Site-Specific Tritium Labeling at the Predefined Internal Position of the Chemically-Modified RNA.

In nucleic acid drug discovery, it is extremely important to develop a technology to understand the distribution in target organs and to trace the degradation process in the body in order to optimize the structure and improve the efficiency of the clinical trial process. Since nucleic acid drugs are essentially metabolically degraded into numerous fragments, labeling at the internal position is preferable to that at the terminus. Due to the high molar specific activity of tritium, various approaches for tritium-labeling have been studied for nucleic acid drugs. Nevertheless, a generally-applicable method for tritium labeling of the internal position of a nucleic acid has not been established. In this study, we have demonstrated a new and efficient method for site-specific tritium labeling of the cytosine base at a predefined internal position in nucleic acid drugs. This method was developed by the chemical modification of the cytosine 4-amino group with the pyridinyl vinyl keto group by the functionality-transfer reaction using the reactive oligodeoxynucleotide (ODN), followed by reduction with NaBT4. Applicability to a variety of chemical structures, such as 5-methyl cytosine, 2'-O-methyl, 2'-fluoro ribose derivatives, Locked/Bridged nucleic acid (LNA/BNA) derivatives, as well as phosphorothioate bonds, has been evidenced using nine oligoribonucleic acid (ORN) substrates. It has been clearly demonstrated that this method is an excellent method for tritium-labeling of nucleic acid with an average conversion efficiency of 74%, an average isolated labeling yield of 60%, and an average specific activity of 61 GBq/mmol. This method is expected to contribute to the preclinical absorption, distribution, metabolism, excretion (ADME) studies of nucleic acid drug candidates.

The Use of Mechanical Ventilation Support at the End of Life in Motor Neurone Disease/Amyotrophic Lateral Sclerosis: A Scoping Review.

There has been an increase in the use of mechanical ventilation (MV) in motor neurone disease (MND) to alleviate symptoms related to hypoventilation. Little is known about its use at the end of life, and the withdrawal of MV is a challenging topic. A scoping review was conducted to map the existing evidence of ventilation use at the end of life in MND. The Joanna Briggs Institute methodological framework was used, and a total of 31 documents were included in the data extraction. Boarder themes around place, planning, cause and comments on death were identified. Our findings show that the focus of the literature has been on the process of the withdrawal of ventilation, creating the subthemes of timing and reason for withdrawal, along with ethical, emotional, and practical issues, medications used and the time until death. There is a foundation of evidence to guide the process and discuss the ethical and emotional issues of withdrawing ventilation. However, there remains limited evidence from patient and family member perspectives. Importantly, there is almost no evidence exploring dying with ventilation in place, the active or passive decisions around this process, how the interface might impact the dying process, or what families think about this.

Application of the Functionality Transfer Oligodeoxynucleotide for the Site-Selective Modification of RNA with a Divers Molecule.

Due to the importance of the RNA chemical modifications, methods for the selective chemical modification at a predetermined site of the internal position of RNA have attracted much attention. We have developed functional artificial nucleic acids that modify a specific site of RNA in a site- and base-selective manner. In addition, the copper-catalyzed azide-alkyne cycloaddition (CuAAC) has been shown to introduce additional molecules on the alkynes attached to the pyridine ring. However, it was found that some azide compounds produced the cycloadduct in lower yields. Therefore, in this study, we synthesized the pyridinyl transfer group with the alkyne attached via a polyethylene glycol (PEG) linker with a different length and optimized its structure for both the transfer and CuAAC reaction. Three new transfer groups were synthesized by introducing an alkyne group at the end of the triethylene (11), tetraethylene (12) or pentaethylen glycol linker (13) at the 5-position of the pyridine ring of (E)-3-iodo-1-(pyridin-2-yl)prop-2-en-1-one. These transfer groups were introduced to the 6-thioguanine base in the oligodeoxynucleotide (ODN) in high yields. The transfer groups 11 and 12 more efficiently underwent the cytosine modification. For the CuAAC reaction, although 7 showed low adduct yields with the anionic azide compound, the new transfer groups, especially 12 and 13, significantly improved the yields. In conclusion, the transfer groups 12 and 13 were determined to be promising compounds for the modification of long RNAs.

Low-temperature n-type doping of insulating ultrathin amorphous indium oxide using H plasma-assisted annealing.

Low-temperature process compatibility is a key factor in successfully constructing additional functional circuits on top of pre-existing circuitry without corrupting characteristics thereof, a technique that typically requires die-to-die (wafer-to-wafer) stacking and interconnecting. And against thermal annealing, which is mandatory and is possible only globally for activating amorphous oxide semiconductors, the selective control of electrical characteristics of the oxide thin-films for integrated circuit applications is challenging. Here, a low-temperature process that enables n-type doping of the designed region of insulating In2O3thin-film is demonstrated. A short hydrogen plasma treatment followed by low-temperature annealing is used to increase interstitial and substitutional hydrogen associated bond states creating shallow donor levels in the insulating In2O3surface to transform the thin-film into an n-type semiconductor. As a result, an In2O3thin-film transistor with a high on/off current ratio (>108), a field-effect mobility of 3.8 cm2V-1s-1, and a threshold voltage of ∼3.0 V has been developed. Compared to performing just thermal annealing, the H-plasma assisted annealing process resulted in an n-type In2O3thin-film transistor showing similar characteristics, while the processing time was reduced by ∼1/3 and the plasma-untreated area still remained insulating. With further development, the hydrogen plasma doping process may make possible a monolithic planar process technology for amorphous oxide semiconductors.

N-Acyltriazinedione; a Novel Acylating Reagent Synthesized from a Triazinone-Type Condensing Reagent.

In this paper, we report the synthesis of N-acyltriazinedione via the unexpected O-N acyl rearrangement of acyloxytriazinone and its utility as an acylating reagent. N-Acyltriazinedione can be isolated by silica gel column chromatography and reacts with amines in the absence of any base to give the corresponding amides in good yields.

Bendable BiVO4-Based Photoanodes on a Metal Substrate Realized through Template Engineering for Photoelectrochemical Water Splitting.

Unlike planar photoelectrodes, bendable and malleable photoelectrodes extend their application to mechanical flexibility beyond conventional rigid structures, which have garnered new attention in the field of photoelectrochemical water splitting. A bendable metal (Hastelloy), which has both bendability and compatibility with various oxide layers, allows high-temperature processes for crystallization; therefore it is far superior as a substrate than a conventional flexible polymer. In this study, we fabricate bendable BiVO4 crystalline thin films on the metal substrates by employing template layers (SrRuO3/SrTiO3) to reduce the structural misfits between BiVO4 and the substrate. The crystallinities were verified through X-ray diffraction and transmission electron microscopy, and photocatalytic performances were examined. The crystallinity of BiVO4 was significantly improved by utilizing similar lattice constants and affinities between BiVO4 and the oxide template layers. We also formed a type II heterojunction by adding a WO3 layer which complements the charge separation and charge transfer as a photoanode. The photocurrent densities of tensile-bent BiVO4/WO3 thin films with a bending radius of 10 mm are comparable to those of pristine BiVO4/WO3 thin film in various aqueous electrolytes. Moreover, photostability tests showed that the tensile-bent crystalline photoanodes retained 90% of their initial photocurrent density after 24 h, which proved their exceptional durability. Our work demonstrates that the bendable photoelectrodes with crystallinity hold great potential in terms of device structure for solar-driven water splitting.

A new solid matrix for preservation of viral nucleic acid from clinical specimens at ambient temperature.

Stabilizing paper matrix methods for retaining nucleic acid from inactivated clinical specimens offer a solution for molecular diagnostics when specimens may be stored or shipped at ambient temperature. We developed cellulose disks (UNEXP) saturated with a total nucleic acid extraction buffer (UNEX) modified from a previously developed lysis buffer for multiple enteric pathogens. Infectivity of hepatitis A virus, adenovirus and poliovirus was destroyed after 2-3 h incubation at room temperature on the UNEXP disks. Norovirus RNA could be detected in UNEXP-eluted nucleic acids by reverse transcription-quantitative PCR (RT-qPCR) from 54 stool samples after 2 weeks storage at room temperature on disks; a subset of seven samples were positive after 3 months storage. Genotyping was successful in 76% of 54 samples tested including six of seven samples stored on the UNEXP disks for up to one month. Comparison of UNEXP with the FTA elute card in a subset of 10 samples demonstrated similar detection and genotyping rates after two weeks of storage at room temperature. UNEXP disks could be useful for epidemiologic investigations of disease outbreaks in resource-limited areas by simplifying specimen transport to regional diagnostic laboratories or shipment to international centers without the need to ship samples on dry ice.

Layered deposition of SnS2 grown by atomic layer deposition and its transport properties.

In this work, we report on the layered deposition of few-layer tin disulfide (SnS2) using atomic layer deposition (ALD). By varying the ALD cycles it was possible to deposit poly-crystalline SnS2 with small variation in layer numbers. Based on the ALD technique, we developed the process technology growing few-layer crystalline SnS2 film (3-6 layers) and we investigated their electrical properties by fabricating bottom-gated thin film transistors using the ALD SnS2 as the transport channel. SnS2 devices showed typical n-type characteristic with on/off current ratio of ∼8.32 × 106, threshold voltage of ∼2 V, and a subthreshold swing value of 830 mV decade-1 for the 6 layers SnS2. The developed SnS2 ALD technique may aid the realization of two-dimensional SnS2 based flexible and wearable devices.

An Instant Donning Multi-Channel EEG Headset (with Comb-Shaped Dry Electrodes) and BCI Applications.

We developed a new type of electroencephalogram (EEG) headset system with comb-shaped electrodes that enables the wearer to quickly don and utilize it in daily life. Two models that can measure EEG signals using up to eight channels have been implemented. The electrodes implemented in the headsets are similar to a comb and are placed quickly by wiping the hair (as done with a comb) using the headset. To verify this headset system, donning time was measured and three brain computer interface (BCI) application experiments were conducted. Alpha rhythm-based, steady-state visual evoked potential (SSVEP)-based, and auditory steady state response (ASSR)-based BCI systems were adopted for the validation experiments. Four subjects participated and ten trials were repeated in the donning experiment. The results of the validation experiments show that reliable EEG signal measurement is possible immediately after donning the headsets without any preparation. It took approximately 10 s for healthy subjects to don the headsets, including an earclip with reference and ground electrodes. The results of alpha rhythm-based BCI showed 100% accuracy. Furthermore, the results of SSVEP-based and ASSR-based BCI experiments indicate that performance is sufficient for BCI applications; 95.7% and 76.0% accuracies were obtained, respectively. The results of BCI paradigm experiments indicate that the new headset type is feasible for various BCI applications.

Antiviral activity of Schizonepeta tenuifolia Briquet against noroviruses via induction of antiviral interferons.

Human noroviruses are the causative agents of non-bacterial gastroenteritis worldwide. The rapid onset and resolution of disease symptoms suggest that innate immune responses are critical for controlling norovirus infection; however, no effective antivirals are yet available. The present study was conducted to examine the antiviral activities of Schizonepeta tenuifolia Briquet extract (STE) against noroviruses. Treatment of human norovirus replicon-bearing HG23 cells with STE at 5 and 10 mg/ml concentrations resulted in the reduction in the viral RNA levels by 77.2% and 85.9%, respectively. STE had no cytotoxic effects on HG23 cells. Treatment of RAW 264.7 cells infected with murine norovirus 1 (MNV-1), a surrogate virus of human noroviruses, with STE at 10 and 20 µg/ml concentrations resulted in the reduction of viral replication by 58.5% and 84.9%, respectively. STE treatment induced the expression of mRNAs for type I and type II interferons in HG23 cells and upregulated the transcription of interferon-ß in infected RAW 264.7 cells via increased phosphorylation of interferon regulatory factor 3, a critical transcription regulator for type I interferon production. These results suggest that STE inhibits norovirus replication through the induction of antiviral interferon production during virus replication and may serve as a candidate antiviral substance for treatment against noroviruses.

Fabrication of high crystalline SnS and SnS2 thin films, and their switching device characteristics.

Representative tin sulfide compounds, tin monosulfide (SnS) and tin disulfide (SnS2) are strong candidates for future nanoelectronic devices, based on non-toxicity, low cost, unique structures and optoelectronic properties. However, it is insufficient for synthesizing of tin sulfide thin films using vapor phase deposition method which is capable of fabricating reproducible device and securing high quality films, and their device characteristics. In this study, we obtained highly crystalline SnS thin films by atomic layer deposition and obtained highly crystalline SnS2 thin films by phase transition of the SnS thin films. The SnS thin film was transformed into SnS2 thin film by annealing at 450 °C for 1 h in H2S atmosphere. This phase transition was confirmed by x-ray diffractometer and x-ray photoelectron spectroscopy, and we studied the cause of the phase transition. We then compared the film characteristics of these two tin sulfide thin films and their switching device characteristics. SnS and SnS2 thin films had optical bandgaps of 1.35 and 2.70 eV, and absorption coefficients of about 105 and 104 cm-1 in the visible region, respectively. In addition, SnS and SnS2 thin films exhibited p-type and n-type semiconductor characteristics. In the images of high resolution-transmission electron microscopy, SnS and SnS2 directly showed a highly crystalline orthorhombic and hexagonal layered structure. The field effect transistors of SnS and SnS2 thin films exhibited on-off drain current ratios of 8.8 and 2.1 × 103 and mobilities of 0.21 and 0.014 cm2 V-1 s-1, respectively. This difference in switching device characteristics mainly depends on the carrier concentration because it contributes to off-state conductance and mobility. The major carrier concentrations of the SnS and SnS2 thin films were 6.0 × 1016 and 8.7 × 1013 cm-3, respectively, in this experiment.

Effect of Power Levels on Inactivation of Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes in Tomato Paste Using 915-Megahertz Microwave and Ohmic Heating.

The effect of power levels on inactivation of Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes in tomato paste was investigated using 915-MHz microwave heating (MW) and ohmic heating (OH). Heating uniformity, pathogen inactivation, and quality aspects were determined with 1.8-, 2.1-, 2.4-, and 3.0-kW MW and corresponding OH. GInaFit was used to analyze pathogen inactivation. The heating uniformity of MW-treated samples was inferior to that of OH-treated samples at low power levels of 1.8 to 2.4 kW but improved as the power level increased. Pathogen inactivation of MW-treated samples was significantly higher than that of OH-treated samples at low power levels of 1.8 to 2.4 kW (P < 0.05) but was not significantly different at the highest power level of 3.0 kW (P > 0.05). Quality aspects (color, pH, and lycopene content), except for L*, of MW-treated samples were not significantly degraded (P > 0.05) by increased power levels. Our results indicate that increasing power levels of MW ensures heating uniformity and microbiological safety and preserves quality aspects of tomato paste.

Spontaneous Marangoni Mixing of Miscible Liquids at a Liquid-Liquid-Air Contact Line.

We investigate the flow patterns created when a liquid drop contacts a reservoir liquid, which has implications on various physicochemical and biochemical reactions including mixing in microfluidic systems. The localized vortical flow spontaneously triggered by the difference of surface tension between the two liquids is studied, which is thus termed the Marangoni vortex. To quantitatively investigate the strength of vortices, we performed particle image velocimetry (PIV) experiments by varying the surface tension difference, the gap of the flow cell, the density and viscosity of the reservoir liquid, and the size of the drop. A scaling law that balances the interfacial energy of the system with the kinetic energy of the vortical flows allows us to understand the functional dependence of the Marangoni vortex strength on various experimental parameters.

Flutter-driven triboelectrification for harvesting wind energy.

Technologies to harvest electrical energy from wind have vast potentials because wind is one of the cleanest and most sustainable energy sources that nature provides. Here we propose a flutter-driven triboelectric generator that uses contact electrification caused by the self-sustained oscillation of flags. We study the coupled interaction between a fluttering flexible flag and a rigid plate. In doing so, we find three distinct contact modes: single, double and chaotic. The flutter-driven triboelectric generator having small dimensions of 7.5 × 5 cm at wind speed of 15 ms(-1) exhibits high-electrical performances: an instantaneous output voltage of 200 V and a current of 60 µA with a high frequency of 158 Hz, giving an average power density of approximately 0.86 mW. The flutter-driven triboelectric generation is a promising technology to drive electric devices in the outdoor environments in a sustainable manner.

Optimization of the elution buffer and concentration method for detecting hepatitis E virus in swine liver using a nested reverse transcription-polymerase chain reaction and real-time reverse transcription-polymerase chain reaction.

The aim of this study was to develop an optimal technique for detecting hepatitis E virus (HEV) in swine livers. Here, three elution buffers and two concentration methods were compared with respect to enhancing recovery of HEV from swine liver samples. Real-time reverse transcription-polymerase chain reaction (RT-PCR) and nested RT-PCR were performed to detect HEV RNA. When phosphate-buffered saline (PBS, pH 7.4) was used to concentrate HEV in swine liver samples using ultrafiltration, real-time RT-PCR detected HEV in 6 of the 26 samples. When threonine buffer was used to concentrate HEV using polyethylene glycol (PEG) precipitation and ultrafiltration, real-time RT-PCR detected HEV in 1 and 3 of the 26 samples, respectively. When glycine buffer was used to concentrate HEV using ultrafiltration and PEG precipitation, real-time RT-PCR detected HEV in 1 and 3 samples of the 26 samples, respectively. When nested RT-PCR was used to detect HEV, all samples tested negative regardless of the type of elution buffer or concentration method used. Therefore, the combination of real-time RT-PCR and ultrafiltration with PBS buffer was the most sensitive and reliable method for detecting HEV in swine livers.

Kinematic analysis of upper extremity movement during drinking in hemiplegic subjects.

BACKGROUND: It is necessary to analyze the kinematic properties of a paralyzed extremity to quantitatively determine the degree of impairment of hemiplegic people during functional activities of daily living (ADL) such as a drinking task. This study aimed to identify the kinematic differences between 16 hemiplegic and 32 able-bodied participants in relation to the task phases when drinking with a cup and the kinematic strategy used during motion with respect to the gravity direction. METHODS: The subjects performed a drinking task that was divided into five phases according to Murphy's phase definition: reaching, forward transport, drinking, backward transport, and returning. We found that the groups differed in terms of the movement times and the joint angles and angular velocities of the shoulder, elbow, and wrist joints. FINDINGS: Compared to the control group, the hemiplegic participants had a larger shoulder abduction angle of at most 17.1° during all the phases, a larger shoulder flexion angle of 7.6° during the reaching phase, and a smaller shoulder flexion angle of 6.4° during the backward transporting phase. Because of these shoulder joint patterns, a smaller elbow pronation peak angle of at most 13.1° and a larger wrist extension peak angle of 12.0° were found in the motions of the hemiplegic participants, as compensation to complete the drinking task. The movement in the gravity direction during the backward transporting phase resulted in a 15.9% larger peak angular velocity for elbow extension in the hemiplegic participants compared to that of the control group. INTERPRETATION: These quantitative kinematic patterns help provide an understanding of the movements of an affected extremity and can be useful in designing rehabilitation robots to assist hemiplegic people with ADL.

Norovirus contamination levels in ground water treatment systems used for food-catering facilities in South Korea.

This study aimed to inspect norovirus contamination of groundwater treatment systems used in food-catering facilities located in South Korea. A nationwide study was performed in 2010. Water samples were collected and, for the analysis of water quality, the temperature, pH, turbidity, and residual chlorine content were assessed. To detect norovirus genotypes GI and GII, RT-PCR and semi-nested PCR were performed with specific NV-GI and NV-GII primer sets, respectively. The PCR products amplified from the detected strains were then subjected to sequence analyses. Of 1,090 samples collected in 2010, seven (0.64%) were found to be norovirus-positive. Specifically, one norovirus strain was identified to have the GI-6 genotype, and six GII strains had the GII, GII-3, GII-4, and GII-17 genotypes. The very low detection rate of norovirus most likely reflects the preventative measures used. However, this virus can spread rapidly from person to person in crowded, enclosed places such as the schools investigated in this study. To promote better public health and sanitary conditions, it is necessary to periodically monitor noroviruses that frequently cause epidemic food poisoning in South Korea.

Sleep/wake estimation using only anterior tibialis electromyography data.

BACKGROUND: In sleep efficiency monitoring system, actigraphy is the simplest and most commonly used device. However, low specificity to wakefulness of actigraphy was revealed in previous studies. In this study, we assumed that sleep/wake estimation using actigraphy and electromyography (EMG) signals would show different patterns. Furthermore, each EMG pattern in two states (sleep, wake during sleep) was analysed. Finally, we proposed two types of method for the estimation of sleep/wake patterns using only EMG signals from anterior tibialis muscles and the results were compared with PSG data. METHODS: Seven healthy subjects and five patients (2 obstructive sleep apnea, 3 periodic limb movement disorder) participated in this study. Night time polysomnography (PSG) recordings were conducted, and electrooculogram, EMG, electroencephalogram, electrocardiogram, and respiration data were collected. Time domain analysis and frequency domain analysis were applied to estimate the sleep/wake patterns. Each method was based on changes in amplitude or spectrum (total power) of anterior tibialis electromyography signals during the transition from the sleep state to the wake state. To obtain the results, leave-one-out-cross-validation technique was adopted. RESULTS: Total sleep time of the each group was about 8 hours. For healthy subjects, the mean epoch-by-epoch results between time domain analysis and PSG data were 99%, 71%, 80% and 0.64 (sensitivity, specificity, accuracy and kappa value), respectively. For frequency domain analysis, the corresponding values were 99%, 73%, 81% and 0.67, respectively. Absolute and relative differences between sleep efficiency index from PSG and our methods were 0.8 and 0.8% (for frequency domain analysis). In patients with sleep-related disorder, our proposed methods revealed the substantial agreement (kappa > 0.61) for OSA patients and moderate or fair agreement for PLMD patients. CONCLUSIONS: The results of our proposed methods were comparable to those of PSG. The time and frequency domain analyses showed the similar sleep/wake estimation performance.

Effect of wash treatments on reducing human norovirus on iceberg lettuce and perilla leaf.

Human noroviruses (NoVs) are major causes of nonbacterial gastroenteritis; they are transmitted by food and water, as well as person-to-person. The consumption of contaminated raw or uncooked food such as vegetables and fruits has been identified as a common source of human NoV outbreaks. In an effort to understand the survival and persistence of human NoVs on fresh produce, the efficacy of washing treatments in the removal of human NoVs from vegetables was evaluated. This study used artificially contaminated vegetables (iceberg lettuce and perilla leaf), and washing was done with tap water for convenience. Wash treatments included immersion in water, rinsing with running water, and a combination of immersion and rinsing (treatments I to III, respectively). The effect of a class I detergent, a commercial product used for washing fruits and vegetables, was also evaluated (treatment IV). After the wash treatments, the remnants of human NoVs on samples were measured via real-time reverse transcriptase PCR. The results varied among treatments and by vegetable. For iceberg lettuce, a reduction of 0.9 log was noted in the treatment III group. The wash treatment was more effective in the perilla leaf samples: each treatment significantly reduced the numbers of human NoVs (0.69- to 1.29-log reduction). These data demonstrated that wash treatments reduced numbers of virus from the surfaces of the vegetables. Therefore, washing would seem to be a basic step in reducing numbers of virus in food preparation and in viral transmission routes.