A relatively simple method for purification of human platelet-derived growth factor (PDGF) was developed. PDGF was purified from clinically outdated, platelet-rich plasma by means of freezing and thawing extraction and successive chromatography on CM-Sephadex, Sephacryl S-200, and Phenyl Sepharose. Further purification of the PDGF obtained showed two silver-stained bands following polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS-PAGE). Amino acid sequence analysis of these two components separated by SDS-PAGE demonstrated that the sequences coincided with those of PDGF A and B chains previously reported and predicted from the nucleotide sequences of the cloned cDNA's of PDGF A and B chain genes. The purified PDGF preparation stimulated a nanogram level of thymidine incorporation into DNA of previously quiescent Balb/c 3T3 cells.
The effects of feeding of an experimental diet consisting of 16 and 32% of gari by weight were compared with those of the gari-free, control diet in rats. The experiment was carried out for a period of six months during which two rats from each group (A, control; B, 16% gari incorporation; and C, 32% gari incorporation) were killed for histological study. Histological examination of the aorta, liver, and kidneys revealed degenerative changes in group B and C rats, as evidenced by vacuolation and accumulation of periportal fat. These manifestations started at the third month of the experiment. At the fourth month there was massive degeneration of the arterial wall linings followed by development of fatty streaks in the lumen of the arteries.
Male Wistar rats were fed an atherogenic diet for a period of 14 weeks to investigate possible diet-induced lipid alterations and Na+-K+ ATPase activity in their brain tissue. Total cholesterol and triglyceride levels were found to increase significantly in both serum and brain, while the phospholipid (PL) level was decreased in both. Serum LDL levels were markedly increased, whereas the HDL levels were decreased. The atherogenic diet caused a significant increase in brain sulphatide levels, but the cerebroside level remained unchanged. Increased spontaneous lipid peroxidation in brain tissue was also observed. Na+-K+ ATPase activity was depressed by about 50% in the brain of rats fed the atherogenic diet.
The protective effect of a novel synthetic zinc-carnosine chelate compound, zinc N-(3-aminopropionyl)-L-histidine (Z-103), on the gastric injury induced by burn stress was studied in rats. Gastric lesions were produced by immersion of half of the animal's body into 80°C water for 10s. The increase in total area of the erosions 3h after the stress and the increase in thiobarbituric acid reactants in the gastric mucosa were significantly inhibited by the administration of Z-103 at doses of 30, 100, and 300mg/kg. The decrease in blood flow in the gastric mucosa was not influenced by the treatment with Z-103. These results suggest that the protective effect of Z-103 against the aggravation of burn-induced gastric mucosal injury may be due to its inhibitory effect on lipid peroxidation.
The adrenal capacity to store cholesterol and to secrete corticosterone, androstenedione, and dehydroepiandrosterone-sulphate (DHEA-S) was investigated following long- and short-term adrenocor-ticotropic hormone (ACTH) stimulation of growing male rats kept on a pantothenic acid-deficient (PAD) diet for about two months. After chronic treatment with ACTH1-24, the pantothenic acid-deficient rats revealed a drastic impairment in adrenal cholesterol storage compared with ad libitum and pair-weight fed control groups. However, corticosteroid plasma responses to short-term as well as to prolonged ACTH stimulation were normal or in the case of DHEA-S, even significantly increased in pantothenic acid deficiency. The elevated secretory capacity for this androgen sulphate was also discernible in an exaggerated renal DHEA-S excretion in PAD rats under ACTH1-24 hyperstimulation. According to the results of a further hormonal study involving metyrapone administration, a PAD-associated increase in specific enzyme activities of the post-pregnenolone pathway of androgen synthesis was excluded as the reason for the DHEA-S hypersecretion. The experimental findings are discussed with regard to a PAD-induced tissue CoA decrease and its consequences for adrenal cholesterol metabolism and steroid hormone secretion. A concept is presented that takes into account the functional differences in the zona fasciculata and the zona reticularis of the adrenal cortex in order to explain the phenomenon of the abnormal capacity of rats to secrete DHEA-S in pantothenic acid deficiency.
A rapid method for the isolation of pure low density lipoprotein (LDL) from normal rabbit serum within 6.5h was developed by a combination of discontinuous density gradient ultracentrifugation in a vertical rotor and Fast Protein, Polypeptide, Polynucleotide Liquid Chromatography (FPLC) system with a Superose 6 column. Analyses by FPLC, agarose gel electrophoresis, and sodium dodecyl sulfatepolyacrylamide gel electrophoresis showed that LDL isolated by the present method is free from the contamination of other lipoproteins and serum proteins, while that by the standard method [Hatch, F.T., and Lees, R.S. (1968) Adv. Lipid Res., 6, 1-68] contained very low density lipoprotein (VLDL) and small amounts of proteins such as albumin. Differing from the case of the standard method, significant lipid peroxidation of the sample does not seem to occur in the present method. This method can also be used for the rapid preparation of VLDL and high density lipoprotein.
The relationships of diet, development, and maturation indices to the status of blood iron and folate indicators were studied among 233 adolescent girls aged 10-16 years. Statistically significant associations between total iron intake and hemoglobin, plasma iron, serum ferritin and transferrin saturation were observed after controlling the confounding effect of other parameters. Iron intake provided by meat was positively related to hemoglobin and serum ferritin levels. Moreover, post-menarcheal girls and girls in their growth spurt peak had a lower serum ferritin level than the others. Plasma and erythrocyte folate levels increased with increasing folate intake from vegetables and fruits. Plasma folate level decreased with decreasing folate from meat. Plasma and erythrocyte folate levels decreased with chronologic age independantly of dietary intake. Among this nutritionally vulnerable group, our results specify the effect of factors known to influence iron and folate status.
In order to clarify the relationship between biliary excretion of antibiotics and bile acid, the concentrations of Cefoperazone (CPZ) and bile acid in bile were measured by HPLC in 8 patients, 4 with T-tube drainage and 4 with percutaneous transhepatic biliary drainage. The biliary excretion rate of CPZ in 6h at the time of jaundice release (more than 3 weeks after the biliary drainage had started) was extremely low, 0.03-1.4% of the intravenously injected dose of 1.0g. The biliary excretion of CPZ after the administration of ursodeoxy cholate rose to 1.1 to 28 times the preadministration level. After reopening of the enterohepatic circulation of bile acid by surgical bypass, it was 1.3 to 41 times the preadministration level. Especially the marked increase was observed in the patients without severe hepatic dysfunction. There was a significant correlation between biliary excretion of total bile acid and the total biliary excretion of CPZ in 6h. We conclude that the concentration of biliary bile acid is one of the factors affecting the transfer of injected CPZ to bile. This fact is important with respect to the treatment of biliary infections.
We report a 50-year-old male with homozygous familial hypercholesterolemia. His plasma cholesterol level was over 600mg/dl. He had suffered from ischemic heart disease since age 20. Receptor assay utilizing skin fibroblasts revealed that he was a homozygote for receptornegative familial hypercholesterolemia. In addition, his fibroblasts synthesized no immunoprecipitable low-density lipoprotein receptor. Considering his disease, his life is extraordinarily long; this is, in part, attributable to his dietary habits.
By using a newly developed assay method for lipid peroxides based on their reaction with a leucomethylene blue derivative, we investigated serum lipid peroxides (LPO) in healthy and in diseased subjects. In 116 healthy subjects, the mean LPO level obtained was 1.45±1.37nmol/ml and showed significant correlations with total bilirubin, direct bilirubin, phospholipid, total cholesterol, high density lipoprotein-cholesterol, and low density lipoprotein-cholesterol. In 12 hypertensive patients, the LPO value was significantly higher (2.60±2.08nmol/ml) compared with the healthy control, and LPO showed a significant correlation with serum globulin. In 16 hyperlipidemic subjects, the value was 1.50±0.86nmol/ml and correlated with fibrinogen, white blood cells, and nonesterified fatty acids. In 19 diabetics, the level was not significantly different with that in the healthy control, and correlated with fasting blood sugar, total protein, and urea nitrogen. In 10 patients with liver dysfunction, LPO was 1.60±1.33nmol/ml and correlated with white blood cells, chloride, and nonesterified fatty acids.