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A new colorimetric ultramicromethod for serum glutamicoxalacetic and glutamic-pyruvic transaminase determination
No full textIn this paper an ultramicromethod for the determination of the serum glutamic-oxalacetic and glutamic-pyruvic transaminase activity is presented; this method is based on the use of glutamate dehydrogenase for the enzymatic estimation of the glutamate formed. The dehydrogenation of the glutamate gives rise to the reduction of a diazonium salt, and it is possible to perform a photometric reading of the colored compound at 520 nm. 20 μl of serum and an incubation time of only 45 min at the temperature of 37° were necessary. The normal values never exceeded 54.5 I.U. for the serum glutamic-oxalacetic transaminase and 52 I.U. for the glutamic-pyruvic transaminase. Under conditions of viral hepatitis values of 390 I.U. for glutamic-pyruvic transaminase and 310 I.U. for serum glutamic-oxalacetic transaminase were obtained. © 1970
A rapid photometric micromethod for serum lipase determination.
No full textA new micromethod for the determination of serum lipase activity is presented. Small quantities of serum are incubated for 30 min at 40° in a substrate consisting of an olive oil suspension in the presence of deoxycholic acid. The pH is 8.5. The hydrolyzed fatty acids are determined with a photometric technique, with a slight modification of the method of Duncombe. The normal values are lower than 20 I.U. © 1972
High light scatter by neutrophils in the Bayer-Technicon H*2 analyzer: a screening test of morphologically defective responsiveness to in vitro chemotactic stimulation
No full textThe Bayer-Technicon H*2 haematological analyser provides differential white blood cell count, including the assay of polymorphonuclear leukocytes by light scattering and the absorbance increase following the cytochemical reaction for myeloperoxidase. The mean value of polymorphonuclear leukocytes scatter, which reflects polymorphonuclear leukocytes volume, is printed in a separate report "for laboratory use only" as a ybar value in arbitrary units. In certain patients neutrophils displayed an unreported correlation between polymorphonuclear leukocytes high ybar basal values (> or = 37.00 arbitrary units) (determined on the H*2) and a defective response in vitro to the chemoattractant, formyl-methionyl-leucyl-phenylalanine (determined by microscopic evaluation of polymorphonuclear leukocytes shape change (polarization)). The patients showing no polymorphonuclear leukocyte response or a defective one to formyl-methionyl-leucyl-phenylalanine were all affected by "Systemic Inflammatory Response Syndrome (SIRS)". Therefore the predictive value of the positive test for SIRS is 100%. On the other hand 8.8% of SIRS patients had polymorphonuclear leukocytes < 37.00 arbitrary units of ybar basal value and a "normal" response to formyl-methionyl-leucyl-phenylalanine; the predictive value of the negative test being 90%. Since we demonstrated in vitro a dose-dependent deactivation of endotoxin or lipopolysaccharide-pretreated polymorphonuclear leukocytes, the "normal" response to formyl-methionyl-leucyl-phenylalanine of the "false negative" cases may occur because the endotoxaemia in these patients is too low to prevent it.(ABSTRACT TRUNCATED AT 250 WORDS
Variazioni sieriche della beta2-microglobulina, del TNF, del recettore solubile dell'IL-2 e degli antigeni solubili CD4 e CD8 nell'infezione da HIV-1
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Assessment of neutrophil aggregation by Coulter STKR and STKS haematological analysers
No full textWe have studied an alternative method to aggregometry for the assessment of human polymorphonuclear (PMN) leucocyte aggregation. This simple, rapid and reliable procedure counts unaggregated cells on both Coulter STKS and STKR haematological analysers by the impedance principle. Aggregation of PMN was induced by 15 min incubation with fresh autologous serum (FAS) after a 10 min phorbol myristate acetate (PMA) activation of neutrophils in small aliquots (0.25 ml) of suspension containing about 4.0 x 10(9) PMN/1. Differences (x 100) between count of resting and PMA+FAS treated neutrophils/count of resting PMN reflect percent aggregation. By this procedure, PMN aggregation did not occur in autologous plasma from EDTA anticoagulated whole blood; it was partially inhibited by hydrocortisone, whereas inactivated or Zymosan activated sera gave values similar to those from FAS induced aggregation. PMA aggregation was dependent on Ca2+ + Mg2+ concentration. Intra-assay analytical variability did not exceed 4% on either instrument. Reference values (n = 20) of percent PMN aggregation were 50.7 +/- 4.7 on STKS and 47.1 +/- 4.8 on STKR. Most probably, the interindividual variance was due to the physiological variability of Mg2+ and/or Ca2+ concentrations in FAS. Thus, this procedure reflects the true PMN aggregability status in a given subject, and in a given electrolyte environment
EDTA-induced platelet aggregation can be avoided by a new anticoagulant also suitable for automated complete blood count
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