NLRP3 Inflammasome Activation Controls Vascular Smooth Muscle Cells Phenotypic Switch in Atherosclerosis

(1) Background: Monocytes and nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) inflammasome orchestrate lipid-driven amplification of vascular inflammation promoting the disruption of the fibrous cap. The components of the NLRP3 inflammasome are expressed in macrophages and foam cells within human carotid atherosclerotic plaques and VSMCs in hypertension. Whether monocytes and NLRP3 inflammasome activation are direct triggers of VSMC phenotypic switch and plaque disruption need to be investigated. (2) Methods: The direct effect of oxLDL-activated monocytes in VSMCs co-cultured system was demonstrated via flow cytometry, qPCR, ELISA, caspase 1, and pyroptosis assay. Aortic roots of VSMCs lineage tracing mice fed normal or high cholesterol diet and human atherosclerotic plaques were used for immunofluorescence quantification of NLRP3 inflammasome activation/VSMCs phenotypic switch. (3) Results: OxLDL-activated monocytes reduced α-SMA, SM22α, Oct-4, and upregulation of KLF-4 and macrophage markers MAC2, F4/80 and CD68 expression as well as caspase 1 activation, IL-1β secretion, and pyroptosis in VSMCs. Increased caspase 1 and IL-1β in phenotypically modified VSMCs was detected in the aortic roots of VSMCs lineage tracing mice fed high cholesterol diet and in human atherosclerotic plaques from carotid artery disease patients who experienced a stroke. (4) Conclusions: Taken together, these results provide evidence that monocyte promote VSMC phenotypic switch through VSMC NLRP3 inflammasome activation with a likely detrimental role in atherosclerotic plaque stability in human atherosclerosis.     

Partitioning of metal species during an enriched fuel combustion experiment. speciation in the gaseous and particulate phases

Combustion processes are the most important source of metal in the atmosphere and need to be better understood to improve flue gas treatment and health impact studies. This combustion experiment was designed to study metal partitioning and metal speciation in the gaseous and particulate phases. A light fuel oil was enriched with 15 organometallic compounds of the following elements: Pb, Hg, As, Cu, Zn, Cd, Se, Sn, Mn, V, Tl, Ni, Co, Cr, and Sb. The resulting mixture was burnt in a pilot-scale fuel combustion boiler under controlled conditions. After filtration of the particles, the gaseous species were sampled in the stack through a heated sampling tube simultaneously by standardized washing bottles-based sampling techniques and cryogenically. The cryogenic samples were collected at -80 degrees C for further speciation analysis by LT/GC-ICPMS. Three species of selenium and two of mercury were evidenced as volatile species in the flue gas. Thermodynamic predictions and experiments suggest the following volatile metal species to be present in the flue gas: H2Se, CSSe, CSe2, SeCl2, Hg(0), and HgCl2. Quantification of volatile metal species in comparison between cryogenic techniques and the washing bottles-based sampling method is also discussed. Concerning metal partitioning, the results indicated that under these conditions, at least 60% (by weight) of the elements Pb, Sn, Cu, Co, Tl, Mn, V, Cr, Ni, Zn, Cd, and Sb mixed to the fuel were found in the particulate matter. For As and Se, 37 and 17%, respectively, were detected in the particles, and no particulate mercury was found. Direct metal speciation in particles was performed by XPS allowing the determination of the oxidation state of the following elements: Sb(V), Tl(III), Mn(IV), Cd(II), Zn(II), Cr(III), Ni(II), Co(II), V(V), and Cu(II). Water soluble species of inorganic Cr, As, and Se in particulate matter were determined by HPLC/ICP-MS and identified in the oxidation state Cr(III), As(V), and Se(IV).     

Atmospheric reactions influence seasonal PAH and nitro-PAH concentrations in the Los Angeles basin

Ambient measurements of polycyclic aromatic hydrocarbons (PAHs) and nitro-PAHs were carried out during August 2002 and January 2003 in Los Angeles, CA, a source site and in Riverside, CA, a downwind receptor site approximately 90 km to the east of Los Angeles. Atmospheric concentrations of PAHs and nitro-PAHs are of interest because both of these compound classes include potent mutagens and carcinogens. To augment our current understanding of atmospheric formation of nitro-PAHs, four sampling periods were employed to study the diurnal variations of these compounds. The PAH concentrations were highest in Los Angeles during January, as a result of traffic input at this source site undertightwintertime atmospheric inversions. In contrast, nitro-PAH levels were highest in Riverside during August, as a result of enhanced summertime photochemistry. Hydroxyl radical-initiated reactions produced nitro-PAHs in both seasons, while in winter little evidence for nitrate radical chemistry was seen. For the August samples, nitrate radical-initiated formation of nitro-PAHs is suggested by nitro-PAH isomer profiles not only at the downwind location as anticipated, but also atthe source site. In southern California, the contribution of atmospheric formation through gas-phase radical-initiated PAH reactions to the ambient burden of nitro-PAHs is dominant, with the semi-volatile nitro-PAHs being the most abundant and 2-nitrofluoranthene being the major particle-associated nitro-PAH.     

Characterization of the "viable but nonculturable" (VBNC) state in the wine spoilage yeast Brettanomyces

Although the viable but not culturable (VBNC) state has been studied in detail in bacteria, it has been suggested that maintenance of viability with loss of culturability also exists in eukaryotic cells, such as in the wine spoilage yeast Brettanomyces. To provide conclusive evidence for the existence of a VBNC state in this yeast, we investigated its capacity to become viable and nonculturable after sulfite stress, and its ability to recover culturability after stressor removal. Sulfite addition induced loss of culturability but maintenance of viability. Increasing the medium pH to decrease the concentration of toxic SO(2) allowed yeast cells to become culturable again, thus demonstrating the occurrence of a VBNC state in Brettanomyces upon SO(2) exposure. Relative to culturable Brettanomyces, VBNC yeast cells were found to display a 22% decrease in size on the basis of laser granulometry. Assays for 4-ethylguaiacol and 4-ethylphenol, volatile phenols produced by Brettanomyces, indicated that spoilage compound production could persist in VBNC cells. These morphological and physiological changes in VBNC Brettanomyces were coupled to extensive protein pattern modifications, as inferred by comparative two-dimensional electrophoresis and mass spectrometric analyses. Upon identification of 53 proteins out of the 168 spots whose abundance was significantly modified in treated cells relative to control, we propose that the SO(2)-induced VBNC state in Brettanomyces is characterized by a reduced glycolytic flux coupled to changes in redox homeostatis/protein turnover-related processes. This study points out the existence of common mechanisms between yeast and bacteria upon entry to the VBNC state.     

Further Exploration of Controlled and Automatic Memory Processes in Early Alzheimer's Disease.

The authors' aim in this study was to explore automatic and controlled processes in Alzheimer's disease (AD) by using a variant of the word-stem completion task that applies the process-dissociation procedure. Several methodological precautions were taken in order to limit problems observed in previous studies (e.g., poor task sensitivity, ceiling and/or floor effects, no control over comprehension of instructions). Our results (a) confirmed the marked deterioration in controlled processes and (b) showed that when psychometric constraints were limited, automatic memory processes were preserved in AD. These data are in line with those from more global studies in suggesting that AD is characterized by an early deterioration in controlled processes and an initial preservation of automatic processes. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Modelling of multiple-mechanism agglomeration in a crystallization process

A three-level agglomeration model coupled with crystal growth is developed. It accounts for Brownian, laminar, and turbulent agglomeration. The desupersaturation profiles, the particle size distributions, the average sizes, and variances (or standard deviations), as well as the instantaneous agglomeration degrees for each mechanism, can be calculated as functions of time. The model is applied to the crystallization of an amorphous solid into a crystalline polymorph in a batch crystallizer. A runaway phenomenon is detected for agglomeration when crystals are switching over from the Brownian regime to the laminar one: this switchover significantly affects the desupersaturation curve and the crystal shapes.     

Self-propagating combustion synthesis of intermetallic matrix composites in the ISS

Combustion Synthesis experiments have been performed on the ISS (International Space Station) during the Belgian taxi-flight mission ODISSEA in November 2002, in the framework of the ESA-coordinated project COSMIC (Combustion Synthesis under Microgravity Conditions). The main objective of the experiments was to investigate the general physico-chemical mechanisms of combustion synthesis processes and the formation of products microstructure. Within the combustion zone, a number of gravity-dependent phenomena occur, while other phenomena are masked by gravity. Under certain conditions, gravity-dependent secondary processes may also occur in the heat-affected zone after combustion. To study the influence of gravity, a specially dedicated reactor ensemble was designed and used in the Microgravity Science Glovebox (MSG) onboard the ISS. In this work, the experiment design is first discussed in terms of the experimental functionality and reactor ensemble integration in the MSG. To investigate microstructure formation, a sample constituted by a cylindrical portion followed by a conical one, the latter being inserted inside a massive copper block, is used. The experiment focused on the synthesis of intermetallic matrix composites (IMCs) based on the Al-Ti-B system. Depending on the composition, different intermetallic compounds (TiAl and TiAl₃) can be formed as matrix phase while TiB2 represents the reinforcing particulate phase. During the ISS mission, six samples with a relatively high green density of 65%TD have successfully been processed. The influence of the composition on the combustion process will be examined.     

Increased biological activity of a recombinant factor IX variant carrying alanine at position +1

In attempts to improve the post-translational modification andprocessing of recombinant factor IX (FIX) we have altered thecDNA sequence encoding pre-pro-FIX using site-directed mutagenesisand have expressed the variant cDNAs in BHK21 cells using avaccinia-virus-derived vector. We find that substitution ofthe tyrosine residue at +1 for an alanine increases the biologicalactivity of the recombinant molecules 2–fold. On the otherhand, substitution of the proline at –3 for a valine resultsin no significant change to the specific activity of the protein.Other alterations to the N-terminus of the FIX proteins, inattempts to mimic other vitamin-K-dependent proteins, resultin the failure to produce a secreted polypeptide. N-terminalsequence analysis of purified recombinant molecules revealsa correlation between specific activity and the efficiency ofcorrect pro-sequence cleavage. -Carboxylation analysis of purifiedrecombinant proteins indicates that each molecule includingunmutated FIX is completely -carboxylated in this system. Thusthe observed increase in biological activity of FIX variantscontaining an alanine at position +1 is not due to increased-carboxylation but, at least in part, to more efficient pro-peptidecleavage.     

Photo-Fenton-peroxide process using FE (II)-embedded composites based on activated carbon: Characterization of catalytic tests

Cost-efficient Fenton-like catalysts for the removal of organic molecules in aqueous solutions were elaborated by a simple process. Porous activated carbons (ACs) were directly impregnated with a precursor solution using the wet impregnation method. Their efficiency as Fenton-like catalysts was studied. Photo-Fenton tests were performed to establish the performance of the prepared Fe-impregnated activated carbons in relation to the degradation of an organic pollutant (Indigo Carmine) model in aqueous solution, under different conditions. Photo-catalytic tests were carried out by means of a laboratory photo-reactor (UV-Consulting Peschl). The influence of several parameters such as solution pH value, initial concentration of the model pollutant, and hydrogen peroxide dose on the process performance was investigated. The ACs and prepared catalysts were characterized by nitrogen adsorption-desorption isotherms at 77 K, SEM, and thermogravimetric analyses. The total Fe content of the synthesized composites was estimated by the phenanthroline method using UV-vis spectrophotometry. The results show an increase in the degradation rate when the heterogeneous photo-Fenton process is conducted with an ozone generating UV lamp.