF-1000i and analyzed 75 subjects. Particle quantification included the analysis of white blood cells (WBC), red blood cells (RBC), squamous epithelial cells (EC), transitional epithelial cells (TC), casts (CAST) and bacteria (BACT). RESULTS: The upper reference values in subjects tested with the Sysmex UF-100 were 9.7WBC/\u3bcL, 10.1RBC/\u3bcL, 7.5EC/\u3bcL, 2.5TC/\u3bcL, 0.7CAST/\u3bcL and 3090BACT/\u3bcL, whereas the upper reference values in subjects tested with the Sysmex UF-1000i were 10.5WBC/\u3bcL, 8.3RBC/\u3bcL, 7.2EC/\u3bcL, 2.9TC/\u3bcL, 0.7CAST/\u3bcL and 48BACT/\u3bcL. No statistically significant differences between genders were found in the value distribution of any of the parameters tested. Similarly, no statistically significant differences were observed between the two urine analyzers, except for BACT. CONCLUSIONS: Automated analysis of urine particles appears a suitable means to optimize the workflow of routine urinalysis of children specimens. The upper reference limits for pediatric subjects obtained in this study were comparable to those previously reported in the literature, with no significant differences between genders and analyzers

Evaluation of reference intervals for complete blood count on Sysmex XN 9000

We aimed to define reference intervals for complete blood count in 240 apparently healthy Italian adults (120 males and 120 females) using Sysmex XN 9000 platform, as recommended by international standards. The recruitment criteria for reference individuals were based on negative anamnesis and physiological serum concentrations of glucose, creatinine, transaminases, ferritin and C-reactive protein. The results were comparable to those previously generated on different European populations. Interesting results were found for some research parameters, for which limited information was available in Italian populations so far. With regard to reticulocyte-related parameters, a significant gender difference was found for reticulocyte hemoglobin, highly fluorescent reticulocyte fraction and delta-He, which represents the difference in hemoglobin content between reticulocytes and erythrocytes. The results also support the reference interval of immature platelet fraction observed in previous studies, except for the absolute value for which larger range and higher values were found. For leukocyte morphological and functional parameters, we were able to define reference intervals in the whole study population as well as in male and female subgroups, since gender-related differences were observed for some parameter