Advanced glycation alters expression of the 67kDa laminin receptor in retinal microvascular endothelial cells

The 67kDa laminin receptor (67LR) plays an important role in vascular cell function and dysfunction. The present study has examined 67LR expression in retinal microvascular endothelial cells after exposure to AGEs. Retinal microvascular endothelial cells were exposed to either AGE-BSA, or were grown on methylglyoxal-modified laminin or Matrigel (TM) and expression of 67LR analysed by Western Blotting and RT-PCR/Southern blotting. Western blotting of plasma membrane and RT-PCR/Southern blotting revealed a significant upregulation of 67LR protein/mRNA expression after exposure to AGEs (p

Obatoclax induces Atg7-dependent autophagy independent of beclin-1 and BAX/BAK

Direct pharmacological targeting of the anti-apoptotic B-cell lymphoma-2 (BCL-2) family is an attractive therapeutic strategy for treating cancer. Obatoclax is a pan-BCL-2 family inhibitor currently in clinical development. Here we show that, although obatoclax can induce mitochondrial apoptosis dependent on BCL-2 associated x protein/BCL-2 antagonist killer (BAX/BAK) consistent with its on-target pharmacodynamics, simultaneous silencing of both BAX and BAK did not abolish acute toxicity or loss of clonogenicity. This is despite complete inhibition of apoptosis. Obatoclax dramatically reduced viability without inducing loss of plasma membrane integrity. This was associated with rapid processing of light chain-3 (LC3) and reduction of S6 kinase phosphorylation, consistent with autophagy. Dramatic ultrastructural vacuolation, not typical of autophagy, was also induced. Silencing of beclin-1 failed to prevent LC3 processing, whereas knockout of autophagy-related (Atg) 7 abolished LC3 processing but failed to prevent obatoclax-induced loss of clonogenicity or ultrastructural changes. siRNA silencing of Atg7 in BAX/BAK knockout mouse embryonic fibroblasts did not prevent obatoclax-induced loss of viability. Cells selected for obatoclax resistance evaded apoptosis independent of changes in BCL-2 family expression and displayed reduced LC3 processing. In summary, obatoclax exhibits BAX- and BAK-dependent and -independent mechanisms of toxicity and activation of autophagy. Mechanisms other than autophagy and apoptosis are blocked in obatoclax resistant cells and contribute significantly to obatoclax's anticancer efficacy. Cell Death and Disease (2010) 1, e108; doi:10.1038/cddis.2010.86; published online 16 December 2010

THE DETECTION OF HOT ETHANOL IN G34.3+0.15

Hot molecular cores in star-forming regions are known to have gas-phase chemical compositions determined by the evaporation of material from the icy mantles of interstellar grains, followed by subsequent reactions in the gas phase. Current models suggest that the evaporated material is rich in hydrogenated species, such as water, methane and methanol. In this paper, we report the detection of 14 rotational transitions of ethanol in the submillimetre spectrum of the molecular cloud associated with the ultra-compact H II region G34.3+0.15. We derive a rotation temperature of 125 K and a beam-averaged column density of 2.0x10(15) cm(-2), corresponding to a fractional abundance on the order of 4x10(-9). This large abundance, which is a lower limit due to the likelihood of beam dilution, cannot be made by purely gas-phase processes, and we conclude that the ethanol must be formed efficiently in the grain surface chemistry. Since it has been argued previously that methanol is formed via surface chemistry, it appears that alcohol formation may be a natural by-product of surface reactions.

ON THE ORIGIN OF NH IN DIFFUSE INTERSTELLAR CLOUDS

The observation by Meyer & Roch of NH in the interstellar clouds towards zeta Per and HD 27778 cannot be explained with conventional gas-phase chemistry models. A simple non-equilibrium model for the zeta Per cloud, which incorporates the grain-surface production of NH and OH or, alternatively, NH3 and H2O, is able to reproduce the abundances of all observed species (except CH+) quite accurately. Moreover, chemical models which include grain-surface reactions can reproduce the observed abundance not only of NH but also of CN, which is efficiently formed at low temperatures, initiated by the reaction of NH with C+. Pure gas-phase models and cloud interface models, in which NH and CH+ are formed in a warm and tenuous environment, fail to explain the observed high abundance of CN. Hence the observation of NH in zeta Per and HD 27778 provides evidence for the presence of grain-surface reactions leading to molecules other than H-2. It is predicted that NH2 and NH3 should have abundances not much below that of NH if NH3 instead of NH is formed on grains. With or without surface reactions, the column densities of H2O and C2H are expected to be about 10(13) cm-2, and these molecules may be detectable in the zeta Per cloud.

CHEMICAL PROCESSES IN DARK INTERSTELLAR CLOUDS - THE EFFECTS OF CNO DEPLETION

We have investigated the effects of depletion of the elements C, N and O on the chemical composition of dark clouds, using both isothermal and isochoric cloud models. Our work differs from previous approaches in that we have considered a much larger range of CNO depletions. We have included the chemistry of the ortho-and para-forms of H2 and the exothermic reaction between N+ and ortho-H2, which synthesizes NH3. In the isothermal models, the ortho:para ratio is very small at large depletions, but NH3 formation is still efficient owing to reactions between He+ and CN or HCN. In the isochoric models, the equilibrium temperature of the gas is larger, and a thermal ortho:para ratio, which is large enough to drive NH3 formation, results. In all cases, the fractional abundance of NH3 is close to 10(-8) and this may help to explain the puzzling observation that, in dark clouds, the column density of NH3 is always close to 10(15) cm-2.

High contrast plasma mirror: spatial filtering and second harmonic generation at 10(19)Wcm(-2)

Recently, the use of plasma optics to improve temporal pulse contrast has had a remarkable impact on the field of high- power laser-solid density interaction physics. Opening an avenue to previously unachievable plasma density gradients in the high intensity focus, this advance has enabled researchers to investigate new regimes of harmonic generation and ion acceleration. Until now, however, plasma optics for fundamental laser reflection have been used in the sub-relativistic intensity regime (10(15) - 10(16)Wcm(-2)) showing high reflectivity (similar to 70%) and good focusability. Therefore, the question remains as to whether plasma optics can be used for such applications in the relativistic intensity regime (> 10(18)Wcm(-2)). Previous studies of plasma mirrors (PMs) indicate that, for 40 fs laser pulses, the reflectivity fluctuates by an order of magnitude and that focusability of the beam is lost as the intensity is increased above 5 x 10(16)Wcm(-2). However, these experiments were performed using laser pulses with a contrast ratio of similar to 10(7) to generate the reflecting surface. Here, we present results for PM operation using high contrast laser pulses resulting in a new regime of operation - the high contrast plasma mirror (HCPM). In this regime, pulses with contrast ratio > 10(10) are used to form the PM surface at > 10(19)Wcm(-2), displaying excellent spatial filtering, reflected near- field beam profile of the fundamental beam and reflectivities of 60 +/- 5%. Efficient second harmonic generation is also observed with exceptional beam quality suggesting that this may be a route to achieving the highest focusable harmonic intensities. Plasma optics therefore offer the opportunity to manipulate ultra-intense laser beams both spatially and temporally. They also allow for ultrafast frequency up-shifting without detrimental effects due to group velocity dispersion (GVD) or reduced focusability which frequently occur when nonlinear crystals are used for frequency conversion.

The plasma mirror - A subpicosecond optical switch for ultrahigh power lasers

Plasma mirrors are devices capable of switching very high laser powers on subpicosecond time scales with a dynamic range of 20–30 dB. A detailed study of their performance in the near-field of the laser beam is presented, a setup relevant to improving the pulse contrast of modern ultrahigh power lasers ~TW–PW!. The conditions under which high reflectivity can be achieved and focusability of the reflected beam retained are identified. At higher intensities a region of high specular reflectivity with rapidly decreasing focusability was observed, suggesting that specular reflectivity alone is not an adequate guide to the ideal range of plasma mirror operation. It was found that to achieve high reflectivity with negligible phasefront distortion of the reflected beam the inequality csDt,lLaser must be met (cs : sound speed, Dt: time from plasma formation to the peak of the pulse!. The achievable contrast enhancement is given by the ratio of plasma mirror reflectivity to cold reflectivity.

Indirect-drive inertial confinement fusion using highly supersonic, radiatively cooled, plasma slugs

We present a new approach to indirect-drive inertial confinement fusion which makes use of highly supersonic, radiatively cooled, slugs of plasma to energize a hohlraum. 2D resistive magnetohydrodynamic simulations of slug formation in shaped liner Z -pinch implosions are presented along with 2D-radiation-hydrodynamic simulations of the slug impacting a converter foil and 3D-view-factor simulations of a double-ended hohlraum. Results for the Z facility at Sandia National Laboratory indicate that two synchronous slugs of 250 kJ kinetic energy could be produced, resulting in a capsule surface temperature of similar to225 eV .

Note on the single-shock solutions of the Korteweg-de Vries-Burgers equation

The well-known shock solutions of the Korteweg-de Vries-Burgers equation are revisited, together with their limitations in the context of plasma (astro)physical applications. Although available in the literature for a long time, it seems to have been forgotten in recent papers that such shocks are monotonic and unique, for a given plasma configuration, and cannot show oscillatory or bell-shaped features. This uniqueness is contrasted to solitary wave solutions of the two parent equations (Korteweg-de Vries and Burgers), which form a family of curves parameterized by the excess velocity over the linear phase speed.

The development of effective biomarkers for Alzheimer's disease:a review

OBJECTIVE: There is a widely recognised need to develop effective Alzheimer's disease (AD) biomarkers to aid the development of disease-modifying treatments, to facilitate early diagnosis and to improve clinical care. This overview aims to summarise the utility of key neuroimaging and cerebrospinal fluid (CSF) biomarkers for AD, before focusing on the latest efforts to identify informative blood biomarkers. DESIGN: A literature search was performed using PubMed up to September 2011 for reviews and primary research studies of neuroimaging (magnetic resonance imaging, magnetic resonance spectroscopy, positron emission tomography and amyloid imaging), CSF and blood-based (plasma, serum and platelet) biomarkers in AD and mild cognitive impairment. Citations within individual articles were examined to identify additional studies relevant to this review. RESULTS: Evidence of AD biomarker potential was available for imaging techniques reflecting amyloid burden and neurodegeneration. Several CSF measures are promising, including 42 amino acid ß-amyloid peptide (Aß(42) ); total tau (T-tau) protein, reflecting axonal damage; and phosphorylated tau (P-tau), reflecting neurofibrillary tangle pathology. Studies of plasma Aß have produced inferior diagnostic discrimination. Alternative plasma and platelet measures are described, which represent potential avenues for future research. CONCLUSIONS: Several imaging and CSF markers demonstrate utility in predicting AD progression and determining aetiology. These require standardisation before forming core elements of diagnostic criteria. The enormous potential available for identifying a minimally-invasive, easily-accessible blood measure as an effective AD biomarker currently remains unfulfilled. Copyright © 2012 John Wiley & Sons, Ltd.

Effects of modified LDL and HDL on retinal pigment epithelial cells: a role in diabetic retinopathy?

Aims/hypothesis: Blood–retina barrier leakage in diabetes results in extravasation of plasma lipoproteins. Intra-retinal modified LDLs have been implicated in diabetic retinopathy (DR), but their effects on retinal pigment epithelial (RPE) cells and the added effects of extravasated modified HDLs are unknown.

Methods: In human retinas from individuals with and without diabetes and DR, immunohistochemistry was used to detect ApoB, ApoA1 and endoplasmic reticulum (ER) stress markers. In cell culture, human RPE cells were treated with native LDL (N-LDL) or heavily-oxidised glycated LDL (HOG-LDL) with or without pretreatment with native HDL (N-HDL) or heavilyoxidised glycated HDL (HOG-HDL). Cell viability, oxidative stress, ER stress, apoptosis and autophagy were assessed by Cell Counting Kit-8 assay, dichlorofluorescein assay, western blotting, immunofluorescence and TUNEL assay. In separate
experiments, RPE cells were treated with lipid oxidation products, 7-ketocholesterol (7-KC, 5–40 µmol/l) or 4-hydroxynonenal (4-HNE, 5–80 µmol/l), with or without pretreatment with N-HDL or HOG-HDL.

Results: ApoB, ApoA1 staining and RPE ER stress were increased in the presence of DR. HOG-LDL but not N-LDL significantly decreased RPE cell viability and increased reactive oxygen species generation, ER stress, apoptosis and autophagy. Similarly, 4-HNE and 7-KC decreased viability and induced ER stress. Pretreatment with N-HDL mitigated these effects, whereas HOG-HDL was less effective by most, but not all, measures.

Conclusions/interpretation: In DR, extravascular modified LDL may promote RPE injury through oxidative stress, ER stress, autophagy and apoptosis. N-HDL has protective effects, but HOG-HDL is less effective. Extravasation and modification of HDL may modulate the injurious effects of extravasated modified LDL on the retinal pigment epithelium.

Glycosylation of low-density lipoprotein enhances cholesteryl ester synthesis in human monocyte-derived macrophages

Glucose can react with the lysine residues of low-density lipoproteins (LDLs) and convert the lipoprotein to a form with a receptor-mediated uptake by cultured cells that is impaired. However, in contrast to other modified lipoproteins taken up by both murine and human macrophages via the scavenger-receptor pathway that may induce the formation of foam cells, glycosylated LDL is not recognized by murine macrophages, and thus far, it has not been shown to lead to marked intracellular accumulation of cholesterol in human macrophages. This study illustrates that glycosylated LDL incubated with human monocyte-derived macrophages, at a concentration of 100 micrograms LDL/ml medium, stimulates significantly more cholesteryl ester (CE) synthesis than does control LDL (10.65 +/- 1.5 vs. 4.8 +/- 0.13 nmol.mg-1 cell protein.20 h-1; P less than .05). At LDL concentrations similar to those of plasma, the rate of CE synthesis in macrophages incubated with glycosylated LDL is more markedly enhanced than that observed in cells incubated with control LDL (3-fold increase). The marked stimulation of CE synthesis in human macrophages exposed to glycosylated LDL is paralleled by a significant increase in CE accumulation in these cells (P less than .001). The increase in CE synthesis and accumulation seem to be mediated by an increase in the degradation of glycosylated LDL by human macrophages. Glycosylated LDL enters the macrophages and is degraded by the classic LDL-receptor pathway in slightly smaller amounts than control LDL, but its degradation by pathways other than the classic LDL receptor or scavenger receptor is markedly enhanced.(ABSTRACT TRUNCATED AT 250 WORDS)

Glycosylation of low density lipoprotein in patients with type 1 (insulin-dependent) diabetes:correlations with other parameters of glycaemic control

Glycosylation of low density lipoproteins obtained from 16 patients with Type 1 (insulin-dependent) diabetes and from 16 age-, sex-, and race-matched controls, was determined. The diabetic patients were normolipaemic and were in good or fair glycaemic control. Eleven patients performed home blood glucose monitoring. Glycosylation of low density lipoproteins in the diabetic patients was significantly higher (p less than 0.001) than in the control subjects, and was significantly correlated with haemoglobin A1c, (p less than 0.01), glycosylation of plasma proteins, (p less than 0.001), and mean home blood glucose, (p less than 0.01). This study confirms that, in diabetic patients, increased glycosylation of low density lipoprotein occurs to an extent which correlates closely with other commonly used indices of glycaemic control.