Novel KCND3 Variant Underlying Nonprogressive Congenital Ataxia or SCA19/22 Disrupt KV4.3 Protein Expression and K+ Currents with Variable Effects on Channel Properties

KCND3 encodes the voltage-gated potassium channel K_V4.3 that is highly expressed in the cerebellum, where it regulates dendritic excitability and calcium influx. Loss-of-function K_V4.3 mutations have been associated with dominant spinocerebellar ataxia (SCA19/22). By targeted NGS sequencing, we identified two novel KCND3 missense variants of the K_V4.3 channel: p.S347W identified in a patient with adult-onset pure cerebellar syndrome and p.W359G detected in a child with congenital nonprogressive ataxia. Neuroimaging showed mild cerebellar atrophy in both patients. We performed a two-electrode voltage-clamp recording of K_V4.3 currents in Xenopus oocytes: both the p.G345V (previously reported in a SCA19/22 family) and p.S347W mutants exhibited reduced peak currents by 50%, while no K+ current was detectable for the p.W359G mutant. We assessed the effect of the mutations on channel gating by measuring steady-state voltage-dependent activation and inactivation properties: no significant alterations were detected in p.G345V and p.S347W disease-associated variants, compared to controls. K_V4.3 expression studies in HEK293T cells showed 53% (p.G345V), 45% (p.S347W) and 75% (p.W359G) reductions in mutant protein levels compared with the wildtype. The present study broadens the spectrum of the known phenotypes and identifies additional variants for KCND3-related disorders, outlining the importance of SCA gene screening in early-onset and congenital ataxia.     

Unusually strong positive cooperativity in binding of peptides to latent membrane protein-1 DNA fragments of the Epstein-Barr viral gene

The DNA-binding preferences of two oligopeptide amides, (His-Pro-Arg-Lys)(3)NH(2) (HR-12) and (Ser-Pro-Arg-Lys)(3)NH(2) (SP-12), have been examined by quantitative DNase I footprinting studies. Two different DNA fragments were investigated: a pair of 5'-(32)P-labeled duplexes from pBR322 with one or other of the complementary strands labeled and a corresponding pair of 5'-(32)P-labeled duplexes representing fragments of the latent membrane protein (LMP-1) gene from a pathogenic Epstein-Barr virus variant derived from nasopharyngeal carcinoma. The major objective was to examine molecular recognition and cooperative features associated with sequence-selective binding of synthetic peptides to the LMP-1 fragments. At various binding sites on the pBR322 fragments, Hill coefficients (n(H)) ranging from 1.9-2.2 were observed; these results indicate modest positive cooperativity between binding sites for both peptides. By contrast, unusually high values of n(H), ranging from 4.0-9.3, were observed at various binding sites on the LMP-1 fragments. Allosteric models can be constructed to interpret the observed cooperative interactions between different DNA recognition sites in the LMP-1 gene upon binding of the peptide ligands. It is noteworthy that these models feature a novel network of cooperativity interconnecting multiple DNA allosteric sites. The evidence of sequence selectivity and strong cooperativity discovered in this work may prove to be a general feature of peptide interactions with some nucleic acids.     

ECG triggering and gating for ultrasonic small animal imaging

Echocardiography (ECG) is routinely used in the clinical diagnosis of cardiac function. The anatomy of the mouse is similar to that of the human, and thus murine ECG has become an effective tool for the assessment of small animal models of human cardiac diseases. Unfortunately, clinical ultrasonic imaging systems are not suitable for murine cardiac imaging due to their limited spatial and temporal resolutions. Murine ECG requires a spatial resolution better than 100 pim, which mandates the use of high-frequency, ultrasonic imaging (i.e., >20 MHz). High-frequency transducer arrays currently are not available, and so such systems use the mechanical scanning of a single-element transducer for which the frame rate is insufficient for directly monitoring the rapid beating of a mouse heart, and thus retrospective image reconstruction is necessary. This paper presents a high-frequency, ultrasonic imaging system for murine cardiac imaging. Two scanning methods have been developed. One is based on ECG triggering and is called the block scanning mode, in which the murine cardiac images from the isovolumic contraction and isovolumic relaxation phases are retrospectively reconstructed within a relatively short data acquisition time using the ECG R-wave as the trigger to the imaging system. The other method is the line scanning mode based on ECG gating, in which both ECG and ultrasound scan lines are continuously acquired over a longer time, enabling images during the entire cardiac cycle to be obtained. It is demonstrated here that the effective frame rate is determined by the pulse repetition frequency and can be up to 2 kHz in the presented system.     

Promotion of generation of hydrogen by splitting of water by CNT over Pt/TiO2 catalysts

The generation of hydrogen over CNT/Pt/TiO2 catalysts by the splitting of water under irradiation with UV light is studied. The maximum rate of evolution of hydrogen was 2300 micromolg(-1)h(-1) on 0.06 wt% Pt/TiO2 (sol-gel) and reached a stable value of approximately 2000 micromolg(-1)h(-1) when the Pt loading exceeded at a Pt loading of over 0.06 wt%. Single wall carbon nanotubes (SWCNTs) were applied to enhance the hydrogen generation activity. The evolution rate of hydrogen on 0.06 wt% Pt/0.02 wt% SWCNT-TiO2 (sol-gel) was 3836 micromolh(-1)g(-1). 0.1 M NaCI yielded more hydrogen than any other tested salt. The XRD spectra show that the crystal lattices of commercial TiO2 (ST-21) and self-made TiO2 (sol-gel method) are of the anatase form. However, the TEM images and other catalytic activity data show that the SWCNTs act as wires for the transmission of electrons.     

Implementing Fourier algorithm by linear modulation method for distance relaying application

Abstract

The Fourier algorithm, due to its good frequency response characteristics, is frequently used in calculating the peak value of a distorted wave for real time measuring applications. A new method, using the linear modulation method, is proposed to implement the algorithm. It improves the defect of a large number of arithmatic operations during implementation of the algorithm. This paper describes the structure and theory of the proposed linear modulated Fourier algorithm. Test results, including the required arithmatic operations and the numerical accuracy, are reported. Moreover, application to the digital impedance relay was simulated, and the simulation results show that the proposed algorithm is suitable for relaying applications.     

Environmentally assisted cracking behavior of dissimilar metal weldments in simulated BWR coolant environments

The environmentally assisted cracking behavior of dissimilar metal (DM) welds, including Alloy 52-A 508 and Alloy 82-A508, under simulated BWR coolant conditions was studied. Effects of postweld heat treatment and sulfur content of the base metal on the corrosion fatigue and SCC growth rates of DM welds were evaluated. The crack growth rates for the DM weld heat-treated at 621 °C for 24 h were observed to be faster than those for the as-welded. But the DM weld heat-treated at 621 °C for 8 h + 400 °C for 200 h showed better SCC resistance than the as-welded. The longer the heat treatment at 621 °C, the higher the chromium carbides density along the grain boundary was observed. Sulfur could diffuse out of the base metal and segregate along the grain boundaries of the dilution zone, leading to weakening the grain boundary strength and the SCC resistance of the Alloy 52-A508 weld.     

Constructing an anticipation formula for fire loss in factory-type

The before fire assessment data from the ARC fire risk assessment system and the financial loss from post-fire damage report for factory-type buildings located in Taiwan were collected. The correlation between assessment data and degree of fire loss (DFL) was calculated by three regression analyses – linear, power, and exponential equations – to produce an anticipation formula. The results revealed that there is more a believable prediction when the fire loss is bigger, regardless of the amount of fire loss or the DFL, while the latter is more related to the assessment grade. By providing proprietors and insurance companies detailed fire risk analysis showing predictable financial loss, it is advantageous for budget management and fire protection, enforcement and should result in the reduction of fire risk and subsequent damage to factory-type buildings.