Mitochondrial creatine kinase from human heart muscle: purification and characterization of the crystallized isoenzyme.

An isoenzyme of creatine kinase [ATP: creatine N-phosphotransferase, EC 2.7.3.2] was isolated in pure, crystalline form from mitochondria of human heart muscle. The enzyme has a molecular weight of 84,000 and consists of two subunits with identical molecular weight, each containing two reactive sulfhydryl groups. The enzyme has a specific activity of 15 +/- 2 U/mg in the direction of creatine phosphate synthesis at optimum pH of 8.7 and of 45 +/- 5 U/mg in the direction of ATP synthesis at optimum pH of 6.7. Mitochondrial creatine kinase has Michaelis constants of 1.70 mM for MgATP2-, 8.00 mM for creatine, 0.15 mM for MgADP-, and 3.00 mM for creatine phosphate. The mitochondrial enzyme differs from the other creatine kinase isoenzymes, i.e. from muscle (CK-MM), brain (CK-BB), and their hybrid form (CK-MB) in (I) its amino acid composition and its amino terminal amino acid sequence, (II) its electrophoretic mobility, and (III) its immunological properties. It thus constitutes a fourth isoenzyme of creatine kinase. By analogy to CK-MM, CK-BB, and CK-MB, the mitochondrial isoenzyme was designated as CK-MiMi.

Physical performance and serum cholesterol fractions in healthy young men.

In 293 healthy, 18--30-year-old male adults (241 well trained athletes with different training types: (A) endurance, (B) mixed, and (C) power training; (D) 52 untrained students), serum total cholesterol and the cholesterol fractions (HDL-, VLDL-, LDL-cholesterol) were investigated. In comparison with the untrained students, lower LDL- and VLDL-cholesterol concentrations were found in group (A) (-0.23 mmol/l, p < 0.01; -0.08 mmol/l, p < 0.001), lower HDL-cholesterol concentrations were found in group (C) (-0.33 mmol/l, p < 0.001). Increased HDL-cholesterol values and decreased VLDL- and LDL-cholesterol values were found in athletes with a rising oxygen-uptake; a significant, age-related increase in HDL-cholesterol occurred only in endurance-trained persons. There was no significant correlation between relative body weight and the HDL-cholesterol in the subjects examined.

Serumlipoprotein cholesterol in sedentary and trained male patients with coronary heart disease.

To demonstrate the influence of exercise training on the lipoprotein cholesterol fractions (high-density lipoprotein (HDL), low-density lipoprotein (LDL), and very low-density lipoprotein (VLDL) cholesterol) in patients with coronary heart disease (CHD), 65 male patients were examined for cardiovascular function and lipid metabolism, before starting or after having participated in a coronary training group for more than one year. There were notable improvements not only in the physical performance data of the trained patients in reference to the tested maximal performance capacity per body weight (MPC/W) as well as the calculated heart volume performance ratio (HVPR) (MPC/W, + 26.0%; HVPR, -18.0%), but also in the lipoprotein cholesterol values and the calculated risk quotient of HDL and total cholesterol (HDL cholesterol, +5.1 mg/dl; LDL cholesterol, -23.3 mg/dl; VLDL cholesterol, -12.5 mg/dl; HDL/total cholesterol, +0.038). The results indicate the positive effect of physical training on lipoprotein cholesterol concentrations, and also in CHD patients in coronary training groups.

Exploitation of plant cells for the production of natural compounds.

A four-step strategy is presented which allows the establishment of plant cell cultures producing high yields of secondary plant products. The application of suitable methods (radioimmunoassay, fluorescence screening) for the selection of overproducing differentiated plants and cell colonies is stressed. By precursor feeding and hormone application, plant cell cultures can greatly be influenced in their production behavior. A highly sensitive, selective regulatory mechanism for the uptake and storage of alkaloids in Catharanthus vacuoles is demonstrated. Overproducing variant cell strains are so far the most promising tool for the future biotechnological application of the plant cell culture method.

The appearance of D-galactosamine-induced hepatitis and generalized edema in adrenalectomized rats.

In adrenalectomized female rats a single dose of 375 mg D-galactosamine.HCl per kg of body weight produces both hepatitis and generalized edema with ascites. These alterations depend upon the dose and the time interval after injection of the aminosugar. The effect is specific for D-galactosamine; 2-deoxy-galactose produces only edema and no hepatitis, whereas D-glucosamine and D-galactose are without any measurable effect. In male adrenalectomized animals D-galactosamine produced hepatitis alone; fluid extravasation occurs only after additional orchiectomy. Glucocorticoids given before or simultaneously with D-galactosamine are able to prevent the animals from gettin edema and to ameliorate hepatitis, while mineralcorticoids do not show any effect on these alterations. It is evident that the effects of D-galactosamine on the hepatocyte and on the endothelial cells are independent from each other. This leads to the conclusion that D-galactosamine acts at least upon two different target organs, the liver and the reticulo-endothelial system in general.

[Anabolic steroids: damages, effect on performance, and on metabolism (author's transl)]. / Anabole Hormone: Schädigung, Leistungsfähigkeit und Stoffwechsel

10 normal persons (age: x=22 years) and 15 weight lifters (age: x=27 years) were studied before and three months after taking Nandrolone danoate (ND). Strength exercises, hear rate during ergometer work, physical working capacity and certain biochemical variables were measured in the experimental group, as well as in a controll group of 7 weight lifters. Six weight lifters in the experimental group who had been taking ND for at least 3 years were also studied to determine whether there were any deleterious effects on their health. In addition, 75 athletes who had been taking anabolic steroids were investigated to determine the possible effects. It was found that: 1. In spite of an 7% increase in performance, the maximal heart rate was not higher when the subjects took ND. During submaximal work loads the heart rate (p less than 0.025) and the blood lactate level (p less than 0.05) were lower. During physical work, there was essentially no effect of ND on glucose or total lipids in serum. 2. The results from the three-months study, from the weight lifters taking ND for 3 years, as well as from 26 of the 57 athletes who had been taking ND showed no evidence of a deleterious effect of ND (based on 26 biochemical measurements). It would appear, therefore, that the general suggestion of a detrimental effect of anabolic hormones is not justified. 3. Damages or functional disturbances were found in 31 athletes and 3 weight lifters of the experimental group after oral application of anabolic alkyl-steroids. After a period of time without alkyl-steroid administration, all investigated biochemical parameters returned to normal levels; thus it appears that the aforementioned pathological findings are reversible changes in liver function.

[Controlled desferrioxamine treatment of congenital anaemia and transfusion siderosis (author's transl)]. / Kontrollierte Desferrioxamin-Therapie bei kongenitaler Anämie und Transfusionssiderose.

A 19-year-old patient with congenital Blackfan-Diamond anaemia has been maintained on a regular red cell transfusion schedule since he was two months old. From the age of four he has received desferrioxamine injections at regular intervals with different dose levels. In spite of the treatment with the chelating agent secondary siderosis developed with typical endocrinological abnormalities and cardiac arrhythmias. Increasing the dose of desferrioxamine to 16 g/24 h resulted in an iron excretion of 184 mg/24 h. Such an intensification of treatment finally produced a negative iron balance and the cardiac arrhythmias disappeared. Desferrioxamine therapy should be done under controlled circumstances only, an iron balance is mandatory, and the dose should be adjusted to the results of the balance.

The Site of the Inhibition of the Shikimate Pathway by Glyphosate: II. INTERFERENCE OF GLYPHOSATE WITH CHORISMATE FORMATION IN VIVO AND IN VITRO.

In the presence of the nonselective herbicide glyphosate (N-[phosphonomethyl]glycine), buckwheat (Fagopyrum esculentum Moench) hypocotyls and cultured cells of Galium mollugo L. accumulate an organic acid, which was identified as shikimate by mass-spectroscopy of its methyl ester. After growth in 0.5 millimolar glyphosate for 10 days, G. mollugo cells contained shikimate in amounts of up to 10% of their dry weight. Synthesis of chorismate-derived anthraquinones in G. mollugo was blocked by glyphosate. Chorismate and o-succinylbenzoate (an anthraquinone precursor) alleviated the inhibition. The conclusion drawn from these experiments, that glyphosate inhibits a step in the biosynthetic sequence from shikimate to chorismate, was substantiated by the finding that glyphosate is a powerful inhibitor of the conversion of shikimate to chorismate in cell-free extracts from Aerobacter aerogenes 62-1.