尾崎 行男

It is known that incidence and short-term mortality rate of acute myocardial infarction (AMI) tend to be higher in the cold season. The aim of our study was to investigate the association of onset-season with patient characteristics and long-term prognosis of AMI. This was a prospective, multicenter, Japanese investigation of 3,283 patients with AMI who were hospitalized within 48 h of symptom onset between July 2012 and March 2014. Patients were divided into 3 seasonal groups according to admission date: cold season group (December-March), hot season group (June-September), and moderate season group (April, May, October, and November). We identified 1356 patients (41.3%) admitted during the cold season, 901 (27.4%) during the hot season, and 1026 (31.3%) during the moderate season. We investigated the seasonal effect on patient characteristics and clinical outcomes. Baseline characteristics of each seasonal group were comparable, with the exception of age, Killip class, and conduction disturbances. The rates of higher Killip class and complete atrioventricular block were significantly higher in the cold season group. The 3-year cumulative survival free from major adverse cardiac events (MACE) rate was the lowest in the cold season (67.1%), showing a significant difference, followed by the moderate (70.0%) and hot seasons (72.9%) (p < 0.01). Initial severity and long-term prognoses were worse in patients admitted during the cold season. Our findings highlight the importance of optimal prevention and follow-up of AMI patients with cold season onset.

This study evaluated whether radial access intervention had a lower risk of post-treatment adverse events in acute coronary syndrome (ACS) even in Japan where the use of a strong antithrombotic regimen was not approved. We retrospectively analyzed a large nation-wide registry in Japan to compare the incidence of post-treatment adverse events according to the types of vessel access (trans-radial; TRI vs. trans-femoral; TFI) among ACS cases (n = 76,835; 43,288 TRI group and 33,547 TFI group). Primary outcome was a composite of in-hospital death, myocardial infarction associated with percutaneous coronary intervention, bleeding complication requiring transfusion, and stent thrombosis during in-hospital stay. Propensity score matching (PS) and instrumental variable (IV) analyses were used to account for treatment selection. The incidence of post-treatment adverse events was lower in the TRI group by 0.95% compared to the TFI group with PS (p < 0.001) and by 0.34% with IV (p = 0.127). A significantly lower risk for access site bleeding was observed by 0.34% with PS (p < 0.001) and by 0.53% with IV (p < 0.001). Radial access was related to a significantly lower risk for access site bleeding compared with femoral access, even without strong antithrombotic drugs for ACS in Japan, and may also relate to lower risk for a wider set of post-treatment adverse events.

Soluble forms of platelet membrane proteins are released upon platelet activation. We previously reported that soluble C-type lectin-like receptor 2 (sCLEC-2) is released as a shed fragment (Shed CLEC-2) or as a whole molecule associated with platelet microparticles (MP-CLEC-2). In contrast, soluble glycoprotein VI (sGPVI) is released as a shed fragment (Shed GPVI), but not as a microparticle-associated form (MP-GPVI). However, mechanism of sCLEC-2 generation or plasma sCLEC-2 has not been fully elucidated. Experiments using metalloproteinase inhibitors/stimulators revealed that ADAM10/17 induce GPVI shedding, but not CLEC-2 shedding, and that shed CLEC-2 was partially generated by MMP-2. Although MP-GPVI was not generated, it was generated in the presence of the ADAM10 inhibitor. Moreover, antibodies against the cytoplasmic or extracellular domain of GPVI revealed the presence of the GPVI cytoplasmic domain, but not the extracellular domain, in the microparticles. These findings suggest that most of the GPVI on microparticles are induced to shed by ADAM10; MP-GPVI is thus undetected. Plasma sCLEC-2 level was 1/32 of plasma sGPVI level in normal subjects, but both soluble proteins significantly increased in plasma of patients with acute coronary syndrome. Thus, sCLEC-2 and sGPVI are released by different mechanisms and released in vivo upon platelet activation.

A 14-year-old boy collapsed suddenly after a basketball game and was transported to our hospital after recovering from ventricular fibrillation by an automated external defibrillator. He had experienced loss of consciousness twice and has been examined for suspected long-QT syndrome at another hospital. The 12-lead electrocardiogram on admission revealed a prolonged QTc interval of 480 milliseconds. After the patient recovered without any sequelae, computed tomography revealed an anomalous left coronary artery arising from the opposite sinus of Valsalva and coursing between the aorta and the pulmonary artery. Furthermore, genetic testing identified a KCNE1-D85N abnormality. An anomalous coronary artery is one of the major causes of sudden death in young people; therefore, surgical revascularization is recommended for left coronary arteries arising from the contralateral sinus and coursing between the aorta and the pulmonary artery, regardless of myocardial ischemia. Transient myocardial ischemia may have exaggerated the instability from the arrhythmic substrate, even though KCNE1-D85N abnormalities alone are not thought to cause fatal arrhythmias. Besides routine electrocardiography, further examinations, including imaging and genetic testing, can characterize the pathophysiology of fatal cardiac disease.

Blood choline has been proposed as a predictor of acute coronary syndrome (ACS), however different testing procedures might affect the choline concentration because the lysophospholipase D activity of autotaxin (ATX) can convert lysophosphatidylcholine to lysophosphatidic acid (LPA) and choline in human blood. Although the influences of ATX on LPA levels are well known in vivo and in vitro, those on choline have not been elucidated. Therefore, we established suitable sampling conditions and evaluated the usefulness of plasma choline concentrations as a biomarker for ACS. Serum LPA and choline concentrations dramatically increased after incubation depending on the presence of ATX, while their concentrations in plasma under several conditions were differently modulated. Plasma choline levels in genetically modified mice and healthy human subjects, however, were not influenced by the ATX level in vivo, while the plasma LPA concentrations were associated with ATX. With strict sample preparation, the plasma choline levels did not increase, but actually decreased in ACS patients. Our study revealed that ATX increased the choline concentrations after blood sampling but was not correlated with the choline concentrations in vivo therefore, strict sample preparation will be necessary to investigate the possible use of choline as a biomarker.

The platelet activation receptor C-type lectin-like receptor 2 (CLEC-2) interacts with podoplanin on the surface of certain types of tumor cells, and this interaction facilitates tumor metastasis. CLEC-2 is also involved in thrombus formation and its stabilization. Because CLEC-2-depleted mice are protected from experimental lung metastasis and thrombus formation and do not show increased bleeding time, CLEC-2 may serve as a good target for antimetastatic or antithrombotic drugs. We screened 6770 compounds for their capability to inhibit CLEC-2-podoplanin binding using an enzyme-linked immunosorbent assay. In the first screening round, 63 compounds were identified and further evaluated by flow cytometry using CLEC-2-expressing cells. We identified protoporphyrin IX (H2-PP) as the most potent inhibitor and modified its hematoporphyrin moiety to be complexed with cobalt (cobalt hematoporphyrin [Co-HP]), which resulted in an inhibitory potency much stronger than that of H2-PP. Surface plasmon resonance analysis and molecular docking study showed that Co-HP binds directly to CLEC-2 at N120, N210, and K211, previously unknown podoplanin-binding sites; this binding was confirmed by analysis of CLEC-2 mutants with alterations in N120 and/or K211. Co-HP at a concentration of 1.53 μM inhibited platelet aggregation mediated through CLEC-2, but not that mediated through other receptors. IV administration of Co-HP to mice significantly inhibited hematogenous metastasis of podoplanin-expressing B16F10 cells to the lung as well as in vivo arterial and venous thrombosis, without a significant increase in tail-bleeding time. Thus, Co-HP may be a promising molecule for antimetastatic and antiplatelet treatment that does not cause bleeding tendency.

C-type lectin-like receptor 2 (CLEC-2) has been identified on the surface of platelets as a receptor for a platelet activating snake venom, rhodocytin/aggretin. CLEC-2 belongs to a C-type lectin superfamily and binds to a sialoglycoprotein, podoplanin, in vivo. Platelets play a crucial role in hemostasis and thrombosis, but recent studies have uncovered multiple roles of platelets beyond hemostasis in physiology and pathology. The interaction between platelet CLEC-2 and podoplanin is the key to several roles of platelets beyond hemostasis. The spatial and temporal expression patterns of podoplanin regulate vascular/lymphatic development, maintenance of vascular integrity, tissue regeneration, and some pathological processes including tumor metastasis and thromboinflammation. CLEC-2 facilitates blood/lymphatic vessel separation during embryonic development by binding to podoplanin on lymphatic endothelial cells. The leakage of platelets from hyperpermeable vessels for maintaining vascular integrity during inflammation depends on CLEC-2. During wound healing, the expression of podoplanin in keratinocytes is upregulated, which helps in the process. Podoplanin is expressed on the surface of tumor cells and facilitates hematogenous metastasis by inducing platelet aggregation through CLEC-2. During thrombotic processes, such as development of deep vein thrombosis, podoplanin is upregulated on unknown cells in the vessel wall in the area of inflammation, facilitates thrombus formation, and promotes further inflammation by binding to CLEC-2. In this article, the roles of platelets beyond hemostasis are comprehensively reviewed.

We investigated the resistance to stem blight disease (Phomopsis asparagi (Sacc.)) in the progeny of two combinations of interspecific crosses between Asparagus officinalis (sensitive) and Asparagus A. kiusianus (resistant) in an effort to produce resistant cultivars. The progeny showed different degrees of disease severity, depending on the combination of crosses. Most of the hybrids derived from AO0060 (A. officinalis) × AK0501 (A. kiusianus) showed high disease resistance comparable to that of A. kiusianus. The results indicate that disease resistance could be introduced from A. kiusianus into A. officinalis, and that the selection of an appropriate cross combination is important for the production of disease-resistant cultivars. We analyzed the parents and hybrids of reciprocal crosses between A. officinalis and A. kiusianus using derived cleaved amplified polymorphic sequence markers to investigate the inheritance of the chloroplast genome, whose inheritance and genetic characteristics are not yet known. The chloroplast DNAs were inherited from the maternal parent, indicating that no major genes related to stem blight resistance were found in the chloroplast DNA.

Effects of photoperiod and temperature on rhizome enlargement (dormancy induction) and accompanied dormancy depth were investigated in this study. Nine-day-old seedlings were transplanted from 26 July at 1 week intervals, and they were grown under a natural photoperiod for 5 weeks in an unheated greenhouse in Fukuoka Prefecture, Japan. Although subterranean stems elongated in the plants grown until 30 August or 6 September, enlarged rhizomes were formed in those grown until 13 September or 20 September. It was revealed from these results that the lotus recognizes a natural photoperiod after 6 September as a short day. When 9 treatments of day length combinations (LD0+SD8–LD8+SD0) were applied to the seedlings, the plants grown under short day after long day treatment of 0 (LD0+SD8), 1 (LD1+SD7), 2 (LD2+SD6), 3 (LD3+SD5), 4 (LD4+SD4), 5 (LD5+SD3), 6 (LD6+SD2), or 7 (LD7+SD1) weeks formed enlarged rhizomes from the fifth, fifth, sixth, seventh, eighth, tenth, twelfth, and fourteenth internodes, respectively. Although photoperiodic treatment in the first week was different between LD0+SD8 and LD1+SD7 treatments, subterranean stems began to enlarge from the same internode (fifth internode) in both treatments. This indicates that photoperiod treatments for the first week do not affect morphology of subterranean stems. Seven treatments of day length combinations (LD2+SD0+LD6–LD2+SD6+LD0) were applied to seedlings after long day treatment for 2 weeks. Enlarged subterranean stems were observed in the plants grown under short day for 6 weeks (LD2+SD6+LD0), but not in those under long day for 6 weeks (LD2+SD0+LD6). On the other hand, subterranean stems elongated again after rhizome enlargement under a subsequent long day following 1 (LD2+SD1+LD5), 2 (LD2+SD2+LD4), 3 (LD2+SD3+LD3), or 4 (LD2+SD4+LD2) weeks of short day. This clarified that morphogenesis in subterranean stems is completely dependent on photoperiod. Further, it is expected that such growth resumption may be attributed to a weak dormant state in the enlarged rhizome. The enlarged rhizomes were exposed to natural low temperatures to examine environmental factors for deepening dormancy. Rhizomes sprouted in all treatments irrespective of exposure to low temperatures when they were transferred to ideal conditions. Rapid growth in leaves and subterranean stems was particularly observed by exposure to low temperature. It was suggested that low temperature is an environmental factor for releasing dormancy, but not for deepening dormancy. It is proposed from these results that subterranean stem growth is completely dependent on photoperiod, and that enlarged rhizomes show weak dormancy.

Fujitsu is working on developing an autonomous robot system in order to realize Monozukuri (Japanese way of manufacturing) that can adapt itself quickly and flexibly to changes in manufacturing products and in production requirements, such as variation of parts and aged deterioration of equipment. The deployment of robots on production lines requires programming by expert engineers, and the robots need to be taught the supply position and assembly position of parts beforehand. However, such a development process necessitates a significant amount of work. Meanwhile in mass production, certain changes in the production environment, such as variation of parts, often cause robots to stop. Therefore, we have developed technology that is capable of automatically generating programs that control robot motions. The technology also makes it easier to automatically convert the programs into applicable data formats, and transmit them to the robots. We are further working to develop technologies that allow robots to "sense," "think," and "act" so that we can achieve autonomous and cooperative control. By enabling flexible response to the above mentioned changes and fluctuations with these technologies, the changeovers and rearrangements of production can be minimized, thereby eliminating the barriers to robot introduction in production lines. This paper describes this autonomous robot system development, which can be applied to variable product and variable volume production for enhanced production efficiency.

Asparagus kiusianus, an important wild relative of cultivated asparagus (A. officinalis), exhibits resistance to stem blight disease caused by Phomopsis asparagi. However, the mechanisms underlying this resistance are not understood and no transcriptomic or genetic resources are available for this species. De novo transcriptome sequencing of A. officinalis and A. kiusianus stems was performed 24 h after inoculation with P. asparagi. In total, 35,259 and 36,321 transcripts were annotated in A. officinalis and A. kiusianus, respectively. 1,027 up-regulated and 752 down-regulated transcripts were differentially expressed in the two Asparagus species. RNA sequencing data were validated using quantitative real-time reverse transcription PCR. Several defense-related genes including peroxidase 4, cationic peroxidase SPC4-like, pathogenesis-related protein-1-like, and jasmonic acid biosynthesis and signaling-related genes including phospholipase D alpha 1, 12-oxophytodienoate reductase and jasmonate-induced protein 23 KD were up-regulated in A. kiusianus relative to A. officinalis. In addition, infected A. kiusianuns exhibited a substantial increase in jasmonic acid and methyl jasmonate relative to A. officinalis. Peroxidase activity was significantly elevated in infected A. kiusianus compared with infected A. officinalis. Our transcriptomic database provides a resource for identifying novel genes and molecular markers-associated with Phomopsis disease resistance and will facilitate breeding and improvement of cultivated asparagus varieties.

Harvesting of asparagus spears is hard work because farmers have to harvest spears in a crouching posture. We previously developed electric long-shafted shears and a large-wheeled cart to improve the harvest posture, and we reported that the visibility of spears can be improved by modified branch training using string, without any yield or quality reduction. In the present study, we investigated the harvest efficiency, harvest posture, and subjective burden with electric long-shafted shears and a large-wheeled cart under modified branch training methods (the new standing harvest method) for labor-saving asparagus spear harvesting. Two male subjects in their 30s and 60s harvested spears for 60 min using (standing harvest) and without using (control) the new standing harvest method. The harvest efficiency of the new standing harvest method was 18% lower than that of the control. In the standing harvest, subjects had to pull the shears up to take the harvested spear from the shears with their left hand after each spear was cut ("take"). The subjects needed time for the "take" action, and the time necessary to harvest a spear with the electric long-shafted shears was longer than that required for the control. We also evaluated the harvest posture using the Ovako Working Posture Analyzing System (OWAS). For the subject in his 30s, action category(AC)2 and AC3 accounted for 72% and 14% of the postures recorded in the control, respectively, whereas in the standing harvest, AC1 and AC2 accounted for 57% and 41%, respectively. For the subject in his 60s, AC2 and AC3 accounted for 41% and 58% of the postures recorded in the control, respectively, whereas in the standing harvest, AC1 and AC2 accounted for 27% and 70%, respectively. Unfavorable postures during harvest were reduced by the new standing harvest method. The subjective physical burden on the subjects was evaluated using a modified Borg scale after 60 min of harvesting using and without using the new standing harvest method. The subjective physical burden in both subjects was highest (&gt; 8) at the waist in the control, whereas the scores were lower than 3 in the standing harvest. Thus, the new standing harvest method reduced harvest efficiency, but resulted in substantial improvements in posture and reduced the physical burden on the subjects.

Dioecy is a plant mating system in which individuals of a species are either male or female. Although many flowering plants evolved independently from hermaphroditism to dioecy, the molecular mechanism underlying this transition remains largely unknown. Sex determination in the dioecious plant Asparagus officinalis is controlled by X and Y chromosomes; the male and female karyotypes are XY and XX, respectively. Transcriptome analysis of A.officinalis buds showed that a MYB-like gene, Male Specific Expression 1 (MSE1), is specifically expressed in males. MSE1 exhibits tight linkage with the Y chromosome, specific expression in early anther development and loss of function on the X chromosome. Knockout of the MSE1 orthologue in Arabidopsis induces male sterility. Thus, MSE1 acts in sex determination in A.officinalis.

Platelets play a key role in the pathophysiological processes of hemostasis and thrombus formation. However, platelet functions beyond thrombosis and hemostasis have been increasingly identified in recent years. A large body of evidence now exists which suggests that platelets also play a key role in inflammation, immunity, malignancy, and furthermore in organ development and regeneration, such as the liver. We have recently identified CLEC-2 on the platelet membrane, which induces intracellular activation signals upon interaction of a snake venom, rhodocytin. Later we discovered that podoplanin, present in renal podocytes and lymphatic endothelial cells, both of which are not accessible to platelets in blood stream, is an endogenous ligand for CLEC-2. In accord with our expectation, platelet-specific CLEC-2 knockout mice have a phenotype of edema, lymphatic vessel dilatation, and the presence of blood cells in lymphatic vessels. It is suggested that lymphatic/blood vessel separation during the developmental stage is governed by cytokines released from platelets activated by the interaction between platelet CLEC-2 and podoplanin present on lymphatic endothelial cells. Recombinant CLEC-2 bound to early atherosclerotic lesions and normal arterial walls, co-localizing with vascular smooth muscle cells (VSMCs). Flow cytometry and immunocytochemistry showed that recombinant CLEC-2, but not an anti-podoplanin antibody, bound to VSMCs, suggesting that CLEC-2 ligands other than podoplanin are present in VSMCs. Protein arrays and Biacore analysis were used to identify S100A13 as a CLEC-2 ligand in VSMCs. S100A13 was released upon oxidative stress, and expressed in the luminal area of atherosclerotic lesions. Megakaryopoiesis is promoted through the CLEC-2/podoplanin interaction in the vicinity of arterioles, not sinusoids or lymphatic vessels. There exist podoplanin-expressing bone-marrow (BM) arteriolar stromal cells, tentatively termed as BM fibroblastic reticular cell (FRC)-like cells, and megakaryocyte colonies were co-localized with periarteriolar BM FRC-like cells in the BM. CLEC-2/podoplanin interaction induces BM FRC-like cells to secrete CCL5 to facilitate proplatelet formation. These observations indicate that a reciprocal interaction with between CLEC-2 on megakaryocytes and podoplanin on BM FRC-like cells contributes to the periarteriolar megakaryopoietic microenvironment in mouse BM.

High temperature has been an obstacle to grow Rhododendron plants, and there are few reports on anatomical changes of Rhododendron plants under heat stress. To identify the mechanism of anatomical adjustment under high temperature, three Rhododendron species with different sensitivity to high temperature stress were selected to investigate the effect of heat stress on the anatomical structure. Heat stress resulted in stomatal closure and the decrease of stomatal density, however, the limited stomatal adjustment reached by R. fortune and R. mariesii may not prevent water loss effectively, of which the leaf thickness was reduced and obvious plasmolysis was observed in the mesophyll cells. R. simsii with smaller stomata and higher stomatal density had greater heat resistance than R. fortune and R. mariesii. Heat stress injured cell membrane structure seriously, and it was found that the nuclear membrane was digested and the nucleolus disappeared. The response of chloroplasts to high temperature was most sensitive, in which thylakoid lamellas became blurred, even degraded in heat sensitive Rhododendron species. The thermal endurances were sequenced as follows: R. simsii, R. mariesii and R. fortunei. Greater heat resistance of R. simsii may be associated with stabilizing anatomical structure under high temperature. The results offered cytological evidence of adaptation for heat stress in heat resistant Rhododendron species.

Background: It has been shown that toll-like receptor (TLR) 2- and TLR4-stimulating abilities of supragingival plaque (SPP) are associated with periodontal conditions. It is hypothesized that SPP might affect the periodontium through its influence on subgingival plaque (SBP). This study investigates relationships between TLR2- and TLR4-stimulating abilities of SBP and periodontal conditions. Methods: One hundred thirteen SBP samples were collected from the deepest pockets in patients with chronic periodontitis. TLR2- and TLR4-stimulating abilities were measured using genetically engineered nuclear factor-kappa B reporter cells. Numbers of Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans in each plaque sample were determined by real-time polymerase chain reaction. Peripheral blood mononuclear cells (PBMCs) were stimulated with SBP samples in presence or absence of TLR4 or TLR2 inhibitor. Production of tumor necrosis factor (TNF)-alpha and interleukin (IL)-8 was analyzed by enzyme-linked immunosorbent assay. Results: TLR4-stimulating ability of SBP was associated with plaque index (PI), but not with other clinical parameters at sampling sites. TLR2-stimulating ability of SBP was associated with none of the parameters. Number of P. gingivalis and A. actinomycetemcomitans in each plaque sample was not associated with TLR2- or TLR4-stimulating ability of SBP. PBMCs stimulated with SBP samples produced TNF-alpha and IL-8, which was inhibited by TLR4 but not by TLR2 inhibitor. Conclusion: TLR4-but not TLR2-stimulating ability of SBP is associated with PI. Enhanced TLR4-stimulating ability at sites with accumulated plaque may mediate gingival inflammation.

We report the case of a 66 year-old woman with chronic atrial fibrillation, hypertrophic cardiomyopathy (HCM), and spinocerebellar atrophy (SCA). Her mother and first-born son had died of heart disease at the ages of 65 and 16 years, respectively. Four of her 8 siblings had died suddenly of unknown cause or of heart disease, and 2 others of cerebral infarction by the 7th decade. Genetic testing revealed that she had a novel mutation (c. 482C &gt; A, p. Alal6lAsp) in the troponin I gene (TNNI3), and no abnormality of the GAA repeat in the frataxin gene. Her older brother with SCA but without HCM was also analyzed, with no abnormality noted in either gene. The Ala161Asp mutation in TNNB was implicated in the pathogenesis of her HCM, though an association between HCM and SCA was not revealed.