Apolipoprotein A-I but not high-density lipoproteins are internalised by RAW macrophages: roles of ATP-binding cassette transporter A1 and scavenger receptor BI

Accumulation of lipid-loaded macrophages (foam cells) within the vessel wall is an early hallmark of atherosclerosis. High-density lipoproteins (HDL) and apolipoprotein A-I (apoA-I) can efficiently promote cholesterol efflux from macrophages. Therefore, the interaction of HDL and apoA-I with macrophages appears to be important in the initial steps of reverse cholesterol transport, i.e. the transport of excess cholesterol from foam cells to the liver. However, although several cellular apoA-I and HDL receptors and transporters have been identified, it is as yet controversial how these interactions lead to cholesterol efflux from foam cells. In this study, we show that RAW264.7 macrophages bind HDL and apoA-I in a competable manner. Furthermore, cell surface biotinylation experiments revealed that apoA-I but not HDL is specifically internalised. Binding of HDL to macrophages is decreased by reducing the expression of scavenger receptor BI (SR-BI) with cyclic adenosine monophosphate (cAMP), acetylated low-density lipoprotein (acLDL) or RNA interference. In contrast, apoA-I cell association and internalisation is modulated in parallel with ATP-binding cassette transporter A1 (ABCA1) expression which is altered by stimulating cells with cAMP and acLDL or expressing short hairpin RNA (shRNA) against ABCA1. Consistent with this, cell surface trapping of ABCA1 with cyclosporin A (CsA) results in increased apoA-I binding but reduced internalisation. Furthermore, blocking apoA-I uptake inhibits cholesterol efflux to apoA-I but not to HDL. Taken together, these data suggest that apoA-I- but not HDL-mediated cholesterol efflux may involve retroendocytosi

Decoding Functional High-Density Lipoprotein Particle Surfaceome Interactions

High-density lipoprotein (HDL) is a mixture of complex particles mediating reverse cholesterol transport (RCT) and several cytoprotective activities. Despite its relevance for human health, many aspects of HDL-mediated lipid trafficking and cellular signaling remain elusive at the molecular level. During HDL's journey throughout the body, its functions are mediated through interactions with cell surface receptors on different cell types. To characterize and better understand the functional interplay between HDL particles and tissue, we analyzed the surfaceome-residing receptor neighborhoods with which HDL potentially interacts. We applied a combination of chemoproteomic technologies including automated cell surface capturing (auto-CSC) and HATRIC-based ligand-receptor capturing (HATRIC-LRC) on four different cellular model systems mimicking tissues relevant for RCT. The surfaceome analysis of EA.hy926, HEPG2, foam cells, and human aortic endothelial cells (HAECs) revealed the main currently known HDL receptor scavenger receptor B1 (SCRB1), as well as 155 shared cell surface receptors representing potential HDL interaction candidates. Since vascular endothelial growth factor A (VEGF-A) was recently found as a regulatory factor of transendothelial transport of HDL, we next analyzed the VEGF-modulated surfaceome of HAEC using the auto-CSC technology. VEGF-A treatment led to the remodeling of the surfaceome of HAEC cells, including the previously reported higher surfaceome abundance of SCRB1. In total, 165 additional receptors were found on HAEC upon VEGF-A treatment representing SCRB1 co-regulated receptors potentially involved in HDL function. Using the HATRIC-LRC technology on human endothelial cells, we specifically aimed for the identification of other bona fide (co-)receptors of HDL beyond SCRB1. HATRIC-LRC enabled, next to SCRB1, the identification of the receptor tyrosine-protein kinase Mer (MERTK). Through RNA interference, we revealed its contribution to endothelial HDL binding and uptake. Furthermore, subsequent proximity ligation assays (PLAs) demonstrated the spatial vicinity of MERTK and SCRB1 on the endothelial cell surface. The data shown provide direct evidence for a complex and dynamic HDL receptome and that receptor nanoscale organization may influence binding and uptake of HDL

Altered Distribution of Unesterified Cholesterol among Lipoprotein Subfractions of Patients with Diabetes Mellitus Type 2

Biomarkers are important tools to improve the early detection of patients at high risk for developing diabetes as well as the stratification of diabetic patients towards risks of complications. In addition to clinical variables, we analyzed 155 metabolic parameters in plasma samples of 51 healthy volunteers and 66 patients with diabetes using nuclear magnetic resonance (NMR) spectrometry. Upon elastic net analysis with lasso regression, we confirmed the independent associations of diabetes with branched-chain amino acids and lactate (both positive) as well as linoleic acid in plasma and HDL diameter (both inverse). In addition, we found the presence of diabetes independently associated with lower concentrations of free cholesterol in plasma but higher concentrations of free cholesterol in small HDL. Compared to plasmas of non-diabetic controls, plasmas of diabetic subjects contained lower absolute and relative concentrations of free cholesterol in all LDL and HDL subclasses except small HDL but higher absolute and relative concentrations of free cholesterol in all VLDL subclasses (except very small VLDL). These disbalances may reflect disturbances in the transfer of free cholesterol from VLDL to HDL during lipolysis and in the transfer of cell-derived cholesterol from small HDL via larger HDL to LDL

Mapping the dynamic high-density lipoprotein synapse

Background and aims Heterogeneous high-density lipoprotein (HDL) particles, which can contain hundreds of proteins, affect human health and disease through dynamic molecular interactions with cell surface proteins. How HDL mediates its long-range signaling functions and interactions with various cell types is largely unknown. Due to the complexity of HDL, we hypothesize that multiple receptors engage with HDL particles resulting in condition-dependent receptor-HDL interaction clusters at the cell surface. Methods Here we used the mass spectrometry-based and light-controlled proximity labeling strategy LUX-MS in a discovery-driven manner to decode HDL-receptor interactions. Results Surfaceome nanoscale organization analysis of hepatocytes and endothelial cells using LUX-MS revealed that the previously known HDL-binding protein scavenger receptor B1 (SCRB1) is embedded in a cell surface protein community, which we term HDL synapse. Modulating the endothelial HDL synapse, composed of 60 proteins, by silencing individual members, showed that the HDL synapse can be assembled in the absence of SCRB1 and that the members are interlinked. The aminopeptidase N (AMPN) (also known as CD13) was identified as an HDL synapse member that directly influences HDL uptake into the primary human aortic endothelial cells (HAECs). Conclusions Our data indicate that preformed cell surface residing protein complexes modulate HDL function and suggest new theragnostic opportunities

DFT calculations and electrochemical studies on azulene ligands for heavy metal ions detection using chemically modified electrodes

A computational study on three related derivatives of 5-[(azulen-1-yl)methylene]-2-thioxoimidazolidin-4-one was conducted using density functional theory by calculating a series of molecular descriptors and properties of their optimized geometries (electrostatic and local ionization potentials, molecular frontier orbitals, etc.). Thermodynamic properties (zero-point energy, enthalpy, constant volume heat capacity, entropy and Gibbs energy) for these derivatives have been calculated and related to ligands electrochemical behavior. Reduction and oxidation potentials have been correlated to their calculated energy levels for LUMO and HOMO orbitals. Chemically modified electrodes based on these derivatives have been tested in view of heavy metal ions recognition, and their detection limits have been correlated to the calculated values of electron affinity

El concepto jurídico de precios habituales del delito de acaparamiento y el régimen económico constitucional del estado peruano

La presente investigación tuvo como objetivo general analizar la manera en que el concepto jurídico de precios habituales del delito acaparamiento se relaciona con el Régimen Económico Constitucional del Estado peruano, de allí que, la pregunta general de investigación fue: ¿De qué manera el concepto jurídico de precios habituales del delito acaparamiento se relaciona con el Régimen Económico Constitucional del Estado peruano?, asimismo la hipótesis general fue: El concepto jurídico de precios habituales del delito acaparamiento se relaciona de manera negativa con el Régimen Económico Constitucional del Estado peruano, en consecuencia, nuestra investigación con respecto al método de investigación es de naturaleza cualitativa teórica e iuspositivista, por tal motivo se utilizó la técnica del análisis documental y su procesamiento se llevó a cabo mediante la argumentación jurídica a través de los instrumentos de recolección de datos siendo este la ficha textual y de resumen cuya finalidad fue recabar información relevante. El resultado más importante fue que: La descripción típica qué se establecía en nuestro Código Penal de 1991 sobre el acaparamiento al respecto refería al sujeto que acapara o de cualquier manera sustrae del comercio, bienes de producción o consumo, con el objetivo de elevar los precios y consigo provocar la escasez de bienes, así como obtener lucros monetarios. La conclusión más relevante fue que: El concepto jurídico de precios habituales del delito acaparamiento se relaciona de manera negativa con el Régimen Económico Constitucional del Estado peruano. Finalmente, la recomendación fue: Derogación del artículo 233 del Código Penal

Cholesterol Efflux Capacity Associates with the Ankle-Brachial Index but Not All-Cause Mortality in Patients with Peripheral Artery Disease

BACKGROUND: Cholesterol efflux is an important mechanism by which high-density lipoproteins (HDLs) protect against cardiovascular disease. As peripheral artery disease (PAD) is associated with high mortality rates, mainly due to cardiovascular causes, we investigated whether cholesterol efflux capacity (CEC) of apolipoprotein B (apoB)-depleted plasma, a widely used surrogate of HDL function, may serve as a predictive marker for mortality in this patient population. METHODS: In this prospective single-center study (median follow-up time: 9.3 years), apoB-containing lipoproteins were precipitated from plasma of 95 patients with PAD and incubated with J744-macrophages, which were loaded with radiolabeled cholesterol. CEC was defined as the fractional radiolabel released during 4 h of incubation. RESULTS: Baseline CEC was lower in PAD patients that currently smoked (p = 0.015) and had a history of myocardial infarction (p = 0.011). Moreover, CEC showed a significant correlation with HDL-cholesterol (p = 0.003) and apolipoprotein A-I levels (p = 0.001) as well as the ankle-brachial index (ABI, p = 0.018). However, CEC did not differ between survivors and non-survivors. Neither revealed Kaplan-Meier and Cox regression analyses any significant association of CEC with all-cause mortality rates. CONCLUSION: Taken together, CEC is associated with ABI but does not predict all-cause mortality in patients with PAD

EUNADICS-AV early warning system dedicated to supporting aviation in the case of a crisis from natural airborne hazards and radionuclide clouds

The purpose of the EUNADICS-AV (European Natural Airborne Disaster Information and Coordination System for Aviation) prototype early warning system (EWS) is to develop the combined use of harmonised data products from satellite, ground-based and in situ instruments to produce alerts of airborne hazards (volcanic, dust, smoke and radionuclide clouds), satisfying the requirement of aviation air traffic management (ATM) stakeholders (https://cordis.europa.eu/project/id/723986, last access: 5 November 2021). The alert products developed by the EUNADICS-AV EWS, i.e. near-real-time (NRT) observations, email notifications and netCDF (Network Common Data Form) alert data products (called NCAP files), have shown significant interest in using selective detection of natural airborne hazards from polar-orbiting satellites. The combination of several sensors inside a single global system demonstrates the advantage of using a triggered approach to obtain selective detection from observations, which cannot initially discriminate the different aerosol types. Satellite products from hyperspectral ultraviolet–visible (UV–vis) and infrared (IR) sensors (e.g. TROPOMI – TROPOspheric Monitoring Instrument – and IASI – Infrared Atmospheric Sounding Interferometer) and a broadband geostationary imager (Spinning Enhanced Visible and InfraRed Imager; SEVIRI) and retrievals from ground-based networks (e.g. EARLINET – European Aerosol Research Lidar Network, E-PROFILE and the regional network from volcano observatories) are combined by our system to create tailored alert products (e.g. selective ash detection, SO2 column and plume height, dust cloud, and smoke from wildfires). A total of 23 different alert products are implemented, using 1 geostationary and 13 polar-orbiting satellite platforms, 3 external existing service, and 2 EU and 2 regional ground-based networks. This allows for the identification and the tracking of extreme events. The EUNADICS-AV EWS has also shown the need to implement a future relay of radiological data (gamma dose rate and radionuclides concentrations in ground-level air) in the case of a nuclear accident. This highlights the interest of operating early warnings with the use of a homogenised dataset. For the four types of airborne hazard, the EUNADICS-AV EWS has demonstrated its capability to provide NRT alert data products to trigger data assimilation and dispersion modelling providing forecasts and inverse modelling for source term estimate. Not all of our alert data products (NCAP files) are publicly disseminated. Access to our alert products is currently restricted to key users (i.e. Volcanic Ash Advisory Centres, national meteorological services, the World Meteorological Organization, governments, volcano observatories and research collaborators), as these are considered pre-decisional products. On the other hand, thanks to the EUNADICS-AV–SACS (Support to Aviation Control Service) web interface (https://sacs.aeronomie.be, last access: 5 November 2021), the main part of the satellite observations used by the EUNADICS-AV EWS is shown in NRT, with public email notification of volcanic emission and delivery of tailored images and NCAP files. All of the ATM stakeholders (e.g. pilots, airlines and passengers) can access these alert products through this free channel.Peer ReviewedArticle escrit per 46 autors/es: Hugues Brenot Nicolas Theys Lieven Clarisse Jeroen van Gent Daniel Hurtmans Sophie Vandenbussche Nikolaos Papagiannopoulos Lucia Mona Timo Virtanen Andreas Uppstu Mikhail Sofiev Luca Bugliaro Margarita Vázquez-Navarro Pascal Hedelt Michelle Maree Parks Sara Barsotti Mauro Coltelli William Moreland Simona Scollo Giuseppe Salerno Delia Arnold-Arias Marcus Hirtl Tuomas Peltonen Juhani Lahtinen Klaus Sievers Florian Lipok Rolf Rüfenacht Alexander Haefele Maxime Hervo Saskia Wagenaar Wim Som de Cerff Jos de Laat Arnoud Apituley Piet Stammes Quentin Laffineur Andy Delcloo Robertson Lennart Carl-Herbert Rokitansky Arturo Vargas Markus Kerschbaum Christian Resch Raimund Zopp Matthieu Plu 1 Vincent-Henri Peuch Michel van Roozendael Gerhard WotawaPostprint (author's final draft

Expression of O6-methylguanine-DNA methyltransferase in childhood medulloblastoma

Medulloblastomas (MB) are the most common malignant brain tumors in childhood. Alkylator-based drugs are effective agents in the treatment of patients with MB. In several tumors, including malignant glioma, elevated O6-methylguanine-DNA methyltransferase (MGMT) expression levels or lack of MGMT promoter methylation have been found to be associated with resistance to alkylating chemotherapeutic agents such as temozolomide (TMZ). In this study, we examined the MGMT status of MB and central nervous system primitive neuroectodermal tumor (PNET) cells and two large sets of primary MB. In sevenMB/PNET cell lines investigated, MGMT promoter methylation was detected only in D425 human MB cells as assayed by the qualitative methylation-specific PCR and the more quantitative pyrosequencing assay. In D425 human MB cells, MGMT mRNA and protein expression was clearly lower when compared with the MGMT expression in the other MB/PNET cell lines. In MB/PNET cells, sensitivity towards TMZ and 1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea (CCNU) correlated with MGMT methylation and MGMT mRNA expression. Pyrosequencing in 67 primary MB samples revealed a mean percentage of MGMT methylation of 3.7-92% (mean: 13.25%, median: 10.67%). Percentage of MGMT methylation and MGMT mRNA expression as determined by quantitative RT-PCR correlated inversely (n=46; Pearson correlation r 2=0.14, P=0.01). We then analyzed MGMT mRNA expression in a second set of 47 formalin-fixed paraffin-embedded primary MB samples from clinically well-documented patients treated within the prospective randomized multicenter trial HIT'91. No association was found between MGMT mRNA expression and progression-free or overall survival. Therefore, it is not currently recommended to use MGMT mRNA expression analysis to determine who should receive alkylating agents and who should no