[Determination of kallikrein in gel-filtered urine using the chromogenic substrate D-valyl-L-leucyl-L-arginyl-4-nitroanilide].
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Biomedical subjects
Publications and source records attributed to K Jung.
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The incorporation of the stable isotope 15N in plasma proteins and blood cells after oral application of 3 g 15NH4Cl (95 At% 15N) per 70 kg body weight was followed up in 11 patients with ischemic heart disease or peripheral arteriosclerosis and in 7 healthy control subjects. Preliminary results indicate that the turnover of plasma protein, especially fibrin, is elevated in patients with arteriosclerosis. Investigations of platelets intimate a decreased turnover of platelet protein in patients with arteriosclerosis as compared to control subjects. Possible reasons for these alterations are discussed.
The sequence of the incorporation of the stable isotope 15N in plasma protein and fibrinogen has been followed by an 15N labelling test in 11 patients with arteriosclerosis and a control group of 7 persons. The investigation showed a higher turnover of plasma protein and especially of fibrinogen in the patients with arteriosclerosis.
A simple microalbuminuria assay using bromphenol blue/glycine reagent is described. Urine samples were prepared using gel filtration on Sephadex G-50 minicolumns and absorbance was measured at 610 nm 20 s after mixing 10 parts of eluate and 1 part of reagent. The detection limit of this method was 3 mg/l; within-run and between-run precision was between 0.5 and 4.1% for borderline and raised albumin concentrations. The recovery of albumin added to samples was 98.7 +/- 2.5%. Results obtained by this method correlated closely with values obtained by radial immunodiffusion (r = 0.987). The test is cheap (reagent costs about 5 cents) and suitable for the non-specialist laboratory.
Growth of Escherichia coli 044 K 74 in liquid medium of raised osmotic strength was stimulated by exogenous L-carnitine, crotonobetaine and gamma-butyrobetaine, respectively. L-Carnitine was accumulated within the cells in dependence on the salt concentration of the media. Osmotic stress during aerobic or anaerobic growth with glucose triggered the L-carnitine uptake in E. coli 044 K 74 whereas L-carnitine uptake by cells of this organism grown anaerobically on glycerol/fumarate was only slightly modified. Synthesis of the enzymes metabolizing L-carnitine to gamma-butyrobetaine in glycerol/fumarate growing bacteria was found to be completely repressed by high NaCl-concentrations. Together, these results indicate that most likely the L-carnitine metabolization sequence does not play a role in osmoregulation in E. coli 044 K 74.
The uptake of L-carnitine by Escherichia coli 044 K74 which is able to metabolize L-carnitine to gamma-butyrobetaine under anaerobic conditions was studied. The uptake system of E. coli was induced in the presence of L-carnitine or crotonobetaine. The optimum influx of L-carnitine was found to take place at 37 degrees C in phosphate buffer, pH 7.5. On the basis of the temperature dependence of the uptake rate an activation energy of 53.5 kJ/mol was estimated. The transport of L-carnitine was energy dependent. The kinetics of L-carnitine entry followed the Michaelis-Menten relationship yielding a Km value of 5.3 x 10(-4) M and a Vmax of 154 nmol/min.mg protein. The transport system showed considerable structural specificity for trimethylamine carboxylic molecules. The results suggest that E. coli possesses an inducible, active and carrier-mediated uptake system for L-carnitine.
During an ultra-long-distance race (1000 km in 20 days) the influence of running was examined on the enzymes aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (AP), gamma-glutamyl-transferase (GGT), and glutamate dehydrogenase (GLDH) with regard to their release from the liver cells or their induction. Furthermore the liver synthetic capacity was assayed by measuring the enzyme activity of cholinesterase and the concentration of serum albumin during the race. Of the 110 participants, 55 finished the race and only the results of these runners were used in our study. AP increased continuously from day 0 (mean = 102 U/L) to day 19 (mean = 120 U/L). A fivefold increase of AST and a twentyfold increase of CK up to day 3 was followed by a significant decrease towards the end of the race. ALT rose as well up to day 6 from a mean value of 8 U/L to 24 U/L but remained at this level. Surprising was the individual increase of the enzymes GLDH (up to twentyfold) and GGT (up to sixfold) in more than half of the finishers on various days indicating liver cell injuries. The activity of CHE and the concentration of serum albumin decreased during the race, both were significantly correlated.
The role of hypothermic storage of rat kidneys on lipid peroxidation compared to its effect on mitochondrial respiratory parameters has been investigated. Rat kidneys were flushed with cold solutions of isotonic sodium chloride; Euro-Collins'; preservation solution containing histidine buffer, tryptophane, and alpha-ketoglutarate (HTK); or hypertonic citrate and then stored for 20 hr at 4 degrees C. After storage, the endogenous contents of malondialdehyde as well as the chemically (by Fe2+/ascorbic acid) and enzymatically (by Fe3+/ADP/NADPH) induced generation of malondialdehyde were measured in cortical homogenates and partly in mitochondria and microsomes by the thiobarbituric-acid reaction. Compared to the values measured in fresh, unstored kidneys, the levels of malondialdehyde were significantly higher in kidneys preserved in solutions of isotonic sodium chloride or HTK. This stimulating effect of the HTK solution on the generation of lipid peroxidation products could also be established when homogenate was incubated with this solution. Euro-Collins' and hypertonic citrate solution did not change the endogenous contents of malondialdehyde in kidneys during hypothermic storage. Both the chemically and enzymatically induced lipid peroxidation increased after hypothermic storage of kidneys in all solutions investigated. No direct relationship between the contents of malondialdehyde and respiratory mitochondrial parameters was detectable. The results demonstrate that the extent of lipid peroxidation does not correlate with preservation effectiveness.
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A rapid latex agglutination test, Culturette Brand CDT from Marion Laboratories, was evaluated and compared to a tissue culture assay (TCA) and isolation of Clostridium difficile in 380 faecal specimens from 226 patients with clinically suspected Clostridium difficile-associated diarrhoea. The sensitivity and specificity of the latex test compared with the TCA were 83% and 80% respectively, and the positive and negative predictive values were 55% and 94% respectively. In patients with repeated sampling the sensitivity increased to 95%. The latex test may be useful as a screening test for negative specimens in laboratories where TCA is not available, but positive specimens have to be confirmed by TCA.
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Urinary excretion of alanine aminopeptidase, alkaline phosphatase, gamma-glutamyltransferase and N-acetyl-beta-D-glucosaminidase was determined in gel-filtered samples of morning random urine specimens of 442 subjects of various ages (5 days to 58 years). Enzyme excretion related to urinary creatinine (enzyme/creatinine ratio; U/mmol creatinine) significantly decreased with increasing age. Sex-related differences of some enzyme excretions were found in age groups over 6 years. From these investigations, we calculated upper reference intervals (97.5 percentiles) for 5 age-dependent groups of children and adolescents and for one group of adults.
alpha 1-Microglobulin was purified from urine to a purity of 97.7% in a yield of 25.8%, and was used to produce antibodies in sheep. These antibodies, purified by affinity chromatography, were used to develop a rapid one-step and a two-step immunoenzymometric assay (IEMA). The equilibrium in the reaction between solid phase-adsorbed antibodies and antigen and between the antigen and enzyme-labelled antibodies was attained within 30 and 100 min, respectively. The one-step IEMA permits a good differentiation of low alpha 1-microglobulin concentrations after 30 min reaction time. Its detection limit is 0.35 micrograms/l, and its measurement range is between 0.5 and 100 micrograms/l. The IEMA correlates well with radial immunodiffusion (r = 0.973). The mean alpha 1-microglobulin serum concentration in women is insignificantly lower (33.2 mg/l) than in men (36.1 mg/l). In both sexes the alpha 1-microglobulin concentration increases with age. HIV-infected symptomless men have a significantly lower (15.9 mg/l) alpha 1-microglobulin concentration in serum than normal persons, whereas in AIDS patients it is significantly higher (45.5 mg/l).
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The Na-K-ATPase is the enzymatic basis of a energy-dependent transport of sodium and potassium through the cell membrane and is therefore of importance to the survival of organs during periods of decreased energy supply. In the present paper the influence of various preservation solutions on the activity of this enzyme was examined. It was shown that Na-K-ATPase activity in rat kidneys did remain unchanged after 48 hour preservation in various preservation solutions (Bretschneider, Ross, Collins, Sacks, Belzer). Therefore, its estimation is no valuable for evaluation of preservation solutions.
In 37 women with long-term use of oral contraceptives (average use 13.4 years) and 12 women who had not taken oral contraceptives for at least 5 years we determined the serum enzymes ALAT, ASAT, Gamma-GT and conducted the 15N-ammonium test as a marker of partial metabolic performance of the liver. In addition we studied total protein, protein fractions in the serum and haematologic parameter. 18 out of 37 women showed a morbid result in the 15N-ammonium test. In 4 of these 18 women morphological changes representing hepatic damage caused by contraceptives could be detected. On the average ALAT, Gamma-GT and 15N-ammonium test in both groups are clearly different. Because the enzymic levels are being in the normal range, only the 15N ammonium test enables to give a hepatotoxic evidence. It is possible to conclude that long-term use of oral contraceptives influences the liver metabolism.