Prediction of coronary heart disease based on clinical suspicion, age, total cholesterol, and triglyceride.

Our results, based on the definition of coronary heart disease by cinearteriography, show that definite relationships exist among incidence of coronary heart disease (CHD), age, total cholesterol (TC), and total triglycerides, with less definite ones between free cholesterol, and phospholipids for a specific group of 450 male patients referred to the Cleveland Clinic because of suspected coronary heart disease. Equations and graphs of the association of incidence of CHD with these conditions are presented. The relationships were strong enough to be of aid in discriminating CHD from normal; the most useful being age, TC, and TG. Incidence determinations were presented in terms of total incidence and incidence associated with the variable itself. Coronary angiography demonstrated 350 CHD and 100 normals or a ratio of 3.5 to 1. This ratio is a reflection of the high accuracy with which physicians suspect CHD. Age and TC were good discriminators in these patients suspected of CHD. An improved relationship and better discrimination was obtained by relating incidence simultaneously to age, TC, and TG. The probability of having CHD is the total incidence corresponding to the patient's age, TC, and TG. The probability level of 0.90 gave 95% assurance of a correct prediction for the physician-selected group of patients. Twenty-nine per cent of them had probabilities above 0.90.

The effect of relaxin on collagen metabolism in the nonpregnant rat pubic symphysis: the influence of estrogen and progesterone in regulating relaxin activity.

The aim of this study was to examine the effects of relaxin on collagen content, solubility, and composition in the rat pubic symphysis. Nonpregnant, female Sprague-Dawley rats were bilaterally ovariectomized and either unprimed or primed with estrogen or progesterone alone, or a combination of estrogen and progesterone. One week later these animals were given increasing doses of a synthetic human (gene-2) relaxin (0-100 micrograms) before being killed 16 h later. Their pubic symphysial tissues were then removed and analyzed for collagen content and solubility, whereas collagen composition was determined by SDS-PAGE. Relaxin administration significantly increased the length (140 +/- 6%) and weight (170 +/- 9%) of the interpubic fibrocartilage in estrogen-primed rats (n = 15). At the same time, it decreased the total collagen content by 68 +/- 6%, without altering the proportions of collagen types, which were predominantly type I (85%) and type II collagen (15%). Relaxin administered alone reduced the total collagen content by 64 +/- 4% but had no effect on collagen solubility or composition. Progesterone abolished the effects of relaxin in estrogen-primed rats. It is concluded that relaxin has a potent effect on the amount of collagen in the rat pubic symphysis that is enhanced by estrogen and antagonized by progesterone. The changes in the extracellular matrix within the pubic symphysis induced by relaxin may be important in the modifications that this tissue undergoes during pregnancy.

Prolonged low-dose dexamethasone, in early gestation, has no long-term deleterious effect on normal ovine fetuses.

Low-dose dexamethasone (D) treatment is used in pregnancies where the fetus is suspected to be at risk of congenital/virilizing adrenal hyperplasia. To study if this treatment had any immediate or long-term effects in normal fetuses, pregnant ewes were treated with D (20 microg/kg maternal body weight x d) or saline (S), from d 25-45 of gestation. Tissue was collected from fetuses killed at 45 d (S = 6; D = 8), 130 d (S = 8; D = 8), or lambs at 2 months of age (S = 6; D = 6) and mRNA levels measured using real-time PCR. D treatment reduced adrenal wt at 45 d (S, 12.2 +/- 0.7 mg; D, 6.3 +/- 0.4 mg) and significantly decreased adrenal mRNA for P(450scc). At 130 d, fetuses from the D treatment were growth retarded (S, 3.2 +/- 0.1 kg; D, 2.5 +/- 0.1 g), but the adrenals were appropriate for the body weight. mRNA levels of angiotensinogen, the AT(1) receptor and mineralocorticoid receptor (MR) and GR were similar in kidney and brain (hypothalamus, hippocampus, medulla oblongata) except for hippocampal expression of MR and GR, which was significantly decreased by D treatment. By 2 months, BW and hippocampal MR and GR mRNA levels were similar, and lambs were normotensive (S, 83 +/- 3 mm Hg; D, 78 +/- 3 mm Hg). Thus, there were no persistent, long-term effects of prolonged low-dose D treatment in normal ovine fetuses.

Autoimmune hyperlipidemia in a patient. Atherosclerotic course and chaning immunoglobulin pattern during 21 years of study.

A 48 year old male patient presented with xanthomatosis, hyperbeta lipoproteinemia and hyper-IgA globulinemia; these two serum components occurred as a "complex." The patient has subsequently been studied for 22 years (1952 to 1974). His serum cholesterol and triglyceride levels have been consistently and excessively high despite efforts to regulate them by means of diet or diet and drugs. Serum immunoglobulin A (IgA) concentration ranged from 1,400 to 3,400 mg/dl compared with a normal value of 156 plus or minus 92 mg/dl. The metabolism of lipoproteins, judged by vitamin A turnover studies was slow. Peripheral atherosclerosis became evident 15 years after beginning the study whereas cinecoronary arteriography concurrently demonstrated only minimum changes. Xanthomas exhibited marked regression only during the last 6 years, after 16 years of diet and the addition of clofibrate for 7 years. Beta lipoprotein and IgA globulin determined by immunofluorescent and immunoelectrophoretic technics were demonstrated in the atherosclerotic material obtained from the patient's arterial wall. They were also found in the plasma cells of the bone marrow. The IgA globulin-beta lipoprotein complex in the serum was broken with difficulty. The patient's isolated IgA globulin, free of lipoprotein, formed a firm complex when mixed with beta lipoprotein prepared from normal human serum. Initially, IgA globulin studies showed presence of both kappa and lambda light chains in normal proportion. But after 18 years, the IgA globulin has become monoclonal, type lambda. The plasma cells of the bone marrow have become progressively more atypical and immature. No clinical indications of multiple myeloma have been found. It is concluded that association of lipoproteins with IgA globulin in the serum of this patient with hyperlipidemia, hyper-IgA globulinemia did not prevent the development of atherosclerotic lesions and the deposition of lipids and lipoproteins in the plaques. It is possible that the lipoprotein-immunoglobulin association may have retarded the process, since it became manifest only after many years of known hyperlipidemia.

Dopaminergic modulation of aldosterone secretion?

Metoclopramide (10 mg i.v. injection followed by 10 mg/h i.v. for 2 h) caused a transient rise in blood concentrations of aldosterone in sodium-replete and sodium-depleted sheep. Infusion of metoclopramide into the adrenal artery of sheep with an autotransplanted adrenal gland, at a rate to give a similar concentration of metoclopramide at the adrenal cell level (calculated from rate of infusion and adrenal blood flow), resulted in no alteration in aldosterone secretion rate in either sodium-replete or sodium-deplete animals, even though intravenous metoclopramide caused transient stimulation of aldosterone secretion in the same sheep when sodium replete. Dopamine administered either into the adrenal arterial blood supply or intravenously had no significant effect on aldosterone secretion and did not reverse the stimulatory effects of angiotensin II on aldosterone secretion in the adrenal transplant. The data do not support the suggestion that direct dopaminergic elements play a tonic inhibitory role in aldosterone secretion. It is possible that the agonist effect of metoclopramide on aldosterone secretion may occur by some non-dopaminergic mechanism and it is tempting to speculate that the effect is centrally mediated.

Structure and expression of Leydig insulin-like peptide mRNA in the sheep.

We have used PCR to isolate and characterise Leydig insulin-like peptide (Ley I-L) mRNA from sheep ovary. The deduced amino acid sequence of sheep Ley I-L has good homology with the pig and human peptides, having 93% and 77% amino acid identity, respectively. Northern blot analysis revealed abundant expression in both ovary and testis. An examination of ovarian RNA from non-pregnant and pregnant sheep showed that pregnancy did not significantly increase Ley I-L mRNA levels. However mRNA levels did alter depending on whether ovaries contained a corpus luteum. Also ovaries were examined by hybridisation histochemistry to locate the site of expression. Abundant Ley I-L mRNA levels were found in the theca interna cells of the ovary.

Studies on stanniocalcin: characterization of bioactive and antigenic domains of the hormone.

Stanniocalcin (STC) decreases branchial Ca(2+)-uptake in fish. In order to determine its bioactive domain, synthetic fragments (U amino acids (aa) 1-20; V aa 103-136; W aa 202-231) of eel STC were tested for their effect on Ca2+ uptake in tilapia (Oreochromis mossambicus). Ca2+ uptake was inhibited by an N-terminal fragment but not by a midfragment nor a C-terminal fragment of the mature hormone. We provide theoretical and experimental evidence that a midportion of STC, which is included in the synthetic fragment V, is the most antigenic site of the molecule. Polyclonal antibodies against stanniocalcin are directed against this midportion although this region of STC appears not to be essential for signal transduction. These results suggest that the currently available antibodies will recognize inactive STC fragments in the circulation. We conclude that the bioactive portion of STC does not correspond with the major antigenic portion of the hormone. The results imply that studies on plasma STC levels employing a polyclonal antiserum against STC should be interpreted with care.

Ontogeny and regulation of the AT1 and AT2 receptors in the ovine fetal adrenal gland.

The expression and regulation of the receptors for angiotensin II (both AT1 and AT2) were examined in the ovine fetal adrenal gland by RNase protection assay (RPA), in situ hybridisation histochemistry, immunohistochemistry and Western blotting. Both mRNA and protein for the AT1 receptor were present in the zona glomerulosa and zona fasciculata of the cortex, but not in the medulla, from as early as these zonas were distinguishable (60 days of gestation; term is 145-150 days), and even present in the steroidogenic cells of the unzoned gland at 40 days. The mRNA for the AT2 receptor was present in the same locations (but never in the medulla) from 40-130 days, and declined to extremely low levels after 140 days. The infusion of ang II, 1 microg/h, for 3 days, at mid-gestation (76 +/- 2 days) caused a significant decrease in mRNA for AT1 but no change in AT2 levels. Thus, the biologically active receptor (in terms of aldosterone stimulation) is present in the ovine fetal adrenal from very early in development, and can be down-regulated by mid-gestation.

Purification and cloning of a corpuscles of Stannius protein from Anguilla australis.

The kidneys of teleost fish are associated with tissues containing secretory granules--the corpuscles of Stannius (CS). Electron microscopy indicates that the granules are of a proteinaceous nature and may represent hormones or enzymes of unrecognized physiological and biochemical function. In the present study, two-dimensional gel electrophoresis and electroelution was used to purify the major protein to homogeneity; it is approximately 32,000 Da in the reduced form and glycosylated. From the partial NH2-terminal sequence, a 75-mer oligonucleotide probe was synthesized and used to isolate a cDNA clone from which the complete amino acid sequence of the major CS protein was deduced. Polyclonal antibodies raised against CS homogenates were specific for the CS proteins (confirmed by immunohistochemistry). Hybridization histochemistry was used to confirm the location of the mRNA encoding the isolated protein. Incubation of CS homogenate with eel plasma or ovine renin substrate did not result in any angiotensin-like peptides whereas kidney homogenate did.

Plane xanthoma and multiple myeloma with lipoprotein--paraprotein complexing.

Clinicopathologic findings are reported of a woman with generalized plane xanthoma, multiple myeloma (IgG type K), and hyperlipemia with very high levels of serum cholesterol and triglyceride. Complexing of the serum lipoproteins and immunoglobulins had cryoglobulin properties and was separable by ultracentrifugation. Immunofluorescent studies of skin and bone marrow demonstrated deposits of IgG with low density lipoprotein apoprotein and IgG with beta-lipoprotein, respectively. Although immunosuppressive therapy resulted in return of serum IgG, lipid, and lipoprotein levels to normal, the patient died from the myeloma. Serum lipoprotein-paraprotein complexes have been demonstrated in at least 20 other cases of cutaneous xanthomatosis and myeloma. This interaction may result in an autoimmune hyperlipemia.

Depression, immunocompetence, and prostaglandins of the E series.

Plasma prostaglandin E1 and E2, and quantitative and qualitative measures of immune function, were determined in depressed patients and healthy controls. Prostaglandin E2 was significantly elevated in the depressed group, and prostaglandin E1 showed a trend in the same direction. Lymphocyte stimulation responses, as measured by phytohemagglutinin, concanavalin A, and pokeweed mitogen, were significantly lower in the depressed group. Helper and suppressor T cell percentages did not significantly differ in the two populations. In the depressed group, prostaglandin E1 showed a significant inverse correlation with concanavalin A, and prostaglandin E2 showed a similar trend. These preliminary data suggest prostaglandins of the E series may be related to abnormalities of cellular immunity previously documented in depression.

Quantitative and qualitative lipid correlation in experimental endogenous hyperlipemia.

The reversible endogenous hyperlipemia in dogs, elicited by the detergent Triton which was given intravenously, was used to study the interrelations of serum lipids. In the cholesterol ester fraction an increase occurs in both monounsaturated and in saturated fatty acids, excepting myristic; while a decrease occurs in polyunsaturated fatty acids. The fatty acids of cholesterol esters of normal dogs contain 22% oleic acid, and only 24% when serum lipids are increased to almost double their normal value (TC=400-500 mg/100 ml). However there is a critical level above which a rapid rise in oleic acid occurs and, in severe hyperlipemia (TC=1500 +/-430 mg/100 ml), this acid constitutes almost half of the esterified fatty acid component.Since there is no evidence that Triton directly regulates fatty acid synthesis, the lipid fraction-fatty acid interrelationship may be secondary to lipid mobilization from endogenous sources. This concept is supported by the fact that the increased serum fatty acids are only those which can be synthesized by animals. It is suggested, on the basis of a marked increased of endogenously produced fatty acids, that, at critical lipid levels, shortage of polyunsaturated fatty acids from exogenous sources occurs. This might be of sufficient degree to accelerate fatty acid synthesis to meet the need for fatty acids for energy requirements. There may also be need of fatty acid for esterification of chiefly the accumulated free cholesterol split from lipoprotein by Triton.Triton-induced changes in cholesterol ester fatty acids result in patterns which closely resemble those in the adipose tissue of dog and man and in the serum of human endogenous hyperlipemia.

Effects of experimental endogenous hyperlipemia on circulating leukocytes and erythrocytes.

Effects of hyperlipemia on circulating leukocytes and erythrocytes were studied in dogs which were given repeated, intravenous injections of a nonionic detergent, Triton WR-1339.Erythrocyte lipid concentrations increased from 3.6+/-0.9 mg/10(10) cells in control animals to 9.3+/-1.5 mg in the hyperlipemic group. This increase was accompanied by a shift in the distribution of total fatty acids toward a higher percentage of saturated and monounsaturated acids. In contrast to the changes observed in erythrocytes, the leukocyte lipid content remained unaltered in dogs with serum cholesterol levels ranging from 500 to more than 2,000 mg/100 ml. Leukocyte counts rose whereas hematocrit values, hemoglobin concentrations, and erythrocyte counts decreased. Oxygen utilization studies showed no significant metabolic differences between leukocytes which were isolated from hyperlipemic or control animals. Circulating leukocytes in dogs with an endogenously induced hyperlipemia were shown therefore to maintain normal lipid concentrations and did not participate in lipophage formation, as reported for certain diet-induced lipemias.

[Characteristics of the interaction of adrenal lipoamide dehydrogenase with physiological and quinone electron acceptors]. / Zakonomernosti vzaimodeistviia lipoamiddegidrogenazy nadpochechnikov s fiziologicheskimi i khinonovymi aktseptorami élektronov.

Lipoamide dehydrogenase (EC 1.6.4.3) from the ketoglutarate dehydrogenase complex of adrenals catalyzes the oxidation of NADH by lipoamide and quinone compounds according to the "ping-pong" scheme. The catalytic constants of these reactions are equal to 220 and 24 s-1, respectively (pH 7.0). The maximal quinone reductase activity is observed at pH 5.6, whereas the lipoamide reductase activity changes insignificantly at pH 7.5-5.5. The maximal dihydrolipoamide-NAD+ reductase activity is observed at pH 7.8. The oxidative constants of quinone electron acceptors vary from 6 X 10(6) to 4 X 10(2) M-1 s-1 and increase with their redox potential. The patterns of NAD+ inhibition in the quinone reductase reaction differ from that of lipoamide reductase reaction. The quinones are reduced by lipoamide dehydrogenase in the one-electron mechanism.