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.     

[110]-surface strained-SOI CMOS devices

We have newly developed [110]-surface strained-silicon-on-insulator (SOI) n- and p-MOSFETs on [110]-surface relaxed-SiGe-on-insulator substrates with the Ge content of 25%, fabricated by applying the Ge condensation technique to SiGe layers grown on [110]-surface SOI wafers. We have demonstrated that the electron and the hole mobility enhancement of [110]-surface strained-SOI devices amounts to 23% and 50%, respectively, against the mobilities of [110]-surface unstrained MOSFETs. As a result, the electron and the hole mobility ratios of [110]-surface strained-SOI MOSFETs to the universal mobility of (100)-surface bulk-MOSFETs increase up to 81% and 203%, respectively. Therefore, the current drive imbalance between n- and p-MOS can be reduced. Moreover, both the electron and the hole mobilities of the [110]-surface strained-SOIs strongly depend on the drain current flow direction, which is qualitatively explained by the anisotropic effective mass characteristics of the carriers on a [110]-surface Si. As a result, the [110]-surface strained-SOI technology with optimization of the current flow directions of n- and p-MOS is promising for realizing higher speed scaled CMOS.     

Catalytic decomposition of HCFC22 (CHClF2)

The catalytic decomposition of CHClF₂ was studied over various acidic metal oxides in a fixed-bed reactor. The Cr₂O₃ZrO₂ exhibited the highest activity. The presence of water vapor in the reaction system suppresses the transformation of oxides to fluorides, progresses the formation of CO₂, and it improves the catalysts life.     

Occurrence of heterogeneity of N-linked oligosaccharides attached to sycamore (Acer pseudoplatanus L.) laccase after excretion

The N-linked oligosaccharide moieties of sycamore (Acer pseudoplatanus L.) laccase are known to be highly heterogeneous. We confirmed that this oligosaccharide heterogeneity was caused not only during the oligosaccharide biosynthesis in Golgi apparatus, but also after the excretion of laccase protein into a culture medium. The culture medium for the sycamore cells (Acer pseudoplatanus L.) contained beta-galactosidase, alpha-L-fucosidase, beta-N-acetylglucosaminidase, alpha-mannosidase and beta-xylosidase activities. We showed that the largest sugar chain in laccase, oligosaccharide F, [formula: see text] was degraded to [formula: see text] by a crude exoglycosidase mixture in the culture medium.     

Characterization of C-S-H from Highly Reactive β-Dicalcium Silicate Prepared from Hillebrandite

β-dicalcium silicate synthesized by thermal dissociation of hydrothermally prepared hillebrandite (Ca₂(SiO₃)(OH)₂) exhibits extremely high hydration activity. Characterization of the hydrates obtained and investigation of the hydration mechanism was carried out with the aid of trimethylsilylation analysis, ²⁹Si magic angle spinning nuclear magnetic resonance, transmission electron microscopy selected area electron diffraction, and XRD. The silicate anion structure of C-S-H consisted mainly of a dimer and a single-chain polymer. Polymerization advances with increasing curing temperature and curing time. The C-S-H has an oriented fibrous structure and exhibits a 0.73-nm dreierketten in the longitudinal direction. On heating, the C-S-H dissociates to form β-C₂S. The temperature at which βC₂S begins to form decreases with increasing chain length of the C-S-H or as the Ca/Si ratio becomes higher. The high activity of β-C₂S is due to its large specific surface area and the fact that the hydration is chemical-reaction-rate-controlled until its completion. As a result, the hydration progresses in situ and C-S-H with a high Ca/Si ratio is formed.     

Monolithic rat-race mixers for millimeter waves

In this paper, we report on fully monolithic millimeter-wave Schottky-barrier diode (SBD) down-converters with an IF amplifier using heterojunction bipolar transistors (HBT's). A rat-race circuit is used for the mixer, and is analyzed using a harmonic-balance simulator. The measured conversion gain and the isolation are 7.1 and 29 dB in the V-band design, and 8.0 and 25 dB in the W-band design, respectively. The conversion gains are matched well with the circuit simulation     

Crosslinking of amidated polybutadiene with resole. I

Various liquid polybutadiene derivatives were examined as modifiers to improve the brittleness of phenolic resins. Amidated polybutadienes obtained from a maleinated polybutadiene and ammonia had the highest curing tendency when they were reacted with resole under mild conditions. The cured resins were homogeneous and the brittleness of the phenolic resins was remarkably improved. Crosslinking efficiency was closely related to the succinamic acid group of the amidated polybutadiene. Thermal behavior of the succinamic acid group was also related to the effects of curing temperature and curing time on crosslinking. From these observations, cross-condensation between the succinamic acid group of an amidated polybutadiene and resole seemed to have occurred to a great extent than the self-condensation of resole itself and oxidative crosslinking of the amidated polybutadiene.     

Electron-microscopic procedure for acrylic rubber

The electron-microscopic visualization of acrylic rubber dispersed in a heterogeneous structural resin composition consisting of a rubber-modified two-phase plastic comprising essentially a butyl acrylate rubber phase and an acrylonitrile–styrene copolymer phase, respectively, has been accomplished. This procedure consists of the following: The molded resin specimen is treated with hydrazine hydrate solution to produce the acrylic acid hydrazides. Allow the treated specimen to soak in osmium tetroxide solution. The acrylic rubber may be indirectly fixed and stained. Some micrographs of ultrathin sections of two or three resin compositions, cut by an ultramicrotome, are presented.