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[Evaluation of bacterial growth rate by the ATP-bioluminescence method (luciferase assay)--comparison with conventional methods and application to clinical studies].

Bacterial growth rate was evaluated by estimating the increase in ATP content of cultures of bacterial suspensions using luciferase assay, and the method was applied to the estimation of bactericidal activity of serum and anti-bacterial activity of antibiotics. The results obtained were as follows: The bacterial numbers or bacterial growth rate determined by the ATP-Bioluminescence method was in accordance with those by CFU assay and spectrometrical assay by optical density readings. Furthermore this method was confirmed to be non-time-consuming and more simple than other methods. Using this method, anti-bacterial activity of fresh serum was evaluated. The serum suppressed the growth rate of E. coli, Str. fecalis, CBS, Kleb. pneumoniae and Staph. aureus at least for 2 hours. Heat-inactivated serum and absorbed serum with bacteria tended to lose such suppressive effect. Thus the main effective factors seem to be complements and specific antibody. The anti-microbial activity of antibiotics was enhanced by addition of fresh serum to bacterial cultures, and the results were not in accordance with those by the conventional Disc method. Thus the sensitivity test under the influence of fresh serum is recommended in order to know the clinical effect of antibiotics. These results suggest the usefulness of ATP-Bioluminescence method in the clinical laboratory examinations or investigations.

Adenosine Triphosphate

Comparison of Eustachian tube function measured by the microflow method and a new quantitative impedance method.

Eustachian tube function in 25 applicants for flight training was tested during simulated flights using two methods alternately; the integrating microflow method with a stationary pressure chamber, and a new quantitative impedance method in combination with a new mobile pressure chamber. All ears were tested by both methods. The results of the various tests did not differ significantly between methods. The reliability of the measurements was good and almost equal in both methods. Several practical advantages with the new impedance method in combination with the mobile pressure chamber make it preferable.

Acoustic Impedance Tests

[Evaluation of methods to determine the vitamin B6 status of humans. 1. alpha-EGOT: methods and validation].

A semiautomatic method to determine the activation coefficient of the glutamic-oxaloacetic-transaminase in the erythrocytes is described. This method is compared with other methods and possible sources of error are discussed. Colorimetric methods show considerable disadvantages, such as lack of precision or unspecific hydrazone formation. The kinetic tests avoid these problems. This is in part due to the fact that enzyme stimulation by excessive PLP-addition as used in our procedure bears practical and theoretical advantages. Nevertheless, comparison with published data on vitamin B6 status remains difficult because of insufficient standardization of the used methods. To validate our method we performed experiments testing the stability of the samples, short-term physiological changes and influence of the chelating agent EDTA on transamination. Heparinised blood samples can be stored only for one day with changing at 4 degrees C or room temperature. Storing of the erythrocytic suspension at deep-freezing temperature (-18 degrees C) leads to considerable changes of results. When kept under liquid nitrogen there is no change of the alpha-values for at least 45 days. The addition of EDTA to the samples is known to exclude the influences of cations, such as non enzymatic transamination of a direct alteration of the enzyme activity and therefore EDTA should be added to improve standardisation of the method. The described procedure is a fast, reliable and precise way to determine the alpha-EGOT.

Aspartate Aminotransferases

[Critical study of methods of estimation by an enzymatic method of serum triglycerides (author's transl)].

The authors describe a critical study of two entirely enzymatic methods of determination of serum triglycerides which differ mainly by the choice of hydrolytic agents: esterase and hydrolase by the Roche methods, lipase and esterase in the case of the Boehringer method. The estimation of the resulting free glycerol requires 3 sequential enzymatic reactions (glycerol kinase, pyruvate kinase, lactate dehydrogenase, ending by the measurement of the consumed reduced NAD. The study of the kinetics of the analysis in two stages is carried out using standard solutions of glycerol and with human sera of variable concentration; the optimal conditions of the reaction time and temperature are drawn up. The trials of repeatability and reproducibility from day to day were carried out together with evaluation of the precision. The method was compared with Beohringer's method (n = 50) using hydrolysis of alcoholic potash. The authors conclude on the use of the methods studied.

Esterases

Combined gel filtration, biuret/copper method compared with an immunochemical method for urinary protein measurement.

We compared an immunochemical method specific for plasma proteins with a chemical method, in which interfering substances are separated by gel filtration and "total" urinary protein is determined by the biuret reaction followed by reaction of protein-bound copper with diethyldithiocarbamate after a second gel filtration to remove nonprotein-bound copper. More than 250 24-h urine samples were analyzed by each method. There was linear agreement and a correlation of 0.96 between the two methods, but urinary protein values determined by the immunochemical method, especially for patients with multiple myeloma, were lower than by the chemical method.

Chromatography, Gel

Comparative study on two methods for serotyping of Staphylococcus aureus: the Pillet method and the Oeding method. II.-Possible identity between Pillet's antigen 9 and Oeding's antigen h1.

Using two techniques (agglutination and immunoprecipitation), the authors have observed that: -Pillet's strain CK9 and Oeding's strains 670 and 17A are very similar; -antigen 9 is also present in the three strains; the same relation is found in wild strains. Using agglutination and immunoprecipitation, there is a cross reactivity between h1 factor serum, serum 9 and strains CK9, 670 and 17A. A precipitation line involving strain CK9 and serum CK9 is described in the Pillet system. Finally, there is a strong presumption that Oeding's h1 antigen and Pillet's antigen 9 are identical.

Antigens, Bacterial