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At least 37 records · Page 2Linked to original sources
PubMed1975

Radio-enzymic assay of oxo steroids by their reduction with the tritiated form of reduced nicotinamide--adenine dinucleotide.

A method of analysis is proposed based on the enzyme-catalysed transfer of tritium from [3H]NADH to suitable substrates. Its practicability is demonstrated on the examples of oestrone and progesterone with the respective use of the 3 beta, 17 beta- and 3 alpha, 20 beta-hydroxysteroid oxidoreductase. Specificity is tested by application to the analysis of the plasma of pregnant women and measurement of the 3H/14C ratios on purification of the enzymic reduction products.

Estrone↗
PubMed1975

Hormone and enzyme assays in pregnancy. V. A rapid method for measuring the placental cystine-aminopeptidase using 1-cystine-bis-1-cystine-bis-p-nitroanilide-nitroanilide as substrate.

A rapid and simple method for determination of the placental cystine-aminopeptidase (P-CAP) activity in plasma is presented. The enzyme-catalysed hydrolysis of the substrate, 1-cystine-bis-p-nitroanilide, is followed in a spectrophotometer by reading the absorbance of the product p-nitroaniline at 380 nm. The absorbance increases linearily with time after a lag-period of 30 seconds to 5 min. The reaction is followed for about 10 min and the increase in absorbtion per min is calculated from the linear part of the absorbtion curve. The test could also be performed by taking 2 readings at about 5 min intervals. In the studies of the enzyme kinetics a competitive inhibitor of the reaction was found in plasma. The amount of the inhibitory substances seemed to be nearly constant in the plasma samples studied from normal pregnant women as well as in plasma samples with high enzyme activity (twin-pregnancy) and low enzyme activity (severe pre-eclampsia). The normal enzyme activity pattern showed an increase from about 210 days of pregnancy and towards the term. A coefficient of correlation of plus 0.83 with the more time consuming method using 1-cystine-di-beta-naphtylamide as substrate was found. It was concluded that the present method could replace the method described by Babuna & Yenen (1966) at least from about 210 days of pregnancy and where a rapid answer is necessary.

Adult↗
PubMed1976

Late-onset acid maltase deficiency. Detection of patients and heterozygotes by urinary enzyme assay.

Daily urinary excretion of acid maltase (12.78 +/- 2.10 units/24 hr/mg of creatinine, in 11 normal adults) was significantly decreased in ten patients with late-onset acid maltase deficiency (1.33 +/- 0.16 units/24 hr; P less than .001) and 11 heterozygotes (3.27 +/- 0.62 units/24 hr; P less than .001). Maximal inhibition of urinary acid maltase activity by antibodies against human placental enzyme was 53% in controls, 30% in heterozygotes, and virtually absent in patients. Investigation of pH curves and enzyme inhibition by antibodies confirmed the presence in the kidney of an immunologically distinct "extra" maltase enzyme active at acid pH. Whether acid maltase in normal urine originates in the kidney or cells of the lower urinary tract, the enzyme defect seems to be expressed in these cells in late-onset acid maltase deficiency.

Adolescent↗
PubMed1979

Enzymic assay for serum theophylline.

We describe an original procedure for determination of theophylline in serum. The drug is extracted from 0.4 mL of serum at pH 7.4 with chloroform/isopropanol (20/1 by vol) and back-extracted into sodium hydroxide (1 mmol/L). The inhibition of beef-liver alkaline phosphatases by theophylline in this alkaline phase is measured at 25 degrees C, with p-nitrophenyl phosphate as substrate, in 2-amino-2-methyl-1-propanol buffer, pH 9.4- The reciprocal of enzyme activity and theophylline concentration are linearly related in the range 2 to 60 mg/L. The maximum interference to be expected from 3-methylxanthine would increase apparent theophylline concentration by no more than 1 mg/L. The method is accurate, free of interference by other xanthines and often-coadministered drugs, and results correlate well with those by the immunoenzymic assay. Major advantages are reagent stability, low cost, and simplicity of instrumentation.

Alkaline Phosphatase↗
PubMed1977

Viral hepatitis: enzyme assays and serologic procedures in the study of an epidemic.

An epidemic of viral hepatitis beginning in late 1975 in a residence for multiply handicapped children, recognized very early in its course, was investigated prospectively to permit comparison of enzymatic and serologic tests. Thirty-three residents of the institution and 46 full- and part-time employees were studied by the immune adherence hemagglutination procedure for antibody (anti-HAV) to hepatitis A virus (HAV). Of these, 31 residents and 37 staff members were susceptible at the beginning of the epidemic. Nineteen and six, respectively, had anti-HAV seroconversion indicating HAV infection. Thus, 12 children (39%) and 31 staff members (81%) of presumed susceptibles did not have serologic evidence of infection. The subclinical/clinical ratio for the children was 1.1:1; for personnel, it was 1:1. Serum alanine aminotransferase (ALT) levels compatible with viral hepatitis occurred in 21 persons (84%) who had anti-HAV seroconversion; conversely, there were 10 persons who had ALT abnormality without detectable anti-HAV in late specimens among the total of 68 susceptibles. There was no evidence the latter could be attributed to hepatitis B virus infection; therefore, they may represent the endemic occurrence of non-A, non-B agent(s).

Adolescent↗
PubMed1978

The null allele in the horse esterase (Es) system detected by enzyme assay and rocket immunoelectrophoresis in heterozygous animals.

The detection of the recessive null allele of horse serum esterase (Es) is possible in heterozygotes Es+/EsO which by starch gel electrophoresis appear like homozygotes Es+/Es+. Two methods are proposed, the titration of enzymatic activity of esterase and the immunochemical titration of esterase as antigen. These methods can be applied to solve the cases of suspect parentage or in population studies.

Alleles↗