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Biomedical subjects

G J Cardinale

Publications and source records attributed to G J Cardinale.

28 records · Page 2Linked to original sources

Increased collagen synthesis in blood vessels of hypertensive rats and its reversal by antihypertensive agents.

Collagen synthesis is increased in the aortas, mesenteric arteries, and to a lesser extent, in the hearts of rats either made hypertensive with desoxycorticosterone acetate-salt or that are spontaneously hypertensive. Several markers of collagen biosynthesis were shown to be increased, including prolyl hydroxylase (EC 1.14.11.2; proline, 2-oxoglutarate dioxygenase), prolyl hydroxylase-related antigen, total collagen content, and the incorporation of [(3)H]proline into total protein and into collagen. The antihypertensive agents chlorothiazide and reserpine, when administered before the onset of hypertension in the rats treated with desoxycorticosterone acetate-salt, prevented or diminished the increase in collagen biosynthesis. When reserpine was given after the onset of hypertension, prolyl hydroxylase activity was decreased concomitant with the decrease in blood pressure. Treatment with reserpine is particularly effective in diminishing arterial prolyl hydroxylase activity.

Animals↗

Activation of prolyl hydroxylase in L-929 fibroblasts by ascorbic acid.

The addition of ascorbate to the culture medium of early log-phase mouse fibroblasts (L-929 cells) resulted in a 5-fold increase of prolyl hydroxylase activity. Maximal activity was reached within 2 hr after addition of 5 muM ascorbate. The total amount of enzyme-related antigen did not change on treatment with ascorbate and the activation was shown to be independent of RNA and protein synthesis. The increase in activity caused by ascorbate is therefore due to the activation of a preformed precursor. Enzyme (molecular weight 260,000-300,000) and putative precursor (molecular weight 85,000-105,000) were separated by chromatography on DEAE-Sephadex. Treatment of intact cells with dithiothreitol resulted in an almost quantitative conversion of the enzyme to the smaller inactive protein. When these cells were treated with ascorbate or incubated overnight in fresh medium the enzyme reappeared and precursor concentrations decreased proportionately. Ascorbate may act by bringing about aggregation of enzymatically inactive subunits.

Animals↗

Cell-free synthesis of procollagen: L-929 fibroblasts as a cellular model for dermatosparaxis.

A cell-free system that actively synthesizes collagen was prepared from L-929 fibroblasts. Chromatographic and electrophoretic techniques were used to demonstrate that the only collagenous products are pro alpha(1) and pro alpha(2) chains. The collagen synthesized by the cell-free system was also compared to the collagen extracted from the cells. The cellular collagen was composed of aggregates of pro-alpha chains, while no alpha chains were found. Procollagen peptidase activity could not be detected in the cells, and the activity present in the medium was low, comparable to that in dermatosparaxic cell cultures. These properties indicate that L-929 cells may be a model system for dermatosparaxis.

Animals↗

Congenital methylmalonic acidemia: enzymatic evidence for two forms of the disease.

Methylmalonic acidemia is an inherited metabolic disorder thus far found in children and characterized by the excessive excretion of methylmalonate in the urine. Typically these children exhibit vomiting, lethargy, ketoacidosis, and failure to grow. Many of the patients are mentally retarded and die early in life. Two variants of this disease are known. In one, the administration of vitamin B(12) will reverse or prevent these clinical findings, whereas in a second variant vitamin B(12) therapy is of no value. This paper presents the first enzymatic evidence (obtained with cell-free liver extracts) that bears on two important aspects of the disease. It has been found that methylmalonylCoA carbonylmutase activity is essentially absent in the livers of patients suffering from one variant (vitamin B(12)-unresponsive) of the disease. Secondly, it has been found that the livers of patients with the second variant (vitamin B(12)-responsive) of the disease show normal enzymatic behavior in the presence of the coenzyme form of vitamin B(12), but are identical to the vitamin B(12)-unresponsive variant in the absence of the added coenzyme. Thus the enzyme studies fully support the clinical observations that two types of this disease exist.

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