Increased viscosity of erythrocyte suspension upon hemolysis

Viscous properties of partially hemolyzed erythrocyte suspensions were studied with a cone-plate viscometer. A Casson plot was successfully applied for analysis of the viscous properties. When the degree of hemolysis was increased, Casson viscosity ( _c) of the partially hemolyzed erythrocyte suspensions increased, but the substitution of hemoglobin solution with the same O₂-carrying capacity of hemolyzed fraction (containing ghost) reduced the _c remarkably. The yield stress (f _c) of the intact erythrocyte suspension increased with increasing hematocrit, but thef _c of partially hemolyzed erythrocyte suspension was dependent on both hematocrit and the degree of hemolysis. The apparent viscosity ( _a) of erythrocyte suspension was drastically increased with the slight hemolysis, and became maximum at a certain degree of hemolysis which was dependent on both total hemoglobin concentration and shear rate. Such influence of hemolysis on the _a was also evident in the mixture of erythrocyte suspension and hemoglobin solution. These results suggest that the hemoglobin molecule plays a certain role in erythrocyte-erythrocyte interaction in their suspension, which modifies the viscosity.     

Information theory of miniaturization advantages of column liquid chromatography for quantitative analysis

Summary The influences of decreasing column diameter and length on two analytical goals, the precision and solvent efficiency, in reversed-phase liquid-chromatography are studied. Low solvent consumption is one of analytical advantages of miniaturized columns and is shown to be quantitatively evaluated by the precision and solvent efficiency which are defined to be the total Shannon mutual information obtained from an assay and the information obtained in a unit solvent volume, respectively. Analysis of paraben food additives on a microbore column is taken as an example. In trace analysis, the relative standard deviation (RSD) of measurements theoretically derived from the precision is shown to approach to the observed ones.     

Identification of Shiga toxins in Shiga toxin-producing Escherichia coli using immunoprecipitation and high-performance liquid chromatography-electrospray ionization mass spectrometry

We developed a rapid and reliable identification method for Shiga toxins in Shiga toxin-producing Escherichia coli (STEC) using immunoprecipitation and high-performance liquid chromatography-electrospray ionization mass spectrometry (HPLC-ESI-MS). Polyclonal antisera specific for Shiga toxin 1 (Stx1) and Shiga toxin 2 (Stx2) were raised in rabbits so as to be used for the immunoprecipitation. The immunoprecipitaion was carried out by mixing sample solutions with 50 microl each of the antisera to Stx1 and Stx2 followed by allowing the mixed solutions to stand for 30 min. The quantity required to obtain the immunoprecipitate was more than 0.5 microg of Shiga toxins. HPLC-ESI-MS analysis of the resulting immunoprecipitates provided accurate molecular weight information on Shiga toxins, leading to direct evidence for the presence of these toxins. It requires at most two days to perform our procedure from toxin extraction to measurement of HPLC-ESI-MS whereas the previous method using isolation procedures required about two weeks to complete. The usefulness of the present method has been demonstrated by identifying Stx1, Stx2 and a variant of Stx2 (Stx2e) in the immunoprecipitates prepared from STEC strains.