Mechanisms underlying the atherosclerosis risk associated with apolipoprotein E isoforms in humans

Apolipoprotein E (apoE) is a ligand for the low density lipoprotein receptor (LDLR), both of which are well recognized determinants of atherosclerosis risk in humans. In humans the APOE gene is polymorphic, with three alleles APOE2, APOE3, and APOE4 coding for the protein apoE2, apoE3, and apoE4. Despite increased LDLR affinity, apoE4 is associated with elevated and apoE2 reduced cholesterol concentration compared to apoE3. Mice that have had their endogenous apoE replaced with the human apoE's do not replicate their human counterparts, and mice with apoE3 and apoE4 are similar to wildtype mice, yet those with apoE2 have drastically elevated lipids, a phenotype observed in only 5% of APOE2 homozygous humans. Either by adenovirus-mediated or global overexpression, elevated LDLR decreased plasma cholesterol in mice with apoE2, but led to a dramatic accumulation of cholesterol-rich VLDL in mice with apoE4 on a western-type high fat diet. In addition to the liver, where both apoE and LDLR are highly expressed and contribute to plasma lipoprotein clearance, they are expressed in vascular cells and macrophages. Increases in LDLR expression in macrophages significantly increased cholesterol uptake in culture, more prominently with lipoproteins containing apoE4 than those containing apoE3. In LDLR-deficient mice expressing the human apoE4 isoform, the replacement of bone marrow cells with those expressing LDLR increased atherosclerotic lesions in a dose-dependent manner compared with mice transplanted with cells having no LDLR. Thus apoE4, but not apoE3, in macrophages enhances atherosclerotic plaque development in mice in an LDLR-dependent manner and this interaction may contribute to the association of apoE4 with an increased cardiovascular risk in humans. Higher LDLR expression decreased the secretion of apoE4 and increased its degradation in both macrophages and hepatocytes. ApoE localization using apoE-GFP fusion proteins expressed by adenovirus in the liver of apoE-deficient mice revealed that apoE4 accumulated in the space of Disse, but apoE2 did not. Using several genetic approaches, apoE4s association with increased atherosclerosis risk can be replicated in mice. ApoE4 binding to the LDLR in macrophages and localization in hepatocytes both contribute to apoE4 pathogenesis

Dynamical population synthesis: Constructing the stellar single and binary contents of galactic field populations

[abridged] The galactic field's late-type stellar single and binary population is calculated on the supposition that all stars form as binaries in embedded star clusters. A recently developed tool (Marks, Kroupa & Oh) is used to evolve the binary star distributions in star clusters for a few Myr so that a particular mixture of single and binary stars is achieved. On cluster dissolution the population enters the galactic field with these characteristics. The different contributions of single stars and binaries from individual star clusters which are selected from a power-law embedded star cluster mass function are then added up. This gives rise to integrated galactic field binary distribution functions (IGBDFs) resembling a galactic field's stellar content (Dynamical Population Synthesis). It is found that the binary proportion in the galactic field of a galaxy is larger the lower the minimum cluster mass, the lower the star formation rate, the steeper the embedded star cluster mass function and the larger the typical size of forming star clusters in the considered galaxy. In particular, period-, mass-ratio- and eccentricity IGBDFs for the Milky Way are modelled. The afore mentioned theoretical IGBDFs agree with independently observed distributions. Of all late-type binaries, 50% stem from M<300M_sun clusters, while 50% of all single stars were born in M>10^4M_sun clusters. Comparison of the G-dwarf and M-dwarf binary population indicates that the stars formed in mass-segregated clusters. In particular it is pointed out that although in the present model all M-dwarfs are born in binary systems, in the Milky Way's Galactic field the majority ends up being single stars. This work predicts that today's binary frequency in elliptical galaxies is lower than in spiral and in dwarf-galaxies. The period and mass-ratio distributions in these galaxies are explicitly predicted.Comment: 14 pages, 9 figures, accepted for publication in MNRA

Apolipoprotein E4 in Macrophages Enhances Atherogenesis in a Low Density Lipoprotein Receptor-dependent Manner

Apolipoprotein E (apoE) and the low density lipoprotein receptor (LDLr) are well recognized determinants of atherosclerosis. In addition to hepatocytes, where both are highly expressed and contribute to plasma lipoprotein clearance, they are expressed in vascular cells and macrophages. In this study, we examined the effects of human apoE isoforms and LDLr levels in atherogenic pathways in primary macrophages ex vivo and atherosclerosis development after bone marrow transfer in vivo using mice expressing human apoE isoforms and different levels of LDLr expression. Increases in LDLr expression significantly increased cholesterol delivery into macrophages in culture, and the effects were more prominent with lipoproteins containing apoE4 than those containing apoE3. Conversely, increased LDLr expression reduced cholesterol efflux in macrophages expressing apoE4 but not in macrophages expressing apoE3. Furthermore, apoE3 protected VLDL from oxidation in vitro more than did apoE4. In LDLr-deficient mice expressing the human apoE4 isoform, Apoe4/4 Ldlr−/−, the replacement of bone marrow cells with those expressing LDLr increased atherosclerotic lesions in a dose-dependent manner compared with mice transplanted with cells having no LDLr. In contrast, atherosclerosis in Apoe3/3 Ldlr−/− mice, expressing the human apoE3 isoform, did not differ by the levels of macrophage LDLr expression. Our results demonstrate that apoE4, but not apoE3, in macrophages enhances atherosclerotic plaque development in mice in an LDLr-dependent manner and suggests that this interaction may contribute to the association of apoE4 with an increased cardiovascular risk in humans

Cross-validation of cut-points in preschool children using different accelerometer placements and data axes

The present study cross-validated various cut-points to assess physical activity and sedentary behaviour in preschoolers, using hip- and wrist-worn accelerometers and both vertical axis and vector magnitude data. Secondly, we examined the influence of epoch length on time estimates of physical activity and sedentary behaviour. Sixty-four preschoolers (34 girls) wore two accelerometers, on their right hip and dominant wrist, during 1 hour of free play. Preschoolers’ activities were observed by two trained researchers. Area under the curve (AUC) was calculated for the receiving operating characteristic (ROC) curves as a measure of precision. AUC ranges were 0.603–0.723 for sedentary behaviour, 0.472–0.545 for light physical activity and 0.503–0.661 for moderate-to-vigorous physical activity (MVPA), indicating poor to fair precision. Percentage of time classified as sedentary behaviour, light or MVPA according to observation and accelerometer data varied largely between cut-points, accelerometer placements and axes. The influence of epoch length on time estimates was minimal across cut-points, except for one hip-based vector magnitude cut-point. Across all accelerometer placements and data axes, no set of cut-points demonstrated adequate precision for sedentary behaviour, light physical activity and MVPA. The highly variable and omnidirectional activity pattern of preschoolers may explain the lack of adequate cut-points

Doubling Expression of the Low Density Lipoprotein Receptor by Truncation of the 3′-Untranslated Region Sequence Ameliorates Type III Hyperlipoproteinemia in Mice Expressing the Human ApoE2 Isoform

The primary receptor mediating clearance of apolipoprotein (apo)E- and apoB100-containing lipoproteins from the circulation is the low density lipoprotein (LDL) receptor. Reduced expression of the LDLR is believed to be a precipitating factor in the pathogenesis of type III hyperlipoproteinemia (HLP) in some humans homozygous for the apoE2 allele (APOE*2). To test the effect of genetic changes in LDL receptor expression on the pathogenesis of type III HLP, we have generated a variant allele at the endogenous mouse Ldlr locus that expresses the human LDL receptor transcript. Transcription of the human LDLR minigene is regulated by the endogenous mouse promoter sequence, but a truncation of 3'-untranslated region results in increased mRNA stability. Consequently, in liver of heterozygotes, steady state levels of mouse and human LDLR transcripts are 50 and 180% the levels of total transcript in wild type mice, respectively. Overall, the 2.3-fold normal level of LDLR message in heterozygotes completely ameliorates type III HLP caused by the homozygosity for the human APOE*2 allele, normalizing their plasma lipoprotein profile. We conclude that a modest increase in expression of the LDLR through message stabilization is sufficient to prevent precipitation of type III HLP in mice

Permeation of macromolecules into the renal glomerular basement membrane and capture by the tubules

Human kidneys contain ∼2 x 106 glomeruli that produce ∼180 L per day of primary filtrate. Downstream tubules reabsorb most of the water, salt, and desirable low-molecular weight substances, leaving 1 to 2 L per day of urine containing undesirable waste products. Currently, most investigators think that the primary filtrate is low in protein because fluid exiting the glomerulus passes through slits spanned by a diaphragm that acts as a low-porosity molecular sieve. Our experiments challenge this view; they show that size-dependent permeation into the glomerular basement membrane and into a gel-like coat that covers the slits, together with saturable tubular reabsorption, determines which macromolecules reach the urine. The slit diaphragm is essential for capillary structure but may not directly determine glomerular size selectivity

The Extreme Hosts of Extreme Supernovae

We use GALEX ultraviolet (UV) and optical integrated photometry of the hosts of seventeen luminous supernovae (LSNe, having peak M_V < -21) and compare them to a sample of 26,000 galaxies from a cross-match between the SDSS DR4 spectral catalog and GALEX interim release 1.1. We place the LSNe hosts on the galaxy NUV-r versus M_r color magnitude diagram (CMD) with the larger sample to illustrate how extreme they are. The LSN hosts appear to favor low-density regions of the galaxy CMD falling on the blue edge of the blue cloud toward the low luminosity end. From the UV-optical photometry, we estimate the star formation history of the LSN hosts. The hosts have moderately low star formation rates (SFRs) and low stellar masses (M_*) resulting in high specific star formation rates (sSFR). Compared with the larger sample, the LSN hosts occupy low-density regions of a diagram plotting sSFR versus M_* in the area having higher sSFR and lower M_*. This preference for low M_*, high sSFR hosts implies the LSNe are produced by an effect having to do with their local environment. The correlation of mass with metallicity suggests that perhaps wind-driven mass loss is the factor that prevents LSNe from arising in higher-mass, higher-metallicity hosts. The massive progenitors of the LSNe (>100 M_sun), by appearing in low-SFR hosts, are potential tests for theories of the initial mass function that limit the maximum mass of a star based on the SFR.Comment: 8 pages, 3 figures, 2 tables, accepted to ApJ, amended references and updated SN designation

Apolipoprotein E knock-out and knock-in mice: atherosclerosis, metabolic syndrome, and beyond: Fig. 1.

Given the multiple differences between mice and men, it was once thought that mice could not be used to model atherosclerosis, principally a human disease. Apolipoprotein E-deficient (apoEKO) mice have convincingly changed this view, and the ability to model human-like plaques in these mice has provided scientists a platform to study multiple facets of atherogenesis and to explore potential therapeutic interventions. In addition to its well-established role in lipoprotein metabolism, recent observations of reduced adiposity and improved glucose homeostasis in apoEKO mice suggest that apoE may also play a key role in energy metabolism in peripheral organs, including adipose tissue. Finally, along with apoEKO mice, knockin mice expressing human apoE isoforms in place of endogenous mouse apoE have provided insights into how quantitative and qualitative genetic alterations interact with the environment in the pathogenesis of complex human diseases