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Starch gel electrophoresis for galactose-1-phosphate uridylyl-transferase applied to dried filter paper blood specimens.

Starch gel electrophoresis of galactose-1-phosphate uridylyl transferase has been adapted for use on dried filter paper blood specimens submitted for the purposes of routine newborn screening for galactosemia and other inborn errors of metabolism. Selected for study were those specimens with reduced transferase activity, as determined by the Beutler enzyme spot screening test. Clinically benign low activity enzyme variants were readily identified, thus reducing significantly the number of requested additional blood specimens. Transferase-deficient specimens with potential clinical significance had too little activity for transferase visualization and thus could be separated from the benign variants. This technique would facilitate routine newborn screening for galactosemia.

Carrier State

[Blood proteolytic enzyme inhibitors and their clinical study].

Data on biochemistry, physiological significance and clinical investigations of two main inhibitors of proteinases in human blood serum (alpha 1-antitrypsin and alpha 2-macroglobulin) are reviewed. Physico-chemical properties of the inhibitors, mechanisms of their interaction with proteinases and methods for determination are discussed. Modification of the specificity of some proteinases by alpha 2-macroglobulin, which probably plays a significant role in the regulation of enzymatic systems of proteolysis in blood, is considered. The use of the inhibitors of proteolysis for diagnostics of pathological processes is discussed. Data on the alpha 1-antitrypsin deficiency and its relation to the development of degenerative diseases in lungs are reviewed.

Arthritis, Rheumatoid

Predictive tests for malignant hyperpyrexia.

Four carriers of the abnormal trait for malignant hyperpyrexia have been studied to determine the value of the various methods of carrier detection. Tests included enzyme estimations, histochemistry, electronmicroscopy, biochemistry and in vitro exposure of muscle strips to halothane. By combining these procedures it is possible to detect all the asymptomatic carriers of this disease.

Adenosine Triphosphatases

Significance of elevated liver alkaline phosphatase in serum.

The serum alkaline phosphatase was fractionated by polyacrylamide gel electrophoresis in 317 patients with elevated serum alkaline phosphatase activity. In 253 patients the source of the elevation was the isoenzyme of presumed liver origin, band L. In 87 of these patients, there was either no obvious liver disease or the alkaline phosphatase elevation was inappropriately high. In 19 of the 87, liver disease was further excluded by liver biopsy or by laparotomy. Because of this, biochemical studies were done to verify the hepatic origin of band L. Band L and alkaline phosphatase extracted from human liver migrated together on polyacrylamide gel electrophoresis before and after digestion with Vibrio cholerae neuraminidase. They had identical pH optima, sedimentation coefficients, Michaelis constants, and rates of inactivation at 55.5 degrees C. They had different rates of inactivation in 3 M urea. Over-all, the data indicate that band L is of liver origin, and that elevation of the hepatic alkaline phosphatase isoenzyme may be a nonspecific finding in certain patients.

Alkaline Phosphatase

Localization of urinary tract infection.

Both the antibody-coating and LDH isoenzyme techniques theoretically fulfill the criteria for an ideal localization test. Both are noninvasive. They are relatively easy to perform and well within the capability of most clinical laboratories. Both can be done rapidly and the results can be in the clinician's hands in a matter of a few hours. The results of the antibody-coating technique do not appear to be valid for children. Since only one controlled study with comparison with other localization techniques has been done in adults, further evaluation needs to be done. The LDH isoenzyme pattern has not been studied in adults but appears to be accurate in children. A prospective comparison of both procedures in adults using either the Fairly bladder washout technique or the Stamey ureteral catheterization method to localize definitively the site of infection would be most helpful. Definitive noninvasive localization techniques could lead to therapy being initiated with the site of infection defined. This would facilitate earlier and more intensive antibiotic therapy in those patients at risk for renal parenchymal damage from upper tract infections.

Antibody-Coated Bacteria Test, Urinary

[The specificity of the CPK MB enzyme kinetic test for the diagnosis of acute myocardial infarction (author's transl)].

The clinical usefullness of the CPK MB test (spectophotometric method) was evaluated on 139 patients admitted to a Cardiovascular Diseases Department with a diagnosis of suspected myocardial infarction. Serial determinations of serum MB isoenzyme creatine kinase, total creatine kinase, lacate dehydrogenase and hydrossibutirrate dehydrogenase were made at 1st, 2th, 3th, 4th, 5th, 6th, 7th day. Incidence of CPK MB false positive and false negative data were also determined and correlated with the electrocardiogram pattern and serum levels of other standard enzymes. Results indicate that the CPK MB kinetic test is highly specific (94%) and promptly available in the early diagnosis of acute myocardial infarction.

Acute Disease

Lactase deficiency--a comparative study of diagnostic methods.

The diagnostic value of 1-14C-lactose breath test was compared with the standard lactose tolerance test and lactase assay in jejunal biopsies in 16 control subjects, 14 patients with lactase deficiency (LD) proven by lactase assay and 20 patients with irritable bowel syndrome (IBS). 14CO2 specific activity in the 2-hr breath collection after administration of 1-14C-lactose (5 muCi) provided a satisfactory separation between the control and LD group. Values were 7.0 +/- 2.0% dose administered/mmoles 14CO2 X 10(-3) (mean +/- SD) in the control group versus 2.1 +/- 1.5 in LD (P less than 0.001) versus 4.9 +/- 2.3 in IBS (P less than 0.01). 1-14C-lactose breath test was superior to standard lactose tolerance test in specificity (P less than 0.05) and provided a satisfactory correlation between 14C-lactose absorption and lactase assay (r = 0.77). The prevalence of LD in IBS was 40% by the breath test and 35% by lactase assay, suggesting that lactose malabsorption may play a role in the symptoms in the population of some patients with IBS. It appears that 1-14C-lactose breath test is a sensitive, specific and accurate method for the diagnosis of LD in clinical practice and suitable for large scale epidemiological surveys.

Breath Tests