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Spinal cord injury (SCI) is a devastating mechanical trauma. Although locomotion of model animals that mimic contusion SCI was actively examined, locomotion after penetrating SCI caused by sharp objects was not extensively studied. Severity of walking difficulty after partial transection of the spinal cord including penetrating SCI likely depends on the regions affected. Therefore, we compared beam walking and overground walking between mice after penetrating SCI at inner spinal cord region and mice with the injury at the outer region. Mice with the both penetrating SCIs did not display changes in beam walking. When appearance and movements of hindlimbs during overground walking was rated using Basso Mouse Scale for locomotion (BMS), however, mice with inner penetrating SCI showed low score shortly after the SCI. However, the score became high at later time points, as seen in contusion SCI mice. By contrast, BMS score did not decrease shortly after the outer penetrating SCI. However, the score became low 3 weeks after the SCI. As quantitative values during overground walking, movement duration in an open field were shorter at 1 day after the two penetrating SCIs. However, slower moving speed and fewer number of movement at 1 day were specific to mice with inner and outer penetrating SCIs, respectively. Moreover, BMS score was correlated with walking distance in open field only in mice with inner penetrating SCI. Thus, inner and outer penetrating SCI cause difficulty in overground walking with different severity and progress.

Polyglutamine (polyQ) diseases are devastating neurological disorders that cannot be effectively treated. Repeat-associated non-AUG (RAN) translation has been documented in transcripts in polyQ diseases. RAN products include proteins with polyleucine (polyL) tracts. Similar to polyQ, polyL tends to aggregate, which is toxic to cells and mice. Irradiation with a free electron laser (FEL) tuned at mid-infrared wavelengths can dissociate polyQ aggregates in cultured cells. However, whether FEL dissociates the polyL is unclear. It is also unclear whether brain dysfunction caused by polyL aggregates in mice can be ameliorated by FEL irradiated polyL. Here, we show that FEL at approximately 6 μm can destroy polyL aggregates, as evidenced by scanning electron microscopy, atomic force microscopy, and dot blot analyses. Although polyL aggregates induced low viability and aberrant morphology of cultured astrocytes, FEL irradiated polyL exhibited mild defects. Likewise, the toxicity of polyL-containing microglia in vitro was ameliorated by FEL irradiation. In vivo, mice administered polyL aggregates in the cerebellum induced loss of Purkinje cells, which was ameliorated when FEL irradiated polyL was injected. These results justify the clearing of aggregates by approaches using molecular chaperones, laser irradiation, and ultrasound as a general therapeutic strategy to correct brain dysfunction by the RAN products.

Perilesional T1 hyperintensity on magnetic resonance imaging (MRI) of intra-axial brain masses is an unusual feature of the perilesional area, characteristic of cavernous malformations (CMs) and metastatic brain tumors (METs). Here, we report a case of primary diffuse glioma with a perilesional T1 hyperintense area (HIA) on MRI. A 61-year-old woman with transient aphasia visited our hospital. Radiological examination revealed an intra-axial mass with acute/subacute hemorrhaging and calcification in the left frontal lobe. It was presumed to be a CM because of the perilesional T1 HIA. Gross total resection of the tumor was performed, and the pathological diagnosis was anaplastic oligodendroglioma, not otherwise specified by World Health Organization 2016 classification. Histopathological findings in the perilesional T1 HIA indicated hemorrhage involvement in the surrounding white matter. No recurrence appeared after radio-chemotherapy. Perilesional T1 HIAs, characteristic of CMs and METs, are also seen in primary diffuse gliomas. Therefore, caution should be taken when using this sign for the differential diagnosis of intracranial masses.

Whether fibril formation increases or decreases cytotoxicity remains unclear. Aggregation of human islet amyloid polypeptide (hIAPP), a pivotal regulator of glucose homeostasis, impairs the function and viability of pancreatic β cells. Evidence suggests that low-order oligomers of hIAPP are more toxic to β cells than fibril. However, it remains unclear whether non-fibril form of hIAPP specifically alters brain functions. This study produced fibril and non-fibril forms from a single hIAPP 8-20 peptide. The non-fibril form-injected mice showed changes in spontaneous motor activities, preference for location in the open field and social behavior. In contrast, the fibril-injected mice showed no changes in these behavioral tests. In line with the behavioral changes, the non-fibril form led to impaired neurite outgrowth of cultured neuron-like cells and the loss of neurons in the mouse hippocampus. These findings suggest that non-fibril form but not fibril form of hIAPP changes brain functions.

Abstract Repeat-associated non-AUG translation (RAN translation) is observed in transcripts that are causative for polyglutamine (polyQ) diseases and generates proteins with mono amino acid tracts such as polyalanine (polyA), polyleucine (polyL) and polyserine (polyS) in neurons, astrocytes and microglia. We have previously shown that microglia with aggregated polyQ led to defective differentiation and degeneration of neuron-like cells. However, it has not been determined whether only microglia containing a specific RAN product, but not other RAN products, is harmful in vitro and in vivo. Here we show that polyL-incorporating microglia specifically led to altered startle response in mice. Aggregated polyA, polyS and polyL induced aberrant differentiation of microglia-like BV2 cells. Differentiated PC12 cells treated with conditioned medium (CM) of polyS- and polyL- but not polyA-incorporating microglia-like BV2 cells showed retraction of neurites and loss of branch of neurites. Injection of the polyL-CM, but not polyA-CM and polyS-CM, into the lateral ventricle lowered startle response in mice. Consistently, polyL induced the highest expression of CD68 in BV2 cells. The lowered startle response was replicated in mice given the polyL-CM in the caudal pontine reticular nucleus (PnC), the key region of startle response. Thus, endogenous RAN proteins having polyL derived from polyQ diseases-causative genes in microglia might specifically impair startle response.

Amyloidosis is known to be caused by the deposition of amyloid fibrils into various biological tissues; effective treatments for the disease are little established today. An infrared free-electron laser (IR-FEL) is an accelerator-based picosecond-pulse laser having tunable infrared wavelengths. In the current study, the irradiation effect of an IR-FEL was tested on an 11-residue peptide (NFLNCYVSGFH) fibril from β2-microglobulin (β2M) with the aim of applying IR-FELs to amyloidosis therapy. Infrared microspectroscopy (IRM) and scanning electron microscopy showed that a fibril of β2M peptide was clearly dissociated by IR-FEL at 6.1 µm (amide I) accompanied by a decrease of the β-sheet and an increase of the α-helix. No dissociative process was recognized at 6.5 µm (amide II) as well as at 5.0 µm (non-specific wavelength). Equilibrium molecular dynamics simulations indicated that the α-helix can exist stably and the probability of forming interchain hydrogen bonds associated with the internal asparagine residue (N4) is notably reduced compared with other amino acids after the β-sheet is dissociated by amide I specific irradiation. This result implies that N4 plays a key role for recombination of hydrogen bonds in the dissociation of the β2M fibril. In addition, the β-sheet was disrupted at temperatures higher than 340 K while the α-helix did not appear even though the fibril was heated up to 363 K as revealed by IRM. The current study gives solid evidence for the laser-mediated conversion from β-sheet to α-helix in amyloid fibrils at the molecular level.

Functional brain architecture underlying autonomic adjustments to exercise has remained unknown. Here, the authors reveal a subcortical pathway that relays volitional motor signals to drive locomotor activities and sympathetic cardiovascular responses.

Abstract Repeat-associated non-AUG (RAN) translation of mRNAs/transcripts responsible for polyglutamine (polyQ) diseases may generate peptides containing different mono amino acid tracts such as polyserine (polyS) and polyleucine (polyL). The propagation of aggregated polyQ from one cell to another is also an intriguing feature of polyQ proteins. However, whether the RAN translation-related polyS and polyL have the ability to propagate remains unclear, and if they do, whether the exogenous polyS and polyL exert toxicity on the recipient cells is also not known yet. In the present study, we found that aggregated polyS and polyL peptides spontaneously enter neuron-like cells and astrocytes in vitro. Aggregated polyS led to the degeneration of the differentiated neuron-like cultured cells. Likewise, the two types of aggregates taken up by astrocytes induced aberrant differentiation and cell death in vitro. Furthermore, injection of each of the two types of aggregates into the ventricles of adult mice resulted in their behavioral changes. The polyS-injected mice showed extensive vacuolar degeneration in the brain. Thus, the RAN translation-related proteins containing polyS and polyL have the potential to propagate and the proteins generated by all polyQ diseases might exert universal toxicity in the recipient cells.

Polyalanine (polyA) disease-causative proteins with an expansion of alanine repeats can be aggregated. Although curative treatments for polyA diseases have not been explored, the dissociation of polyA aggregates likely reduces the cytotoxicity of polyA. Mid-infrared free electron laser (FEL) successfully dissociated multiple aggregates. However, whether the FEL dissociates polyA aggregates like other aggregates has not been tested. Here, we show that FEL at 6.1 μm experimentally weakened the extent of aggregation of a peptide with 13 alanine repeats (13A), and the irradiated 13A exerted lesser cytotoxicity to neuron-like cells than non-irradiated 13A. Then, we applied molecular dynamics (MD) simulation to follow the dissociation process by FEL. We successfully observed how the intermolecular β-sheet of polyA aggregates was dissociated and separated into monomers with helix structures upon FEL irradiation. After the dissociation by FEL, water molecules inhibited the reformation of polyA aggregates. We recently verified the same dissociation process using FEL-treated amyloid-β aggregates. Thus, a common mechanism underlies the dissociation of different protein aggregates that cause different diseases, polyA disease and Alzheimer's disease. However, MD simulation indicated that polyA aggregates are less easily dissociated than amyloid-β aggregates and require longer laser irradiation due to hydrophobic alanine repeats.

Abstract In polyalanine (PA) diseases, the disease-causing transcription factors contain an expansion of alanine repeats. While aggregated proteins that are responsible for the pathogenesis of neurodegenerative disorders show cell-to-cell propagation and thereby exert toxic effects on the recipient cells, whether this is also the case with expanded PA has not been studied. It is also not known whether the internalized PA is toxic to recipient cells based on the degree of aggregation. In this study, we therefore prepared different degrees of aggregation of a peptide having 13 alanine repeats without flanking sequences of PA disease-causative proteins (13A). The aggregated 13A was spontaneously taken up by neuron-like cultured cells. Functionally, strong aggregates but not weak aggregates displayed a deficit in neuron-like differentiation in vitro. Moreover, the injection of strong but not weak 13A aggregates into the ventricle of mice during the neonatal stage led to enhanced spontaneous motor activity later in life. Thus, PA in the extracellular space has the potential to enter adjacent cells, and may exert toxicity depending on the degree of aggregation.

Adaptive thermogenesis is regulated by central neuronal circuits. Here, the authors show that microRNA-33 in the brain contributes to the maintenance of brown adipose tissue thermogenesis and whole-body energy balance via enhanced sympathetic nerve tone, and regulating the expression of GABAa receptor subunits.

Precise evaluation of motor functions using simple and reproducible tests for mouse models of spinal cord injury (SCI) are required. Overground walking of SCI mice has been tested by Basso Mouse Scale for locomotion (BMS). In contrast, only a few works quantify walking performances of SCI mice on narrow beams, a different task. Here, we established a novel scoring system using a single beam walking apparatus for SCI mice. The scoring system uses binary judgments of values such as retention, moving forward and reaching the goal on a beam for rating. In addition, high score was given to SCI mouse when the mouse efficiently used hindlimbs for locomotion on the beam. A high rate of concordance of the score derived from positions of hindlimbs between two observers was obtained. Mice displayed the lowest total score on the beam immediately after the SCI, then the score gradually increased like time course of BMS score. Furthermore, the total scores reflected gradation of severity of SCI in 2 strains of mice. The beam walking score proved to be strongly correlated with that of BMS score, indicating that performances between overground walking and beam walking are partly correlated in SCI mice. Collectively, the novel scoring system offers an opportunity to easily evaluate motor performances of mice with SCI.

The Japanese National Action Plan on Antimicrobial Resistance (AMR) was adopted to strengthen AMR surveillance and monitoring in companion animals. The Japanese Veterinary Antimicrobial Resistance Monitoring (JVARM) system monitors the sale of veterinary antimicrobial drugs by pharmaceutical companies, and the sale of human drugs by principal wholesale companies to companion animal (dogs and cats) clinics. However, the data do not include sales by local drug suppliers and personal importation to companion animal clinics in Japan. The purposes of this study were to estimate total antimicrobial drug use by companion animal clinics in Japan and to identify the factors associated with their use. In 2018, questionnaires gathering data on attributes of the clinic and volumes of antimicrobial drugs used were sent to 212 clinics across Japan by the Japan Veterinary Medical Association. Out of the clinics, 170 valid questionnaires were returned (80.2% response rate). Antimicrobial drugs were categorized first as human, veterinary, or imported drugs and then further categorized as important drugs (critically important drugs for humans and second-choice veterinary drugs) or others. Total antimicrobial drug use was estimated based on the number of clinics reported in 2016. The relationships between antimicrobial drug use and various questionnaire items were analyzed using non-parametric regression analysis. Total antimicrobial drug use was estimated at 29.9t, which was 2.1 times higher than reported by the JVARM survey on the sales of antimicrobial drugs. In terms of total use, important drugs and human drugs accounted for 12.6 and 61.8%, respectively. Clinic income per veterinarian was associated with total antimicrobial use per veterinarian. The proportion of important drugs among all antimicrobial drugs used in a clinic was high in recently established clinics with middle-aged and older directors.

Importing necessary metabolites into the mitochondrial matrix is a crucial step of fuel choice during stress adaptation. Branched chain-amino acids (BCAAs) are essential amino acids needed for anabolic processes, but they are also imported into the mitochondria for catabolic reactions. What controls the distinct subcellular BCAA utilization during stress adaptation is insufficiently understood. The present study reports the role of SLC25A44, a recently identified mitochondrial BCAA carrier (MBC), in the regulation of mitochondrial BCAA catabolism and adaptive response to fever in rodents. We found that mitochondrial BCAA oxidation in brown adipose tissue (BAT) is significantly enhanced during fever in response to the pyrogenic mediator prostaglandin E2 (PGE2) and psychological stress in mice and rats. Genetic deletion of MBC in a BAT-specific manner blunts mitochondrial BCAA oxidation and non-shivering thermogenesis following intracerebroventricular PGE2 administration. At a cellular level, MBC is required for mitochondrial BCAA deamination as well as the synthesis of mitochondrial amino acids and TCA intermediates. Together, these results illuminate the role of MBC as a determinant of metabolic flexibility to mitochondrial BCAA catabolism and optimal febrile responses. This study also offers an opportunity to control fever by rewiring the subcellular BCAA fate.

Relationships between wheat (Triticum æstivum L.) yield components and growing season weather conditions (e.g. temperature, precipitation, and sunshine duration) were investigated for a rice-wheat rotation paddy system situated in Northern Kyushu, Japan over a 17-year period (2000–2016). A 1.0 mm increase in precipitation from 21 to 32 days after sowing decreased wheat yield by about 27 kg ha−1 at early seedling stage with one to two leaves (Zadoks (Z) growth stage Z1.1–1.2). Number of tillers and spikes showed significant negative correlations to the amount of precipitation during the period. An hour increase in mean daily sunshine hours from 94 to 111 days after sowing resulted in an 328 kg ha−1 rise in grain yield at stem elongation stage with the first and second node detection (Z3.1–3.2). The grain number per square meter also showed strong positive correlations to sunshine hours during the same period. However, this positive effect of sunshine was negated when precipitation exceeded about 30 mm during the early seedling stage. Consequently, precipitations at early seedling stage and sunshine hours at stem elongation stage mostly determined wheat yield in Northern Kyushu paddy field. Increasing precipitations during the early seedling stage highlights the pressing need for effective paddy field drainage management from wheat sowing through harvest. Abbreviations: CP: Chikushi Plains; FARC: Fukuoka prefectural Agriculture and Forestry Research Center; SARC: Saga prefectural Agricultural Research Center; DAS: days after sowing

Wheat (Triticum aestivum L.) grain yield is predicted to decrease in the future because of an increase in air temperature globally. To clarify the effects of the vernalization response gene in wheat to warmer winters, we compared dry matter production and grain yield between spring wheat ‘Asakazekomugi’ and its winter-type near-isogenic line (NIL) carrying different alleles of the vernalization response gene Vrn-D1 under early-, standard-, and late-sowing conditions. Under early-sowing conditions, dry matter production of the NIL carrying the winter allele of Vrn-D1, named Asa (Vrn-D1b), exceeded that of ‘Asakazekomugi’ from mid-March (after stem elongation in Asa (Vrn-D1b)) when the temperatures rose. Tiller number and leaf area index under early-sowing conditions were consistently higher in Asa (Vrn-D1b) than in ‘Asakazekomugi’ from mid-March onward. It was suggested that the early-sown ‘Asakazekomugi’ could not effectively absorb solar radiation to produce dry matter because of the acceleration of stem elongation caused by the Vrn-D1 gene during the cold season. The grain yield of Asa (Vrn-D1b) with early sowing was higher than with standard sowing. In contrast, the grain yield of ‘Asakazekomugi’ was lower in the early-sown crop than in the crop sown at the standard date. These results suggested that the higher grain yield of Asa (Vrn-D1b) than that of ‘Asakazekomugi’ under early-sown conditions could be due to Asa (Vrn-D1b) maintaining high dry matter production after the jointing stage by suppressing acceleration of growth caused by warm conditions after sowing. Abbreviations CGR: crop growth rate; HI: harvest index; LAI: leaf area index; NIL: near-isogenic line; SNP: single-nucleotide polymorphism

Intracranial epidermoids are rare lesions accounting for 0.2%-1.8% of all intracranial tumors. They commonly develop in the cerebellopontine angle and the parasellar region and can appear with atypical neuroimaging features due to intracystic hemorrhages which complicate diagnosis. The authors present a case of a 62-year-old woman with a frontal epidermoid cyst with a hemorrhage and tumor growth. A series of atypical radiological findings showed gradual changes in the lesion appearance that were confirmed with surgery and histopathology. To avoid surgical complications such as chemical meningitis, it is important to remember that epidermoid cysts occasionally bleed, leading to atypical MRI and/or CT findings and diagnostic difficulties. Development of epidermoid cysts in atypical locations in the brain may result in challenges to accurate diagnosis. Keywords: Epidermoid cyst, Intracystic hemorrhage, Atypical radiological findings

Expanded polyglutamine-containing proteins in neurons intrinsically contributes to neuronal dysfunctions and neuronal cell death in polyglutamine (polyQ) diseases. In addition, an expanded polyQ-containing protein in microglia also leads to apoptosis of neurons. However, detailed morphological analysis of neurons exposed to conditioned medium (CM) derived from polyQ-containing microglia has not been essentially carried out. Here, we introduced aggregated peptide with 69 glutamine repeat (69Q) into BV2 microglial cells. The 69Q-containing BV2 cells showed shorter branches. The CM from 69Q-containing microglia (69Q-CM) induced neurite retraction and fewer number of branch point of neurites of differentiated PC12 cells. Likewise, the 69Q-CM induces disturbed differentiation of PC12 cells with shorter total length of neurites and fewer number of branch point of neurites. Thus, the factor(s) released from polyQ-containing microglia affect both differentiation and degeneration of neuron-like cells.

Supernormal antithrombin (AT) activity is rare in patients with sepsis. This study compared mortality rate of patients with sepsis and supernormal AT activity with that of other patients. This retrospective study included patients with sepsis from 42 intensive care units (ICUs) in Japan. Patients were included if their AT activity was measured on ICU admission, and if they did not receive AT concentrate. They were categorized into low, normal, and supernormal with respective AT activity of ≤70%, >70% to ≤100%, and >100%. The primary outcome was hospital in-patient mortality. Nonlinear regression analysis showed that mortality risk gradually increased with AT activity in the supernormal range, but without statistical significance. Survival rate was significantly lower in low (67%) and supernormal (57%) AT groups than in the normal AT group (79%; P < .001 and P = .008, respectively). After adjusting for disease severity and AT activity on day 2, supernormal AT activity was the only independent predictor of mortality. Sepsis with supernormal AT activity associated with high mortality, independent of disease severity, might be a predictor of in-hospital mortality.

Milling properties and flour color are essential selection criteria in soft wheat breeding. However, high phenotypic screening costs restrict selection to relatively few breeding lines in late generations. To achieve marker-based selection of these traits in early generations, we performed genetic dissection of quality traits using three doubled haploid populations that shared the high-quality soft wheat variety Kitahonami as the paternal parent. An amplicon sequencing approach allowed effective construction of well-saturated linkage maps of the populations. Marker-based heritability estimates revealed that target quality traits had relatively high values, indicating the possibility of selection in early generations. Taking advantage of Chinese Spring reference sequences, joint linkage maps of the three populations were generated. Based on the maps, multifamily quantitative trait locus (QTL) analysis revealed a total of 86 QTLs for ten traits investigated. In terms of target quality traits, 12 QTLs were detected for flour yield, and 12 were detected for flour redness (a* value). Among these QTLs, six for flour yield and nine for flour a* were segregating in more than two populations. Some relationships among traits were explained by QTL collocations on chromosomes, especially group 7 chromosomes. Ten different ideotypes with various combinations of favorable alleles for the flour yield and flour a* QTLs were generated. Phenotypes of derivatives from these ideotypes were predicted to design ideal genotypes for high-quality wheat. Simulations revealed the possibility of breeding varieties with better quality than Kitahonami.

SCR systems are mainly used in exhaust systems as an emission control device for diesel engines with excellent thermal efficiency. In this paper we explored the design of mixing devices which are an important element in the system. The optimum shape for various conditions and designs to disperse ammonia from a urea solution was optimized with CAE and design space exploration, and validated with visualization experiments for correlation.

Abstract Thermoregulatory behaviour, such as migration to a comfortable thermal environment, is a representative innate animal behaviour and facilitates effective autonomic regulation of body temperature with a reduced cost of resources. Here we determine the central thermosensory ascending pathway that transmits information on environmental temperature from cutaneous thermoreceptors to elicit thermoregulatory behaviour. To examine the contribution of the spinothalamocortical pathway, which is known to mediate thermosensory transmission for perception of skin temperature, we lesioned thalamic regions mediating this pathway in rats. Thalamic-lesioned rats showed compromised electroencephalographic responses in the primary somatosensory cortex to changes in skin temperature, indicating functional ablation of the spinothalamocortical pathway. However, these lesioned rats subjected to a two-floor innocuous thermal plate preference test displayed intact heat- and cold-avoidance thermoregulatory behaviours. We then examined the involvement of the lateral parabrachial nucleus (LPB), which mediates cutaneous thermosensory signaling to the thermoregulatory center for autonomic thermoregulation. Inactivation of neurons in the LPB eliminated both heat- and cold-avoidance thermoregulatory behaviours and ablated heat defense. These results demonstrate that the LPB, but not the thalamus, mediates the cutaneous thermosensory neural signaling required for behavioural thermoregulation, contributing to understanding of the central circuit that generates thermal comfort and discomfort underlying thermoregulatory behaviours.

The extent of spontaneous recovery in patients with a spinal cord injury (SCI) has not been thoroughly investigated. It is essentially not known whether SCI animals exhibit full recovery from both motor and sensory deficits as well. Here, we developed an appropriate condition to produce a mild SCI in mice. Mice given a mild contusion SCI showed transient low performances in the Basso Mouse Scale for locomotion (BMS), rotarod and beam walking tests after the SCI, which was followed by complete restoration in a short time. The SCI mice also showed functional full recovery from low sensitivity to light touch using dynamic touch test. Nevertheless, the fully-recovered SCI mice still exhibited significant loss of myelin in the spinal cord. These results suggest a high potential of adaptation of motor and sensory systems in mice and might provide insight into the prognoses of SCI patients.

Torrential and long-lasting rainfall often causes long-duration floods in flat and lowland areas in data-scarce Nyaungdon Area of Myanmar, imposing large threats to local people and their livelihoods. As historical hydrological observations and surveys on the impact of floods are very limited, flood hazard assessment and mapping are still lacked in this region, making it hard to design and implement effective flood protection measures. This study mainly focuses on evaluating the predicative capability of a 2D coupled hydrology-inundation model, namely the Rainfall-Runoff-Inundation (RRI) model, using ground observations and satellite remote sensing, and applying the RRI model to produce a flood hazard map for hazard assessment in Nyaungdon Area. Topography, land cover, and precipitation are used to drive the RRI model to simulate the spatial extent of flooding. Satellite images from Moderate Resolution Imaging Spectroradiometer (MODIS) and the Phased Array type L-band Synthetic Aperture Radar-2 onboard Advanced Land Observing Satellite-2 (ALOS-2 ALOS-2/PALSAR-2) are used to validate the modeled potential inundation areas. Model validation through comparisons with the streamflow observations and satellite inundation images shows that the RRI model can realistically capture the flow processes (R2 ≥ 0.87; NSE ≥ 0.60) and associated inundated areas (success index ≥ 0.66) of the historical extreme events. The resultant flood hazard map clearly highlights the areas with high levels of risks and provides a valuable tool for the design and implementation of future flood control and mitigation measures.

Development of wheat cultivars that achieve high yields despite the short growing season is essential for increasing wheat production in southwestern Japan. The objectives of this study were to assess the genetic progress in grain yield and to clarify yield-attributing traits of high-yielding wheat lines in southwestern Japan. We conducted field experiments for two growing seasons (2012–2013 and 2013–2014) using three commercial wheat cultivars (Shiroganekomugi, Chikugoizumi, and Iwainodaichi) and four high-yielding wheat lines including Hakei W1380 developed in southwestern Japan. In an ancillary field experiment, we compared a commercial cultivar, Shiroganekomugi, and a high-yielding line, Hakei W1380, in the 2014–2015 season. Across the two seasons, grain yield of high-yielding lines was generally higher than commercial cultivars. Hakei W1380 achieved the highest grain yield across the two seasons, and successfully produced more than 900 g m−2 in the 2013–2014 season. Correlation analysis showed that recent yield progress of wheat lines in southwestern Japan was derived from enhanced biomass production and grain number m−2. Larger numbers of grains m−2 in high-yielding lines than in commercial cultivars were associated with higher crop growth rate at the pre-anthesis stage, and therefore higher spike dry weight m−2 at anthesis. Genotypic differences in crop growth rate from jointing to anthesis resulted mainly from differences in leaf area index. These results indicate that further improvements in grain yield in southwestern Japan could be achieved by increasing the amount of radiation intercepted at the pre-anthesis stage and grain number m−2.

The present experimental study deals with a pulsatile turbulent flow simulating the exhaust flow of an automotive engine. In the experiments, a four-cylinder engine is used as an exhaust-flow generator to realize flow conditions close to those in an engine environment. Particle image velocimetry (PIV) measurements visualize the flow field in an S-shaped double-bend duct at a Reynolds number of 48,000 and a Womersley number of 70.9. Stereo PIV, which is classified as a two-dimensional three-component measurement, is conducted in the duct cross sections located downstream of the bends. The stereo PIV system is synchronized with the engine operation to enable phase-locked measurements at particular phases, and the phase-averaged results show the large-scale vortical structures and the duct axial velocity distribution. Downstream of the first bend, the secondary flow consists of vortices that circulate as Dean-type vortices. Downstream of the second bend, by contrast, vortices that circulate in opposite directions to the Dean-type vortices, so-called Lyne-type vortices, form in the core of the cross section. These secondary flows persist without significant changes in their large-scale vortical structures over time. Time-resolved PIV is conducted to track the temporal evolution of the flow in the bend planes. The results show that the flow reverses locally along the inner wall of the bends during flow deceleration. Snapshot proper orthogonal decomposition (POD) is used on the time-resolved PIV data to extract the significant flow structure from the instantaneous field. We propose POD as a good post-processing tool for the instantaneous data of pulsatile cases.

Objective. In most cases, about the voice of the patient with female-to-male/gender identity disorder (FTM/GID), hormone therapy makes the voice low-pitched. In success cases, there is no need for phonosurgery. However, hormone therapy is not effective in some cases. We perform type 3 thyroplasty in these cases. Method. Hormone therapy was started in 2008 but did not lower the speaking fundamental frequencies (SFFs). We therefore performed TP3 under local anesthesia. Results. In our case, the SFF at the first visit was 146 Hz. The postoperative SFF was 110 Hz. Conclusions. TP3 was performed under local anesthesia in a patient with FTM/GID in whom hormone therapy proved ineffective. With successful conversion to a lower-pitched voice, the patient could begin to live daily life as a male. QOL improved significantly with TP3. If hormone therapy proves ineffective, TP3 may be selected as an optional treatment and appears to show few surgical complications and was, in this case, a very effective treatment.

Therapeutics that target ERBB2, such as lapatinib, often provide initial clinical benefit, but resistance frequently develops. Adaptive responses leading to lapatinib resistance involve reprogramming of the kinome through reactivation of ERBB2/ERBB3 signaling and transcriptional upregulation and activation of multiple tyrosine kinases. The heterogeneity of induced kinases prevents their targeting by a single kinase inhibitor, underscoring the challenge of predicting effective kinase inhibitor combination therapies. We hypothesized that, to make the tumor response to single kinase inhibitors durable, the adaptive kinome response itself must be inhibited. Genetic and chemical inhibition of BET bromodomain chromatin readers suppresses transcription of many lapatinib-induced kinases involved in resistance, including ERBB3, IGF1R, DDR1, MET, and FGFRs, preventing downstream SRC/FAK signaling and AKT reactivation. Combining inhibitors of kinases and chromatin readers prevents kinome adaptation by blocking transcription, generating a durable response to lapatinib, and overcoming the dilemma of heterogeneity in the adaptive response.

A pulsatile turbulent flow within an S-shaped double bend pipe is experimentally and numerically studied to characterize the flow field in conditions resembling an automotive engine environment. Particle image velocimetry (PIV) measurements were carried out to measure streamwise and secondary flow velocities. The flows are accelerated around the inner side walls of both bends. The secondary flow, after passing through the second bend, is directed toward the inner side in the core of the cross section, and, as a result, Lyne-type vortices, which are not consistent with the second bend curvature, are formed. A numerical simulation is performed under the same condition as the experiments with computational fluid dynamics software. The numerical simulation gives qualitative results in comparison with the experimental data though there is some deviation, and shows the cause of the Lyne-type vortex formation in the second bend. After passing through the first bend, the high-speed region appearing around the inner side shifts in accordance with the Dean-type secondary flow formed in the first bend, and thus the non-uniform flow enters the second bend. In the second bend, the low-velocity region in which the centrifugal force is not strong enough to direct the flow toward the outer side, appears in the core of the cross section. Details of the Lyne-type vortex formation are discussed by considering the driving forces of the secondary flow.

The recurrent network composed of excitatory and inhibitory neurons is fundamental to neocortical function. Inhibitory neurons in the mammalian neocortex are molecularly diverse, and individual cell types play unique functional roles in the neocortical microcircuit. Recently, vasoactive intestinal polypeptide-positive (VIP+) neurons, comprising a subclass of inhibitory neurons, have attracted particular attention because they can disinhibit pyramidal cells through inhibition of other types of inhibitory neurons, such as parvalbumin- (PV+) and somatostatin-positive (SOM+) inhibitory neurons, promoting sensory information processing. Although VIP+ neurons have been reported to receive synaptic inputs from PV+ and SOM+ inhibitory neurons as well as from cortical and thalamic excitatory neurons, the somatodendritic localization of these synaptic inputs has yet to be elucidated at subcellular spatial resolution. In the present study, we visualized the somatodendritic membranes of layer (L) 2/3 VIP+ neurons by injecting a newly developed adeno-associated virus vector into the barrel cortex of VIP-Cre knock-in mice, and we determined the extensive ramification of VIP+ neuron dendrites in the vertical orientation. After immunohistochemical labeling of presynaptic boutons and postsynaptic structures, confocal laser scanning microscopy revealed that the synaptic contacts were unevenly distributed throughout the perisomatic (< 100 µm from the somata) and distal-dendritic compartments (≥ 100 µm) of VIP+ neurons. Both corticocortical and thalamocortical excitatory neurons preferentially targeted the distal-dendritic compartment of VIP+ neurons. On the other hand, SOM+ and PV+ inhibitory neurons preferentially targeted the distal-dendritic and perisomatic compartments of VIP+ neurons, respectively. Notably, VIP+ neurons had few reciprocal connections. These observations suggest different inhibitory effects of SOM+ and PV+ neuronal inputs on VIP+ neuron activity; inhibitory inputs from SOM+ neurons likely modulate excitatory inputs locally in dendrites, while PV+ neurons could efficiently interfere with action potential generation through innervation of the perisomatic domain of VIP+ neurons. The present study, which shows a precise configuration of site-specific inputs, provides a structural basis for the integration mechanism of synaptic inputs to VIP+ neurons.

Selective sodium glucose cotransporter-2 inhibitor (SGLT2i) treatment promotes urinary glucose excretion, thereby reducing blood glucose as well as body weight. However, only limited body weight reductions are achieved with SGLT2i treatment. Hyperphagia is reportedly one of the causes of this limited weight loss. However, the effects of SGLT2i treatment on systemic energy expenditure have not been fully elucidated. Herein, we investigated the acute effects of dapagliflozin, a SGLT2i, on systemic energy expenditure in mice. Eighteen hours after dapagliflozin treatment oxygen consumption and brown adipose tissue (BAT) expression of ucp1, a thermogenesis-related gene, were significantly decreased as compared to those after vehicle treatment. In addition, dapagliflozin significantly suppressed norepinephrine (NE) turnover in BAT and c-fos expression in the rostral raphe pallidus nucleus (rRPa) which contains the sympathetic premotor neurons responsible for thermogenesis. These findings indicate that the dapagliflozin-mediated acute decrease in energy expenditure involves a reduction in BAT thermogenesis via decreased sympathetic nerve activity from the rRPa. Furthermore, common hepatic branch vagotomy abolished the reductions in ucp1 expression and NE contents in BAT and c-fos expression in the rRPa. In addition, alterations in hepatic carbohydrate metabolism, such as decreases in glycogen contents and upregulation of phosphoenolpyruvate carboxykinase, manifested prior to the suppression of BAT thermogenesis, e.g. 6 hours after dapagliflozin treatment. Collectively, these results suggest that SGLT2i treatment acutely suppresses energy expenditure in BAT via regulation of an inter-organ neural network consisting of the common hepatic vagal branch and sympathetic nerves.

Pedigree analysis was conducted for early maturing wheat cultivars developed in Japan. Materials used for this analysis were mainly developed at Nagano Agricultural Experiment Station (Tozan lines). In a recently released Tozan line, the maximum number of generations traced in the pedigree, total number of ancestors in the pedigree and total number of ancestors except common ones was 11, 222 and 94, respectively. Chunaga contributed 24.0% of the genetic background of Tozan lines. Seven ancestors, collectively, contributed 51.5% to the gene pool. Hiyokukomugi had the highest mean coefficient of parentage to Tozan lines and the mean value was 0.216, followed by Kinuiroha (0.213), Norin 61 (0.206), Mikunikomugi (0.205) and Tokai 80 (0.194). The mean coefficient of parentage between Tozan lines and cultivars in the Kanto-Tokai region, Kinki-Chugoku-Shikoku region and Kyushu region was 0.165, 0.155 and 0.157, respectively. Tozan lines more related to cultivars in the Tohoku region tended to be late heading and more cold-tolerant. Tozan lines more related to Ayahikari or Kinuazuma tended to be early maturing. Fukuhokomugi, which was a high yield cultivar and often used as a cross parent, did not contribute to high flour protein. KS831957 showed a positive effect on the crude protein content of flour. In general, Tozan lines had no specific cultivars used extensively as a cross parent having significant influence on yield or flour quality.

Predicting the response to immunosuppressive therapy could provide useful information to help the clinician define treatment strategies for patients with aplastic anemia. In our current study, we evaluated the relationship between telomere length of lymphocytes at diagnosis and the response to immunosuppressive therapy in 64 children with aplastic anemia, using flow fluorescence in situ hybridization. Median age of patients was ten years (range 1.5–16.2 years). Severity of the disease was classified as very severe in 23, severe in 21, and moderate in 20 patients. All patients were enrolled in multicenter studies using antithymocyte globulin and cyclosporine. The response rate to immunosuppressive therapy at six months was 52% (33 of 64). The probability of 5-year failure-free survival and overall survival were 56% (95% confidence interval (CI): 41–69%) and 97% (95%CI: 87–99%), respectively. Median telomere length in responders was −0.4 standard deviation (SD) (−2.7 to +3.0 SD) and −1.5 SD (−4.0 to +1.6 (SD)) in non-responders (P

Parotid gland carcinoma is extremely rare in children. We report a case of pediatric parotid gland carcinoma with extensive infiltration into surrounding tissues including the skin and temporomandibular joint capsule at initial examination. Total resection of the parotid gland was conducted together with skull base surgery and mandibular dissection. The patient was a 14-year-old girl. In addition to the skin and temporomandibular joint, infiltration into the anterior wall of the external auditory meatus and masseter muscle was also seen, and T4N0M0 stage IV parotid carcinoma was diagnosed. Skin was resected together with the pinna, and temporal craniotomy and skull base surgery were performed to resect the temporomandibular joint capsule and external auditory meatus en bloc, and mandible dissection was conducted. Facial nerves were resected at the same time. Level I to level IV neck dissection was also conducted. A latissimus dorsi myocutaneous flap was used for reconstruction. The postoperative permanent pathology diagnosis was high-grade mucoepidermoid carcinoma with a low-grade component. Postoperatively, radiotherapy at 50 Gy alone has been conducted, with no recurrence or metastasis observed for over 4 years.

The winter wheat variety Kitahonami shows a superior flour yield in comparison to other Japanese soft wheat varieties. To map the quantitative trait loci (QTL) associated with this trait, association mapping was performed using a panel of lines from Kitahonami's pedigree, along with leading Japanese varieties and advanced breeding lines. Using a mixed linear model corrected for kernel types and familial relatedness, 62 marker-trait associations for flour yield were identified and classified into 21 QTLs. In eighteen of these, Kitahonami alleles showed positive effects. Pedigree analysis demonstrated that a continuous pyramiding of QTLs had occurred throughout the breeding history of Kitahonami. Linkage analyses using three sets of doubled haploid populations from crosses in which Kitahonami was used as a parent were performed, leading to the validation of five of the eight QTLs tested. Among these, QTLs on chromosomes 3B and 7A showed highly significant and consistent effects across the three populations. This study shows that pedigree-based association mapping using breeding materials can be a useful method for QTL identification at the early stages of breeding programs.

Heterozygosity for dominant-negative STAT1 mutations underlies autosomal dominant Mendelian susceptibility to mycobacterial diseases. Mutations conferring Mendelian susceptibility to mycobacterial diseases have been identified in the regions of the STAT1 gene encoding the tail segment, DNA-binding domain and SH2 domain. We describe here a new heterozygous mutation, Y701C, in a Japanese two-generation multiplex kindred with autosomal dominant Mendelian susceptibility to mycobacterial diseases. This mutation affects precisely the canonical STAT1 tyrosine phosphorylation site. The Y701C STAT1 protein is produced normally, but its phosphorylation is abolished, resulting in a loss-of-function for STAT1-dependent cellular responses to interferon-γ or interferon-α. In the patients’ cells, the allele is dominant-negative for γ-activated factor-mediated responses to interferon-γ, but not for interferon-stimulated gene factor-3-mediated responses to interferon-α/β, accounting for the clinical phenotype of Mendelian susceptibility to mycobacterial diseases without severe viral diseases. Interestingly, both patients displayed multifocal osteomyelitis, which is often seen in patients with Mendelian susceptibility to mycobacterial diseases with autosomal dominant partial IFN-γR1 deficiency. Multifocal osteomyelitis should thus prompt investigations of both STAT1 and IFN-γR1. This experiment of nature also confirms the essential role of tyrosine 701 in human STAT1 activity in natura.

Primary submandibular gland oncocytic carcinoma is a rare pathology, with only 10 cases being reported to date. We encountered a case of primary submandibular gland oncocytic carcinoma and report it herein. The patient was a 69-year-old man who came to our hospital with right submandibular cancer as the main complaint. Based on the results of computed tomography and magnetic resonance imaging, submandibular gland tumor was diagnosed. Preoperative cytodiagnosis suggested class III oncocytic carcinoma. Resection of the right submandibular tumor was performed along with right neck dissection. Postoperative histopathological diagnosis was oncocytic carcinoma. As of 3 years following surgery, no recurrence has been identified.

Malignant melanoma of the sphenoid sinus is a very rare disease, and only 6 cases have previously been reported. The present case involved a 74-year-old woman who was examined for visual disturbance of the left eye. Computed tomography revealed a soft tissue shadow, but only mucosal hypertrophy was found on opening the sphenoid sinus under general anesthesia. One month postoperatively, visual disturbance of the right eye and paresis of cranial nerve III appeared. Malignant melanoma was diagnosed from biopsy. Multiple bone metastases were identified, but the patient declined active treatment. As a result, palliative care was provided and she died 3 months later. When there is no improvement in postoperative visual acuity as in this case, in consideration of the possibility of neoplastic lesions, rigorous followup including monitoring for neurological symptoms is warranted.

Background. Small cell carcinoma (SmCC) in the nasal cavity and paranasal sinuses is very rare, and definitive therapies have not yet been established. Methods. Chemoradiotherapy comprised 60 Gy of external radiation, with the administration of irinotecan intravenously at 60 mg/m2 on days 1, 8, and 15 and cisplatin at 60 mg/m2 on day 1. Results. Case 1 involved a 66-year-old woman with stage III cancer. Adverse events included decreased white blood cells, anemia, and oral mucositis, all Grade 2. The patient remained free of cancer as of 3 years and 6 months after completing the treatment. Case 2 involved a 60-year-old man with stage IV cancer. He also experienced adverse events of decreased white blood cells, anemia, and oral mucositis, all Grade 2. He died after 11 months due to metastases throughout the body. Conclusions. The results suggest that this regimen may be tolerable as a therapy for this type of carcinoma.

Synthesis of IL-1β and TNFα by human monocytesmacrophages was significantly inhibited by eleven bisbenzylisoquinolines and one half-molecule (benzylisoquinoline), with IC50 values in the μM range. The results indicate that these compounds may have value in the therapy of human diseases where these inflammatory cytokines have a central role in pathogenesis.