Analysis of Cerebrospinal Fluid Extracellular Vesicles by Proximity Extension Assay: A Comparative Study of Four Isolation Kits

There is a lack of reliable biomarkers for disorders of the central nervous system (CNS), and diagnostics still heavily rely on symptoms that are both subjective and difficult to quantify. The cerebrospinal fluid (CSF) is a promising source of biomarkers due to its close connection to the CNS. Extracellular vesicles are actively secreted by cells, and proteomic analysis of CSF extracellular vesicles (EVs) and their molecular composition likely reflects changes in the CNS to a higher extent compared with total CSF, especially in the case of neuroinflammation, which could increase blood–brain barrier permeability and cause an influx of plasma proteins into the CSF. We used proximity extension assay for proteomic analysis due to its high sensitivity. We believe that this methodology could be useful for de novo biomarker discovery for several CNS diseases. We compared four commercially available kits for EV isolation: MagCapture and ExoIntact (based on magnetic beads), EVSecond L70 (size-exclusion chromatography), and exoEasy (membrane affinity). The isolated EVs were characterized by nanoparticle tracking analysis, ELISA (CD63, CD81 and albumin), and proximity extension assay (PEA) using two different panels, each consisting of 92 markers. The exoEasy samples did not pass the built-in quality controls and were excluded from downstream analysis. The number of detectable proteins in the ExoIntact samples was considerably higher (~150% for the cardiovascular III panel and ~320% for the cell regulation panel) compared with other groups. ExoIntact also showed the highest intersample correlation with an average Pearson’s correlation coefficient of 0.991 compared with 0.985 and 0.927 for MagCapture and EVSecond, respectively. The median coefficient of variation was 5%, 8%, and 22% for ExoIntact, MagCapture, and EVSecond, respectively. Comparing total CSF and ExoIntact samples revealed 70 differentially expressed proteins in the cardiovascular III panel and 17 in the cell regulation panel. To our knowledge, this is the first time that CSF EVs were analyzed by PEA. In conclusion, analysis of CSF EVs by PEA is feasible, and different isolation kits give distinct results, with ExoIntact showing the highest number of identified proteins with the lowest variability.     

Methyltransferase Contingencies in the Pathway of Everninomicin D Antibiotics and Analogues

Everninomicins are orthoester oligosaccharide antibiotics with potent activity against multidrug-resistant bacterial pathogens. Everninomicins act by disrupting ribosomal assembly in a distinct region in comparison to clinically prescribed drugs. We employed microporous intergeneric conjugation with Escherichia coli to manipulate Micromonospora for targeted gene-replacement studies of multiple putative methyltransferases across the octasaccharide scaffold of everninomicin effecting the A₁, C, F, and H rings. Analyses of gene-replacement and genetic complementation mutants established the mutability of the everninomicin scaffold through the generation of 12 previously unreported analogues and, together with previous results, permitted assignment of the ten methyltransferases required for everninomicin biosynthesis. The in vitro activity of A₁- and H-ring-modifying methyltransferases demonstrated the ability to catalyze late-stage modification of the scaffold on an A₁-ring phenol and H-ring C-4’ hydroxy moiety. Together these results establish the potential of the everninomicin scaffold for modification through mutagenesis and in vitro modification of advanced biosynthetic intermediates.     

Formation and wear mechanism of nickel titanium intermetallics during heat treatment of nickel coating on Ti-6Al-4V substrate

In the present work, inter-diffusion of nickel and titanium and formation of Ni-Ti intermetallic compounds on Ti-6Al-4V substrate have been studied. Initially, nickel was electrodeposited on the alloy using a modified Watts bath solution at a current density of 2 A dm^?2 for 1?h. The coated specimens were then heat treated for different durations at 750, 800 and 850 °C under argon atmosphere. The effects of temperature and time on the characteristics, hardness and wear resistance of intermetallic phases were investigated. The results showed that a multilayer structure was formed after heat treatment, an outer layer of residual nickel, an area of intermetallic layers with different compositions followed by a solid solution of Ni-Ti. It was also observed that an increase in time or temperature at first led to the formation of thicker intermetallic layers; however, after passing a critical point, the intermetallic layers seem to dissolve into the substrate. Furthermore, the wear rates of the diffusion treated samples were four times lower compared to Ti-6Al-4V alloy when sliding against AISI 52100 hardened steel.     

Influence of SiC content on the oxidation of carbon fibre reinforced ZrB2/SiC composites at 1500 and 1650 °C in air

The microstructure and the oxidation resistance in air of continuous carbon fibre reinforced ZrB₂–SiC ceramic composites were investigated. SiC content was varied between 5–20?vol.%, while maintaining fibre content at ～40?vol.%. Short term oxidation tests in air were carried out at 1500 and 1650?°C in a bottom-up loading furnace. The thickness, composition and microstructure of the resulting oxide scale were analysed by SEM-EDS and X-Ray diffraction. The results show that contents above 15?vol.% SiC ensure the formation of a homogeneous protective borosilicate glass that covers the entire sample and minimizes fibre burnout. The scale thickness is ～90?μm for the sample containing 5?vol.% SiC and decreases with increasing SiC content.     

Preparation of CaO granules using the granulation method

ABSTRACT

Improving the hydration resistance of CaO particle in manufacturing and application of free CaO-containing materials has practical significance. In this study, CaO granules was made from Ca(OH)₂ particles, which were fabricated by the granulation method. The results showed that the hydration resistance of the CaO granules which was prepared under 1700?r?min^?1 was the best, the CaO granules was sintered well in calcination process, the shell of CaO granules was relatively dense, which improves the hydration resistance of CaO granules, and the rate of hydration weight increment was 0.58% after placed in the air for 20 days under a temperature of 10–14°C and a relative humidity of 57–81%.     

Studying the effect of organo-modified nanoclay loading on the thermal stability,flame retardant,anti-corrosive and mechanical properties of polyurethane nanocomposite for surface coating

Clay polyurethane nanocomposite (CPN) coating films were fabricated by uniformly dispersing nanoclay, organically modified with 25–30 wt.% octadecylamine in varying concentrations up to 5 wt.%, in a commercial two component, glossy, acrylic aliphatic polyurethane using ultrasonication. Organo-modified nanoclay was characterized by X-ray diffraction (XRD). The dispersion of the nanoclay into the matrix was investigated by scanning electron microscopy (SEM). CPN coating films were characterized by thermogravimetric analysis (TGA), and flame retardant, corrosion resistance and mechanical properties were also investigated. The XRD measurement indicated that, the organo-modified nanoclay particles were mainly constituted of montmorillonite with traces of quartz and calcite also found to be present. The SEM analysis showed that the nanoclay layers were dispersed and intercalated into the polyurethane coating. Thermogravimetric analysis showed that incorporating 5 wt.% organo-nanoclay into polyurethane considerably enhanced the thermal stability and increased the char residue to 14.11 wt.% relative to 4.58 for the sample without organo-nanoclay (blank polyurethane). The limiting oxygen index (LOI) test revealed that incorporation of organo-nanoclay led to a further increase in LOI values, which indicate an improvement in flame retardancy properties. The corrosion resistance also improved and this improvement increases with increase nanoclay wt.%. The mechanical resistance measurements demonstrated that the gloss of the CPN coating films slightly decreased, although hardness, adhesion and impact resistance of the CPN coating films improved with the incorporation of the organo-nanoclay.     

Effect of in-situ formation of columnar mullite and pore structure refinement on high temperature properties of corundum casatbles

The effects of in-situ synthesis columnar mullite and pore structure on the hot modulus of rupture (HMOR), thermal shock resistance and corrosion resistance of corundum castables have been investigated in this paper. When 2% nano silica was added, the pore diameters of castables could be decreased to 15 nm (at 110 °C), 1 μm (1100 °C) and 6 μm (1500 °C), respectively. The corresponding reducing magnitude of pore size is 98.5%, 83.3% and 33.3%. The HMOR of castables fired at 1500 °C increased by 110% to 3.64 MPa. Furthermore, after three thermal shock cycles, the residual strength ratio of castables increased from 5.2% to 15.3%. A large amount of cross-distributed columnar mullite was formed between nano silica and α-Al₂O₃ by the two-dimensional nucleation mechanism, which remarkably enhanced the high temperature properties. The penetration index reduced from 30.86% to 19.88%, suggesting that smaller pore size and higher viscosity had a great influence to the penetration process.     

Corrosion resistance of Al–Cr-slag containing chromium–corundum refractories to slags with different basicity

Al–Cr slag is the solid waste generated by the smelting of Cr metal. It presents a range of environmental hazards. This study addressed the corrosion resistance of Al–Cr slag containing chromium–corundum refractories to slags with different basicity. Herein, we provide suggestions for the use of Cr–corundum of different basicity in kilns. Al–Cr slag, brown fused Al₂O₃, and chrome green were used as the raw materials, with pure calcium aluminate cement being used as a binder. The brick samples, prepared using different blends of chrome green and corundum, were fired at 1600?°C, and subsequently subjected to a slag corrosion test. After corrosion by slag of different basicity, the phase composition and microstructure of the sample were analyzed by X-ray diffraction, energy dispersive spectrometer and scanning electron microscopy. There were two major findings. First, Cr–corundum brick made from Al–Cr slag has a better slag corrosion resistance than that made from Cr₂O₃ and brown fused Al₂O₃. Second, Cr–corundum brick made from Al–Cr slag has superior corrosion resistance to slag with a CaO:SiO₂ ratio of 2:1.