The isoforms distribution of the glycoprotein antithrombin III (ATIII) derived from human plasma was investigated by means of isoelectric focusing (IEF) in polyacrylamide gels with immobilized pH gradients (IPG) and two-dimensional gel electrophoresis (2-DE) as well as capillary electrophoretic methods. It turned out that the presence of high concentrations of chaotropics (urea, thiourea) and zwitterionic detergents (3-[(3-cholamidepropyl)dimethylammonio]-1-propanesulfonate (CHAPS)) was decisive for attaining good resolution of the protein isoforms. Resolution by IPG-IEF was obtained with excellent reproducibility and pI differences down to 0.01 pH units could be distinguished. ATIII-alpha and ATIII-beta-fractions preseparated by heparin affinity chromatography showed an analogous but shifted spot pattern consisting each of one major and three minor isoforms. The main isoforms of ATIII-alpha and ATIII-beta exhibit pI values of 5.18 and 5.32, respectively, both values determined in the presence of high concentrations of urea. The pI difference of 0.14 pH units correspond to the effect of two sialic acids absent in ATIII-beta. The formation and occurrence of ATIII dimers and trimers turned out to be dependent on the sample preparation. The results obtained by 2-DE were compared with those of capillary zone electrophoresis (CZE) and capillary IEF (CIEF). Quantitative analysis regarding the CZE separated isoforms of plasma derived ATIII yielded a content of about 70% ATIII-alpha main isoform and about 6.6% of ATIII-beta. The pI values of ATIII determined by CIEF with internal calibration were in fair agreement with the pI values of the main isoforms achieved with 2-DE.
In this study, liquid egg, albumen, and egg yolk were artificially inoculated with E. coli. Ultrasound equipment (20/40 kHz, 180/300 W; 30/45/60 min) with a circulation cooling system was used to lower the colony forming units (CFU) of E. coli samples. Frequency, absorbed power, energy dose, and duration of sonication showed a significant impact on E. coli with 0.5 log CFU/mL in albumen, 0.7 log CFU/mL in yolk and 0.5 log CFU/mL decrease at 40 kHz and 6.9 W absorbed power level. Significant linear correlation (p < 0.001) was observed between the energy dose of sonication and the decrease of E. coli. The results showed that sonication can be a useful tool as a supplementary method to reduce the number of microorganism in egg products. With near-infrared (NIR) spectra analysis we were able to detect the structural changes of the egg samples, due to ultrasonic treatment. Principal component analysis (PCA) showed that sonication can alter C–H, C–N, –OH and N–H bonds in egg. The aquagrams showed that sonication can alter the properties of H2O structure in egg products. The observed data showed that the absorbance of free water (1412 nm), water molecules with one (1440 nm), two (1462 nm), three (1472 nm) and four (1488 nm) hydrogen bonds, water solvation shell (1452 nm) and strongly bonded water (1512 nm) of the egg samples have been changed during ultrasonic treatment.
Zusammenfassung: Es wird eine reaktionskinetische Glucosebestimmung in Blut, Urin und Liquor mit Hilfe des neuen Eppendorf Analysenautomaten ACP 5040 beschrieben. Dabei entsteht aus Glucose mittels Ghicosedehydrogenäse Gluconsäure, NAD* wird zu NADH reduziert. Die von uns ausgearbeitete Bestimmungsvariante zeichnet sich durch gute Präzision, Richtigkeit und Durchsatzgeschwindigkeit aus. Die Korrelation mit der Hexokinase/Glucose-6-phosphatdehydrogenase-Endpunktmethode ist gut.
The kinetic determination of glucose vv/rft the glucose dehydrogenase method using the Eppendorf automatic analyzer 5040Summary: A kinetic determination of glucose from blood, urine and spinal fluid is described with use of the new automatic analyzer ACP 5040 Eppendorf. The method uses glucose-dehydrogenase which converts glucose to gluconic acid. The NADH formed can be measured by the increase in absorbance at 334 nm. Our variation of test methodology gives good precision, accuracy and a high performance speed. There is a good correlation with the hexokinase-glucose-6-phqsphate-dehydrogenase end point method.
Einführung
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