Effects of physalaemin, a vaso-active peptide from amphibian skin, on the excitability of an identifiable molluscan giant neurone of Achatina fulica Férussac.

We examined effects of several vasoactive peptides (substance P, physalaemin, neurotensin, bradykinin, angiotensin etc.) on the excitability of molluscan giant neurones identified in the subesophageal ganglia of Achatina fulica Férussac. Of these peptides, only physalaemin showed a remarkable excitatory effect on a giant tonically autoactive neurone.     

Effects of physalaemin,a vase-active peptide from amphibian skin,on the excitability of an identifiable molluscan giant neurone ofAchatina fulica Férussac

Summary We examined effects of several vasoactive peptides (substance P, physalaemin, neurotensin, bradykinin, angiotensin etc.) on the excitability of molluscan giant neurones identified in the subesophageal ganglia ofAchatina fulica Férussac. Of these peptides, only physalaemin showed a remarkable excitatory effect on a giant tonically autoactive neurone.     

Somatostatin inhibits in vitro release of luteinizing hormone releasing hormone from rat mediobasal hypothalamic slices

Summary Somatostatin, in concentrations ranging from 10^–10 M to 10^–7 M, induces a dose-dependent inhibition of LHRH release from mediobasal hypothalamic slices incubated in vitro. In contrast, VIP, secretin, glucagon, substance P, neurotensin and arginine-vasotocin do not affect spontaneous release of LHRH.     

Substance P-induced fibrinolysis in the forearm of healthy humans

Physiological saline with or without substance P (50 ng/ml) was infused into the humeral artery in 6 healthy males. Indices of fibrinolytic activity (whole blood diluted lysis time, euglobulin lysis time, lysis areas in non-heated fibrin plates produced by plasma or euglobulin precipitate, plasminogen plasma levels, ₂-macroglobulin, Cl-inhibitor, and ₂-antiplasmin) were evaluated in the homolateral antecubital vein before and after 5 min of substance P or saline infusion. After substance P the fibrinolytic activity increased, as can be seen from the shortening of lysis times (p<0.01) and enlargement of the lysis areas (p<0.01). A reduction of plasminogen plasma levels (p<0.01), associated with a decrease in ₂-anitplasmin (p<0.01), was also found. Alpha₂-macroglobulin and Cl-inhibitor were instead unaltered by the peptide. The saline infusion, on the other hand, was unable to modify any of the examined indices. We concluded that exogenous substance P given intra-arterially increases fibrinolytic activity in locally-sampled venous blood through a mechanism which remains to be elucidated.     

Is β-(4-chlorophenyl)-GABA a specific antagonist of substance P on cerebral cortical neurons?

Summary -(4-chlorophenyl)-GABA (Baclofen, Lioresal antagonized the excitant actions of acetylcholine and substance P to comparable extents.L-glutamate-induced excitation was affected to a lesser extent These findings do not support the suggestion that -(4-chlorophenyl)-GABA is a specific substance P antagonist.Acknowledgments. The author is indebted to Ciba-Geigy (Canada) Ltd. for a gift of -COPG.Brenda Robson provided technical assistance. This work was supported by grants from the CanadianM. R. C. and the university of Saskatchewan.     

Interaction and interrelation of P2X7 and P2X4 receptor complexes in mouse lung epithelial cells

P2X4 and P2X7 receptors are ATP-gated ion channels that are co-expressed in alveolar epithelial type I cells. Both receptors are localized to the plasma membrane and partly associated with lipid rafts. Here we report on our study in an alveolar epithelial cell line of the molecular organization of P2X7R and P2X4R receptors and the effect of their knockdown. Native gel electrophoresis reveals three P2X7R complexes of ~430, ~580 and ~760 kDa. The latter two correspond exactly in size to signals of Cav-1, the structural protein of caveolae. Interestingly knockdown of P2rx7 affects protein levels, the intracellular distribution and the supramolecular organization of Cav-1 as well as of P2X4R, which is mainly detected in a complex of ~430 kDa. Our data suggest upregulation of P2X4R as a compensatory mechanism of P2X7R depletion.     

Alfalfa seeds: Effects on cholesterol metabolism

Summary Plasma cholesterol concentrations were reduced in 3 human volunteers during ingestion of diets containing alfalfa seeds (AS) for 3 weeks. No signs of toxicity were detected through serum determinations of multiple parameters. The ingestion of AS in rats decreased the concentration of plasma cholesterol, reduced intestinal absorption of exogenous and endogenous cholesterol, and increased fecal biliary excretion.The work described in this article, Publication No. 1082 of the Oregon Regional Primate Research Center, was supported with grants RR-00163 and HL-16587 from the National Institutes of Health.     

Effects of bromocriptine on plasma testosterone and gonadotropin levels and testicular lipid fractions in adult rats

Summary Bromocriptine treatment of adult male rats resulted in a decrease in testicular testosterone (T) content and a reduction in plasma T levels. This was accompanied by increase in testicular total lipids and cholesterol and depletion of testicular phospholipids.Acknowledgments. We thank Dr R. L. Elton of Sandoz Pharmaceuticals for provision of CB-154. This work was supported by the Rockefeller Foundation, New York, NY, and by NIH grants HD 12642 and P-30 HD 10202 (Radioimmunoassay Core).     

Is beta-(4-chlorophenyl)-GABA a specific antagonist of substance P on cerebral cortical neurons?

Beta-(4-chlorophenyl)-GABA (Baclofen, Lioresal) antagonized the excitant actions of acetylcholine and substance P to comparable extents. L-glutamate-induced excitation was affected to a lesser extent. These findings do not support the suggestion that beta-(4-chlorophenyl)-GABA is a specific substance P antagonist.     

Plasma lecithin: cholesterol acyltransferase activity in high-and low-responding rhesus monkeys

Summary The initial rate of esterification of plasma cholesterol by lecithin: cholesterol acyltransferase (LCAT) was measured in high- and low-responding rhesus monkeys fed a moderately high cholesterol (0.15 mg/kcal) diet. The results show that the rate of esterification of cholesterol in the plasma of the high-responders was significantly (p<0.025) higher than that of the low-responding animals. In view of known relationships between LCAT activity and plasma lipoprotein metabolism, it is suggested that the lipoprotein metabolism in the high-responders would differ from that in the low-responders.Acknowledgments. Financed by USPHS research grant HL-08974 from the National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, Md.     

Lack of platelet factor-3 activation after incubation of platelet-rich plasma with kaolin in the rat.

Stypven times, measured in rat platelet-rich plasma (P.R.P.) after incubation with kaolin, did not shorten as incubation proceeded, thus reflecting the lack of development of platelet factor-3 (PF3) availability in this test system. Repeated freezing and thawing of P.R.P. or aggregation with collegan did result in PF-3 availability. Aggregation and PF-3 availability were inhibited by the compound VK774. These findings add another aspect to the list of species differences in platelet function.     

Lack of platelet factor-3 activation after incubation of platelet-rich plasma with kaolin in the rat

Summary Stypven times, measured in rat platelet-rich plasma (P. R. P.) after incubation with kaolin, did not shorten as incubation proceeded, thus reflecting the lack of development of platelet factor-3 (PF₃) availability in this test system. Repeated freezing and thawing of P. R. P. or aggregation with collagen did result in PF-3 availability. Aggregation and PF-3 availability were inhibited by the compound VK774. These findings add another aspect to the list of species differences in platelet function.This study was partly supported by a grant from I. W. O. N. L.     

Comparison of rat,mouse, guinea-pig and human slow reacting substance of anaphylaxis (SRS-A)

Summary Slow reacting substance of anaphylaxis obtained from rat, mouse, guinea-pig and human tissues have exhibited similar biological activity and have reacted in the same way to chemical and enzymatic treatments. It is concluded that they appear to be the same substance or a similar class of compounds.Acknowledgments. The authors are grateful to Le Conseil de Recherche en Santé du Québec and to Fisons Ltd for financial support. This work was also partly supported by a grant to the late Dr R. P. Orange from the Medical Research Council of Canada (MT-4605).     

Role of the ubiquitin–proteasome system in the regulation of P2Y13 receptor expression: impact on hepatic HDL uptake

The protective effect of high density lipoproteins (HDL) against atherosclerosis is mainly attributed to their capacity to transport excess cholesterol from peripheral tissues back to the liver for further elimination into the bile, a process called reverse cholesterol transport (RCT). Recently, the importance of the P2Y₁₃ receptor (P2Y₁₃-R) was highlighted in HDL metabolism since HDL uptake by the liver was decreased in P2Y₁₃-R deficient mice, which translated into impaired RCT. Here, we investigated for the first time the molecular mechanisms regulating cell surface expression of P2Y₁₃-R. When transiently expressed, P2Y₁₃-R was mainly detected in the endoplasmic reticulum (ER) and strongly subjected to proteasome degradation while its homologous P2Y₁₂ receptor (P2Y₁₂-R) was efficiently targeted to the plasma membrane. We observed an inverse correlation between cell surface expression and ubiquitination level of P2Y₁₃-R in the ER, suggesting a close link between ubiquitination of P2Y₁₃-R and its efficient targeting to the plasma membrane. The C-terminus tail exchange between P2Y₁₃-R and P2Y₁₂-R strongly restored plasma membrane expression of P2Y₁₃-R, suggesting the involvement of the intra-cytoplasmic tail of P2Y₁₃-R in expression defect. Accordingly, proteasomal inhibition increased plasma membrane expression of functionally active P2Y₁₃-R in hepatocytes, and consequently stimulated P2Y₁₃-R-mediated HDL endocytosis. Importantly, proteasomal inhibition strongly potentiated HDL hepatic uptake (>200 %) in wild-type but not in P2Y₁₃-R-deficient mice, thus reinforcing the role of P2Y₁₃-R expression in regulating HDL metabolism. Therefore, specific inhibition of the ubiquitin–proteasome system might be a novel powerful HDL therapy to enhance P2Y₁₃-R expression and consequently promote the overall RCT.     

One lipid, multiple functions: how various pools of PI(4,5)P2 are created in the plasma membrane

Phosphatidylinositol 4,5-bisphosphate [PI(4,5)P₂] is a minor lipid of the inner leaflet of the plasma membrane that controls the activity of numerous proteins and serves as a source of second messengers. This multifunctionality of PI(4,5)P₂ relies on mechanisms ensuring transient appearance of PI(4,5)P₂ clusters in the plasma membrane. One such mechanism involves phosphorylation of PI(4)P to PI(4,5)P₂ by the type I phosphatidylinositol-4-phosphate 5-kinases (PIP5KI) at discrete membrane locations coupled with PI(4)P delivery/synthesis at the plasma membrane. Simultaneously, both PI(4)P and PI(4,5)P₂ participate in anchoring PIP5KI at the plasma membrane via electrostatic bonds. PIP5KI isoforms are also selectively recruited and activated at the plasma membrane by Rac1, talin, or AP-2 to generate PI(4,5)P₂ in ruffles and lamellipodia, focal contacts, and clathrin-coated pits. In addition, PI(4,5)P₂ can accumulate at sphingolipid/cholesterol-based rafts following activation of distinct membrane receptors or be sequestered in a reversible manner due to electrostatic constrains posed by proteins like MARCKS.     

Marked elevation of HDL-cholesterol in cold-adapted golden hamsters

Golden hamsters with spontaneous hypercholesterolemia at 22 degrees C developed a further increase in plasma cholesterol when they were maintained at 6 degrees C. This hypercholesterolemia was associated with a redistribution of plasma cholesterol between VLDL and HDL. Plasma cholesterol transported in the VLDL decreased while cholesterol in the HDL increased by 45%. The LDL profile was not significantly modified.     

3-Hydroxy-3-Methylglutaric acid and triton-induced hyperlipidemia in rats

Summary 3-Hydroxy-3-methylglutaric acid (HMG) significantly decreased cholesterol, triglyceride and phospholipid levels in whole serum, serum -lipoproteins and liver of Triton-induced hyperlipidemic rats. Therapeutically 50 mg HMG/kg is equivalent to 200 mg nicotinic acid/kg in lowering all these lipid parameters. HMG may exert its hypolipidemic effect through inhibition of lipoprotein synthesis.Acknowledgments. We are grateful to Dr.M. Saleemuddin for encouragement, Dr.P. E. Schurr, Upjohn Co. USA for generous gift of Triton W. R 1339 and Lady Tata Memorial Trust (India) for financial assistance to one of us (SYKY).     

Marked elevation of HDL-cholesterol in cold-adapted golden hamsters

Summary Golden hamsters with spontaneous hypercholesterolemia at 22°C developed a further increase in plasma cholesterol when they were maintained at 6°C. This hypercholesterolemia was associated with a redistribution of plasma cholesterol between VLDL and HDL. Plasma cholesterol transported in the VLDL decreased while cholesterol in the HDL increased by 45%. The LDL profile was not significantly modified.     

Tangier disease: still more questions than answers

High-density lipoproteins (HDLs) play a central role in transporting cholesterol from peripheral tissues to the liver for elimination from the body. Impairment of HDL-mediated cholesterol transport favors cholesterol deposition in the arterial wall and promotes development of arteriosclerosis. Tangier disease is a severe HDL deficiency syndrome characterized by the accumulation of cholesterol in tissue macrophages and prevalent atherosclerosis. A three-decade search for a culprit in Tangier disease led to the identification of mutations in a cell membrane protein called ABCA1, which mediates the secretion of excess cholesterol from cells into the HDL metabolic pathway. Because of its ability to deplete cells of cholesterol and to raise plasma HDL levels, ABCA1 has become a promising therapeutic target for preventing cardiovascular disease.     

Dissociation of obesity,hypercholesterolemia and diabetes from atherosclerosis in ob/ob mice

Summary Genetically obese, diabetic and hypercholesterolemic C57BL/6J-ob/ob mice were placed on Purina Laboratory Chow containing 2% cholesterol for up to 4 months. They developed higher plasma cholesterol levels and accumulated an increased quantity of cholesterol in the liver but failed to develop atherosclerotic lesions in the aorta as would be expected in an obese, diabetic and hypercholesterolemic human adult.Acknowledgments. We wish to thank Mr. Willis M. Overton for his excellent technical assistance.