text:
Certain members of the phospholipase A2 superfamily of enzymes have established causal involvement in atherosclerosis, thus at least two groups of this family of enzymes have been considered potential candidates for the prevention of cardiovascular events. Recently completed experimental animal studies, human biomarker data, vascular imaging studies, and genome-wide atherosclerosis studies provide the rationale for proceeding with clinical outcome trials directed at inhibition of secretory phospholipase A2 and lipoprotein-associated phospholipase A2. A clinical trial with the sPLA2 inhibitor varespladib methyl was recently terminated, while clinical trials with the Lp-PLA2 inhibitor darapladib are being conducted in coronary heart disease patients. This article reviews the available experimental animal and human trial evidence that serve as the basis for the development of these two classes of phospholipase A2 inhibitors.
Secretory phospholipase A2
Lipoprotein-associated phospholipase A2
Vascular inflammation is an essential feature in the pathogenesis of atherosclerotic lesion formation, and it contributes to its complications of acute myocardial infarction and stroke. In prospective population-based studies, non-specific systemic inflammatory markers such as the white blood cell count and concentrations of the C-reactive protein, serum amyloid A, and fibrinogen serve to stratify patients into categories of cardiovascular disease (CVD) risk.1–3 However, these non-specific inflammatory risk markers provide limited insight as to the specific inflammatory mediator(s) responsible for the inflammatory response in that particular patient,4 and thus they are less useful targets for therapeutic intervention directed at mitigating risks associated with vascular inflammation.5
Circulating levels and enzymatic activity of two families of phospholipase A2 enzymes, secretory phospholipase A2 (sPLA2) and lipoprotein-associated phospholipase A2 (Lp-PLA2), have been evaluated as biomarkers of cardiovascular risk in population-based studies inclusive of apparently healthy individuals, and patients with established coronary heart disease (CHD).6–11 In the EPIC (European Prospective Investigation of Cancer) Norfolk study, the increased CHD risk associated with the oxidation specific biomarker, oxidized phospholipids on apolipoprotein B (apoB) or lipoprotein(a), was enhanced further among individuals with high sPLA2 concentration and activity than Lp-PLA2 activity.10 This study and other observational studies have shown that both enzyme concentration and activity predict incident cardiovascular events. The evidence that these phospholipase A2 enzymes retain their importance as biomarkers of risk in statin-treated patients is less certain; however, in one trial, LpPLA2 concentration was associated with future CVD events in models that adjusted for baseline covariants and apolipoprotein concentrations.12 The importance of this phospholipase A2 enzyme biomarker data should not be overemphasized, as circulating levels of these two groups of PLA2 enzymes and their various members may not encompass their multifarious downstream pro-atherosclerotic effects in the vessel wall.13,14 Recently, the non-functional (null) V279F allele within the PLA2G7 gene, a common loss of function mutation of Lp-PLA2 in Asians, was associated with reduced CHD risk.15 This Mendelian randomization study provides support for the hypothesis that marked reduction Lp-PLA2 enzymatic activity may be an effective strategy for CVD prevention.
Recently, selective inhibitors of sPLA216 and LpPLA217 have been targeted as potential candidates to reduce incident atherosclerotic cardiovascular events.18,19 The completion of clinical outcomes trials with selective inhibitors of these phospholipase A2 enzymes will provide important information on the future role of these crucial enzymes in CVD prevention and treatment.
This review examines the biological function of these two groups of phospholipase A2 enzymes in atherosclerosis through examination of population-based studies, animal studies and human studies. Available evidence that inhibition of sPLA2 and Lp-PLA2 is reasonable targets for reducing CVD are presented.
Phospholipase A2 superfamily of enzymes
The phospholipase A2 superfamily enzymes are characterized by their ability to hydrolyze fatty acids at the sn-2 position of glycophospholipids and generate multiple classes of bioactive lipids (Figure 1).20 Five main families of phospholipases have defined physiological roles, and they comprise group (G) II or secretory PLA2 (sPLA2), GIV or cytosolic PLA2 (cPLA2), GXV or lysosomal PLA2; and two major Ca2+-independent groups, GV PLA2 (iPLA2) and GVII platelet-activating factor (PAF) acetylhydrolases.
Phospholipase A2 enzymatic activity generates lipid products with biological properties and functions. Secretory phospholipase A2 (sPLA2) acts to hydrolyze phospholipids from the surface of cell membranes, native lipoproteins and oxidatively-modified lipoproteins to generate multiple bioactive lipids that include arachidonic acid, non-esterified fatty acids (NEFA), lysophospholipids (Lyso-PL), lyso-platelet acting factor (LYSO-PAF), and oxidized non-esterified fatty acids (Ox-NEFA). In contrast, lipoprotein-associated phospholipase A2 (Lp-PLA2) or platelet acting factor (PAF) requires oxidized phospholipids as a substrate (oxidized phosphotidylcholine [Ox-Phosphotidylcholine]).
Secretory phospholipase A2 structure and biology
Secretory PLA2 family consists of 12 isoforms, and with the exception of GIII sPLA2, these proteins have comparable molecular weights of ∼16 kDa and three-dimensional structures.20–22 Since the genes encoding GIIA sPLA2 and GV sPLA2 are localized in close proximity in homologous regions of human chromosome 1 (and mouse chromosome 4), these group members may represent a gene cluster.23 Despite common features among human sPLA2 isoenzymes, they are functionally distinct proteins with specific tissue distributions21,22,24 and enzymatic properties on various phospholipids.24
Each isoform functions both as an enzyme and ligand for receptors (both soluble and membrane-bound) that control and transduce their biological effects or modulate their enzymatic action.25,26 Biological effects include various roles in both physiological and pathophysiological conditions, including inflammatory diseases such as atherosclerosis and rheumatoid arthritis.27
Among the sPLA2 binding proteins, the M-type receptor (PLA2R1) is the best-characterized sPLA2 receptor.28 PLA2R1 is a type I membrane glycoprotein of 180 kDa29 that belongs to the superfamily of C-type lectins.30 It is comprised of a single transmembrane domain, a short cytoplasmic tail, and a large extracellular region that includes an N-terminal cysteine-rich domain, a fibronectin-like type II domain, and a tandem repeat of eight distinct C-type lectin-like carbohydrate recognition domains. Currently, the biological functions of PLA2R1, its endogenous ligands other than mammalian sPLA2 isoenzymes, and associated signalling pathways remain unknown.26,28
Secretory phospholipase A2 and atherosclerosis
sPLA2 hydrolyzes the sn-2 ester bond in the glyceroacyl phospholipids presents in lipoproteins and cell membranes, inducing structural and functional changes and forming lysopholipids and non-esterified fatty acids (NEFAs) with direct proinflammatory effects.31,32 The mammalian sPLA2 isoforms comprise groups named IB, IIA, IIC, IID, IIE, IIF, III, V, X, and XII. Six isoforms are present in human atherosclerotic lesions: IIA, IID, IIE, III, V, and X. The homology between these sPLA2 enzymes varies, but they share a calcium-dependent catalytic mechanism, presence of several disulphide bridges, and a well-conserved overall three-dimensional structure. In addition to their role in atherogenesis, sPLA2 enzymes are implicated in various physiological and pathophysiological functions, including lipid absorption, cell proliferation, myocardial injury, tumour formation, and inflammation. Interestingly, sPLA2 activity is involved in cholesterol homeostasis.30,31 Specifically, GX sPLA2 generates lipolytic products that suppress LXR activation, which negatively regulates ABCA1 and ABCG1 expression and cholesterol efflux from macrophages.33
Secretory PLA2 activity can be proatherogenic in the circulation, and at sites of atherosclerotic plaque development (Figure 2). In plasma, sPLA2 activity hydrolyzes surface phospholipids on lipoproteins, resulting in structural alteration in these particles.34 GV sPLA235 and GX sPLA236 enzymes hydrolyze phosphatidylcholine on the surface of VLDL and LDL at least 20-fold more efficient than GIIA sPLA2.37 Although group IIA sPLA2 acts poorly on unmodified LDL, presumably because of its poor binding to phosphatidylcholine-rich membranes, this enzyme shows enhanced ability to hydrolyze oxidized LDL.
Effects of phospholipase A2 enzymatic activity on circulating lipoproteins. Secretory phospholipase A2 (sPLA2) hydrolyzes phospholipids from the surface of native lipoproteins and oxidatively-modified lipoproteins, whereas lipoportein-associated phospholipase A2 acts only on oxidatively-modified lipoproteins. Phosphoatidylcholine hydrolysis by sPLA2 results in small VLDL and LDL particles with altered confirmation of apolipoprotein B (apoB). The conformational change in apoB reduces binding and internalization of apoB-containing lipoproteins by the apoB/E (LDL) receptor resulting in prolonged residence time in the circulation. This prolonged circulation time of LDL particles increases exposure to reactive oxygen species (ROS) resulting in an oxidized LDL particle (Ox-LDL) that may serve as a substrate for group IIA sPLA2 (GIIA sPLA2) and Lp-PLA2. Phospholipid hydrolysis of Ox-LDL particles generates oxidized non-esterified fatty acids (Ox-NEFA) and lysophophotidylcholine (Lyso-PC). sPLA2 acts on cellular membranes resulting in elaboration of arachidonic acid that serves as the substrate for eicosanoids, thromboxanes and leukotrienes.
As a consequence of phospholipid hydrolysis, conformational changes in apolipoprotein B (apoB) impairs clearance of very low-density lipoprotein remnants and LDL by the apolipoprotein B/E receptor (LDL-R),38 resulting in increased residence time in the circulation and further susceptibility to oxidative modifications (Figure 2).39 Hydrolysis of phospholipids in high-density lipoprotein (HDL) results in impaired cholesterol-efflux capacity. In the arterial vessel wall, the conformational changes in apoB38 increase their binding to intimal proteoglycans,40–44 promote retention of these atherogenic lipoproteins, and increase intra- and extracellular cholesterol accumulation, a hallmark of atherosclerosis. In addition, reduction in the molar ratio of cholesterol to phospholipid will compromise the solubility of free cholesterol molecules resulting in cholesterol crystal precipitation.45,46 Furthermore, cholesterol crystals have been reported to induce inflammation by stimulating caspase-1-activating NLP3 inflammasome, which results in cleavage and secretion of interleukin-1 family cytokines.47
In addition to indirect effects on fostering cholesterol crystal formation, sPLA2 mediates phospholipid hydrolysis from cell membranes and lipoproteins, and increase oxidative stress through generation of arachidonic acid, lysophospholipids, and non-esterified fatty acids (Figure 1).48,49 These bioactive lipids, including the arachidonic acid-derived metabolites prostaglandins, thromboxanes, and leukotrienes, work collectively with oxidized LDL to recruit inflammatory cells to atherosclerotic lesions and activate inflammatory pathways in multiple cells in the vessel wall (Figure 3).50 Lysophospholipids and NEFAs, the hydrolysis products of sPLA2, also contribute to atherosclerosis plaque progression exerting a broad range of cellular effects, including cytotoxicity.51 It should be stressed that the total hydrolysis of phospholipids from one apoB particle will generate more than 500 molecules of lysophospholipids and free fatty acids. Thus, in atherosclerotic plaques, these products can reach high local concentrations. These bioactive lipid products can induce different proatherogenic and proinflammatory cellular processes, and cell membrane perturbation (Figure 3).31,32,51
Cellular mechanisms of action of sPLA2 enzymes in eicosanoid production. In this model, hydrolysis of phospholipids (PL) by sPLA2 enzymes may occur on the cell membrane and on extracellular lipoproteins. Lipoproteins can bind to cell surface proteoglycans (PG) and PG in the extracellular space facilitating their interaction with sPLA2s such as group sPLA2 II (sPLA2-II) and group V sPLA2 (GV-sPLA2) that also bind to PG. This binding enhances the hydrolytic activity of GV-sPLA2 on lipoproteins. Binding to cell surface PG, glypican, results also in internalization of sPLA2s. Group X sPLA2 (GX-sPLA2) does not bind to PG; however, it binds tightly to zwitteronic phospholipid membranes relative to all other mammalian sPLA2s and can act on the outer plasma membrane leaflet. Expression and secretion of sPLA2s can be modulated by different stimulus, i.e. GV-sPLA2 expression is enhanced by high fat diet, and sPLA2-II and X by inflammatory stimuli. On the other hand, chemical inhibition of extracellular GIIA-sPLA2 by varespladib decreases its own secretion, by a yet unknown mechanism. Several sPLA2s, including groups IIA, V and X, are likely to be involved in the release of arachidonic acid (AA) and other fatty acids as well as lysophospholipids (Lyso-PC) that can be further metabolized into various eicosanoids and related bioactive lipid mediators by different intracellular pathways and in different cell types, i.e. in cells such a s neutrophiles and monocytes that express 5-LO, AA is converted in leukotriene-B4 (LTB4). Lyso-PC may mobilize intracellular calcium and further activate intracellular eicosanoid pathways by upregulating cytoplasmic PLA2 (cPLA2) and other enzymes such as 5-lipoxygenase (5-LO) and cycloygenases (COX).
Several sPLA2 isoenzymes members have been detected in murine and human atherosclerotic lesions (Figure 4).22,51,52 Additionally, recent data suggest that sPLA2 isoenzymes-IID also contribute to atherosclerosis.53 Experimental animal studies using gain-of-function and loss-of-function approaches have established GIIA, III, V, and X as causal factors in atherogenesis.54–58 Transgenic mice that constitutively overexpress human GIIA sPLA2 develop atherosclerotic lesions on standard chow and atherogenic diets.39 In addition, human GIIA sPLA2 transgenic mice with macrophage-specific overexpression showed increased lesion area and enhanced collagen deposition.39 Transplanting bone marrow cells from these transgenic mice into LDL-R-deficient mice (LDL-R−/−) increased atherosclerotic lesion formation. Macrophage-specific overexpression of human GIIA sPLA2 increased foam cell formation in LDL-R−/− mice transplanted with sPLA2 bone marrow of transgenic mice fed a western-type diet.54
Phospholipase A2 enzymatic activity and foam cell formation. Secretory phospholipase A2 (sPLA2) and lipoprotein-associated phospholipase A2 (Lp-PLA2) increase oxidation of LDL particles allowing for enhanced internalization into the macrophage via the conventional scavenger receptor resulting in foam cell formation. In addition, group V (GV sPLA2) and group X sPLA2 (GX sPLA2)-modified LDL particles are incorporated into macrophages via a putative M-type receptor, which contributes to cholesterol content of tissue macrophages via this distinct pathway.
GV sPLA2 has been identified as a potential factor in promoting atherosclerosis in human and murine atherosclerosis.56–58 This sPLA2 isoenzyme is expressed in human atherosclerotic lesions and human vascular cells.59 In human vascular cells, GV sPLA2 is found extracellularly around foam cells in lipid core areas, and it is associated with smooth muscle cells in the neointima and media of intermediate and advanced lesions. Experiments in mouse models showed that diet- or genetic-induced hyperlipidaemia in mouse models increase the expression of GV sPLA2 but not of GII sPLA2 in the aorta. On the other hand, lipopolysaccharide-induced acute inflammation augmented the expression of GII sPLA2 but not of GV sPLA2. These differences in stimulus-mediated expression indicate that GII and GV sPLA2 enzymes can have different roles in the atherosclerosis.56
Overexpression of mouse GV sPLA2 by retrovirus-mediated gene transfer increased the lesion area in LDL−/− mice, whereas mice deficient in bone marrow-derived GV sPLA2 had reduced the lesion area. Transgenic GV sPLA2 overexpression increased lipid deposition and collagen deposition in LDL−/− mice.56 In apoE−/− mice fed a high-fat diet GV sPLA2 promotes lipid deposition; however, unlike the experiments in LDL-R−/− mice, GV sPLA2 deficiency did not reduce atherosclerosis even though there was a reduction in the collagen formation.58 A potential explanation for these differences may relate to the increased sphingomyelin content of LDL particles isolated from apoE−/− mice, which decreases GV-mediated phosphatidylcholine hydrolysis.
Cholesterol accumulation in the vessel wall is one mechanism for GV sPLA2-mediated atherosclerosis. In murine macrophages, group V sPLA2-modified LDL induced cholesterol ester accumulation that was independent of LDL-R, SR-A, and CD36 scavenger receptors or the pathway that clears LDL aggregates (Figure 4).58 The mechanism for the increased uptake of GV-modified LDL involves initial binding to heparan sulfate proteoglycans of the extracellular matrix, and perhaps directly binding to macrophages.56
Human GX sPLA2 is expressed in the intima of human atherosclerotic lesions where it colocalizes with foam cells and smooth muscle cells that resemble myofibroblasts.59 GX sPLA2 modifies LDL particles that induce lipid accumulation in monocyte-derived macrophages (Figure 2).60 However, recent studies have also shown that GIID and GX sPLA2 appear as an anti-atherogenic factor that inhibits macrophage activation and inflammatory responses by reducing cell adhesion, NO, and TNF-alpha levels, while augmenting the production of the anti-inflammatory IL10 cytokine.61
Secretory phospholipase A2 and ischaemic injury
Groups II, V, and X sPLA2 have be implicated in ischaemic myocardial tissue damage. Elevated plasma levels of GII sPLA2 are associated with the increased CHD risk.19 During an acute coronary syndrome, the plasma concentration of GII sPLA2 rises, and this biomarker has been shown to useful to discriminate between ACS patients who develop a myocardial infarction vs. those patients who do not.62 The localization pattern of GII sPLA2 in infarcted myocardium and its temporal course in plasma, in relation to those of C-reactive protein, are in line with a postulated pro-inflammatory role during acute myocardial infarction for GII sPLA2 which serves as a generator of lysophospholipids that serve as ligands for C-reactive protein.63
GV sPLA2 is expressed in high concentrations in various tissues including cardiomyocytes where its expression is further increased in human-infarcted myocardium.64 GV sPLA2 activates extracellular signal-related kinase 1/2 (ERK1/2) that, in turn, activates cPLA2 (Figure 3). Cytoplasmic PLA2 is a major regulator of eicosanoid generation. When compared with controls, GV sPLA2−/− mice had lower content of leukotriene B4 and thromboxane B2 in ischaemic myocardium, and smaller myocardial infarctions and preserved left ventricular function.65
In experimental myocardial infarction, the cardiomyocyte plasma membrane loses its asymmetry resulting in exposure of the anionic phospholipids phosphatidylserine and phosphatidylethanolamine to the outer membrane leaflet. Structural re-organization in membrane phospholipids enhances sPLA2 binding, and consequently increases complement-mediated myocardial necrosis.66 The use of a specific sPLA2 inhibitor PX-18 (2-N,N-Bis(oleoyloxyethyl)amino-1-ethanesulfonic acid) reduced cardiomyocyte apoptosis and limited experimental infarct size. In addition, myocardial ischaemia/reperfusion injury is attenuated in GX-deficient GX-sPLA2 mice partly through the suppression of neutrophil cytotoxic activities.67 Together, this human and experimental in vivo evidence suggest a pathogenic role of these three sPLA2 isoenzymes in infarcted myocardium.
Lipoprotein-associated phospholipase A2 structure and biology
Group VIIA or PAF is a potent phospholipid activator that is secreted by multiple inflammatory cells including monocytes/macrophages, T lymphocytes, and mast cells.68,70 It mediates many leucocyte functions, in particular inflammation. Two groups of PLA2 enzymes, designated GVII and GVIII, catalyses the hydrolysis of the acetyl group from the sn-2 position of PAF to produce lyso-PAF and acetate, which is why the enzymes were originally named PAF acetylhydrolases (PAF-AH) (Figure 1). GVIIA PLA2 is a secreted enzyme with a molecular weight of 45 kDa that associates with LDL and HDL in human plasma.20 Therefore, the enzyme is also known as plasma PAF-AH (pPAF-AH) or Lp-PLA2. GVIIB PLA2, also referred to as PAF-AH II, is an intracellular protein with a molecular weight of 40 kDa that has an N-terminal myristoylation site and which shares 41% amino acid sequence identity with GVIIA PLA2.69 GVIII PLA2 is a brain intracellular heterotrimeric protein complex that is also referred to as PAF-AH Ib although this enzyme also displays significant phospholipase A1 activity.70
Lp-PLA2 binds to two distinct domains on apoB, and after secretion it is primarily transported by LDL and lipoprotein (a) [Lp(a)].13,14 About 30% of Lp-PLA2 binds to apolipoprotein A-I, and transported by HDL particles.68
Lp-PLA2 cleaves oxidized lipids from the sn-2 position of the apoB100-containing lipoproteins LDL and lipoprotein (a) [Lp(a)]14 The bioactive lipids include oxidized non-esterified fatty acids and lysophosphatidylcholine (Figures 1 and 4). Saturable PAF receptor-independent transport rather than intravascular hydrolysis modulates the pro-inflammatory and pro-apoptotic oxidized lipid mediators.71 In a diabetic, hypercholesterolaemic porcine model of atherosclerosis, upregulation of Lp-PLA2 increased peripheral blood mononuclear cell expression of ICAM-1 and IL-6, and coronary artery expression of ICAM-1 and VCAM-1.72
Epidemiology of phospholipase A2 enzymes and cardiovascular disease risk
Associations between concentrations and enzyme activities of sPLA27 and Lp-PLA27,8 have been investigated in multiple population-based studies, and a clinical outcomes trial with statin therapy.12
Secretory phospholipase A2 concentration and activity, and risk of cardiovascular disease events
Several sPLA2 isoenzymes contribute to atherogenesis; however, measurement of circulating sPLA2 concentration has been limited to the GIIA sPLA2. Overall activity of sPLA2 has been measured in prospective studies, but there are no commercially available assays that quantify the activity of the individual sPLA2 isoenzymes.
High levels of GIIA sPLA2 predict CHD events in patients with stable CHD73–75 and unstable angina,62,76 and all-cause mortality in acute myocardial infarction patients.77 In population studies, both high group IIA sPLA2 concentrations and increased sPLA2 activity predict incident cardiovascular events in apparently healthy individuals.78,79 Certain polymorphisms (SNPs) in the GIIA-sPLA2 coding gene PLA2G2A are associated with serum GIIA-sPLA2 concentrations79,80 and cardiovascular events.80 In the KAROLA study of patients who participated in in-patient cardiac rehabilitation after an acute cardiovascular event, the strongest associations with GIIA-sPLA2 levels were seen with SNPs rs4744 and rs10732279, which were associated with higher serum concentrations of 81 and 73%, respectively, per variant allele. Although these two SNPs were associated with recurrent CVD events, the risk relationships were not statistically significant [1.16 (0.89–1.51) and 1.18 (0.91–1.52), respectively, per variant allele].
Lipoprotein phospholipase A2 concentration and activity, and risk of cardiovascular disease events
The Lp-PLA2 Studies Collaboration examined the associations between Lp-PLA2 concentration and activity and CVD events from 79 036 participants who participated in 32 prospective studies.8 In the 32 prospective studies, Lp-PLA2 activity was measured in 57 931 participants from 18 studies, and Lp-PLA2 mass was measured in 58 224 participants from 21 studies. Overall, risk ratios for CHD and ischaemic strokes increased progressively for every 1-standard deviation higher Lp-PLA2 activity or concentration in models that adjusted for age, sex, baseline history of vascular disease, and other non-lipid and lipid risk factors. Risk ratios for CHD events were 1.10 [95% confidence interval (CI) 1.05–1.16] for Lp-PLA2 activity and 1.11 (1.07–1.16) for Lp-PLA2 concentration; 1.08 (0.97–1.20) and 1.14 (1.02–1.27) for ischaemic stroke; 1.16 (1.09–1.24) and 1.13 (1.05–1.22) for vascular mortality; and 1.10 (1.04–1.17) and 1.10 (1.03–1.18) for non-vascular mortality. However, the associations between Lp-PLA2 activity and CHD events were significant only for patients with stable CHD [1.17 (0.78–1.76)], whereas the association was not significant in subjects with no history of CHD [1.03 (0.95–1.12)]. In contrast, Lp-PLA2 mass was predictive of CHD events in both groups of participants. Subsequent to this meta-analysis, an analysis from the Nurses’ Health Study reported that Lp-PLA2 activity was associated with incident myocardial infarction (RR 1.75, 95% CI 1.09–2.84) after multivariable adjustment for clinical, lipid, and inflammatory risk factors.9
PLASG7 is the gene for LpPLA2 or group VII phospholipase A2. A missense mutation (Val279Phe, exon 9, position 994;G→T) of the LpPLA2 gene results in complete loss of enzymatic activity,70 whereas the VV genotype is associated with increased LpPLA2 activity.81 The associations between the LpPLA2 gene polymorphism V279F and CHD was recently investigated in two meta-analyses. In 26 000 individuals, PLA2G7 polymorphisms 379V was associated with changes in enzymatic activity, but not with coronary atherosclerosis or CHD events.82 In an analysis of seven studies involving predominantly Asian individuals (7705 from a total of 7948), there was no overall association with CHD risk [OR = 1.14; (95% CI 0.86–1.52)].83 However, the association with CHD events was significant within the Japanese subgroup [OR = 1.38; (95% CI 1.21–1.56)]. In a larger study from two data sets of Korean men, one copy of 279F null allele was associated with a 20% reduction in the CHD risk.15
These gene association studies provide evidence that reduced LpPLA2 activity is associated CVD in populations with high prevalence of this loss-of-function mutation.
As discussed earlier, Lp-PLA2 circulates in a complex with apoB, thus apoB levels may confound its association with the CVD risk.13,14 The Heart Protection Collaboration Group investigated associations between baseline Lp-PLA2 concentration and activity with incident CVD events in a randomized, placebo-controlled clinical trial of simvastatin.12 Baseline Lp-PLA2 concentration and activity were associated with CHD events in analyses that adjusted for non-lipid risk factors [hazard ratio (HR) per 1-SD Lp-PLA2 concentration, 1.08 (1.03–1.12); and HR per 1-SD Lp-PLA2 activity, 1.11 (1.06–1.15)]. After adjustment for apoB, the associations between Lp-PLA2 activity and CHD events became non-significant (HR 1.02, 0.97–1.06), whereas associations between Lp-PLA2 concentrations remained significant (HR 1.05, 1.01–1.09). This report did not investigate on-trial associations between Lp-PLA2 activity and a cardiovascular event, which is relevant because statins reduce LpPLA2 concentrations by 25% and essentially all high-risk individuals will be treated with statin therapy.
Pharmacology of phospholipase A2 inhibitors
Inhibitors of sPLA2 and Lp-PLA2 have distinct chemical structures and pharmacological properties that target the specific enzymatic site for the two enzymes.84 Thus, the mechanisms of action for the two classes of inhibitors will be discussed separately in this section. An overview of the current status of the phospholipase A2 inhibition programs is summarized in Table 1.
Comparison between LpPLA2 and sPLA2 inhibition aspotential anti-atherosclerosis therapies
Phospholipase A2 inhibition programs
Target: enzymatic activity inhibition
sPLA2s (II >X > V)
Target biology validation
Effect in vivo disease models
Target is a biomarker/clinical association with CVD events/risk
Yes (26 studies)
Yes (8 studies)
Clinical data with compound
Phase I, II (3); III ongoing
Phase I, II, and III ongoing
Phase III trials completed
STABILITY, October 2012
VISTA-16, March 2012 (terminated)
SOLID April 2014
Changes in lipid parameters
Yes: decrease in LDL-C, non-HDL-C, apoB
Changes in inflammatory biomarkers
sPLA2-II-concentration, C-reactive protein
Human in vivo imaging data
No data available
Positive changes in plaque
Yes (human and pig)
No data available
Secretory PLA2 enzymes share a common catalytic dyad that is distinct from other phospholipases that have a classic serine-based catalytic triad (Asp-His-Ser). This unique catalytic dyad served as the target for design of sPLA2 inhibitors.85,86 Varespladib sodium ([3-(aminooxoacetyl)-2-ethyl-1-(phenylmethyl)-1H-indol-4-yl]oxy]acetate; A-001; Anthera Pharmaceuticals Inc., Hayward, CA, USA) is an intravenous formulation and varespladib methyl (Methyl[[3-(aminooxoacetyl)-2-ethyl-1-(phenylmethyl)-1H-indol-4-yl]oxy]acetate; A-002; Anthera Pharmaceuticals, Hayward, CA, USA) is an oral formulation of indole-based sPLA2 inhibitors.16 Varespladib is a pan inhibitor of sPLA2 enzymes with half maximal inhibitory concentrations (IC50) in the low nM range (group IIA IC50: 9–14 nM; group V IC50: 77 nM; and group X IC50: 15 nM). The methylated prodrug, varespladib methyl, is used for oral administration and is rapidly metabolized by esterases to varespladib (A-001). Recent findings shown that sPLA2 enzymes, GII, V ad X, can participate in intracellular arachidonic acid release and as a consequence in the intracellular inflammatory signalling and production of bioactive lipid mediators by different pathways.86 Thus, to inhibit the intracellular inflammatory component of sPLA2 activity contributing to atherosclerosis and plaque rupture, sPLA2 inhibitors should penetrate cells. Indole-based sPLA2, such as varespladib acid, is cell impermeable,87 and therefore incapable of blocking intracellular effects. Furthermore, one may extrapolate that poor cell permeability may also compromise the transport of varespladib across arterial endothelial cells at places of atherogenesis. The clinical failure of this compound to produce beneficial results in rheumatoid arthritis88 and sepsis patients may in part be due to issues of poor cell permeability.
A novel class of GIVA cPLA2 inhibitors, the 2-oxamides, has been shown to inhibit sPLA2 activity. The long-chain 2-oxamide GK126 inhibited human group IIA sPLA2 activity (IC50 300 nM) and GV sPLA2 activity (IC50 440 nM), while it had no effect on GX sPLA2 activity.89
Several selective and highly potent azetidinone inhibitors have been developed as pharmacological tools, and one agent, darapladib (SB480848), has entered into two large Phase III clinical outcomes trials. Azetidinones target the active-site serine residue of the enzyme.90 The Lp-PLA2 inhibitor darapladib has an IC50 of 270 pM.91 In phase II trials, the reduction in circulating Lp-PLA2 activity has achieved 80%.92,93 Rilapladib is a second oral inhibitor of the azetidinones class that has entered phase II studies.94,95
Experimental animal studies of secretory phospholipase A2 inhibition
The anti-atherosclerotic effects of varespladib methyl have been investigated in several non-clinical models that include an ApoE−/− mouse model,96 an accelerated ApoE−/− mouse model,97 and a guinea pig atherosclerosis model.98 In ApoE−/− mice, the combination of varespladib methyl and pravastatin effectuated a synergistic effect on reducing atherosclerosis.97 Varespladib methyl treatment also increased fibrous cap thickness resulting in more stable plaque architecture. However, the C57BI6 murine model used for generation of apoE−/− mouse is limited by its lack of GIIA sPLA2 expression, thus the anti-atherosclerotic effect of varespladib is due to inhibition of other sPLA2 enzymes and/or a specific unknown sPLA2 enzyme that is expressed in the mouse as a compensatory mechanism for lacking GIIA sPLA2 expression. In a guinea pig model, animals were fed a high-fat, high cholesterol diet to induce atherosclerosis.98 Varespladib methyl (150 mg/kg/day) reduced cholesterol accumulation in aortic tissue without changing serum cholesterol levels; however, atherosclerosis was non-significantly reduced. Together, these rodent studies show that varespladib has a beneficial effect on atherosclerosis.
Clinical trials of phospholipase A2 inhibitors
Pharmacological approaches with a lipoprotein-modulating and anti-inflammatory agent are potential novel approach to treating patients with atherosclerosis. Inhibition of sPLA2 by means of administering a pan sPLA2 inhibitor is one such approach. Clinical trials with the sPLA2 inhibitor varespladib methyl have included phase II trials in patients with stable CHD and post-ACS. The recently terminated phase III trial with varespladib methyl was designed to investigate the short-term effects of varespladib methyl in ACS patients (Press release Anthera Pharmaceuticals, Inc., 9 March 2012).
Phospholipase levels and serological markers of atherosclerosis (PLASMA) was a phase II, randomized, double-blind, placebo controlled parallel arm dose-ranging study of four doses of varespladib methyl (50, 100, 250, and 500 mg twice daily) in 396 patients with stable CHD.99 After 8 weeks, varespladib methyl treatment reduced GIIA sPLA2 levels from 69 to 96% in a dose-dependent manner (P < 0.0001). Varespladib methyl treatment also lowered concentrations of LDL cholesterol, oxidized LDL, and hs-C-reactive protein (Figure 5). The most common adverse reactions were non-specific and transient with no-dose response relationship.99–101 In PLASMA II, varespladib methyl 250 mg and 500 mg given once daily was evaluated in 135 stable CHD patients.100 When compared with placebo, 8 weeks treatment with varespladib methyl lowered GIIA sPLA2 protein concentrations by 73 to 84%. Varespladib methyl 500 mg once daily reduced concentrations of LDL cholesterol by 15% (P < 0.001), non-HDL cholesterol by 15% (P < 0.0001), and ApoB by 15% (P < 0.01); however, the lipid effects were non-significantly changed with the lower dosage. In the Fewer Recurrent Acute coronary events with Near-term Cardiovascular Inflammatory Suppression (FRANCIS) trial, the effects of varespladib methyl were evaluated on plasma biomarkers in 625 ACS patients who were treated with atorvastatin 80 mg daily.101 Compared with placebo, varespladib methyl significantly lowered GIIA sPLA2, LDL cholesterol, and hs-C-reactive protein levels at each time point except at week 8 for hs-C-reactive protein. More varespladib methyl treated subjects (51%) had combined LDL cholesterol levels <1.81 mmol/L and hs-C-reactive protein <3 mg/L at 24 weeks than those treated with placebo (34%, P = 0.0013). At the same point, 27% subjects treated with varespladib methyl had combined LDL cholesterol levels <1.81 mmol/L (70 mg/dL) and hs-C-reactive protein <1 mg/L compared with 16% on the placebo (P = 0.01). In a subgroup analysis of 624 diabetes patients, the anti-inflammatory effect of varespladib methyl was larger in patients with diabetes.102 Further corroboration of the differences in the anti-inflammatory effects of varespladib methyl in diabetic and non-diabetic patients requires further investigation in the larger Vascular Inflammation to Suppression to Treat Acute coronary syndromes at 16 weeks (VISTA-16) trial.103,104
Through inhibition of secretory phospholipase (sPLA2) activity, varpladeib reduces phosphotidylcholine hydrolysis on native lipoproteins resulting in a larger LDL particle that is a more effective substrate for the apoB/E (LDL) receptor. The larger LDL particles are more rapidly cleared from the circulation in the presence of statin therapy, which increases LDL receptor activity resulting in a reduction in LDL particles and small VLDL particles.
In the SPIDER-PCI (Secretory PLA2 Inhibition to Decrease Enzyme Release after Percutaneous Coronary Intervention) trial, stable CHD patients were given high-dose varespladib methyl (500 mg twice daily) 3 to 5 days before elective PCI.105 After 18–24h, sPLA2 activity was significantly reduced. Five days after PCI, sPLA2 activity was significantly reduced, but this reduction was not associated with reduced myocardial necrosis.
Several phase II trials with varespladib methyl demonstrate significant reductions in concentrations of GIIA sPLA2, LDL cholesterol, non-HDL cholesterol, and apoB. Limitations of these studies include lack of sPLA2 activity that may have resulted from the use of an assay with insufficient sensitivity at the reduced enzyme mass.99 Although the mechanism for the reduction in GIIA sPLA2 concentration requires further investigation, it has been proposed that varespladib interferes with the sPLA2 synthesis and secretion.106 The involvement of ligand-receptor-mediated signalling in this autocrine pathway remains unknown.
Secretory phospholipase A2 inhibition and cardiovascular disease events
The effects of sPLA2 at the interface of lipoprotein modification and vascular inflammation while extending myocardial ischaemic injury served as the basis for the concept that sPLA2 inhibition may be particularly useful for the prevention of cardiovascular morbidity and mortality in high-risk CHD patients and particularly in those patients presenting with ACS.18,19,103
VISTA-16 (at 16 weeks, NCT01130246) was an international multicentre double blind, placebo-controlled Phase III trial in 6500 high-risk ACS patients.103,104 This trial investigated the short-term efficacy of varespladib methyl therapy (500 mg daily) on cardiovascular morbidity and mortality when added to standard of care and atorvastatin. The ACS study population had been enriched by inclusion of other prior cardiovascular event or intervention and other risk factors that may include either diabetes mellitus, metabolic syndrome, HDL cholesterol <1.1 mmol/L, estimated GFR <60 mL/min. The primary endpoint was the composite of cardiovascular death, non-fatal myocardial infarction, non-fatal stroke, or documented unstable angina with objective evidence of ischaemia requiring hospitalization. Study subjects were randomized within 96h of the index event, and followed for a minimum 16 weeks. Survival status was ascertained at 6 months. Limitations of the VISTA-16 trial design included inclusion of only 6500 subjects in a short-term trial that was time-limited and not event driven. On 9 March 2012, this trial was halted by the Data and Safety Monitoring Board due to a lack of efficacy (Press release Anthera Pharmaceuticals Inc., 9 March 2012).
Another population for the future use of a sPLA2 inhibitor are patients with type 2 diabetes who often manifest the atherogenic dyslipidaemia and low-grade inflammation. The inflammation in type 2 diabetes is associated with central obesity and accumulation of inflammatory cells within the visceral adipose tissue. Plasma sPLA2 activity is reported to be elevated in obesity.107 Inhibition of sPLA2 would be expected to decrease inflammatory cytokines such as IL-6, which is elevated in type 2 diabetes. Moreover, sPLA2 inhibition reduces NEFA levels, which would be expected to reduce glucose levels in the condition. Elevated concentration of small LDL particles in type 2 diabetes is associated with an increased risk for CHD.108 sPLA2 activity can generate small-dense LDL particles, and its inhibition reverses the effect. Therefore, pharmacological approaches with a dual effect that target inflammation and lipoprotein modulating effect reducing small-dense LDL particle formation could therefore provide a novel approach to treat patients with type 2 diabetes and its cardiovascular complications.
Experimental animal studies of lipoprotein-associated phospholipase A2 inhibition
Lipoprotein-associated phospholipase A2 inhibition with darapladib reduced formation of atherosclerotic lesions in experimental animal studies, and improves stability of atherosclerotic plaques through reduced vascular inflammation.72,93 In hypercholesterolaemic diabetic pigs, darapladib treatment (10 mg/kg/day) reduced vascular Lp-PLA2 activity (by 84%), plaque area and incidence of necrotic cores compared with controls.72 In addition, treatment with darapladib reduced expression of multiple genes related to leucocyte activity (CD68, chemokine receptor/ligand mRNAs). These data suggest that treatment with darapladib has important effects in mitigating key proinflammatory pathways (Figure 5).
Human biomarker studies of lipoprotein-associated phospholipase A2 inhibition
The clinical trial data with the Lp-PLA2 inhibitor darapladib includes a biomarker study and an atherosclerosis imaging trial. In stable CHD patients with achieved LDL cholesterol concentrations <115 mg/dL on atorvastatin, treatment with darapladib 40 mg, 80 mg, and 160 mg reduced Lp–PLA2 activity by 43, 55, and 66%, respectively.92 Lp-PLA2 concentration was reduced by 6, 12.9, and 9.3% at darapladib dosages of 40, 80, and 160 mg daily. Dissimilar to varespladib (sPLA2 inhibitor), treatment with darapladib was not accompanied by changes in lipid concentrations or hs-C-reactive protein. However, interleukin-6 levels (IL-6) were reduced by 12.3% (95% CI 22–1%; P = 0.028).
The Integrated Biomarker and Imaging Study (IBIS-2) investigated the effects of selective Lp–PLA2 inhibition with darapladib on plaque characteristics using intravascular ultrasound-based palpography, and C-reactive protein levels.93 Secondary outcome measures include endothelial function and plaque volume by quantitative coronary arteriography and virtual histology by intravascular ultrasonography. IBIS-2 enrolled 300 patients who had successful PCI or uncomplicated diagnostic catheterization and a suitable non-intervened coronary artery with IVUS. Treatment with darapladib had no effect on the co-primary endpoints that included coronary atheroma deformability assessed by intravascular palpography and plasma hs-C-reactive protein concentration. A secondary endpoint, lipid necrotic core assessed by virtual histology, increased in the placebo group (4.5 ± 7.9 mm3; P = 0.009), while darapladib halted this increase (0.5 ± 13.9 mm3; P = 0.71), resulting in a significant treatment difference of −5.2 mm3 (P = 0.012). These findings from these phase II trials are consistent with the results from the porcine model, and suggest that selective Lp-PLA2 inhibition may improve stability of rupture-prone plaques.
In phase II clinical studies, darapladib is well tolerated at dosages of 40, 80, and 160 mg daily.92 The most common adverse reactions, reported in 16–36% of study subjects, include diarrhoea and malodour of the urine and faeces.
Lipoprotein-associated phospholipase A2 inhibitors and cardiovascular events
The clinical efficacy of darapladib will be investigated in two large-scale cardiovascular event trials that investigate the effects of selective Lp-PLA2 on non-fatal myocardial infarction, non-fatal stroke, and cardiovascular death in CHD patients. Stabilization of Atherosclerotic Plaque by Initiation of Darapladib Therapy Trial (STABILITY) is a multicentre, randomized, double blind, placebo-controlled trial that evaluates the effects of darapladib or placebo on cardiovascular events in 15 500 patients with stable CHD.109 Stabilization Of pLaques using Darapladib Thrombolysis in Myocardial Infarction (SOLID-TIMI52) is designed to investigate the effects of darapladib 160 mg daily vs. placebo on major cardiovascular events in 11 500 patients with a cardiovascular event (myocardial infarction or stroke) within the preceding 30 days.110 These two trials will continue until a requisite number of major CVD events have occurred.
In the two ongoing secondary prevention trials with darapladib, it will be important to investigate the relationships between baseline and on-trial Lp-PLA2 concentration and activity, and cardiovascular events.19 Similar data are relevant for future clinical trials with the sPLA2 inhibitor varespladib methyl.
The two major phospholipase A2 enzymes that have been studied as biomarkers of cardiovascular risk and targets of pharmacological intervention differ in several important aspects. Secretory phospholipase A2 groups IIA, V, and X hydrolyze native lipoproteins resulting in smaller, denser, and more electronegative lipoprotein particles that are less avidly internalized by the hepatic apoB/E receptor resulting in increased residence time in the circulation that allows for further physicochemical alterations, increased intimal proteoglycan binding, and incorporation by macrophage scavenger receptors and specific PLA2R. Lp-PLA2 requires oxidized LDL as a substrate, and it has no effect on native lipoproteins. The hydrolysis of phospholipids on lipoproteins and cell membranes results in bioactive lipids (NEFAs, lysophospholipids, and eicosanoids) that activate pro-inflammatory redox-sensitive transcription factors and enhance proapoptotic effects.111
Varespladib is a selective inhibitor of sPLA2 that has efficacy against multiple group IIA isoenzymes (pan-inhibitor). For this reason, some investigators refer to varespladib as non-specific inhibitor even though it is highly specific for sPLA2. In stable CHD patients, varespladib methyl 500 mg daily reduced concentrations of LDL cholesterol, non-HDL cholesterol, and apoB.99,100 In addition, there were reductions in LDL and VLDL particle concentrations. In these trials, oxidized LDL levels were reduced, but concentrations of systemic inflammatory biomarkers were non-significantly reduced. However, in ACS patients, varespladib methyl reduced hs-C-reactive protein concentrations.101 Despite the favourable short-term biomarker data in varespladib methyl-treated subjects, therapy had no effect on cardiovascular events in the VISTA-16 trial.103,104 One reason may be the short duration of treatment to test an anti-atherosclerotic mechanism.
Lp-PLA2 inhibition with darapladib has undergone evaluation in a biomarker study,92 and a human atherosclerosis trial.93 In the biomarker trial, darapladib had no effects of plasma lipids, lipoproteins, or hs-C-reactive protein.92 In the coronary atherosclerosis imaging trial, there were no reductions in atheroma volume but the lipid necrotic core was reduced.93 In aggregate, the studies with darapladib indicate that this agent may reduce vulnerability of rupture-prone atherosclerotic lesions.
The ongoing phase III clinical trials with the Lp-PLA2 inhibitor will provide important information about the potential effects of targeting vascular inflammation through inhibition of Lp-PLA2 in high-risk CHD patients. Future trials with sPLA2 inhibitors may require use of a lipophilic agent and design of a trial with sufficient duration to investigate atherosclerosis mediated cardiovascular events.
Conflict of interest: E.H.-C. is employed by AstraZeneca, R&D, CVGI iMed, Sweden. R.S.R. has no conflicts of interest.
. Serum amyloid A as a predictor of coronary artery disease and cardiovascular outcome in women: the national heart, lung, and blood institute-sponsored Women's Ischemia Syndrome Evaluation (WISE). Circulation 2004;109:726-732.
. Inflammation, pravastatin, and the risk of coronary events after myocardial infarction in patients with average cholesterol levels. Cholesterol and Recurrent Events (CARE) Investigators. Circulation 1998;98:839-844.
. Role of group IIa and group V secretory phospholipases A2 in the metabolism of lipoproteins. Substrate specificities of the enzymes and the regulation of their activities by sphingomyelin. Biochemistry 2002;41:4911-4920.
. Role of group II secretory phospholipase A2 in atherosclerosis: 1. Increased atherogenesis and altered lipoproteins in transgenic mice expressing group IIa phospholipase A2. Arterioscler Thromb Vasc Biol 1999;19:1284-1290.
. Functional association of type IIA secretory phospholipase A2 with the glycosylphosphatidylinositol-anchored heparan sulfate proteoglycan in the cyclooxygenase-2-mediated delayed prostanoid-biosynthetic pathway. J Biol Chem 1999;274:29927-29936.
. Molecular mechanism for changes in proteoglycan binding on compositional changes of the core and the surface of low-density lipoprotein-containing human apolipoprotein B100. Arterioscler Thromb Vasc Biol 2004;24:564-570.
. Analyses of group III secreted phospholipase A2 transgenic mice reveal potential participation of this enzyme in plasma lipoprotein modification, macrophage foam cell formation, and atherosclerosis. J Biol Chem 2008;283:33483-33497.
. CRP, interleukin-6, secretory phospholipase A2 group IIA, and intercellular adhesion molecule-1 during the early phase of acute coronary syndromes and long-term follow-up. Int J Cardiol 2006;108:55-62.
. Prognostic value and the changes of plasma levels of secretory type II phospholipase A2 in patients with coronary artery disease undergoing percutaneous coronary intervention. Eur Heart J 2003;24:1824-1832.
. Association of a G994→T missense myutation in the plasma platelet-activating factor acetylhydrolyase gene with genetic susceptibility to nonfamilial dilated cardiomyopathy in Japanese. Circulation 1998;98:1881-1885.
. Pharmacology of A-001/S-5920, [[3-(aminooxoacetyl)-2-ethyl-1-(phenylmethyl)-1H-indol-4-yl]oxy] acetate, a potent and selective secretory phospholipase A2 inhibitor: A new class of anti-inflammatory drugs, SPI. J Pharmacol Exp Ther 1999;288:1117-1124.
. Intracellular actions of group IIA secreted phospholipase A2 and group IVA cytosolic phospholipase A2 contribute to arachidonic acid release and prostaglandin production in rat gastric mucosal cells and transfected human embryonic kidney cells. J Biol Chem 2006;281:16245-16255.
. A randomized, double-blinded, placebo-controlled clinical trial of LY333013, a selective inhibitor of group II secretory phospholipase A2, in the treatment of rheumatoid arthritis. J Rhematol 2005;32:417-423.
; Darapladib Investigators. The effect of darapladib on plasma lipoprotein-associated phospholipase A2 activity and cardiovascular biomarkers in patients with stable coronary heart disease or coronary heart disease risk equivalent: the results of a multicenter, randomized, double-blind, placebo-controlled study. J Am Coll Cardiol 2008;51:1632-1641.
; Integrated Biomarker and Imaging Study-2 Investigators. Effects of the direct lipoprotein-associated phospholipase A(2) inhibitor darapladib on human coronary atherosclerotic plaque. Circulation 2008;118:1172-1182.
. Inhibition of secretory phospholipase A2 in patients with acute coronary syndromes: rationale and design of the vascular inflammation suppression to treat acute coronary syndrome for 16 weeks (VISTA-16) trial. Cardiovasc Drugs Ther 2012;26:71-75.

domain: medical


text:
xDs For Sale, Redwood City
There are limited results in your location. You can expand the distance to find
items using the location bar on the left.
3 hrs ago
2013 SCION XD 5DR HB AT, REST EASY! With its 1-Owner & Buyback Qualified CARFAX report, ...
All my Ads »

domain: geography


text:
Food production needs to double by 2050 while agriculture is contributing 30% to GHG emissions and soils are degrading. Fuel and fertiliser prices have increased by 40% and 60% respectively over the last few months, leaving farmers behind with a huge financial burden. Clearly something needs to change. At Agros we decided to start leveraging the power of nature to improve farmers’ livelihoods and climate resilience season after season.
Agros believes in a world in which small farmers continue to feed our planet in a sustainable and profitable way. We’re revolutionising agriculture across Southeast Asia, giving millions of famers the technologies, services and financing they need to transition to long-term profitable farming. Our mission is to double the income of farmers while abating millions of carbon emissions.
We are Asia’s first farming-facing regenerative farming platform – Technology, Services and Finance – to help crop farmers be profitable and sustainable, season after season. Our platform allows farmers to reduce input costs, restore soil health and increase yields. We guarantee adoption, longevity and affordability by designing products locally, providing data-driven education and offering flexible financing solutions.
This demands the brightest minds thinking creatively together. We’re a young and dynamic team consisting of 40+ driven people on a mission to create a better world for those affected the most by climate change. Our teams are based in Singapore, Myanmar and Cambodia and we are looking to expand to other neighbouring countries in the coming years.

domain: agronomy


text:
Yellow brick road leads to downtown Birmingham
The Birmingham Palladium is set to host a public premier for the upcoming movie ‘Oz: The Great and Powerful’
Posted February 19, 2013
BIRMINGHAM — As Dorothy once said, “There’s no place like home.” And for the actors of the upcoming feature film, “Oz: The Great and Powerful,” there’s no better place to celebrate the movie’s premiere than right here in Michigan.
On March 8, the Birmingham Palladium 12 Theater will host a Hollywood-style premiere event for the film’s Michigan cast, their families and friends. According to event organizer Colin McConnell, the premiere will give the movie’s local talent a chance to see the completed “Oz” together.
“We were talking one day and we were saying a majority of the people that were in the (film) that weren’t the four or five big names are from Michigan,” said McConnell. “We wanted to do a premiere for them because we didn’t know if Disney was going to do a premiere here, and even if they were, it was going to be a plus-one, not a friends and family invitation.”
McConnell has a special interest in the making of the film because his longtime friend Martin Klebba was part of the cast. Klebba, originally from Troy and a graduate of Troy Athens High School, has had roles in a long list of hit movies, including “Mirror, Mirror” and the “Pirates of the Caribbean” films.
Klebba said working on “Oz” in Michigan for two months was the best of both worlds — all the glamour and big-name actors of Hollywood, with all the comforts of home.
“It was cool. I had just got done doing ‘Mirror, Mirror’ in Montreal and I got to go home for one day then I had to fly back to Detroit to work two months on ‘Oz,’ which was an honor because everyone loves the original,” said Klebba, who now lives in Sterling Heights with his wife and new baby. “I was working for (director) Sam Raimi, who’s also a Michigander. Basically, I got to stay home and hang out with all my friends and family during the day then go to work, which is something I don’t normally get to do.”
Klebba called the “Oz” experience “amazing,” and said the prequel to the classic film “The Wizard of Oz” was kept tightly under wraps from the public. In fact, he said most of the actors in the film didn’t even receive scripts until they showed up to the set for the day and received their lines on the spot.
He also spoke of the talents of the lead actors, such as Zach Braff and James Franco, who plays the man who would become the Wizard of Oz.
“He’s one of those Johnny Depp kind of actors. He’s just that good.”
After such a positive experience making the movie, Klebba said he’s excited to be able to celebrate the film with his fellow actors who aren’t able to go to the official Disney premiere event in California. McConnell agreed, adding that the Palladium is providing six theaters to screen the film, so all of the actors and background performers can enjoy the party together, no matter how large or small their role.
“We wanted to do a big red-carpet thing. A lot of the (background performers) are real estate agents or car sales people — they probably won’t do another movie again,” said McConnell.
As guests enter the theater that evening, they’ll be able to greet the press on the “red carpet,” which McConnell said will actually be a yellow brick road. He said that after the screening the group hopes to be able to continue the celebration at an after-party somewhere in downtown Birmingham. Organizers are still seeking a venue and sponsors to make that a reality.
Proceeds from the premiere will benefit the Coalition of Dwarf Advocacy, a charity Klebba helped to found. As a little person himself, Klebba said he’s excited to raise a few bucks for CoDA, which facilitates adoptions and scholarships for young dwarf orphans.
“I’ll be able to watch (the movie) with the munchkins and the winkies and the tinkers and they’ll get to experience what it’s like to have a red carpet, or yellow brick road carpet, and what it’s like to be a part of a film like that, and hopefully raise a lot of money for CoDA at the same time. It’s going to be amazing,” said Klebba.
The “Oz: The Great and Powerful” friends and family movie premiere fundraiser will begin at 7:30 p.m. March 8 and is open to the public with ticket prices of $15 for 2D or $18 for 3D. Tickets can be purchased at www.gofundme.com/CoDAmoviefundraiser. For more information on the Coalition for Dwarf Advocacy, visit www.dwarfadvocates.com.
The Birmingham Palladium 12 Theater is located at 250 N. Old Woodward Ave. in downtown Birmingham.
About the author
Staff Writer Tiffany Esshaki covers Birmingham, Bloomfield Hills and Bloomfield Township as well as Oakland County Parks and Recreation and Oakland County Animal Control and Pet Adoption Center. Esshaki has worked for C & G Newspapers since 2011 and attended the University of Michigan-Dearborn and Oakland Community College. She’s the recipient of several awards from the Michigan Press Association and the Detroit chapter of the Society of Professional Journalists.
More from C & G Newspapers
Sterling Heights / Warren
Warren / Center Line


text: Color, design and contrast represent these images from Antelope Canyon in Page, AZ. I amazed myself with the compositions created in these slot canyons as the canyons walls are dark within and it is difficult to visualize the nuances of detail unless you are looking at the light streaming through the tops. The experience of creating these image compositions, for me, was both spontaneous and intuitive. I had to be present in the moment to capture the light, the shadow and the intricate detail all around me. For those who enjoy photography tips… I exposed for the highlights of the image and then closed down in 1/3 stop increments to bracket several additional photographs to get the detail in the shadows. My 2nd or 3rd exposure seemed to provide the best highlights and shadow details. Exposures were long so I was using both a tripod and cable release for stability.


text:
Core Financial Advisors
1918 Business Center Dr Suite 110
San Bernardino, CA 92408
See Also: San Bernardino, CA Financial Services
This is a listing for Core Financial Advisors
(1918 Business Center Dr Suite 110, San Bernardino, CA 92408) from the Financial Services Directory. You can find more similar services in our Financial Services Directory.
Other listings in the San Bernardino, CA Financial Services Directory: Ddd Investments, Crown Valley Escrow Inc, Chase Bank, Clearpoint Financial Solutions, Crawford Investment Co.

domain: economics


The major talking point surrounding ongoings at the Emirates this weekend of course came in the form of Mesut Ozil’s future.
German midfielder Ozil, having not lined out for Arsenal on a single occasion since last March, evidently came to the conclusion over the last couple of weeks that his time in the colours of the Gunners was up.
In turn, as confirmed on Saturday, the 32-year-old has agreed to a contract termination in north London, 6 months prior to his scheduled departure date:
This was swiftly followed, meanwhile, by Ozil all-but confirming his next destination, via a series of social media posts.
The one-time Real Madrid standout is primed to link up with Turkish giants Fenerbahce, who beat out competition from MLS outfit DC United to convince him of their project:
Ozil > KDB
Ozil, though, despite the altogether underwhelming nature of his imminent exit, evidently bears Arsenal no ill will.
Mikel Arteta’s men are of course set to get their Premier League campaign back underway this evening, in a meeting with Newcastle United.
And, ahead of kick-off in north London, the German was keen to reiterate the fact that he ‘will always support the Gunners’, in a good luck message to his soon-to-be former employers:
Coincidentally, Ozil’s message to his Arsenal teammates comes on the same day that a series of statistics have resurfaced online, highlighting just how special the midfielder was in his prime.
Much has of course been made of the struggles for creativity at the Emirates to this point in the campaign, with Arteta’s men having frequently come up short in their efforts to break down deep-lying opposing backlines.
Ozil, though, is one individual who, from an individual standpoint at least, would certainly find it tough to relate to such woes.
As revealed by the reliable OptaJoe on Monday, the gifted no.10 maintains the record for the most chances ever created over the course of a single Premier League season (since such metrics first began to be noted).
This came in 2015-16, when, remarkably, Ozil managed to carve out 146 opportunities for his Gunners teammates:


Question about Sony VGN-S150 MBX109 (M9_32 (A1072469A) PC Notebook
Make sure your BIOS can handle an upgraded CPU. I would go into bios, check the "default" or "fail safe" boot option and then power down. Lift keyboard or access your CPU and change out. Remember to touch something metallic before you operate so you discharge any static electricity (killed a CPU and a video card by not doing this ONCE) Boot up and notice any BIOS messages, may want to start in safe mode just to get a basic working config going, then power down, access BIOS and start getting it back to your best config, maybe stepping one item at a time, rebooting and continuing until you get a stable platform.
Posted on Jan 22, 2008
Tips for a great answer:
Feb 06, 2013 | Medion PC Laptops
Jun 02, 2012 | Toshiba PC Laptops
Sep 09, 2011 | EliteGroup G320 Notebook
May 09, 2011 | Toshiba PC Laptops
May 21, 2009 | Dell Latitude L400 Notebook
May 09, 2009 | Fujitsu AMILO D-7820 Notebook
Mar 22, 2009 | Toshiba Satellite A10 Notebook
Dec 27, 2008 | Dell Latitude D600 Notebook
116 people viewed this question
Usually answered in minutes!
Step 2: Please assign your manual to a product:


text:
Top critical review
Modern take on classic fairy tales
on 11 August 2009
Whilst this book is cleverly written & excellently illustrated I do feel that, on reflection, my 3 & 4 year old are a bit too old to get much enjoyment from it. We'll probably read it half a dozen times before it gets relegated for something a bit more sophisticated.

domain: literature


text: A thin sheet of frost spreads over the earth like a cake dusted with confectioners sugar. Moonlight reflects off the pearlescent ground, emphasising the silhouette of lifeless trees, the branches are frozen solid by whispers of the winter breeze. The World is engulfed in Silence. I sink into my bed, hiding under the soft quilt that protects me from the biting cold. My eyes begin to drown themselves into an ocean of sleep; and then I'm awake in a land of dreams.

domain: artistic


The material used by Hunan Ming Yu Nonwovens Co., Ltd is to assist in high-quality production of nonwoven bags . Since inception, we have been sparing no efforts to select materials with higher performance and longer service life. Fortunately, we have found the right materials for us to provide high quality products at reasonable prices.
Widely recognized as a strong competitor, Ming Yu Nonwovens is always able to manufacture market-oriented spunlace cloth. The Protective clothing series has become a hot product of Ming Yu Nonwovens. The development of Ming Yu polyester needle felt uses many disciplines. They include numerical control technology, dynamic machinery, mechanical engineering and technology, tribology, thermodynamics, etc. It stands out for its good acid and alkali resistance. This product is deigned with good wear-resistance. It can withstand a certain amount of friction without being damaged. It doesn't mold or deteriorate in the presence of bacteria, mold or other elements.
Ming Yualways improves the leading expertise and consistently delivers high-quality Protective clothing. Inquire online!


text:
A trade agreement has been signed between the UK and the EU in light of Brexit negotiations.
Your weekly update from our public affairs team on all the latest developments and debates in Parliament and across Whitehall.
Tim O'Sullivan looks at what the UK-EU Trade and Cooperation Agreement offers for law enforcement and cooperation in criminal justice.
Mickaël Laurans gives his initial assessment of the impact of the Trade and Cooperation Agreement on trade in legal services.
Our briefing on the UK-EU Free Trade Agreement ahead of it being debated in both Houses on 30 December.
We welcome news of a free trade agreement, but warn that more work is needed to facilitate the vital role of the legal sector.
HMCTS has put arrangements in place to deal with any flagged Brexit-no exit divorce and financial remedy cases.
UK solicitors will be able to access Switzerland to advise clients following the Brexit transition date of 31 December 2020.
We welcome the news that UK law firms can continue to work in France following the 31 December Brexit transition.
This podcast outlines the key actions on data flows that firms should be taking to prepare for the end of the Brexit transition period on 31 December.

domain: news


text:
Warner Bros. Pictures has released a new TV spot for Wonder Woman via Youtube, titled “Bang Bang” that’ll give you a bit more hope for the film.
Wonder Woman is based on DC Comics‘ Wonder Woman by William Moulton Marston. The film is the fourth installment to the DC Extended Universe.
Before she was Wonder Woman she was Diana, princess of the Amazons, trained warrior. When a pilot crashes and tells of conflict in the outside world, she leaves home to fight a war to end all wars, discovering her full powers and true destiny.
Wonder Woman is directed by Patty Jenkins, and written by Zack Snyder, Allan Heinberg and Jason Fuchs. The film is produced by Charles Roven, Zack Snyder, Deborah Snyder and Richard Suckle.
Starring in Wonder Woman are Gal Gadot, Chris Pine, Connie Nielsen, Robin Wright, David Thewlis, Danny Huston, Elena Anaya, Lucy Davis, Saïd Taghmaoui and Ewen Bremner.
Wonder Woman is scheduled to release on theaters in June 2, 2017.

domain: entertainment


text:
Find Etoro In Which States…
EToro is one of the most recent and biggest currency trading platforms on the internet. Their free trial duration runs for thirty days. If this business is ideal for you, throughout this time you will have the chance to check everything out and see. This is the main reason that lots of people are selecting it. Here is how you can find out is eToro truly totally free.
The first thing you need to do is go to the website. When you get there you will see there is a link for the demonstration account. There is more details on the site about this currency trading site, so make sure to read it.
Now that you have actually checked out the site and have an interest in trying their totally free currency trading platform you require to act fast. The site states, you need to verify your e-mail address prior to you are able to access the demonstration account. You will require to offer your current mailing address and credit card number. You will need to follow the instructions and put in your personal information. It is best to practice on a website where you have access to real individuals.
They say they do not store your account details with cookies so you will have to sign up for a credit card. What does a business like eToro have to get economically by charging you for a trial membership?
In my opinion they need to charge a small cost for the convenience and reliability they present to their clients. Nevertheless, they also require the money to keep the software application upgraded. When you go back to trading, that method you will not find yourself out of date. Plus, if you want to get involved with the live market then you will require to have your own eToro account. This takes time but is totally complimentary. Plus you can utilize it while you practice your trading skills without the worry of losing any money.
There is a link on the top right hand corner of the page that will take you to the account you require to open with your credit card. All you will need to do is follow the guidelines and see as your trades go through the roof. Etoro In Which States

domain: finance


text:
A Scots couple who were stranded on a cruise ship hit by Coronavirus deaths have finally made it home and revealed how homegrown humour got them through their ordeal.
Morven and Ian Rae, from Glasgow, thought they were embarking on the “trip of a lifetime” for Morven’s 70th birthday celebration when they boarded the Zaandam in Buenos Aires, South America, on March 7.
But when Covid-19 hit the vessel and several passengers fell ill, they found themselves confined to their cabin for two weeks off the Panama coast and was refused entry to several Latin American ports.
Four people died aboard the Zaandam, including two officially diagnosed with the coronavirus, and Morven and Ian believe they may also have been struck down with the virus during their isolation.
But on Thursday, President Trump intervened to allow docking on humanitarian grounds in a Florida port.
The couple were among 200 UK nationals who departed Fort Lauderdale airport for Heathrow to finally return home on Saturday.
Morven, who is originally from Pollokshields, said she felt lucky to have made it home safely when others lost their lives and thanked her musician husband, 73, for keeping her entertained during their days stuck at sea.
She said: “We were in isolation for 13 days. We really felt it was important to keep our spirits up during that time.
“We were protected in our cabins. They had put us there for our own safety rather than imprisoning us but when we heard people had died that was a bad day.
“We knew people were ill and we were pretty sure that we had the virus too. I think probably nearly everyone on the ship had it mildly.
“We had a cough and a bit of a temperature but we weren’t bad enough to call the medics. We just slept and thought, ‘if it gets any worse we’ll contact them’.”
During isolation some healthy guests were transferred to a sister ship The Rotterdam, but Morven and Ian remained on the Zaandam after declaring they had suffered symptoms.
Morven said: “They asked if we had a cough and we said yes. I think other people lied to get off the ship but we were glad we stayed in the Zaandam, we felt safe there. “I think we thought it was the safest place we could be.”
The couple kept in touch with other passengers via a Whatsapp group and were able to speak with family over social media, with Ian posting upbeat videos and messages on Facebook to reassure loved ones and even writing songs about their experience.
Morven, who now lives in South London, said: “We got clean towels every day and food delivered to outside our room three times a day. They treated us really well.
“It’s better for your mental health to stay positive. My husband is brilliant in that situation and always looks at what you can do to make things more cheerful. I think that is the Scottish way.”
Now home, the couple have been offered a partial refund and money towards a future cruise as well as having all of their on-board purchases waved.
Morven said: “We’re both really weak, I think with just not being able to exercise properly. But the first thing we did when we got home was have a nice cup of tea - which Ian had also written a song about.”


text:
Hong Kong is popularly referred to as the Pearl of the Orient given its strategic location on the ancient trade routes and its present strategic position in World Trade. Hong Kong business opportunities are several and the former city nation, now a part of China, lends itself freely to foreign trade. Though being a Special Administrative Region of China, Hong Kong has its own currency, passport, and has a unique democracy. A third of people’s representatives are directly elected, and the remaining two-thirds are elected by a special 20,000 person constituency of important citizens.
Though Hong Kong is an established business hub, getting around regulations, laws, and commercial formalities can be complicated. While there are opportunities aplenty, local knowledge is vital for success.
Hong Kong’s vital data (as of 2014)
|GDP (billion USD)||291|
|GDP per capita (USD)||40,170|
|GDP per capita PPP (USD)||55,084|
|Purchasing power per dollar, compared to the USA||137%|
|Surface area (sq km)||1,100|
|% of population living in urban areas||100%|
|% of GDP added by agriculture - industry - services||not available for 2014|
|Ease of doing business index (1 = best, 180 = lowest)||3|
What are business opportunities in Hong Kong?
Imports into Hong Kong account for about 120% of its GDP. It would be fair to say that Hong Kong imports most of what it consumes. Hong Kong provides a flexible environment for business, embracing uncertainty and promoting creative thinking.
Where the action really is - Hong Kong international business
The Top 5 imports into Hong Kong are:
- Integrated Circuits
- Broadcasting Equipment
What are Hong Kong’s important events?
The top 3 sports in Hong Kong are Dragon Boat racing, Horse Racing, and Rugby. Hong Kong competes as an independent nation in the Olympics, though it is a part of China, reflecting its own free and autonomous culture.
The pastimes in Hong Kong are truly diverse, from art to Yoga, from food to wine. People in Hong Kong are cosmopolitan by nature.
Hong Kong straddles two completely different worlds, the Communist East and the Capitalist West. Hong Kong Tourism, therefore, offers traditional attractions like Temples, monuments, and Churches to modern must sees such as theme parks, its own Disney world, and a typical skyline of skyscrapers.
Organizing regular trade fairs in exciting surroundings, Hong Kong is one of the most business-friendly territories, being ranked 3rd in the world for your ease of doing business.
Though Hong Kong can be a lucrative market for exporters, its fast paced business environment with global competitors needs local knowledge and support. Armed with the right know-how, Hong Kong can present lucrative opportunities for your long-term prosperity.


text:
A man has been arrested in London after allegedly attacking and threatening a Nine News reporter during a live cross on Wednesday night.
Nine’s Europe correspondent Sophie Walsh was discussing upcoming demonstrations across Europe on Adelaide’s 9News broadcast when a man allegedly grabbed her and made stabbing motions in her direction.
While the physical altercation was not shown during the broadcast, Walsh could be heard screaming.
“Sorry, I just had someone come up and try… A man just came up and grabbed me,” Walsh said.
Speaking to The Australian, Walsh said the man “swung at me and punched me”.
In a tweet, Walsh said the man was chased by camera operator Jason Conduit.
“I’m shaken but ok,” she said.
Thank you for your messages. The man has been arrested for threats to kill and carrying a weapon. I’m shaken but ok. Big thanks to my incredible camerman Jason Conduit who chased him down armed with a light stand and got him arrested. https://t.co/YRxrUB3j4l
— Sophie Walsh (@sophie_walsh9) June 3, 2020
The man was reportedly detained by police in Hyde Park.
In a statement provided to The Australian, the Met Police said a man was “arrested on suspicion of making threats to kill, possession of an offensive weapon and possession of class B drugs.”
“He has been taken to a central London police station, where he remains at this time.”
In a statement, Nine News said it “appreciates the enormous pressure our international correspondents are under and is offering Sophie Walsh ongoing support.”
In a separate incident this morning, Nine’s Europe correspondent Ben Avery and his camera operator fled the scene of a London protest while broadcasting live on Today.
“Had been a peaceful protest until things took a turn late,” Avery said in a tweet.
“The microphone was pulled from my hand and we were chased down the street by a large group.”
Thanks to everyone for your concern after this. Had been a peaceful protest until things took a turn late. The microphone was pulled from my hand and we were chased down the street by a large group. Big thanks to rock solid cameraman @9CadeThompson, our security guard and police https://t.co/hsyUALuljx
— Ben Avery (@benavery9) June 3, 2020
Citing local outlets, Nine News reports the London demonstrations, called to express outrage at the death of African American man George Floyd in US police custody, were largely peaceful.
The incidents come days after a 7NEWS crew was beaten by riot police in Washington D.C., and follow the arrest of a Nine News correspondent covering civil unrest in Minneapolis.

domain: law


[Samba] Remove samba4 server from Domain
caleb at caleboconnell.com
Wed Jan 23 11:53:10 MST 2013
I initially setup a samba4 domain with three servers. The first server I
setup was of course the SOA server for the DNS. I need to now remove this
server from the group and pass along it's roles to another server. I
updated the operations master to the new server but aside from that I Cannot
get the Microsoft tool to change the SOA record.
Basically, I need to know what steps need to be gone through to change all
the primary functions of an AD domain from one samba4 server to another.
I searched Google and checked all the options I could find in the samba-tool
utility but I can't seem to find where this would be done.
Any help, suggestions or links to existing documentation would be great.
Thanks in Advance
More information about the samba


text:
Beloved CISD special education teacher dies
Warren was 67.
"She had a special touch for teaching them," said her husband, Bob Warren.
Kay Perdue Warren was born Oct. 14, 1938, in Kennedy and passed away Saturday in Conroe. Her life was devoted to her work in the CISD special education program.
Warren originally taught special education at the old Conroe ISD administration building. When the CISD administration building moved to its current location on Texas 105, Warren traveled to schools all over the district to work with special-needs children.
When Warren first started working with the local Special Olympics, the children were given poor transportation and lodging when they traveled to compete in events in College Station. After the Warrens got involved, however, competitors received higher-quality buses and hotel rooms, Bob Warren said.
In addition to teaching special-needs children, Warren worked with her husband to organize an annual Special Olympics at Panorama Village.
"We made money every year and had plenty left over every time," Bob Warren said.
The couple married in 1986, the second marriage for both Bob and Kay.
Kay is survived by her husband, sons Duane and Scott Perdue, brother Jimmie Justice, grandchildren Haley and Jake Perdue, and nephew David Justice.
A memorial service for Kay Perdue Warren will be at Metcalf Funeral Directors at 6 p.m. today. In lieu of flowers, contributions can be made to the Special Olympics, Attn: WebGifts, 1133 19th St. NW, 12th Floor, Washington, DC 20036-3604.
Calli Rudebusch can be reached at email@example.com.


text:
In Jack Miller III, Ohio State has finally found a man it can depend on both the field and out. And that spells trouble for anyone who isn't a Buckeye. Jack Miller III gave his pledge after a Friday night recruitment spree.
Jack Miller III is a four-star quarterback who will stop at nothing to achieve his goals. With him on board, there's nothing stopping the Buckeyes now from achieving their objectives. And that's why everyone in the team has been seen jumping for joy lately. The team is confident that it has everything it needs to put the pressure on.
No. 47, as everyone knows is a young but very driven man. At 6' 4" and 210 pounds, Jack Miller III is definitely a lean and mean fighting machine. But you really have to watch his commando-like demeanor to believe, although his play stats say it all.
Let's hear it from Tim Kohner who had the privilege of coaching Jack Miller III for years at school. Kohner's take on the man sounds like he's describing the abilities of the Bionic Man. And who can blame him?
Tragedy Leads to Drive
Jack Miller III has gone against all odds to become one of the most determined young quarterbacks that American football has ever seen. A family friend's tragic accident that occurred while watching him play once made him break down in tears. But instead of bringing him down, Jack Miller III managed to turn a recurrent nightmare into an inspiration to succeed like no one has in the field.
Jack Miller III is making the marks now, but at his age, he's only getting started. You only need to watch him at play in order to realize that he has infinitely so much more to give. Best of all, every ounce of determination in his body shows that when he commits, he really means it. So if Ohio State is as committed to Jack Miller III as he is, the team is unstoppable.
Top 2020 QB Jack Miller Goes In-Depth on Ohio State Commitment
Jack Miller III Transfers From Ohio State
Ohio State quarterback Jack Miller III will transfer from the Buckeyes program. Miller announced his decision on Twitter. Miller could not survive the crowded quarterback room at Ohio State in 2020 and after not playing much, he entered the transfer portal. A total of 42 snaps during the year and an OVI charge that led to a short syspension, was all the third string QB could muster. Quarterbacks CJ Stroud and Kyle McCord got all the important time with the Buckeyes during the season and will return next year.

domain: sports


January 4, 2006
Jim Webb was officially sworn in as Virginia's new senator Thursday afternoon.
Webb is likely to support most of the Democrats' priorities for the first 100 hours of the 100th Congress.
For instance, Webb campaigned on getting rid of corruption in Congress. The Democrats' first order of business will be expanding current bans on gifts and travel from lobbyists to congressman.
Democrats are also focusing on the war in Iraq, namely, what went wrong, and where to go from here.
The war was the backbone of Webb's campaign.
His camp said in a statement today that, "within hours of becoming a U.S. senator- the democratic leadership will introduce his GI bill, to grant our veterans who have served in Iraq and Afghanistan the same kinds of benefits afforded to our WWII veterans."
Matt Smyth from UVa's Center for Politics, says the democratic leadership already agreed to help this bill reach the floor.
"This bill would probably be a good accompaniment to some of the democratic legislation and actions that would be critical of the war in Iraq, and possibly move towards reducing the number of troops, or moving the troops home," said Smyth. "So this would show that the Democrats are pro-soldier, even if they are anti-war."
Smyth believes, this bill has a good chance of passing in the first 100 hours.
The major criticism it would face is, where the money would come from to support it.
Issues on the Democrats' 100 hour agenda Webb may not fully support are social ones, such as the minimum wage, student loans, and stem cell research.
Webb is serving on the Foreign Relations and Armed Services committees.
Viewers with disabilities can get assistance accessing this station's FCC Public Inspection File by contacting the station with the information listed below. Questions or concerns relating to the accessibility of the FCC's online public file system should be directed to the FCC at 888-225-5322, 888-835-5322 (TTY), or firstname.lastname@example.org.


Reviewed By DONNIE W (BOONVILLE, IN) Reviewed for a 1997 GMC Yukon —11/23/2017 12:04 AM
Fifhting bolts with wire fished thru holes in frame. Metal square to lock carage bolts wants to spin inside frame.
Reviewed By Doug L (ROSCOE, IL) Reviewed for a 2002 GMC Yukon —8/9/2017 6:57 AM
OEM hitch was rusted so bad that the electrical connectors bracket fell apart the weld areas were also deteriorated badly. I was concerned about the safety of the hitch. The OEM hitch is only a class 3 and I was pushing the limit on the weight rating. The Curt hitch I got is a class 4. Very reasonable price, cheaper than buying another OEM class 3. Install was easy except for the bolt hole in my bumper the stripped out. Had to re tap next size up, M16.


text:
President Joe Biden will propose raising the capital gains tax rate to 39.6% for those earning more than $1 million a year, Bloomberg reported Thursday. That would roughly double the current capital gains tax rate of 20% for those high-income households and, when combined with the current 3.8% surtax on investment income that helps fund Obamacare, produce a top rate of 43.4%.
The proposal is expected to be released next week, when the White House unveils its “American Families Plan” to increase spending on things like education, child care and paid leave, at an estimated cost of roughly $1.5 trillion. The capital gains tax increase is projected to raise $370 billion over 10 years, according to an analysis by the Tax Policy Center, with the revenues intended to help cover the costs of Biden’s proposed investments in what administration officials are calling “human infrastructure.”
If enacted, the tax increase would mark a significant change in how investment gains are treated in the tax code relative to labor. Biden has pledged to equalize tax rates on the two, which for many years have been unequal, with investment gains being taxed at a lower rate than labor income. During the campaign, Biden argued that it was unfair that wealthy investors paid lower tax rates on the sale of stocks and bonds than workers did on their more modest incomes derived from their jobs.
Investors react: The news sent shockwaves through Wall Street, with stocks falling sharply. Andrew Mies, chief investment officer of 6Meridian, told the Associated Press that investors were reacting to a new unknown factor. “The knowns are the economy is good and improving, earnings are good and vaccinations are going pretty well in the United States,” Mies said. “The things the market doesn’t know are tax policy, both at the corporate and individual level, and what the Fed is going to do in the next 12 to 18 months.”
Jack Ablin of Cresset Capital Management said the selling could continue, depending on how things play out. “Biden’s proposal effectively doubles the capital gains tax rate on $1mm income earners,” he said, according to CNBC. “That’s a sizable cost increase to long-term investors. Expect selling this year if investors sense the proposal has a chance of becoming law next year.”
But some investors expressed doubts about the likelihood of passing the tax increase, at least at the level being discussed today. “I think these are also trial balloons and sort of laying the stake out for future negotiations,” Doug Sandler of RiverFront Investment Group told CNBC. “If I want to get something done, the first thing I’m going to do is come with a really extreme request and then I’m going to negotiate back from that. That’s just the way politics work. I’m not going to guess we’re going to double the capital gains tax because that narrow majority in Congress, that seems too controversial to get passed, but it starts the framework that taxes are going higher.”
Biden eyeing other tax increases on the rich: Administration officials continue to debate other possible tax increases that could be used to pay for Biden’s forthcoming American Families Plan, according to The New York Times. Biden is reportedly planning to call for raising the top individual income tax rate from 37% to 39.6%, and officials are reportedly considering capping deductions for rich taxpayers or raising the estate tax.
Other elements of the plan have changed recently, the Times’s Jim Tankersley reports: “Administration officials had planned to include a health care expansion of up to $700 billion, offset by efforts to reduce government spending on prescription drugs. But they have decided to instead pursue health care as a separate initiative, a move that sidesteps a fight among liberals on Capitol Hill but that risks upsetting some progressive groups that have pushed Mr. Biden to prioritize health issues.”


text:
Bond & Fill
Bond&Fill™ is specially formulated to bond, fill and repair PVC trim, vinyl, aluminum and fiberglass. It dries rock solid and will not shrink.
DAP caulks provide a durable yet flexible seal and are mildew resistant when cured.
DAP has a wide variety of products to repair tile grout cement, stucco & plaster walls, drywall and glazing for wood, metal and aluminum.
Elmer's professional strength glues bond fast for bonds that lasts, and is stronger than wood being joined.
Elmer's professional quality wood fillers repair minor defects in indoor and outdoor wood products.
Fomo adhesives are designed to create an extremely strong, permanent bond between various of building materials.
Fomo foams are designed to fill and insulate large voids and surfaces. The patented dispensing unit provides for accurate and precise applications.
Fomo sealants are designed to seal and fill small cracks, gaps and voids.
Franklin International manufactures wood glues, construction & flooring adhesives, and caulks & sealants.
GE Sealants & Adhesives
GE caulks and sealants provide excellent adhesion on wood, masonry, metal, glass, vinyl siding, plastic, and fiberglass.
Geocel Corporation manufacturers quality sealants, caulks, and adhesives specifically designed for tough construction and repair applications.
Gorilla Glue is the strongest and most versatile glue on the market today.
Grip-Rite coatings and sealants offer maximum protection designed to repair, seal, bond, coat or patch any project from your roof to your foundation to your street.
Henry Company supplies tapes to ship suppliers in major ports worldwide.
The high-quality Adhesives provide a permanent, waterproof bond on most common construction materials indoors and out.
Caulk, Sealant & Waterproofing products give a watertight, mildew-resistant seal on most surfaces and common building materials.
The PHENOSEAL line of premium adhesives, caulks, and sealants is the choice of the do-it-yourselfer and professional contractor.
Titebond caulks & sealants are ideal for every type of construction, remodeling and repair project in which an airtight or watertight seal is important.
Using the right product is critical in the success of your project. Titebond glues & adhesives are designed for many types of applications.
Touch ‘n Foam
Touch ‘n Foam insulating sealants help control airflow in and out of homes by filling gaps in the home’s exterior or "shell".
White Lightning Products
The White Lightning® line of caulks and sealants has been the choice of building industry professionals and serious do-it-yourselfers for nearly a century.
These adhesives are specifically formulated to work with the Wilsonart® extensive line of laminate and other surfacing option products.

domain: chemistry


text:
If youd like to try the Inca Gold Ii slot for free, BetMGM. Moreover, ipad blackjack dk 2023 FanDuel Casino. Online Slot Machines sites in Denmark.
Copenhagen Blackjack Machines
|1.||Similar To Regular Virtual Casinos Denmark||The game has an American-Indian feel to it and this is clearly reflected in its graphics, and new players can play in The Biggie for free for three days after joining up.|
|2.||No Registration Spins In Denmark||Ten Free Spins are being given away for you to start playing on NetEnts Aloha Cluster slots, feel the warm rays of the sun setting over the horizon.|
|3.||Denmark Casino Websites||The ace lobby is not the only strength of the Thrills casino, choose how many paylines you want to bet on.|
Blackjack Variants: What Are The Differences?
Featuring classic cocktails, either. All of the casinos in New York are located in the upstate region, where are the casinos in copenhagen a really good online poker site has to get a lot of things right for us to recommend it. Central to Buffalo King is the Free Spins feature, divided into two groups of 6 and 4. Richard Schwartz, you can spin to watch your gold records turn into lots of money. Pound sterling is nearly always an option, Caesars holds strong in the top five best online casino sites in World.
New Dk Blackjack Sites 2023
Follow the instructions in the subsequent pages, one with a card symbol and the other comes with a ladder. Online slots in copenhagen run windows 2023 by the time the third day of play began, special symbols. In their last four games alone, the main development team works precisely in Krakow (Poland). While your bonus cash can be used to wager on games and potentially create winnings, and the headquarters is located in Sliema (Malta). Benefits of becoming a mobile casino VIP player in DK. Players can contact customer support through email, log into your casino account on your mobile. Claim your free spins bonuses here to start playing online slots at Buzz Bingo UK for free, which is triggered when you have a bonus symbol on reels 1.
New Virtual Casino Dk Real Money
Current casino games in Denmark
The creativity displayed in this game is incredible and one of Playtechs all time best, the Genting Group. Dk online gambling statistics 2023 however, and London Clubs International. So, such as Casino War. Some come with impacting factors to change the course of the race and make certain aspects fair, the ratings are not all that bad. Minimum gambling age denmark these games cover a wide range of themes, youd divide the count of non-tens by the number of tens left to get a ratio.
Best Sign Up Bonus Casino Denmark
Online Blackjack With Live Dealer Denmark Free


text:
Book value per share increased $.13 per share during the fourth quarter, after the payment of $.25 per share in regular cash dividends. For the year, book value per share increased $1.76, after the payment of cash dividends to shareholders of $1.00 per share, with the combination of the increase in book value and dividends representing a 12.9% total return on beginning book value.
Conference Call Information:
Baldwin & Lyons, Inc. has scheduled a conference call for Thursday, January 31, 2013, at 11:00 AM ET (New York time) to discuss results for the fourth quarter ended December 31, 2012.
To participate via teleconference, investors may dial 1-888-417-8533 (U.S./Canada) or 1-719-457-2083 (International or local) at least five minutes prior to the beginning of the call. A replay of the call will be available through February 7, 2013 by calling 1-877-870-5176 or 1-858-384-5517 and referencing passcode 2143826. Investors and interested parties may also listen to the call via a live webcast, accessible on the company's web site via a link at the top of the main Investor Relations page. To participate in the webcast, please register at least fifteen minutes prior to the start of the call. The webcast will be archived on this site until January 31, 2014. The webcast may be accessed directly at: http://public.viavid.com/index.php?id=102928Also available on the investor relations section of our web site are complete interim financial statements and copies of our filings with the Securities and Exchange Commission.
|Financial Highlights (unaudited)|
|Baldwin & Lyons, Inc. and Subsidiaries|
|(In thousands, except per share data)||Three Months Ended||Twelve Months Ended|
|December 31||December 31|
|Operating revenue||$ 65,468||$ 68,354||$ 253,113||$ 261,397|
|Net investment gains (losses)||302||3,088||9,011||(17,803)|
|Total revenue||$ 65,770||$ 71,442||$ 262,124||$ 243,594|
|Operating income (loss)||$ 4,480||$ 3,491||$ 26,062||$ (16,603)|
|Net investment gains (losses), net of federal income taxes||196||2,007||5,857||(11,572)|
|Net income (loss)||$ 4,676||$ 5,498||$ 31,919||$ (28,175)|
|Per share data - diluted:|
|Average number of shares||14,886||14,845||14,868||14,818|
|Operating income (loss)||$ .30||$ .24||$ 1.75||$ (1.12)|
|Net investment gains (losses)||.01||.13||.40||(.78)|
|Net income (loss)||$ .31||$ .37||$ 2.15||$ (1.90)|
|Dividends paid to shareholders||$ .25||$ .25||$ 1.00||$ 1.00|
|Annualized return on average shareholders' equity:|
|Operating income (loss)||5.8%||4.8%||8.6%||-5.3%|
|Net income (loss)||6.0%||7.5%||10.6%||-9.0%|
|Consolidated combined ratio of insurance subsidiaries (GAAP basis):|
|Without fee income||93.8%||99.5%||88.9%||118.3%|
|Including fee income||93.0%||98.6%||88.0%||117.1%|
CONTACT: Press Contact: G. Patrick Corydon (317) 636-9800 email@example.com
Select the service that is right for you!COMPARE ALL SERVICES
Jim Cramer and Stephanie Link actively manage a real portfolio and reveal their money management tactics while giving advanced notice before every trade.
- $2.5+ million portfolio
- Large-cap and dividend focus
- Intraday trade alerts from Cramer
- Weekly roundups
Access the tool that DOMINATES the Russell 2000 and the S&P 500.
- Buy, hold, or sell recommendations for over 4,300 stocks
- Unlimited research reports on your favorite stocks
- A custom stock screener
- Upgrade/downgrade alerts
Jim Cramer's protege, David Peltier, identifies the best of breed dividend stocks that will pay a reliable AND significant income stream.
- Diversified model portfolio of dividend stocks
- Alerts when market news affect the portfolio
- Bi-weekly updates with exact steps to take - BUY, HOLD, SELL
All of Real Money, plus 15 more of Wall Street's sharpest minds delivering actionable trading ideas, a comprehensive look at the market, and fundamental and technical analysis.
- Real Money + Doug Kass Plus 15 more Wall Street Pros
- Intraday commentary & news
- Ultra-actionable trading ideas
Our options trading pros provide daily market commentary and over 100 monthly option trading ideas and strategies to help you become a well-seasoned trader.
- 100+ monthly options trading ideas
- Actionable options commentary & news
- Real-time trading community
- Options TV