An apolipoprotein A-V gene SNP is associated with marked hypertriglyceridemia among Asian-American patients

Apolipoprotein A-V (apoA-V) is an important regulator of plasma levels of triglyceride (TG) in mice. In humans, APOA5 genetic variation is associated with TG in several populations. In this study, we determined the effects of the p.185Gly>Cys (c.553G>T; rs2075291) polymorphism on plasma TG levels in subjects of Chinese ancestry living in the United States and in a group of non-Chinese Asian ancestry. The frequency of the less common cysteine allele was 4-fold higher (15.1% vs. 3.7%) in Chinese high-TG subjects compared with a low-TG group (Chi-square = 20.2; P < 0.0001), corresponding with a 4.45 times higher risk of hypertriglyceridemia (95% confidence interval, 2.18-9.07; P < 0.001). These results were replicated in the non-Chinese Asians. Heterozygosity was associated, in the high-TG group, with a doubling of TG (P < 0.001), mainly VLDL TG (P = 0.014). All eleven TT homozygotes had severe hypertriglyceridemia, with mean TG of 2,292 +/- 447 mg/dl. Compared with controls, carriers of the T allele had lower postheparin lipoprotein lipase activity but not hepatic lipase activity. In Asian populations, this common polymorphism can lead to profound adverse effects on lipoprotein profiles, with homozygosity accounting for a significant number of cases of severe hypertriglyceridemia. This specific apoA-V variant has a pronounced effect on TG metabolism, the mechanism of which remains to be elucidated.     

Macrophage PLTP is atheroprotective in LDLr-deficient mice with systemic PLTP deficiency

Systemic phospholipid transfer protein (PLTP) is a recognized risk factor for coronary heart disease. In apolipoprotein E-deficient mice, systemic PLTP deficiency is atheroprotective, whereas PLTP overexpression is proatherogenic. As expected, we also observed significantly smaller lesions (P < 0.0001) in hypercholesterolemic double mutant low density lipoprotein receptor-deficient (LDLr(-/-)) PLTP-deficient (PLTP(-/-)) mice compared with LDLr(-/-) mice expressing systemic PLTP. To assess the specific contribution of only macrophage-derived PLTP to atherosclerosis progression, bone marrow transplantation was performed in LDLr(-/-) mice that also lacked systemic PLTP. Groups of double mutant PLTP(-/-)LDLr(-/-) mice were irradiated with 1,000 rad and injected with bone marrow (BM) cells collected from either PLTP(-/-) or wild-type mice. When fed a high-fat diet, BM cell expression of PLTP decreased plasma cholesterol of PLTP(-/-)LDLr(-/-) mice from 878 +/- 220 to 617 +/- 183 mg/dl and increased HDL cholesterol levels from 54 +/- 11 to 117 +/- 19 mg/dl. This decreased total plasma cholesterol and increased HDL cholesterol contributed to the significantly smaller atherosclerotic lesions in both aortas and heart sinus valves observed in these mice. Thus, unlike total systemic PLTP, locally produced macrophage-derived PLTP beneficially alters lipoprotein metabolism and reduces lesion progression in hyperlipidemic mice.     

Origins of intestinal ABCA1-mediated HDL-cholesterol

The origins of cholesterol utilized by intestinal ABCA1 were investigated in the human intestinal cell line Caco-2. Influx of apical membrane cholesterol increases ABCA1 mRNA and mass, resulting in enhanced efflux of HDL-cholesterol. Luminal (micellar) cholesterol and newly synthesized cholesterol are not transported directly to ABCA1 but reach the ABCA1 pool after incorporation into the apical membrane. Depleting the apical or the basolateral membrane of cholesterol by cyclodextrin attenuates the amount of cholesterol transported by ABCA1 without altering ABCA1 expression. Filipin added to the apical side but not the basal side attenuates ABCA1-mediated cholesterol efflux, suggesting that apical membrane "microdomains," or rafts, supply cholesterol for HDL. Preventing cholesterol esterification increases the amount of cholesterol available for HDL. Ezetimibe, a Niemann-Pick C1-like 1 protein inhibitor, does not alter ABCA1-mediated cholesterol efflux. U18666A and imipramine, agents that mimic cholesterol trafficking defects of Neimann-Pick type C disease, attenuate cholesterol efflux without altering ABCA1 expression; thus, intestinal NPC1 may facilitate cholesterol movement to ABCA1. ABCA1-mediated cholesterol efflux is independent of cholesterol synthesis. The results suggest that following incorporation into plasma membrane and rafts of the apical membrane, dietary/biliary and newly synthesized cholesterol contribute to the ABCA1 pool and HDL-cholesterol. NPC1 may have a role in this process.     

Membrane fusogenic activity of the Alzheimer's peptide A beta(1-42) demonstrated by small-angle neutron scattering

Amyloid-β peptide (Aβ) is considered a triggering agent of Alzheimer's disease. In relation to a therapeutic treatment of the disease, the interaction of Aβ with the cell membrane has to be elucidated at the molecular level to understand its mechanism of action. In previous works, we had ascertained by neutron diffraction on stacked lipid multilayers that a toxic fragment of Aβ is able to penetrate and perturb the lipid bilayer. Here, the influence of Aβ(1-42), the most abundant Aβ form in senile plaques, on unilamellar lipid vesicles of phospholipids is investigated by small-angle neutron scattering. We have used the recently proposed separated form factor method to fit the data and to obtain information about the vesicle diameter and structure of the lipid bilayer and its change upon peptide administration. The lipid membrane parameters were obtained with different models of the bilayer profile. As a result, we obtained an increase in the vesicle radii, indicating vesicle fusion. This effect was particularly enhanced at pH 7.0 and at a high peptide/lipid ratio. At the same time, a thinning of the lipid bilayer occurred. A fusogenic activity of the peptide may have very important consequences and may contribute to cytotoxicity by destabilizing the cell membrane. The perturbation of the bilayer structure suggests a strong interaction and/or insertion of the peptide into the membrane, although its localization remains beyond the limit of the experimental resolution.     

Extensive conformational transitions are required to turn on ATP hydrolysis in myosin

Conventional myosin is representative of biomolecular motors in which the hydrolysis of adenosine triphosphate (ATP) is coupled to large-scale structural transitions both in and remote from the active site. The mechanism that underlies such “mechanochemical coupling,” especially the causal relationship between hydrolysis and allosteric structural changes, has remained elusive despite extensive experimental and computational analyses. In this study, using combined quantum mechanical and molecular mechanical simulations and different conformations of the myosin motor domain, we provide evidence to support that regulation of ATP hydrolysis activity is not limited to residues in the immediate environment of the phosphate. Specifically, we illustrate that efficient hydrolysis of ATP depends not only on the proper orientation of the lytic water but also on the structural stability of several nearby residues, especially the Arg238-Glu459 salt bridge (the numbering of residues follows myosin II in Dictyostelium discoideum) and the water molecule that spans this salt bridge and the lytic water. More importantly, by comparing the hydrolysis activities in two motor conformations with very similar active-site (i.e., Switches I and II) configurations, which distinguished this work from our previous study, the results clearly indicate that the ability of these residues to perform crucial electrostatic stabilization relies on the configuration of residues in the nearby N-terminus of the relay helix and the “wedge loop.” Without the structural support from those motifs, residues in a closed active site in the post-rigor motor domain undergo subtle structural variations that lead to consistently higher calculated ATP hydrolysis barriers than in the pre-powerstroke state. In other words, starting from the post-rigor state, turning on the ATPase activity requires not only displacement of Switch II to close the active site but also structural transitions in the N-terminus of the relay helix and the “wedge loop,” which have been proposed previously to be ultimately coupled to the rotation of the converter subdomain 40 Å away.     

Developmentally regulated alternative splicing of densin modulates protein-protein interaction and subcellular localization

Densin is a member of the leucine-rich repeat (LRR) and PDZ domain (LAP) protein family that binds several signaling molecules via its C-terminal domains, including calcium/calmodulin-dependent protein kinase II (CaMKII). In this study, we identify several novel mRNA splice variants of densin that are differentially expressed during development. The novel variants share the LRR domain but are either prematurely truncated or contain internal deletions relative to mature variants of the protein (180 kDa), thus removing key protein–protein interaction domains. For example, CaMKIIα coimmunoprecipitates with densin splice variants containing an intact C-terminal domain from lysates of transfected HEK293 cells, but not with variants that only contain N-terminal domains. Immunoblot analyses using antibodies to peptide epitopes in the N- and C- terminal domains of densin are consistent with developmental regulation of splice variant expression in brain. Moreover, putative splice variants display different subcellular fractionation patterns in brain extracts. Expression of green fluorescent protein (GFP)-fused densin splice variants in HEK293 cells shows that the LRR domain can target densin to a plasma membrane-associated compartment, but that the splice variants are differentially localized and have potentially distinct effects on cell morphology. In combination, these data show that densin splice variants have distinct functional characteristics suggesting multiple roles during neuronal development.     

On the causes of density banding in skeletons of corals of the genus Montastraea

To obtain a reliable climate reconstruction from coral skeletons it is first necessary to understand the way these grow and incorporate information. Thickness of skeletal elements (exothecal and endothecal dissepiments, costae, septa, and theca-wall) as well as the spacing between exothecal and endothecal dissepiments of the four extant Atlantic species of Montastraea (M. annularis, M. faveolata, M. franksi and M. cavernosa) were measured through high and low density bands. Our results show that growth periodicity, controlled by the effect of temperature, is expressed in changes in thickness of costae and exothecal dissepiments in the four studied Montastraea species, with no changes in endothecal elements and theca-wall thickness which, in turn, has implications for research on inclusive records using these species. Spacing between both exothecal and endothecal dissepiments resulted without changes along the high and low density bands, and we found evidence that there is a rhythmical formation of these structures linked somehow to lunar cycles.     

Waterfowl Nesting in Fall-Seeded and Spring-Seeded Cropland in Saskatchewan

Abstract: Waterfowl nesting in annual croplands has remained a little-known aspect of waterfowl nesting ecology because of the inability of many studies to systematically search this habitat through the nesting season. Where searches have been conducted, they are generally restricted to the period prior to seeding, and many nests found are destroyed by the seeding operation. Consequently, fall-seeded crops have been promoted as an alternative cropping practice that could increase nest survival of waterfowl nesting in croplands. During 1996–1999, we conducted 3–4 complete nest searches on 4,274 ha of cropland, including spring-seeded wheat and barley, winter wheat, and fall rye in southern Saskatchewan, Canada. Using suites of predictive models, we tested hypotheses regarding relative nest abundance and nest survival among crop types and tested the influence of several landscape-scale covariates on these metrics. Apparent nest densities were higher in fall-seeded crops (winter wheat: 0.39 nests/ha, fall rye: 0.25 nests/ha) than in spring-seeded crops (0.03 nests/ha), and nest density in spring-seeded croplands increased with percent cropland and percent wetland habitat in the surrounding landscape. Nest survival was higher in winter wheat (38%) than in either fall rye (18%) or spring-seeded crops (12%), and nest survival in spring-seeded crops increased with relative nest initiation date. Nest survival was unaffected by surrounding landscape characteristics but tended to be higher in years of average wetness. Based on our findings, winter wheat and fall rye have the potential to provide productive nesting habitat for ≥7 species of upland nesting ducks and fall-seeded crops are a conservation tool well suited to highly cropped landscapes.     

Estimation of Black Bear Abundance Using a Discrete DNA Sampling Device

Abstract We developed a snare for collection of black bear (Ursus americanus) hair that obtained a unique hair sample at each snare site, improved the quantity of collected hair compared to barbed-wire corrals, and was easy to deploy over a wide range of topographical features and habitat conditions. This device allowed us to implement intensive sampling methodology needed in mark-recapture experiments with minimal effort. By improving the quantity of hair collected, we also lowered the potential for bear identification errors at the lab. During 2003–2004, bears in 2 study areas triggered snares 1,104 times, which resulted in the collection of 981 hair samples. Of the samples we collected, 79% (775) produced valid genetic data. In 2003, 454 samples identified 79 genetically distinct individuals, and 321 samples identified 86 genetically distinct individuals in 2004. Analysis of capture-recapture data indicated that capture probabilities were affected by heterogeneity among individuals and behavioral responses, but showed little evidence of time effects. Consequently, we used the Pollock and Otto (1983) estimator for model M_bh to estimate abundance with reasonably good precision (CV: 12–14%). Density on the Steamboat and Toketee, Oregon, USA, study areas over the 2-year period averaged 19 bears/100 km² and 22 bears/100 km², respectively. Average capture and recapture probabilities over the 2 years of the study were 30% and 63%, respectively, indicating a trap-prone behavioral response. Knowledge of bear densities on the Steamboat and Toketee study areas will enable managers to set hunting quotas, advise land management agencies on habitat issues, and create a baseline database to assist in the long-term monitoring of bear trends in a changing landscape.