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.     

Effects of SO2 on selective catalytic reduction of NO with NH3 over a TiO2 photocatalyst

The effect of SO₂ gas was investigated on the activity of the photo-assisted selective catalytic reduction of nitrogen monoxide (NO) with ammonia (NH₃) over a TiO₂ photocatalyst in the presence of excess oxygen (photo-SCR). The introduction of SO₂ (300 ppm) greatly decreased the activity of the photo-SCR at 373 K. The increment of the reaction temperature enhanced the resistance to SO₂ gas, and at 553 K the conversion of NO was stable for at least 300 min of the reaction. X-ray diffraction, FTIR spectroscopy, thermogravimetry and differential thermal analysis, x-ray photoelectron spectroscopy (XPS), elemental analysis and N₂ adsorption measurement revealed that the ammonium sulfate species were generated after the reaction. There was a strong negative correlation between the deposition amount of the ammonium sulfate species and the specific surface area. Based on the above relationship, we concluded that the deposition of the ammonium sulfate species decreased the specific surface area by plugging the pore structure of the catalyst, and the decrease of the specific surface area resulted in the deactivation of the catalyst.     

Anti‐Sway Crane Control Considering Wind Disturbance and Container Mass

A method for estimating the sway angle using an observer has already been proposed. The state observer estimates the sway angle accurately and must use the detected sway angle value. However, the estimated sway angle has an error owing to rope length error, friction force, and wind. Moreover, the container mass cannot be determined, and therefore the observer parameter is not suitable. We already proposed robust antisway control for overcoming rope length error without adding a new sensor. Further, we designed a friction disturbance observer to cancel out the influence of the friction force. In this paper, we first propose a container mass estimation method when a crane system performs rolling up control. The observer parameter can be selected using the estimated mass value. Second, in crane parallel shift control, we propose a robust antisway control even when there is a wind disturbance. We design a wind disturbance observer and propose a wind disturbance estimator to separate the friction observer output from the wind disturbance observer output. We confirm through experiments that the proposed method can reduce vibration.     

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.     

Tunneling Conductance Dependence of the Tunneling Spectra in Bi2Sr2CaCu2O8

We have measured the tunneling spectra in Bi₂Sr₂CaCu₂O₈ with a scanning tunneling microscope(STM) at various tip-sample distances by changing the tunneling conductance in a controlled manner. When the tunneling conductance is increased from 1×10^–9 to 1×10^–5 S, the spectra do not show changes in particular. However, the gap value decreases steeply and the asymmetric back ground density of states turns inverted V-shaped one above 6×10^–4 S. The changes in the tunneling spectra at the high tunneling conductances are explained by the enhancement of the local carrier density induced by the pressure that the STM tip applied to the sample.     

An on-chip active decoupling circuit to suppress crosstalk in deep-submicron CMOS mixed-signal SoCs

A decoupling circuit using an operational amplifier is proposed to suppress substrate crosstalk in mixed-signal system-on-chip (SoC) devices. It overcomes the parasitic inductance problem of on-chip capacitor decoupling. The effect of the proposed decoupling circuit is not limited by parasitic fine impedance. A 0.13-/spl mu/m CMOS test chip showed that substrate noise at frequencies from 40 MHz to 1 GHz was incrementally suppressed by sequentially activating three of the proposed circuits in parallel. The power dissipation of each circuit was 3.3 mW at a 1.0-V power supply. The test chip measurement showed that the proposed decoupling reduced crosstalk by 31% at 200 MHz, whereas it was reduced by 4.4% with capacitor decoupling. This 7:1 ratio, or 17 dB, corresponds to the gain of the opamp. Design of the opamp and its feedback loop for active decoupling is simple, making the opamp useful for SoC applications.     

Biotransformation of three aromadendrane‐type sesquiterpenoids by Aspergillus wentii

BACKGROUND: The biotransformation of sesquiterpenoids, which are a large class of naturally occurring compounds, using microorganisms as a biocatalyst to produce useful novel organic compounds was investigated. The biotransformation of sesquiterpenoids, (+)‐aromadendrene ( 1 ), (−)‐alloaromadendrene ( 2 ) and (+)‐ledene ( 3 ) has been investigated using Aspergillus wentii as a biocatalyst. Results: Compound 1 was converted to (−)‐(10S,11S)‐10,13,14‐trihydroxyaromadendrane ( 4 ). Compound 2 was converted to (+)‐(1S,11S)‐1,13‐dihydroxyaromadendrene ( 5 ) and (−)‐5,11‐epoxycadin‐1(10)‐en‐14‐ol ( 6 ). Compound 3 was converted to compound 6 , (+)‐(10R,11S)‐10,13‐dihydroxyaromadendr‐1‐ene ( 7 ) and (+)‐(10S,11S)‐10,13‐dihydroxyaromadendr‐1‐ene ( 8 ). The structure of the metabolic products has been elucidated on the basis of their spectral data. CONCLUSION: Compound 1 gave only one product that was hydroxylated at C‐10, C‐13 and C‐14. By contrast, compounds 2 and 3 gave a number of products, one of which was common. The differences in oxidation of 1–3 are due to the configuration of the C‐1 position. Compounds 4–8 were new compounds. Copyright © 2008 Society of Chemical Industry     

Strengthening of Preloaded RC Beams Using Hybrid Carbon Sheets

Reinforced concrete (RC) beams subject to service loads of 40 or 60% of steel yielding were strengthened using hybrid continuous carbon fiber sheets. The hybrid systems were made of high-strength and high-modulus carbon sheets, and compared with systems using only high-strength carbon. It was found that the use of high-modulus carbon sheets in hybrid systems could increase the yielding load, the flexural stiffness, the postyielding ductility, and reduce the crack opening in concrete. The slope changes on load-deflection curves at steel yielding are not noticeable in hybrid systems. The tensile strains developed in hybrid sheets after the fracture of high-modulus carbon are higher in magnitude and distributed in a larger area, leading to an ultimate carbon fracture with concrete crushing. These unique features are attributed to the high stiffness and low ultimate tensile strain of the high-modulus carbon fibers which stiffen the structures, avoid or delay the fiber-reinforced polymer debonding, and facilitate the deformability during their subsequent breakdown.