A Novel LRRK2 Variant p.G2294R in the WD40 Domain Identified in Familial Parkinson’s Disease Affects LRRK2 Protein Levels

Leucine-rich repeat kinase 2 (LRRK2) is a major causative gene of late-onset familial Parkinson’s disease (PD). The suppression of kinase activity is believed to confer neuroprotection, as most pathogenic variants of LRRK2 associated with PD exhibit increased kinase activity. We herein report a novel LRRK2 variant—p.G2294R—located in the WD40 domain, detected through targeted gene-panel screening in a patient with familial PD. The proband showed late-onset Parkinsonism with dysautonomia and a good response to levodopa, without cognitive decline or psychosis. Cultured cell experiments revealed that p.G2294R is highly destabilized at the protein level. The LRRK2 p.G2294R protein expression was upregulated in the patient’s peripheral blood lymphocytes. However, macrophages differentiated from the same peripheral blood showed decreased LRRK2 protein levels. Moreover, our experiment indicated reduced phagocytic activity in the pathogenic yeasts and α-synuclein fibrils. This PD case presents an example wherein the decrease in LRRK2 activity did not act in a neuroprotective manner. Further investigations are needed in order to elucidate the relationship between LRRK2 expression in the central nervous system and the pathogenesis caused by altered LRRK2 activity.     

Hydrogen production through dry reforming of biogas using a porous electrochemical cell: Effects of a cobalt catalyst in the electrode and mixing of air with biogas

This paper reports the performance of porous Gd-doped ceria (GDC) electrochemical cells with Co metal in both electrodes (cell No. 1) and with Ni metal in the cathode and Co metal in the anode (cell No. 2) for CO₂ decomposition, CH₄ decomposition, and the dry reforming reaction of a biogas with CO₂ gas (CH₄ + CO₂ → 2H₂ + 2CO) or with O₂ gas in air (3CH₄ +?1.875CO₂ +?1.314O₂ → 6H₂ +?4.875CO +?0.7515O₂). GDC cell No. 1 produced H₂ gas at formation rates of 0.055 and 0.33?mL-H₂/(min?m²-electrode) per 1?mL-supplied gas/(min?m²-electrode) at 600?°C and 800?°C, respectively, by the reforming of the biogas with CO₂ gas. Similarly, cell No. 2 produced H₂ gas at formation rates of 0.40?mL-H₂/(min?m²) per 1?mL-supplied gas/(min?m²) at 800?°C from a mixture of biogas and CO₂ gas. The dry reforming of a real biogas with CO₂ or O₂ gas at 800?°C proceeded thermodynamically over the Co or Ni metal catalyst in the cathode of the porous GDC cell. Faraday's law controlled the dry reforming rate of the biogas at 600?°C in cell No. 2. This paper also clarifies the influence of carbon deposition, which originates from CH₄ pyrolysis (CH₄ → C + 2H₂) and disproportionation of CO gas (2CO → C + CO₂), on the cell performance during dry reforming. The dry reforming of a biogas with O₂ molecules from air exhibits high durability because of the oxidation of the deposited carbon by supplied air.     

Synthesis of thermoresponsive block and graft copolymers via the combination of living cationic polymerization and RAFT polymerization using a vinyl ether-type RAFT agent

A novel vinyl ether-type RAFT agent, benzyl 2-(vinyloxy)ethyl carbonotrithioate (BVCT) was synthesized for various block copolymers via the combination of living cationic polymerization of vinyl ethers and reversible addition−fragmentation chain transfer (RAFT) polymerization. The novel BVCT–trifluoroacetic acid adduct play an important role to produce well-defined block copolymers, which is both as a cationogen under EtAlCl₂ initiation system in the presence of ethyl acetate for living cationic polymerization and a RAFT agent for blocks by RAFT polymerization. The resulting polymer, poly(vinyl ether)s, by living cationic polymerization had a high number average α-end functionality (≥0.9) as determined by both ¹H NMR and MALDI-TOF-MS spectrometry. In addition, this poly(vinyl ether)s worked well as a macromolecular chain transfer agent for RAFT polymerization. The RAFT polymerization of radically polymerizable monomers was conducted in toluene using 2,2′-azobis(isobutyronitrile) at 70 °C. For example, a double thermoresponsive block copolymer (MOVE₆₁-b-NIPAM₁₅₀) consisting of 2-methoxyethyl vinyl ether (MOVE) and N-isopropylacrylamide (NIPAM) was prepared via the combination of living cationic polymerization and RAFT polymerization. The block copolymer reversibly formed and deformed micellar assemblies above the phase separation temperature (T_ps) of poly(NIPAM) block in water. This BVCT is not only functioned as an initiator, but also acted as a monomer. When BVCT was copolymerized with MOVE by living cationic polymerization, followed by graft copolymerization with NIPAM via RAFT polymerization, well-defined graft copolymers (MOVE_n-co-BVCT_m)-g-NIPAM_x (n = 62–73, m = 1–9, x = 19–214) were successfully obtained. However, no micelle formed in water above T_ps of poly(NIPAM) graft chain unlike the case of block copolymers.     

The study of metal ion release and cytotoxicity in Co–Cr–Mo and Ti–Al–V alloy in total knee prosthesis – scanning electron microscopic observation

We surgically retrieved two cobalt(Co)–chromium(Cr)–molybdenum(Mo) and five titanium(Ti)–aluminum(Al)–vanadium(V) alloy knee prostheses from patients because of mechanical failure and pain. We examined the distribution of the small particles which were released from the Co–Cr–Mo and Ti–Al–V alloys using a backscattered scanning electron microscopy (SEM). In addition we analyzed the metals in the artificial knee joints and the tissues adjacent to them using energy dispersive X-ray spectroscopy (EDS). We demonstrated that a myriad of fine particles, produced by the abrasion of both Co–Cr–Mo and Ti–Al–V alloys, accumulated in the synovial cells. As Co–Cr–Mo alloys disintegrate easily in the cells, Co dissolves from the peripheral areas of them, although Cr remains within the cells. In contrast Ti–Al–V alloys are very stable in the synovial cells. From these findings we conclude that the Co–Cr–Mo alloys are hazardous to the body as the alloys release Co which enters the body. In contrast the Ti–Al–V alloys are very stable and are patently safer. Artificial joints, however, are still in considerable need of improvement.     

Facile one-step route to polyaniline–silver nanocomposite particles and their application as a colored particulate emulsifier

Polyaniline–silver nanocomposites were synthesized in the form of colloidal particles by the facile one-step aqueous chemical oxidative dispersion polymerization of aniline using silver nitrate as an oxidant and poly(vinyl alcohol) as a colloidal stabilizer. Aniline monomer was oxidized by silver ions, yielding polyaniline and elemental Ag simultaneously. The synthesized nanocomposite particles were colloidally stable over 2 years and transmission electron microscopy studies indicated the production of spherical, plate and rod-shaped polyaniline–silver nanocomposite particles with a silver core–polyaniline shell morphology. The conductivity of a pressed pellet of the nanocomposite particles using the conventional four-point probe technique was 1.4 × 10^?2 S/cm at 25 °C. The nanocomposite particles behaved as a ‘colored’ particulate emulsifier for the stabilization of transparent oil-in-water emulsions.     

Connection, a microcomputer program for storing and analyzing structural properties of neural circuits

The application of a microcomputer-based system (the Connection system) designed to deal with neuroanatomical information commonly analyzed by researchers and involved in the study of structural properties of neural circuits is presented. This system can be employed at first as a readily-accessible database containing physiological and anatomical data from nuclei of the central nervous system which define a network with up to 45 elements and their subdivisions and connections. Once the database from a specific network is built and stored in a file, routines of this system can be used to classify the nuclei in term of their afferents and efferents and also to display all possible pathways linking any pair of nuclei and their respective length (number of synapses). The role of such a system as an auxiliary tool in neuroanatomical and electrophysiological research is discussed by presenting the results obtained from the analysis of the neural circuits involved in cardiovascular function control in higher vertebrates.     

Reduction of in vitro allergenicity of buckwheat Fag e 1 through the Maillard-type glycosylation with polysaccharides

A major allergenic protein of buckwheat, Fag e 1 prepared from common buckwheat (Fagopyrum esculentum), was covalently linked with food-grade polysaccharides, arabinogalactan or xyloglucan through the controlled dry-heating at 60 °C under 65% relative humidity. The introduction of polysaccharide chain onto the molecular surface of Fag e 1 reduced the allergenicity of Fag e 1. The results revealed that the Maillard-type glycosylation of Fag e 1 with polysaccharides brought about a drastic reduction of the reactivity against human sera of buckwheat-allergy subjects, using immuno dot-blotting, QCM analysis and ELISA. In addition, the glycosylation of Fag e 1 yielded a great improvement of its surface functionality. Solubility of Fag e 1 at the neutral pH was substantially increased up to 13.5 times and 9.6 times by the conjugation with arabinogalactan and xyloglucan, respectively. Emulsifying properties of Fag e 1 were also improved by the glycosylation, of which both emulsifying activity and emulsion stability were more than 6 times higher than those of the native protein.     

Field effect surface passivation of SiO2/Si interfaces by heat treatment with high-pressure H2O vapor

We investigated a simple field effect passivation of the silicon surfaces using the high-pressure H₂O vapor heating. Heat treatment with 2.1×10⁶ Pa H₂O vapor at 260°C for 3 h reduced the surface recombination velocity from 405 cm/s (before the heat treatment) to 38 cm/s for the thermally evaporated SiO_x film/Si. Additional deposition of 140 nm-SiO_x films (x<2) with a high density of fixed positive charges on the SiO₂/Si samples further decreased the surface recombination velocity to 22 cm/s. We also demonstrated the field effect passivation for n-type silicon wafer coated with thermally grown SiO₂. Additional deposition of 210 nm SiO_x films on both the front and rear surfaces increased the effective lifetime from 1.4 to 4.6 ms. Combination of thermal evaporation of SiO_x film and the heat treatment with high-pressure H₂O vapor is effective for low-temperature passivation of the silicon surface.     

Monomer arrangement in HSP90 dimer as determined by decoration with N and C-terminal region specific antibodies

Electron microscopy using the low-angle rotary shadowing replica method showed that the HSP90 dimer consists of four globular domains aligning in a tandem fashion. When decorated with two monoclonal antibodies against epitopes mapped on the N-terminal region of HSP90, these antibodies bound to both ends of the HSP90 dimer. A C-terminal region specific antibody was shown to bind to the side of HSP90. These results support a model for HSP90 dimer whereby two HSP90 monomers are arranged in an antiparallel fashion and dimerize through the C-terminal domain. Treatment of HSP90 at elevated temperatures or with ATP at room temperature, though not with ADP, induces molecular transformation of the linear HSP90 dimer into an O-ring-shaped structure.