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Ribosome collisions are recognized by E3 ubiquitin ligase Hel2/ZNF598, leading to RQC (ribosome-associated quality control) and to endonucleolytic cleavage and degradation of the mRNA termed NGD (no-go decay). NGD in yeast requires the Cue2 endonuclease and occurs in two modes, either coupled to RQC (NGDRQC+) or RQC uncoupled (NGDRQC−). This is mediated by an unknown mechanism of substrate recognition by Cue2. Here, we show that the ubiquitin binding activity of Cue2 is required for NGDRQC− but not for NGDRQC+, and that it involves the first two N-terminal Cue domains. In contrast, Trp122 of Cue2 is crucial for NGDRQC+. Moreover, Mbf1 is required for quality controls by preventing +1 ribosome frameshifting induced by a rare codon staller. We propose that in Cue2-dependent cleavage upstream of the collided ribosomes (NGDRQC−), polyubiquitination of eS7 is recognized by two N-terminal Cue domains of Cue2. In contrast, for the cleavage within collided ribosomes (NGDRQC+), the UBA domain, Trp122 and the interaction between Mbf1 and uS3 are critical.

Intense, quasi-monochromatic, polarized $\gamma$-ray beams produced by Compton scattering of laser photons against relativistic electrons are used for fundamental studies and applications. Following a series of photoneutron cross section measurements in the Giant Dipole Resonance energy region performed at the NewSUBARU synchrotron radiation facility, we have developed the eliLaBr Monte Carlo simulation code for characterization of the scattered $\gamma$-ray photon beams. The code is implemented using Geant4 and is available on the GitHub repository (https://github.com/dan-mihai-filipescu/eliLaBr). Here we report the validation of the eliLaBr code on NewSUBARU LCS $\gamma$-ray beam flux and spectral distribution data and two applications performed with it for asymmetric transverse emittance profiles electron beams, characteristic for synchrotrons. The first application is based on a systematic investigation of transverse collimator offsets relative to the laser and electron beam axis. We show that the maximum energy of the LCS $\gamma$-ray beam is altered by vertical collimator offsets, where the edge shifts towards lower energies with the increase in the offset. Secondly, using the eliLaBr code, we investigate the effect of the laser polarization plane orientation on the properties of the LCS $\gamma$-ray beams produced with asymmetric emittance electron beams. We show that: 1. The use of vertically polarized lasers contributes to the preservation of the LCS $\gamma$-ray beam maximum energy edge by increasing the precision in the vertical collimator alignment. 2. Under identical conditions for the electron and laser beams phase-space distributions, the energy spectrum of the scattered LCS $\gamma$-ray beam changes with the laser beam polarization plane orientation: the use of vertically polarized laser beams slightly deteriorates the LCS $\gamma$-ray beam energy resolution.

The mouse bioassay (MBA) for paralytic shellfish toxins (PSTs) in bivalves has been used as an official method in Japan. It is necessary to develop an alternative method to animal experiments in PSTs assay because 3Rs (Replacement, Reduction, and Refinement) of animal experiments are required from the animal welfare point of view. Various methods such as HPLC-FL, receptor binding assay, LC-MS/MS and ELISA have been established to detect PSTs without performing animal experiments. The present study was undertaken to develop a screening method using oligonucleotide lateral flow immunoassay (OLFIA) for detecting PSTs in bivalves. The screening level was defined as positive at 2 MU/g of MBA that is the half regulation limit of PSTs monitoring in Japan. All 20 positive (equal to or more than 2 MU/g) samples judged from MBA showed a positive reaction in the OLFIA. No positive samples resulted in a false negative reaction. The OLFIA exhibited high accuracy at 2 MU/g of screening criteria. The authors demonstrated here that the OLFIA can be useful for rapid detection of PSTs in bivalves.

An undulator generating a magnetic field whose longitudinal profile is arbitrarily varied has been developed, which is one of the key components in a number of proposed new concepts in free-electron lasers. The undulator is composed of magnet modules, each of which corresponds to a single undulator period, and is driven by a linear actuator to change the magnetic gap independently. To relax the requirement on the actuator, the mechanical load on each module due to magnetic force acting from opponent and adjacent modules is reduced by means of two kinds of spring systems. The performance of the constructed undulator has been successfully demonstrated by magnetic measurement and characterization of synchrotron radiation.

eIF2α phosphorylation-mediated translational regulation is crucial for global repression of translation by various stresses, including the unfolded protein response (UPR) in eukaryotes. Although translational control during UPR has not been extensively investigated in S. cerevisiae, Hac1-mediated production of long transcripts containing uORFs was shown to repress the translation of histidine triad nucleotide-binding 1 (HNT1) mRNA. The present study showed that uORF3 is required for HNT1 expression, as well as down-regulating HNT1 translation. Translation initiation by uORF3 is inefficient, with uORF3 having a strong Kozak sequence efficiently repressing the translation of HNT1. We propose that leaky scanning of uORF3 is responsible for the downregulation of HNT1 during UPR.

Ribosome stalling triggers the ribosome-associated quality control (RQC) pathway, which targets collided ribosomes and leads to subunit dissociation, followed by proteasomal degradation of the nascent peptide. In yeast, RQC is triggered by Hel2-dependent ubiquitination of uS10, followed by subunit dissociation mediated by the RQC-trigger (RQT) complex. In mammals, ZNF598-dependent ubiquitination of collided ribosomes is required for RQC, and activating signal cointegrator 3 (ASCC3), a component of the ASCC complex, facilitates RQC. However, the roles of other components and associated factors of the ASCC complex remain unknown. Here, we demonstrate that the human RQC-trigger (hRQT) complex, an ortholog of the yeast RQT complex, plays crucial roles in RQC. The hRQT complex is composed of ASCC3, ASCC2, and TRIP4, which are orthologs of the RNA helicase Slh1(Rqt2), ubiquitin-binding protein Cue3(Rqt3), and zinc-finger type protein yKR023W(Rqt4), respectively. The ATPase activity of ASCC3 and the ubiquitin-binding activity of ASCC2 are crucial for triggering RQC. Given the proposed function of the RQT complex in yeast, we propose that the hRQT complex recognizes the ubiquitinated stalled ribosome and induces subunit dissociation to facilitate RQC.

Read-through or mutations of a stop codon resulting in translation of the 3'-UTR produce potentially toxic C-terminally extended proteins. However, quality control mechanisms for such proteins are poorly understood in mammalian cells. Here, a comprehensive analysis of the 3'-UTRs of genes associated with hereditary diseases identified novel arrest-inducing sequences in the 3'-UTRs of 23 genes that can repress the levels of their protein products. In silico analysis revealed that the hydrophobicity of the polypeptides encoded in the 3'-UTRs is correlated with arrest efficiency. These results provide new insight into quality control mechanisms mediated by 3'-UTRs to prevent the production of C-terminally extended cytotoxic proteins.

Objective Preoperative diagnosis and successful management of acute torsion of a subserosal fibroid by using appropriate imaging modalities and single-port laparoscopic surgery. Case report A 44-year-old nulliparous woman presented with lower abdominal pain. Computed tomography and magnetic resonance imaging with contrast enhancement revealed a tumor in the pouch of Douglas with a low contrast at the center and thin-rim enhancement. Torsion of a uterine subserosal fibroid was diagnosed preoperatively. Laparoscopic single-port surgery by pneumoperitoneum was performed. Torsion of the pedicle attached to the uterine wall was excised by bipolar coagulation and cut with scissors. The extirpated fibroid was extracted from the umbilical wound. The pneumoperitoneum single-port laparoscopic surgery was completed as a gynecologic emergency operation. Conclusion Torsional uterine fibroids are difficult to diagnose preoperatively as symptoms are nonspecific and need emergent surgical management as an acute abdomen. Preoperative diagnosis using appropriate imaging modalities is important to perform single-port laparoscopic surgery.

Methionine adenosyltransferase (MAT) catalyzes the synthesis of S-adenosylmethionine (SAM). As the sole methyl-donor for methylation of DNA, RNA, and proteins, SAM levels affect gene expression by changing methylation patterns. Expression of MAT2A, the catalytic subunit of isozyme MAT2, is positively correlated with proliferation of cancer cells; however, how MAT2A promotes cell proliferation is largely unknown. Given that the protein synthesis is induced in proliferating cells and that RNA and protein components of translation machinery are methylated, we tested here whether MAT2 and SAM are coupled with protein synthesis. By measuring ongoing protein translation via puromycin labeling, we revealed that MAT2A depletion or chemical inhibition reduced protein synthesis in HeLa and Hepa1 cells. Furthermore, overexpression of MAT2A enhanced protein synthesis, indicating that SAM is limiting under normal culture conditions. In addition, MAT2 inhibition did not accompany reduction in mechanistic target of rapamycin complex 1 activity but nevertheless reduced polysome formation. Polysome-bound RNA sequencing revealed that MAT2 inhibition decreased translation efficiency of some fraction of mRNAs. MAT2A was also found to interact with the proteins involved in rRNA processing and ribosome biogenesis; depletion or inhibition of MAT2 reduced 18S rRNA processing. Finally, quantitative mass spectrometry revealed that some translation factors were dynamically methylated in response to the activity of MAT2A. These observations suggest that cells possess an mTOR-independent regulatory mechanism that tunes translation in response to the levels of SAM. Such a system may acclimate cells for survival when SAM synthesis is reduced, whereas it may support proliferation when SAM is sufficient.

Comprehensive genome-wide analysis has revealed the presence of translational elements in the 3′ untranslated regions (UTRs) of human transcripts. However, the mechanisms by which translation is initiated in 3′ UTRs and the physiological function of their products remain unclear. This study showed that eIF4G drives the translation of various downstream open reading frames (dORFs) in 3′ UTRs. The 3′ UTR of GCH1, which encodes GTP cyclohydrolase 1, contains an internal ribosome entry site (IRES) that initiates the translation of dORFs. An in vitro reconstituted translation system showed that the IRES in the 3′ UTR of GCH1 required eIF4G and conventional translation initiation factors, except eIF4E, for AUG-initiated translation of dORFs. The 3′ UTR of GCH1-mediated translation was resistant to the mTOR inhibitor Torin 1, which inhibits cap-dependent initiation by increasing eIF4E-unbound eIF4G. eIF4G was also required for the activity of various elements, including polyU and poliovirus type 2, a short element thought to recruit ribosomes by base-pairing with 18S rRNA. These findings indicate that eIF4G mediates translation initiation of various ORFs in mammalian cells, suggesting that the 3′ UTRs of mRNAs may encode various products.

Summary: Ribosome movement is not always smooth and is rather often impeded. For ribosome pauses, fundamental issues remain to be addressed, including where ribosomes pause on mRNAs, what kind of RNA/amino acid sequence causes this pause, and the physiological significance of this attenuation of protein synthesis. Here, we survey the positions of ribosome collisions caused by ribosome pauses in humans and zebrafish using modified ribosome profiling. Collided ribosomes, i.e., disomes, emerge at various sites: Pro-Pro/Gly/Asp motifs; Arg-X-Lys motifs; stop codons; and 3′ untranslated regions. The electrostatic interaction between the charged nascent chain and the ribosome exit tunnel determines the eIF5A-mediated disome rescue at the Pro-Pro sites. In particular, XBP1u, a precursor of endoplasmic reticulum (ER)-stress-responsive transcription factor, shows striking queues of collided ribosomes and thus acts as a degradation substrate by ribosome-associated quality control. Our results provide insight into the causes and consequences of ribosome pause by dissecting collided ribosomes.

Aims Fractional flow reserve (FFR), assessed using distal coronary pressure/aortic pressure (Pd)/(Pa) ratio, functionally evaluates coronary stenosis. An assessment method without vasodilators would be helpful. A single intracoronary bolus of saline decreases Pd because of the speculated low-viscosity effect. We hypothesised that saline-induced Pd/Pa ratio (SPR) could functionally evaluate coronary stenosis. This study aimed to test the accuracy and utility of SPR for predicting FFR ≤0.80. Methods and results In 137 coronary lesions with over 50% angiographic diameter stenosis, SPR was assessed using an intracoronary bolus of saline (2 mL/s) for five heartbeats (SPR-5) and three heartbeats (SPR-3). FFR was obtained after intravenous adenosine infusion (140 µg/kg/min). There was a strong correlation between FFR and SPR-5 or SPR-3 (R=0.941 and R=0.933, respectively). Receiver operating characteristic (ROC) curve analysis demonstrated good accuracy (86.3%) for SPR-5, with a cut-off of ≤0.84 for predicting FFR ≤0.80 (area under ROC curve 0.96, specificity 94.3, sensitivity 79.9). Thirty-three lesions (24%) were located in the "grey zone" (SPR 0.83-0.88). No complications were observed in 673 SPR measurements. Conclusions SPR may accurately predict FFR and can limit adenosine use to one in four lesions. Further studies are needed to confirm the validity of SPR.

The high-angular-resolution electron backscatter diffraction (HR-EBSD) technique has been utilized to evaluate the elastic strain distribution of grinding-induced damage layers in silicon carbide wafers. HR-EBSD analysis, along with transmission electron microscopy observation, revealed that the damage layers formed beneath the wafer surface when the surface was ground by diamond abrasives; the layers were classified hierarchically based on the distribution of elastic strain and lattice defects. In particular, very large elastic strain formed in a defective region of roughly 0.6 μm in thickness, just beneath the ground wafer surface, where lattice defects such as dislocations, stacking faults, and microcracks were introduced inhomogeneously by abrasive interaction and related plastic deformation and fracture. Based on this inhomogeneity, the defective region was itself classified into two types: one, a highly defective region with very large, complicated strain and high defect density; and the other, a basal plane dislocation (BPD)-glide region, with small strain and few BPDs or stacking faults. Beneath the defective region, a strain gradient region of roughly 1.8 μm in thickness, which was unambiguously identified by HR-EBSD strain analysis alone, revealed a monotonic gradient in the dominant compressive strain component, with no grinding-induced defects. Overall, HR-EBSD analysis revealed a nanometer-scale, hierarchical elastic strain distribution in the grinding-induced damage layers.

We analyse the performance of a gas time projection chamber (TPC) as a high-performance gamma-ray telescope and polarimeter in the e$^+$e$^-$ pair creation regime. We use data collected at a gamma-ray beam of known polarisation. The TPC provides two orthogonal projections $(x,z)$ and $(y,z)$ of the tracks induced by each conversion in the gas volume. We use a simple vertex finder in which vertices and pseudo-tracks exiting from them are identified. We study the various contributions to the single-photon angular resolution using Monte Carlo simulations and compare them with the experimental data and find that they are in excellent agreement. The distribution of the azimutal angle of pair conversions shows a bias due to the non-cylindrical-symmetric structure of the detector. This bias would average out for a long duration exposure on a space mission, but for this pencil-beam characterisation we have ensured its accurate simulation by a double systematics control scheme, data taking with the detector rotated at several angles with respect to the beam polarisation direction and systematics control with a non-polarised beam. We measure, for the first time, the polarisation asymmetry of a linearly polarised gamma-ray beam in the low energy pair creation regime. This sub-GeV energy range is critical for cosmic sources as their spectra are power laws which fall quickly as a function of increasing energy. This work could pave the way to extending polarised gamma-ray astronomy beyond the MeV energy regime., Comment: Submitted to Astroparticle Physics

We conducted a retrospective chart-review study, examining predictors of the repetition of short-term self-harm (

Evaluation of surface damage layers formed by mechanical grinding processes is indispensable in epi-ready SiC wafer preparation. As well as microstructure, the analysis of local strain distribution in the damage layers gives a clue on control of the wafer quality. Advanced electron backscatter diffraction (EBSD) technique is applied to evaluate the strain distribution of the damage layers. It is revealed that the elastic strain distribution can be classified into a hierarchy of three regions with respect to depth from the surface. Combining EBSD analysis with TEM observation, large compressive elastic strain and misorientation are introduced in the highly-defective region underneath the ground wafer surface. In addition, the gradient distribution of the strain is observed clearly below the highly-defective region. The knowledge of correlating between strain distribution and microstructure is promising to control the damage layer for the wafer preparation.

Current {\gamma}-ray telescopes based on photon conversions to electron-positron pairs, such as Fermi, use tungsten converters. They suffer of limited angular resolution at low energies, and their sensitivity drops below 1 GeV. The low multiple scattering in a gaseous detector gives access to higher angular resolution in the MeV-GeV range, and to the linear polarisation of the photons through the azimuthal angle of the electron-positron pair. HARPO is an R&D program to characterise the operation of a TPC (Time Projection Chamber) as a high angular-resolution and sensitivity telescope and polarimeter for {\gamma} rays from cosmic sources. It represents a first step towards a future space instrument. A 30 cm cubic TPC demonstrator was built, and filled with 2 bar argon-based gas. It was put in a polarised {\gamma}-ray beam at the NewSUBARU accelerator in Japan in November 2014. Data were taken at different photon energies from 1.7 MeV to 74 MeV, and with different polarisation configurations. The electronics setup is described, with an emphasis on the trigger system. The event reconstruction algorithm is quickly described, and preliminary measurements of the polarisation of 11 MeVphotons are shown., Comment: Proceedings VCI2016

In protein synthesis, ribosome movement is not always smooth and is rather often impeded for numerous reasons. Although the deceleration of the ribosome defines the fates of the mRNAs and synthesizing proteins, fundamental issues remain to be addressed, including where ribosomes pause in mRNAs, what kind of RNA/amino acid sequence causes this pause, and the physiological significance of this slowdown of protein synthesis. Here, we surveyed the positions of ribosome collisions caused by ribosome pausing in humans and zebrafish on a genome-wide level using modified ribosome profiling. The collided ribosomes, i.e., disomes, emerged at various sites: the proline-proline-lysine motif, stop codons, and the 3′ untranslated region (UTR). The number of ribosomes involved in a collision is not limited to two, but rather four to five ribosomes can form a queue of ribosomes. In particular, XBP1, a key modulator of the unfolded protein response, shows striking queues of collided ribosomes and thus acts as a substrate for ribosome-associated quality control (RQC) to avoid the accumulation of undesired proteins in the absence of stress. Our results provide an insight into the causes and consequences of ribosome slowdown by dissecting the specific architecture of ribosomes.

This paper proposes a methodology of model predictive control for alleviating shallow drowsiness of office workers and thus improving their productivity. The methodology is based on dynamically scheduling setting values for air conditioning and lighting to minimize drowsiness level of office workers on the basis of a prediction model that represents the relation between future drowsiness level and combination of indoor temperature and ambient illuminance. The prediction model can be identified by utilizing state-of-the-art drowsiness estimation method. The proposed methodology was evaluated in regard to a real routine task (performed by six subjects over five workdays), and the evaluation results demonstrate that the proposed methodology improved the processing speed of the task by 8.3% without degrading comfort of the workers., International Conference of the IEEE Engineering in Medicine and Biology Society (EMBC) 2019 - accepted

AIM Suicide attempters have a high risk of repeated suicide attempts and completed suicide. There is evidence that assertive case management can reduce the incidence of recurrent suicidal behavior among suicide attempters. This study evaluated the effect of an assertive-case-management training program. METHODS This multicenter, before-and-after study was conducted at 10 centers in Japan. Participants were 274 medical personnel. We used Japanese versions of the Attitudes to Suicide Prevention Scale, the Gatekeeper Self-Efficacy Scale, the Suicide Intervention Response Inventory (SIRI), and the Attitudes Toward Suicide Questionnaire. We evaluated the effects with one-sample t-tests, and examined prognosis factors with multivariable analysis. RESULTS There were significant improvements between pre-training and post-training in the Attitudes to Suicide Prevention Scale (mean: -3.07, 95% confidence interval [CI]: -3.57 to -2.57, P

The authors have researched and developed Building Energy Management Systems (BEMS) to promote energy-savings in small and medium-sized office buildings. In order to accelerate the installation of BEMS into these buildings, it is important to develop BEMS which meets the following two requirements;1) To achieve the system configuration through the ease of installation.2) To have functions of inducing energy-saving behaviors of people in the office.For the purpose of developing practical BEMS, it is essential to examine the above from a viewpoint of the human side. In this paper, a concept of the system developed by the authors is briefly introduced, and followed by the discussion on users' energy-saving behaviors induced by the information providing method. First, the outline and features of the system are introduced. Next, the research on the information providing method for inducing energy-saving behaviors is explained. Finally, based on these researches, we propose a design policy for the information providing method to induce energy-saving behaviors. It is to provide information that increases the motivation before actions are taken and to reduce the occurrences of dissatisfaction after taking such actions. Currently, the effects of the system are under validation regarding whether it maintains and induces energy-saving behaviors.