Cardiometabolic disease in South Asians: A global health concern in an expanding population.

Cardiovascular disease (CVD) is one of the main causes of mortality and morbidity worldwide. As an emerging population, South Asians (SAs) bear a disproportionately high burden of CVD relative to underlying classical risk factors, partly attributable to a greater prevalence of insulin resistance and diabetes and distinct genetic and epigenetic influences. While the phenotypic distinctions between SAs and other ethnicities in CVD risk are becoming increasingly clear, the biology of these conditions remains an area of active investigation, with emerging studies involving metabolism, genetic variation and epigenetic modifiers (e.g., extracellular RNA). In this review, we describe the current literature on prevalence, prognosis and CVD risk in SAs, and provide a landscape of translational research in this field toward ameliorating CVD risk in SAs.

Glutathione peroxidase potentiates the inhibition of platelet function by S-nitrosothiols.

GSH peroxidase (Px) catalyzes the reduction of lipid hydroperoxides (LOOH), known metabolic products of platelets and vascular cells. Because interactions between these cells are modulated by nitric oxide (NO) and LOOH inactivate NO, we investigated the effect of GSH-Px on the inhibition of platelet function by the naturally occurring S-nitrosothiol, S-nitroso-glutathione (SNO-Glu). Concentrations of SNO-Glu that alone did not inhibit platelet function (subthreshold inhibitory concentrations) were added to platelet-rich plasma together with GSH-Px (0.2-20 U/ml); this led to a dose-dependent inhibition of platelet aggregation with an IC50 of 0.6 U/ml GSH-Px. In the presence of subthreshold inhibitory concentrations of SNO-Glu, the LOOH, 5-hydroperoxy-6,8,11,14-eicosatetraenoic acid, increased platelet aggregation, an effect reversed by GSH-Px. Glutathione and SNO-Glu were equally effective as cosubstrates for GSH-Px. Incubation of SNO-Glu with GSH-Px for 1 min led to a 48.5% decrease in the concentration of SNO-Glu. Incubation of SNO-Glu with serum albumin led to the formation of S-nitroso-albumin, an effect enhanced by GSH-Px. These observations suggest that GSH-Px has two functions: reduction of LOOH, thereby preventing inactivation of NO, and metabolism of SNO-Glu, thereby liberating NO and/or supporting further transnitrosation reactions.

Nitric oxide released from activated platelets inhibits platelet recruitment.

Vessel injury and thrombus formation are the cause of most ischemic coronary syndromes and, in this setting, activated platelets stimulate platelet recruitment to the growing thrombus. Recently, a constitutive nitric oxide synthase (NOS) has been identified in human platelets. To further define the capacity of platelets to produce nitric oxide (NO), as well as to study the role of this NO in platelet recruitment, we adapted a NO-selective microelectrode for use in a standard platelet aggregometer, thereby permitting simultaneous measurement of platelet aggregation and NO production. Treatment of platelets with the NO synthase inhibitor -NG-nitroarginine methyl ester (L-NAME), reduced NO production by 92+/-8% in response to 5 microM ADP compared to control but increased aggregation by only 15+/-2%. In contrast, L-NAME had a more pronounced effect on platelet recruitment as evidenced by a 35+/-5% increase in the extent of aggregation, a 33+/-3% decrease in cyclic GMP content, and a 31+/-5% increase in serotonin release from a second recruitable population of platelets added to stimulated platelets at the peak of NO production. To study platelet recruitment accurately, we developed an assay that monitors two platelet populations simultaneously. Nonbiotinylated platelets were incubated with L-NAME or vehicle and activated with ADP. At peak NO production, biotinylated platelets were added. As measured by three-color flow cytometry, there was a 56+/-11% increase in the number of P selectin- positive platelets in the nonbiotinylated population treated with L-NAME as compared to control. When biotinylated platelets were added to the L-NAME-treated nonbiotinylated population, the number of P selectin positive biotinylated plate-lets increased by 180+/-32% as compared to biotinylated platelets added to the control. In summary, stimulated platelets produce NO that modestly inhibits platelet activation but markedly inhibits additional platelet recruitment. These data suggest that platelet-derived NO may regulate platelet recruitment to a growing thrombus.

Decreased platelet inhibition by nitric oxide in two brothers with a history of arterial thrombosis.

Highly reactive oxygen species rapidly inactivate nitric oxide (NO), and endothelial product which inhibits platelet activation. We studied platelet inhibition by NO in two brothers with a cerebral thrombotic disorder. Both children had hyperreactive platelets, as determined by whole blood platelet aggregometry and flow cytometric analysis of the platelet surface expression of P-selectin. Mixing experiments showed that the patients'platelets behaved normally in control plasma; however, control platelets suspended in patient plasma were not inhibited by NO. As determined by flow cytometry, in the presence of plasma from either patient there was normal inhibition of the thrombin-induced expression of platelet surface P-selectin by prostacyclin, but not NO. Using a scopoletin assay, we measured a 2.7-fold increase in plasma H2O2 generation in one patient and a 3.4-fold increase in the second patient, both compared woth control plasma. Glutathione peroxidase (GSH-Px) activity was decreased in the patients' plasmas compared with control plasma. The addition of exogenous GSH-Px led to restoration of platelet inhibition by NO. These data show that, in these patients' plasmas, impaired metabolism of reactive oxygen species reduces the bioavailability of NO and impairs normal platelet inhibitory mechanisms. These findings suggest that attenuated NO-mediated platelet inhibition produced by increased reactive oxygen species or impaired antioxidant defense may cause a thrombotic disorder in humans.

Opioid receptor modulation of a metabolically sensitive ion channel in rat amygdala neurons.

We have used single-channel patch-clamp recordings to study opiate receptor effects on freshly dissociated neurons from the rat amygdalohippocampal area (also called the posterior nucleus of the amygdala), an output nucleus of the amygdala implicated in appetitive behaviors. Dissociated cells included a distinct subpopulation that was 30-40 micrometer in diameter, multipolar or pyramidal in shape, and immunoreactive for neuron-specific enolase, mu opioid receptors, and galanin. In whole-cell perforated-patch recordings, these cells responded to low concentrations of mu opioid agonists with a hyperpolarization. In cell-attached single channel recordings, these cells expressed a large variety of K(+)-permeable ion channels, including 20-100 pS inward rectifiers and 150-200 pS apparent Ca(2+)-activated K(+) channels, none of which appeared sensitive to the presence of opioid drugs. In contrast, a 130 pS inwardly rectifying channel was selectively activated by mu opioid receptors in this same subpopulation of cells and was active only in the presence of opioid agonists, and inhibited in the presence of antagonists. Channels identical to the 130 pS channel in conductance and voltage sensitivity were activated in the absence of opioids, when the cells were treated with glucose-free medium or with the metabolic inhibitor rotenone. The sulfonylurea drug tolbutamide inhibited 130 pS channel openings elicited by opioids. Thus, a subpopulation of amygdala projection neurons expresses a metabolically sensitive ion channel that is selectively modulated by opiate receptors. This mechanism may allow opioid neurotransmitters to regulate ingestive behaviors, and thus, opiate drugs to influence reward pathways.

Select flavonoids and whole juice from purple grapes inhibit platelet function and enhance nitric oxide release.

BACKGROUND: Moderate red wine consumption is inversely associated with coronary ischemia, and both red wine and purple grape juice (PGJ) contain flavonoids with antioxidant and antiplatelet properties believed to be protective against cardiovascular events. Acute cardiac events are also associated with decreased platelet-derived nitric oxide (NO) release. In this study, the effects of PGJ and PGJ-derived flavonoids on platelet function and platelet NO production were determined. METHODS AND RESULTS: Incubation of platelets with dilute PGJ led to inhibition of aggregation, enhanced release of platelet-derived NO, and decreased superoxide production. To confirm the in vivo relevance of these findings, 20 healthy subjects consumed 7 mL. kg(-1). d(-1) of PGJ for 14 days. Platelet aggregation was inhibited after PGJ supplementation, platelet-derived NO production increased from 3.5+/-1.2 to 6.0+/-1.5 pmol/10(8) platelets, and superoxide release decreased from 29.5+/-5.0 to 19.2+/-3.1 arbitrary units (P<0.007 and P<0.05, respectively). alpha-Tocopherol levels increased significantly after PGJ consumption (from 15.6+/-0.7 to 17.6+/-0.9 micromol/L; P<0.009), and the plasma protein-independent antioxidant activity increased by 50.0% (P<0.05). Last, incubation of platelets with select flavonoid fractions isolated from PGJ consistently attenuated superoxide levels but had variable effects on whole-blood aggregation, platelet aggregation, and NO release. CONCLUSIONS: Both in vitro incubation and oral supplementation with PGJ decrease platelet aggregation, increase platelet-derived NO release, and decrease superoxide production. These findings may be a result of antioxidant-sparing and/or direct effects of select flavonoids found in PGJ. The suppression of platelet-mediated thrombosis represents a potential mechanism for the beneficial effects of purple grape products, independent of alcohol consumption, in cardiovascular disease.

Low plasma ascorbic acid independently predicts the presence of an unstable coronary syndrome.

OBJECTIVES: This study sought to investigate the relations between plasma antioxidant status, extent of atherosclerosis and activity of coronary artery disease. BACKGROUND: Previous studies indicate that increased antioxidant intake is associated with decreased coronary disease risk, but the underlying mechanisms remain controversial. METHODS: Plasma samples were obtained from 149 patients undergoing cardiac catheterization (65 with stable angina, 84 with unstable angina or a myocardial infarction within 2 weeks). Twelve plasma antioxidant/oxidant markers were measured and correlated with the extent of atherosclerosis and the presence of an unstable coronary syndrome. RESULTS: By multiple linear regression analysis, age (p < 0.001), diabetes mellitus (p < 0.001), male gender (p < 0.001) and hypercholesterolemia (p = 0.02) were independent predictors of the extent of atherosclerosis. No antioxidant/oxidant marker correlated with the extent of atherosclerosis. However, lower plasma ascorbic acid concentration predicted the presence of an unstable coronary syndrome by multiple logistic regression (odds ratio [OR] 0.59, 95% confidence interval [CI] 0.40 to 0.89, p = 0.01). The severity of atherosclerosis also predicted the presence of an unstable coronary syndrome (OR 1.7, 95% CI 1.14 to 2.47, p = 0.008) when all patients were considered. When only patients with significant coronary disease were considered (at least one stenosis >50%), ascorbic acid concentration (OR 0.56, 95% CI 0.37 to 0.85, p = 0.008) and total plasma thiols (OR 0.52, 95% CI 0.34 to 0.80, p = 0.004) predicted the presence of an unstable coronary syndrome, whereas the extent of atherosclerosis did not. CONCLUSIONS: These data are consistent with the hypothesis that the beneficial effects of antioxidants in coronary artery disease may result, in part, by an influence on lesion activity rather than a reduction in the overall extent of fixed disease.

The role of circulating platelet transcripts.

Our understanding of platelets, anucleate cells with a traditional role in hemostasis and inflammation, has developed greatly over the last decade. Platelets' role in the systemic response of the body to vascular injury, inflammation, and infection has expanded as has our understanding of their importance to the body's regulation of these processes. One recently explored mechanism by which platelets regulate the body's inflammatory and immune response is through its endogenous RNA. Platelets' messenger RNA (mRNAs) and microRNA (miRNAs) profiles have been shown to reflect disease and disease risk factors and have been correlated with select human clinical phenotypes. Developing an understanding of platelet transcripts in the circulation elucidates how platelets function in both their traditional thrombotic role and non-traditional functions and may have widespread implications in several fields including thrombosis, infection, cancer, and systemic inflammation.

Nitric oxide and its relationship to thrombotic disorders.

Nitric oxide (NO) is released by the endothelium preventing platelet adhesion to the vessel wall. When released by platelets, NO inhibits further recruitment of platelets to a growing thrombus. Modulation of endogenous NO release may be a mechanism by which the thrombotic response can be regulated as suggested by several clinical diseases associated with impaired bioactive NO. Diseases including atrial fibrillation and coronary atherothrombotic disease have been associated with impaired NO release or decrease in NO bioavailability.

Soluble adhesion molecules and unstable coronary artery disease.

Leukocyte adhesion and transendothelial migration, prerequisites in the development of atherosclerosis, are largely mediated by adhesion molecules. In addition, unstable coronary syndromes usually involve platelet activation and thrombus formation at the site of atherosclerotic plaque. Therefore, we compared plasma levels of soluble P-selectin, a measurement of platelet activation, as well as E-selectin, intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) in patients with atherosclerosis undergoing coronary angiography (n=76). Soluble P-selectin levels, as measured by ELISA, were significantly elevated in patients with unstable (n=44) vs stable (n=32) atherosclerotic disease (73.0 +/- 2.5 ng/ml vs 52.3 +/- 3.0 ng/ml, respectively, P<0.01). By logistic regression analysis, plasma level of soluble P-selectin was an independent predictor of an unstable coronary syndrome (OR 4.2, CI 1.4-12.9, P<0.01). Soluble E-selectin level, a marker of endothelial activation, was associated with extent of atherosclerosis but did not correlate with disease stability. Interestingly, soluble P-selectin was inversely correlated with plasma levels of the antioxidant alpha-tocopherol (R=-0.443, P<0.001), a known inhibitor of platelet function. In summary, amongst the soluble adhesion molecules, only P-selectin is significantly increased in patients with unstable coronary syndromes. This study suggests that platelet activation persists in patients with unstable coronary syndromes despite concurrent aspirin therapy. In addition, the beneficial effects of alpha-tocopherol in patients with cardiovascular disease may be related to inhibition of platelet function.

Dual effects of the snake venom polypeptide vipoxin on receptors for acetylcholine and biogenic amines in Aplysia neurons.

Vipoxin, a 13,000-dalton polypeptide component of Russell's viper venom, has a dual pattern of effects on the responses of voltage-clamped Aplysia neurons to acetylcholine and biogenic amines. Application of low doses of vipoxin by pressure ejection reversibly antagonized all three types of ionic response to acetylcholine and carbachol. The blockade by vipoxin of acetylcholine responses was not prevented by eserine. The order of susceptibility of acetylcholine responses to blockade by vipoxin was Na+ greater than K+ greater than Cl-. Low doses of vipoxin also produced a reversible potentiation of excitatory responses to dopamine with a slower time course of onset and recovery. Inhibitory responses to dopamine (Cl-, K+) and both excitatory and inhibitory responses to histamine and 5-hydroxytryptamine were little affected by vipoxin. Higher doses of vipoxin directly evoked current responses which were always of the same ionic type as that evoked by acetylcholine or carbachol. Responses to cholinergic agonists and vipoxin were both blocked by cholinergic antagonists but not by antagonists of biogenic amine receptors, which reversibly antagonized the responses to amines on the same cell. These results suggest that vipoxin, which has no demonstrated actions on vertebrate acetylcholine receptors, acts as a partial agonist at all three types of acetylcholine receptor in Aplysia neurons. Our observations thus provide evidence for some degree of phylogenetic difference between vertebrate and molluscan acetylcholine receptors.

Platelets and plasminogen activation.

Platelets serve as a site for assembly of the proteins of the plasminogen activator system. Once bound to the platelet surface, tissue-type plasminogen activator manifests enhanced catalytic activity. Plasmin, once formed, also binds to the platelets surface and, at low concentrations, renders the platelet dysfunctional by cleaving glycoprotein IIIa selectively in the presence of bound fibrinogen. At higher concentrations (approximately 1 caseinolytic unit/ml), plasmin activates the platelet directly. Activated platelets also bind plasminogen and tissue-type plasminogen activator, and manifest enhanced catalytic efficiency of plasminogen activation. These observations suggest that plasminogen activation by tissue-type plasminogen activator is an autocatalytic process on the platelet surface, and that unique reciprocating mechanisms govern the interaction between platelets and the components of the plasminogen activator system.

Endothelial dysfunction and atherothrombotic occlusive disease.

The endothelium plays a crucial role in the regulation of vascular function through the release of locally important molecular effectors such as endothelium-derived relaxing factor (EDRF) and prostacyclin. Endothelial cells also regulate vascular patency and tissue perfusion by inhibiting platelet aggregation and thrombosis, suppressing intimal proliferation, and maintaining vascular tone. Disturbances of the regulatory functions of the endothelium contribute to the pathophysiology of various disease states, including cardiovascular disease and stroke. Most studies focused on endothelial control of vasomotion and, in particular, on the action of EDRF; many studies have also emphasised that altered endothelial control of fibrinolysis and intimal growth influence the clinical expression of atherothrombotic disease. Importantly, understanding the pathophysiological role of endothelial dysfunction may lead to new therapeutic approaches for disease states caused by vascular disease.

Mastoparan activation of dopamine-modulated K+ channels on rat striatal neurons.

Coupling mechanisms between a D2-like dopamine receptor and an 85 pS K+ channel on freshly dissociated rat caudate-putamen neurons were studied using cell-attached patch-clamp electrophysiology. Channel currents were observed in the absence of dopamine receptor agonists when mastoparan, an activator of certain guanyl nucleotide binding proteins (G-proteins), was applied via the patch pipette. Channel current-voltage relationships and open probabilities observed with mastoparan were indistinguishable from those obtained with dopaminergic agonists. Alamethicin, which mimics the membrane-perturbing but not the G-protein activating properties of mastoparan, did not activate the channel, suggesting that non-specific effects of mastoparan were unlikely to account for this effect. Our results indicate that coupling between the D2 dopamine receptor and this K+ channel may involve a mastoparan-sensitive G-protein.

Vipoxin both activates and antagonizes three types of acetylcholine response in Aplysia neurons.

The effects of vipoxin, a 13,000 Dalton protein component of Russell's viper venom on responses of voltage-clamped Aplysia neurons to acetylcholine (ACh) and monoamines has been studied. At low doses vipoxin reversibly antagonizes all 3 types of ionic response to ACh or carbachol, the order of susceptibility to blockade being Na+ greater than K+ greater than Cl-. High doses of vipoxin directly evoke the same ionic response on a given cell as that evoked by ACh. Responses to vipoxin are reversibly antagonized by cholinergic antagonists (e.g. hexamethonium, tetraethylammonium), but not by monoamine antagonists (e.g. bufotenine, ergometrine, cimetidine). In addition to activation of cholinergic responses, high doses of vipoxin also produce a reversible potentiation of responses to dopamine and 5-hydroxytryptamine on some cells. In contrast to its effects on Aplysia neurons, vipoxin has neither agonist nor antagonist actions at the frog neuromuscular junction. These results suggest that this venom protein acts as a partial agonist at molluscan ACh receptors and provides evidence for some phylogenetic difference between molluscan and vertebrate ACh receptors.

Morphine and clonidine activate different K+ channels on rat amygdala neurons.

In cell-attached patch-clamp recordings from freshly dissociated neurons of the rat amygdalohippocampal area, clonidine principally activated a 95-pS (picosiemens) inwardly rectifying K+-permeable channel, whereas morphine acting at mu-opioid receptors activated a different 130-pS channel. Clonidine's effects were largely antagonized by the alpha(2)-adrenoceptor antagonist idazoxan, but were poorly mimicked by agmatine. These results partially contradict the prevailing hypothesis that alpha(2) and opioid receptors act through the same ion channel transduction pathways.

Quinine potently blocks single K+ channels activated by dopamine D-2 receptors in rat corpus striatum neurons.

In single channel recordings from acutely dissociated neurons of the rat corpus striatum, a membrane K+ channel which is activated by dopamine D-2 receptors was blocked by nanomolar concentrations of quinine. An intermittent partial blockade was observed at 4-10 nM quinine, with a voltage dependence consistent with quinine binding to the channel near the extracellular surface of the membrane. A nearly complete blockade of channel current was observed at 100 nM quinine and above. Such concentrations are known to be too low to block various other ion channels, and may be attained in human brain at antimalarial dosages of quinine. Blockade of this channel by quinine may provide a useful experimental probe of dopaminergic function, as an alternative to D-2 receptor binding site blockade by neuroleptics.

Idazoxan (RX 781094) selectively antagonizes alpha 2-adrenoceptors on rat central neurons.

We have studied the alpha-adrenoceptor antagonist effects of idazoxan (RX 781094) in extracellular recordings from single locus coeruleus and dorsal raphe neurons of chloral hydrate-anesthetized rats. Idazoxan blocked alpha 2 responses with an ED50 of 14 +/- 8 micrograms/kg i.v., while the potency at alpha 1-receptors was only 420 +/- 190 micrograms/kg. A similar alpha 2 selectivity was seen when idazoxan was applied microiontophoretically. Idazoxan at doses which blocked alpha 1-receptors had little or no effect on locus coeruleus responses to morphine or dorsal raphe responses to LSD. When sodium pentobarbital was used as the anesthetic, systemically administered idazoxan slowed the firing rate of locus coeruleus cells, but iontophoretic experiments showed this to be an interaction with the anesthetic and not a direct agonist effect. We conclude that in rat brain idazoxan is a pure antagonist and that it has a selectivity for alpha 2- over alpha 1-receptors markedly superior to piperoxane, yohimbine, or rauwolscine.

U-37883A potently inhibits dopamine-modulated K+ channels on rat striatal neurons.

An 85 pS K+ channel of rat caudate-putamen neurons, which is activated by dopamine D2 receptors and inhibited by sulfonylurea drugs, was studied using cell-attached patch-clamp electrophysiology. This channel was inhibited by externally-applied U-37883A (4-morpholinecarboximidine-N-1-adamantyl-N'-cyclohexyl hydrochloride), a blocker of vascular ATP-sensitive K+ channels, with a half-maximal effect at a concentration of approximately 0.1 microM. Channel inhibition occurred in a time-dependent fashion when U-37883A was applied to the membrane from a back-filled patch pipette. Inhibition was associated with a decrease in fractional open time, but was voltage-insensitive and did not alter channel conductance, suggesting an effect on channel gating at a site largely insensitive to the electrical field of the channel. U-37883A was about 50 times more potent at inhibiting this channel than was the sulfonylurea drug glibenclamide. This relative potency, opposite to that found in pancreatic tissue, indicates that U-37883A is a useful tool to distinguish amongst different subtypes of sulfonylurea-sensitive K+ channels.

The putative dopamine D3 receptor agonist 7-OH-DPAT: lack of mesolimbic selectivity.

7-Hydroxy-N,N-di-n-propyl-2-aminotetralin (7-OH-DPAT), an agonist with relative selectivity for the dopamine D3 receptor, was examined in several electrophysiological assays to determine whether it exhibits preferential effects in the mesolimbic versus nigrostriatal dopamine systems. Extracellular single unit activities of substantia nigra pars compacta (A9) and ventral tegmental area (A10) dopamine neurons, and caudate-putamen and nucleus accumbens neurons, were recorded in male rats anesthetized with chloral hydrate. Intravenous (+/-)-7-OH-DPAT potently and completely inhibited the firing of both A9 and A10 dopamine neurons (ED50's: 3.5 +/- 0.7 micrograms/kg and 3.9 +/- 0.9 micrograms/kg, respectively). The active enantiomer, (+)-7-OH-DPAT, was 2 to 3 times more potent than the racemic drug (ED50's: 1.2 +/- 0.3 micrograms/kg and 1.7 +/- 0.4 micrograms/kg for A9 and A10 cells, respectively). There were no significant differences in potency for either form in inhibiting A9 and A10 dopamine neurons. In other studies, iontophoretically applied (+)-7-OH-DPAT caused current-dependent inhibitions of spontaneously active or glutamate-driven caudate-putamen and nucleus accumbens neurons (I50 values, 6.5 and 7.9 nA, respectively). Again, no difference in potency between cell populations was noted. Finally, in cell-attached patch-clamp recordings from freshly dissociated rat caudate-putamen neurons, an 85 pS K+ channel known to be activated by dopamine and the "D2-like" agonist quinpirole was also observed with (+/-)-7-OH-DPAT (0.2-1 microM) applied in the patch pipette.(ABSTRACT TRUNCATED AT 250 WORDS)