Lipase-based quantitation of triacylglycerols in cellular lipid extracts: Requirement for presence of detergent and prior separation by thin-layer chromatography

A protocol, based on the use of Pseudomonas lipase, is presented to measure quantitatively the amount of triacylglycerols in extracts from cultured cells or tissues. Since the lipase also acts on di- and monoacylglycerols, separation of the extracts by thin-layer chromatography is recommended. In order to allow the lipase-catalyzed hydrolysis to proceed efficiently, lipid extracts or eluates from silica scraping were mixed with the detergent Thesit [dodecylpoly(ethylene glycol ether)], prior to drying. After dissolution of the dried residues in water, the amount of triacylglycerols was quantified using Pseudomonas sp. lipase, glycerol kinase, glycerol-phosphate oxidase, and peroxidase. The activity of the latter enzyme was followed either colorimetrically in the presence of 4-aminoantipyrine and 2,4,6-tribromo-3-hydroxybenzoic acid or fluorimetrically in the presence of homovanillic acid.     

Effects of cutting angle,edge preparation,and nano-structured coating on milling performance of a gamma titanium aluminide

Gamma titanium aluminides are intermetallic alloys. Recently, they have been evaluated as important contenders for structural applications in the automotive and aerospace sectors. This is due to their excellent high-temperature performances and their significantly lower density compared to nickel-based superalloys. In this paper, an analysis of machinability of a gamma TiAl obtained via an electron beam melting (EBM) process is presented. The effects of tool geometry modifications, in terms of cutting tool angles and cutting edge preparation, were investigated. The reduction of radial rake angle and the drag finishing process for cutting edge preparation resulted in an increase of the tool life of the carbide end mills. Nanogradient tool coatings were also observed to affect tool wear during milling tests, and the results highlight that AlSiTiN coating performs better compared to CrAlSiN coating. A post-coating polishing treatment was also taken into account, and it allowed a further reduction of tool wear. The overall results indicate that the machinability of this difficult-to-cut material can be significantly improved by an adjustment of the cutting edge geometry, and by using an AlSiTiN coating system.     

Characterization and reactivity of MoO3/SiO2 catalysts in the selective catalytic oxidation of ammonia to N2

Silica-supported MoO₃ catalysts with MoO₃ loadings up to 21% w/w were prepared, characterized and tested in the selective catalytic oxidation (SCO) of ammonia to N₂ under dilute conditions. It is found that the catalysts are active and selective in the reaction, and that the catalytic performance increases on increasing the Mo loading. Crystalline MoO₃, detected over the silica support, is supposed to be the active species in the reaction. The reactivity of the catalysts is depressed by water addition to the feed at low temperatures and is enhanced by the presence of selected promoters, like Bi and Pb.     

Identification of the role of surface acidity in the deactivation of TiO2 in the selective photo-oxidation of cyclohexane

Anatase-structured Ti_1−xZr_xO₂ materials with x = 0.00, 0.01 and 0.06, were prepared by a reverse microemulsion method, characterized, and tested as catalysts for the selective photo-oxidation of cyclohexane to cyclohexanone. In situ ATR-FTIR spectroscopy was used to evaluate the reaction. Zr incorporation into the anatase lattice enhances the surface acidity of TiO₂ without causing any significant structural or electronic modification. As expected, also the stability of surface adsorbed water, i.e. the hydrophilicity, was enhanced. The increase in the Brønsted acidity, together with the higher hydrophilicity, is shown to be detrimental for performance (selectivity and stability) in the selective photo-oxidation of cyclohexane. Apparently potential intrinsic catalytic advantages of having higher acidity are outweighed by (i) the enhanced number of water born OH radicals, inducing non-selective reactions, and (ii) enhanced hydrophilicity leading to slow desorption and consecutive oxidation of cyclohexanone.     

Highly bright and photostable cyanine dye-doped silica nanoparticles for optical imaging: Photophysical characterization and cell tests

Spherical silica nanoparticles containing fluorescent trimethine indocyanine dyes (λ_abs = 547 nm, λ_em = 570 nm) were prepared using a water-in-oil microemulsion method. The nanoparticles were of 50 nm diameter and were almost monodispersed in aqueous solution at pH 5.5. Entrapment of dye molecules in the silica matrix stabilised photoemission over several hours of continuous irradiation. The photoemission intensity of the indocyanine was increased 13-fold over that recorded in solution. As each nanoparticle contained 110 dye molecules, the photoemission brightness of each particle was enhanced by three orders of magnitude. The fluorescent nanoparticles have been tested as imaging tools in in vitro tests. As an example of non-macrophagic cells, a highly differentiated neuronal cell line (GT1-7) was used and the results showed that the prepared nanoparticles can be incorporated into these cells with no apparent toxicity for up to three days.     

High Mobility Group Box 1 Protein in Cerebral Thromboemboli

High-mobility group box 1 protein (HMGB1) is a damage-associated molecular pattern (DAMP) involved in neutrophil extracellular trap (NET) formation and thrombosis. NETs are regularly found in cerebral thromboemboli. We here analyzed associated HMGB1 expression in human thromboemboli retrieved via mechanical thrombectomy from 37 stroke patients with large vessel occlusion. HMGB1 was detected in all thromboemboli, accounting for 1.7% (IQR 0.6–6.2%) of the total thromboemboli area and was found to be colocalized with neutrophils and NETs and in spatial proximity to platelets. Correlation analysis revealed that the detection of HMGB1 was strongly related to the number of neutrophils (r = 0.58, p = 0.0002) and platelets (r = 0.51, p = 0.001). Our results demonstrate that HMGB1 is a substantial constituent of thromboemboli causing large vessel occlusion stroke.     

A priori performance assessment of line-less mobile assembly systems

Present assembly systems are often based on rigid, line-based approaches and are hindered in their reconfiguration capability. Line-less Mobile Assembly Systems (LMAS) are a novel approach for assembly organization. They improve flexibility through mobile resources, permitting spatiotemporal freedom in scheduling and resource assignment. This paper presents a method for a priori assessment of LMAS during the early stages of the assembly system design process. The method applies a modified, extended mean value analysis to a closed queuing network representation of LMAS to estimate performance. The method is validated model analysis and comparison on two use cases indicating plausible model behavior.     

Doxorubicin Impairs Smooth Muscle Cell Contraction: Novel Insights in Vascular Toxicity

Clinical and animal studies have demonstrated that chemotherapeutic doxorubicin (DOX) increases arterial stiffness, a predictor of cardiovascular risk. Despite consensus about DOX-impaired endothelium-dependent vasodilation as a contributing mechanism, some studies have reported conflicting results on vascular smooth muscle cell (VSMC) function after DOX treatment. The present study aimed to investigate the effects of DOX on VSMC function. To this end, mice received a single injection of 4 mg DOX/kg, or mouse aortic segments were treated ex vivo with 1 μM DOX, followed by vascular reactivity evaluation 16 h later. Phenylephrine (PE)-induced VSMC contraction was decreased after DOX treatment. DOX did not affect the transient PE contraction dependent on Ca²⁺ release from the sarcoplasmic reticulum (0 mM Ca²⁺), but it reduced the subsequent tonic phase characterised by Ca²⁺ influx. These findings were supported by similar angiotensin II and attenuated endothelin-1 contractions. The involvement of voltage-gated Ca²⁺ channels in DOX-decreased contraction was excluded by using levcromakalim and diltiazem in PE-induced contraction and corroborated by similar K⁺ and serotonin contractions. Despite the evaluation of multiple blockers of transient receptor potential channels, the exact mechanism for DOX-decreased VSMC contraction remains elusive. Surprisingly, DOX reduced ex vivo but not in vivo arterial stiffness, highlighting the importance of appropriate timing for evaluating arterial stiffness in DOX-treated patients.     

Pt–Ba–Al2O3 for NOx storage and reduction: Characterization of the dispersed species

Pt–Ba–Al₂O₃ active and selective for NO_x storage and selective reduction to N₂ has been prepared and tested. Characterization of the parent Al₂O₃, Pt–Al₂O₃ and Ba–Al₂O₃ materials, as well as of Pt–Ba–Al₂O₃ catalyst in the oxidized, reduced and sulphated state has been performed by FT-IR spectroscopy of low-temperature adsorbed carbon monoxide and of adsorbed acetonitrile. XRD, TEM and XPS analyses have also been performed. Evidence for the predominance of Ba species, which are highly dispersed on the alumina support surface, and may be carbonated or sulphated, has been provided. Competitive interaction of Pt and Ba species with the surface sites of alumina has also been found.     

Enabling Electrocatalytic Fischer–Tropsch Synthesis from Carbon Dioxide Over Copper-based Electrodes

We report on the discovery that paraffins and olefins up to C₆ hydrocarbons can be obtained in CO₂ electroreduction at room temperature and atmospheric pressure by application of a commercially available Cu-electrode (Eurofysica), provided pretreatment by electropolishing is avoided. The product distribution follows the Schultz–Flory distribution and, depending on the applied potential, the chain growth probability (α) ranges from 0.23 to 0.31, values lower than those obtained in Fischer–Tropsch synthesis over heterogeneous Co- or Fe-based catalysts.