Comparative reactivity of the myeloperoxidase-derived oxidants HOCl and HOSCN with low-density lipoprotein (LDL): Implications for foam cell formation in atherosclerosis

© 2015 Elsevier Inc. All rights reserved. Abstract Atherosclerosis is characterised by the accumulation of lipids within macrophages in the artery wall. Low-density lipoprotein (LDL) is the source of this lipid, owing to the uptake of oxidised LDL by scavenger receptors. Myeloperoxidase (MPO) released by leukocytes during inflammation produces oxidants that are implicated in atherosclerosis. Modification of LDL by the MPO oxidant hypochlorous acid (HOCl), results in extensive lipid accumulation by macrophages. However, the reactivity of the other major MPO oxidant, hypothiocyanous acid (HOSCN) with LDL is poorly characterised, which is significant given that thiocyanate is the favoured substrate for MPO. In this study, we comprehensively compare the reactivity of HOCl and HOSCN with LDL, and show key differences in the profile of oxidative damage observed. HOSCN selectively modifies Cys residues on apolipoprotein B100, and oxidises cholesteryl esters resulting in formation of lipid hydroperoxides, 9-hydroxy-10,12-octadecadienoic acid (9-HODE) and F2-isoprostanes. The modification of LDL by HOSCN results macrophage lipid accumulation, though generally to a lesser extent than HOCl-modified LDL. This suggests that a change in the ratio of HOSCN:HOCl formation by MPO from variations in plasma thiocyanate levels, will influence the nature of LDL oxidation in vivo, and has implications for the progression of atherosclerosis

Phosphorus rates on yield and quality of lettuce seeds.

Because of lack of information about phosphorus fertilization in lettuce from the standpoint of seed production, this study was undertaken. The work was carried out in Botucatu, São Paulo state, Brazil, from September 25, 2003 to February 19, 2004, in order to study the influence of crescents phosphorus rates on yield and quality of lettuce seeds, cultivar Verônica. The experimental design was randomized blocks with five treatments (0; 200; 400; 600 and 800 kg ha-1 of P2O5) and five replications. The following characteristics were evaluated: seed production per plant (g plant-1), number of seeds per plant and quality (percentage of germination and seed vigor). A linear increasing was observed on production and number of seeds per plant of lettuce with phosphorus rates, but these rates did not affect the seed physiological quality evaluated on weight of 1,000 seeds (average = 0.91 g), first count of germination (98%), standard germination test (99%), seed germination speed index (68.5), seedling emergence speed index (97.0) and emerged seedlings (49.3%)

Nanogap structures for molecular nanoelectronics

This study is focused on the realization of nanodevices for nano and molecular electronics, based on molecular interactions in a metal-molecule-metal (M-M-M) structure. In an M-M-M system, the electronic function is a property of the structure and can be characterized through I/V measurements. The contact between the metals and the molecule was obtained by gold nanogaps (with a dimension of less than 10 nm), produced with the electromigration technique. The nanogap fabrication was controlled by a custom hardware and the related software system. The studies were carried out through experiments and simulations of organic molecules, in particular oligothiophenes

Singular Location and Signaling Profile of Adenosine A2A-Cannabinoid CB1 Receptor Heteromers in the Dorsal Striatum

The dorsal striatum is a key node for many neurobiological processes such as motor activity, cognitive functions, and affective processes. The proper functioning of striatal neurons relies critically on metabotropic receptors. Specifically, the main adenosine and endocannabinoid receptors present in the striatum, ie, adenosine A2A receptor (A2AR) and cannabinoid CB1 receptor (CB1R), are of pivotal importance in the control of neuronal excitability. Facilitatory and inhibitory functional interactions between striatal A2AR and CB1R have been reported, and evidence supports that this cross-talk may rely, at least in part, on the formation of A2AR-CB1R heteromeric complexes. However, the specific location and properties of these heteromers have remained largely unknown. Here, by using techniques that allowed a precise visualization of the heteromers in situ in combination with sophisticated genetically-modified animal models, together with biochemical and pharmacological approaches, we provide a high resolution expression map and a detailed functional characterization of A2AR-CB1R heteromers in the dorsal striatum. Specifically, our data unveil that the A2AR-CB1R heteromer (i) is essentially absent from corticostriatal projections and striatonigral neurons, and, instead, is largely present in striatopallidal neurons, (ii) displays a striking G protein-coupled signaling profile, where co-stimulation of both receptors leads to strongly reduced downstream signaling, and (iii) undergoes an unprecedented dysfunction in Huntington’s disease, an archetypal disease that affects striatal neurons. Altogether, our findings may open a new conceptual framework to understand the role of coordinated adenosine-endocannabinoid signaling in the indirect striatal pathway, which may be relevant in motor function and neurodegenerative diseases

Identification of NUB1 as a Suppressor of Mutant Huntingtin Toxicity via Enhanced Protein Clearance

Huntington’s disease is caused by expanded CAG in HTT, conferring toxic gain of function to mutant HTT (mHTT) protein. Reducing mHTT levels is postulated as a strategy for therapeutic intervention. We conducted genome-wide RNAi screens for genes modifying mHTT levels and identified 13 hits. Ten were tested in vivo in a Drosophila Huntington’s disease model and 6 exhibited activity consistent with in vitro screening. Among these, NUB1 overexpression lowered mHTT in neuronal models, and rescued mHTT-induced death. NUB1 reduces mHTT level by enhancing poly-ubiquitination and proteasomal degradation of mHTT protein. The process requires CUL3 and the ubiquitin-like protein NEDD8 necessary for CUL3 activation. As a potential approach to modulate NUB1 for treatment, interferon beta (IFNβ) lowered mHTT and rescued neuronal toxicity via induction of NUB1. Thus, we have identified genes modifying endogenous mHTT using high-throughput screening and demonstrate NUB1 as an exemplar entry point for therapeutic intervention of Huntington’s disease