尾崎 行男

We examined the effects of exogenous ethylene on rhizome transition to storage organ in lotus (Nelumbo nucifera) by using seed derived plants. The rhizomes elongated in long days and enlarged in short days with ethephon treatments as well as those in the control. It is suggested that ethylene is not involved in rhizome swelling in lotus plants.

CLEC-2 has been described recently as playing crucial roles in thrombosis/hemostasis, tumor metastasis, and lymphangiogenesis. The snake venom rhodocytin is known as a strong platelet activator, and we have shown that this effect is mediated by CLEC-2 (Suzuki-Inoue, K., Fuller, G. L., Garc a, A., Eble, J. A., Pohlmann, S., Inoue, O., Gartner, T. K., Hughan, S. C., Pearce, A. C., Laing, G. D., Theakston, R. D., Schweighoffer, E., Zitzmann, N., Morita, T., Tybulewicz, V. L., Ozaki, Y., and Watson, S. P. (2006) Blood 107, 542-549). Podoplanin, which is expressed on the surface of tumor cells, is an endogenous ligand for CLEC-2 and facilitates tumor metastasis by inducing platelet aggregation. Mice deficient in podoplanin, which is also expressed on the surface of lymphatic endothelial cells, show abnormal patterns of lymphatic vessel formation. In this study, we report on the generation and phenotype of CLEC-2-deficient mice. These mice are lethal at the embryonic/neonatal stages associated with disorganized and blood-filled lymphatic vessels and severe edema. Moreover, by transplantation of fetal liver cells from Clec-2(-/-) or Clec-2(+/+) embryos, we were able to demonstrate that CLEC-2 is involved in thrombus stabilization in vitro and in vivo, possibly through homophilic interactions without apparent increase in bleeding tendency. We propose that CLEC-2 could be an ideal novel target protein for an anti-platelet drug, which inhibits pathological thrombus formation but not physiological hemostasis.

Objectives Stent fracture (SF) of sirolimus-eluting stents (SES) has emerged recently in the literature and shown to be associated with an increased risk of restenosis; however, little is known regarding SF after bare-metal stent implantation. We sought to assess whether the use of SES was associated with an increased risk of SF compared with its bare-metal platform, the Bx-velocity stent (BX-BMS). Methods A total of 478 lesions in 416 patients undergoing SES implantation and subsequent angiography 6-9 months after the index procedure were compared with 152 lesions in 142 consecutive patients treated with BX-BMS. Stented lesions with total stent-length greater than 40 mm were excluded. Results There were no significant differences in overall baseline clinical and anatomic features between the SES and BX-BMS groups, or in SF frequencies at 6-9 month follow-up (4.4% for SES and 1.3% for BX-BMS, P = 0.078). In-stent restenosis was observed more often in SF lesions versus non-SF lesions (34.8 vs. 7.7%, P < 0.001) in association with a higher 3-year adverse events rate (27.3 vs. 13.6%, P = 0.076). The risk of SF at 6-9 months was independently associated with total stent length [odds ratio (OR), 2.13; 95% confidence interval (CI), 1.18-3.83; P = 0.012], angulated lesions (OR, 4.25; 95% CI, 1.80-10.00; P = 0.001), and right coronary artery lesions (OR, 3.55; 95% CI, 1.46-8.62; P = 0.005) but not with SES use. Conclusion Stent implantation in right coronary artery lesions, tortuous lesions, and/or longer lesions covered with longer stents, and not SES versus BX-BMS use, may be associated with increased likelihood of SF. Coron Artery Dis 21: 298-303 (C) 2010 Wolters Kluwer Health vertical bar Lippincott Williams & Wilkins.

Background: The Japan Assessment of Pitavastatin and Atorvastatin in Acute Coronary Syndrome (JAPAN-ACS) trial has found that early aggressive statin therapy in patients with acute coronary syndrome (ACS) significantly reduces the plaque volume (PV) of non-culprit coronary lesions. The purpose of the present study was to evaluate clinical factors that have an impact on plaque regression using statin therapy. Methods and Results: Serial intravascular ultrasound observations over 8-12 months were performed in 252 ACS patients receiving pitavastatin or atorvastatin. Linear regression analysis identified the presence of diabetes mellitus (DM) and PV at baseline as inhibiting factors, and serum remnant-like particle-cholesterol level at baseline as a significant factor significantly affecting the degree of plaque regression. Significant correlation between % change of PV and low-density lipoprotein cholesterol (LDL-C) level was found in patients with DM (n=73, P<0.05, r=0.4), whereas there was no significant correlation between the 2 parameters in patients without DM (n=178). Conclusions: The regression of coronary plaque induced by statin therapy after ACS was weaker in diabetic patients than their counterparts. Moreover, vigorous reduction of the LDL-C levels might induce a greater degree of plaque regression in ACS patients with DM. (Circ J 2010; 74: 1165-1174)

Pigment and molecular analyses were carried out to elucidate how the white marginal picotee in the petals of Camellia japonica 'Tamanoura' is expressed. HPLC analyses showed that two major anthocyanins (cyanidin 3-glucoside and cyanidin 3-galactoside) were accumulated in the red part of the petals of 'Tamanoura', as found in those of wild type C. japonica, whereas no anthocyanins were detected in the white picotee part, indicating that the anthocyanin biosynthetic pathway might be blocked at some steps in the white part. Transcriptional levels of the genes involved in anthocyanin biosynthesis were investigated by RT-PCR. Most genes were equally expressed in both red and white parts of 'Tamanoura' petals, but the expression of chalcone synthase (CHS) was strongly suppressed only in the white picotee part. Full-length cDNA sequence of CjCHS was determined using the 5' and 3' RACE approach. The deduced amino acid sequence of CjCHS shared high homology with those of several woody plants. cDNA RT-PCR and genomic DNA PCR revealed no length differences in PCR products between red and white picotee parts in the petals of 'Tamanoura', suggesting that no insertion of transposable elements and DNA rearrangement occurred in CjCHS in the white part.

Partial cDNA sequences of three anthocyamin biosynthetic genes (FSH, flavanone 3-hydroxylase, DFR, dihydroflavonol 4-reductase, ANS, anthocyanidin synthase) were isolated from the petals of Camellia japonica. Then deduced partial amino acid sequences shared high homologies with those of woody plant species (CjF3Ha, 98.0%, CjF3Hb, 91.2% and CjDFR, 99.0% with Camellia sinensus, CjANS, 90.3% with Rhododendron x pulchrum). Some important amino acid residues for enzymatic activities were also conserved in the isolated clones, suggesting that the genes we identified in this study were the homologues of C. japonica. Gene-specific primer pairs were designed basect on each partial cDNA sequence. The application of these primer pairs to RT-PCR analyses tested.

Aim: Platelet-derived microparticles (PDMPs) play roles in normal hemostatic responses to vascular injury because they possess prothrombinase activity. Although the most widely used method for studying PDMP is flow cytometry, we previously developed an enzyme-linked immunosorbent assay (ELISA) method as an easier and more reproducible PDMP assay. The purpose of this study was to use various clinical settings to verify whether this ELISA method can produce equivalent results to flow cytometry for PDMP. Methods: We performed a large-scale clinical study for various thrombotic and subatherothrombotic diseases using an ELISA kit. The study group included 692 patients with cerebral infarction, heart failure, acute coronary syndrome or diabetes mellitus. Results: When baseline PDMP values in the various diseases were compared with those in healthy controls, significant differences were noted in all cases. There were significantly elevated levels of PDMPs in diabetic patients with complications but no thrombosis. When baseline PDM[P values in cerebral infarction were compared within the subclassifications, atheroma and other types of infarction exhibited significantly elevated PDMP levels compared with lacunar infarction. Cerebral infarction exhibited a significant change in PDMPs after therapy compared with the baseline (before therapy), but not in acute coronary syndrome and heart failure. The ELISA method exhibited results almost identical to flow cytometry for PDMP in various atherothrombotic diseases. Conclusion: Although further examinations to evaluate the therapeutic usefulness of these diseases are necessary, ELISA kits possibly represent a new tool for PDMP related to atherothrombosis.

Background-Although there is a statistically significant association between modestly raised baseline plasma C-reactive protein (CRP) values and future cardiovascular events, the debate is still unsettled in regard to whether CRP plays a causal role in the pathogenesis of atherosclerosis. Methods and Results-We generated 2 lines of transgenic (Tg) rabbits expressing human CRP (hCRP). The plasma levels of hCRP in hCRP-Tg-1 and hCRP-Tg-2 rabbits were 0.4 +/- 0.13 (n=14) and 57.8 +/- 20.6 mg/L (n=12), respectively. In addition, hCRP isolated from Tg rabbit plasma exhibited the ability to activate the rabbit complement. To define the role of hCRP in atherosclerosis, we compared the susceptibility of hCRP-Tg rabbits to cholesterol-rich diet-induced aortic and coronary atherosclerosis with that of non-Tg rabbits. After being fed with a cholesterol-rich diet for 16 weeks, Tg and non-Tg rabbits developed similar hypercholesterolemia and lesion sizes in both aortic and coronary arteries. Immunohistochemical staining and Western blotting revealed that hCRP was indeed present in the lesions but did not affect macrophage accumulation and smooth muscle cell proliferation of the lesions. Conclusions-Neither high nor low plasma concentrations of hCRP affected aortic or coronary atherosclerosis lesion formation in hCRP-Tg rabbits. (Circulation. 2009;120:2088-2094.)

We have reported in this journal in vitro susceptibilities of clinical isolates to antibiotics every year since 1992. In this paper, we report the results of an analysis of in vitro susceptibilities of 12,919 clinical isolates from 72 centers in Japan to selected antibiotics in 2007 compared with the results from previous years. The common respiratory pathogens, Streptococcus pyogenes, Streptococcus pneumoniae, Moraxella catarrhalis and Haemophilus influenzae maintained a high susceptibility to fluoroquinolones (FQs). The resistance of S. pyogenes to macrolides has been increasing every year and this was especially clear this year. Most strains of Enterobacteriaceae except for Escherichia coli showed a high susceptibility to FQs. Almost 30% of E. coli strains were resistant to FQs and the resistance increased further this year. FQs resistance of methicillin-resistant Staphylococcus aureus (MRSA) was approximately 95% with the exception of 45% for sitafloxacin (STFX). FQs resistance of methicillin-susceptible S. aureus (MSSA) was low at about 10%. FQs resistance of methicillin-resistant coagulase negative Staphylococci (MRCNS) was higher than that of methicillin-susceptible coagulase negative Staphylococci (MSCNS), but it was lower than that of MRSA. However, FQs resistance of MSCNS was higher than that of MSSA. FQs resistance of Enterococcus faecalis was 22.5% to 29.6%, while that of Enterococcusfaecium was more than 85% except for STFX (58.3%). In clinical isolates of Pseudomonas aeruginosa derived from urinary tract infections, FQs resistance was 21-27%, which was higher than that of P. aeruginosa from respiratory tract infections at 13-21%, which was the same trend as in past years. Multidrug resistant strains accounted for 5.6% in the urinary tract and 1.8% in the respiratory tract. Acinetobacter spp. showed high susceptibility to FQs. The carbapenem resistant strains, which present a problem at present, accounted for 2.7%. Neisseria gonorrhoeae showed high resistance of 86-88% to FQs. The results of the present survey indicated that although methicillin-resistant Staphylococci, Enterococci, E. coli, P. aeruginosa, and N. gonorrhoeae showed resistance tendencies, and other species maintained high susceptibility rates more than 90% against FQs, which have been used clinically for over 15 years.

T cells play important roles in bone destruction and osteoclastogenesis and are found in chronic destructive bone lesions. Lipopolysaccharide (LPS) is one of several pathological factors involved in inflammatory bone destruction. We previously described the importance of T cells in the inflammatory bone resorption that occurs after repeated LPS administration. However, whether local or systemic T cells are important for inflammatory bone resorption and whether immunization of host animals influences bone resorption remain unclear. The present study examines the effects of local extant T cells from LPS-immunized mice on LPS-induced bone resorption. T cells from LPS-immunized or non-immunized mice were injected together with LPS into the gingival tissues of mice with severe combined immunodeficiency disease that lack both T and B cells. We histomorphometrically evaluated bone resorption at sites of T cell injections and examined the influence of T cells from LPS-immunized mice on osteoclastogenesis in vitro. We found that locally administered T cells from LPS-immunized but not non-immunized mice accelerated LPS-induced bone resorption in vivo. Moreover, T cells from LPS-immunized mice increased osteoclastogenesis in vitro induced by receptor activator of NF-kappa B ligand and LPS and anti-tumor necrosis factor (TNF)-alpha antibody inhibited this increase. These results demonstrated that local extant T cells accelerate inflammatory bone resorption. Furthermore. T cells from LPS-immunized mice appear to elevate LPS-induced bone resorption using TNF-alpha. (C) 2009 Elsevier Inc. All rights reserved.

Introduction: A simple, validated method to measure platelet function is unavailable for bedside use. Measurement of platelet retention rate using a column of collagen-coated beads and whole blood is a new, simple assay that reflects platelet aggregation. This study was aimed to examine the utility of this assay to assess efficacy of antiplatelet drug therapy. Methods: Citrated whole blood (1.5 ml) in a syringe was passed through a polyvinyl tube packed with collagen-coated beads for 40 seconds using a syringe pump. Platelet retention rate in the column was calculated from platelet counts in blood before and after passage. An increase in the retention rate reflects an increase in platelet activity. This new platelet retention assay and the traditional optical aggregometry assay were performed in 331 patients with stable coronary artery disease (CAD). Results: The retention rate was significantly reduced in patients taking dual antiplatelet therapy (aspirin plus clopidogrel or ticlopidine) compared with aspirin alone. There was a significant linear correlation between the platelet retention rate and platelet aggregability measured by the traditional method (r=0.44, p<0.001). In multivariate Cox proportional hazards analysis, higher platelet retention rate was an independent predictor of future cardiovascular events in patients on dual antiplatelet therapy (hazard ratio 3.9, 95% CI 1.6 to 9.5, p = 0.003). Conclusions: Measurement of the platelet retention rate in a column of collagen-coated beads may be useful for monitoring the efficacy of antiplatelet drug therapy in patients with CAD. (C) 2008 Elsevier Ltd. All rights reserved.

Sphingolipids, including ceramide (Cer), sphingosine (Sph), and sphingosine 1-phosphate (Sph-1-P) have recently emerged as signal-transducing molecules. Functionally, a distinguishing characteristic of these lipids is their apparent participation in pro- or anti-proliferative cell regulation pathways. In this study, we examined the involvement of sphingolipids in the fate of FRTL-5 thyroid follicular cells. We first examined the effects of sphingolipids on FRTL-5 cell viability. Sph and Cer induced apoptosis, as revealed by fluorescence microscopy of TUNEL-positive fragmented nuclei and 180300 bp DNA fragmentation on agarose gel electrophoresis while Sph-1-P was confirmed to prevent FRTL-5 cell apoptosis induced by deprivation of serum and TSH, possibly via cell surface receptors. We then analysed the metabolism of radiolabelled Sph and C(6)-Cer (a synthetic cell-permeable Cer) in FRTL-5 cells by thin layer chromatography, followed by autoradiography. Sph was mainly metabolized to Cer, and then to sphingomyelin, while Sph conversion into Sph-1-P was hardly detected. These changes were not affected by stimulation of the cells with TSH. Our results indicate the involvement of sphingolipid mediators in the fate of FRTL-5 thyroid cells.

Novel synthetic collagen fibers, poly(PHG) made by polycondensation of Pro-Hyp-Gly, spontaneously assume polymeric structure with molecular weights greater than 10(5). Its application for biomaterials has been explored, but that for a platelet agonist has not been investigated. Poly(PHG)-induced platelet aggregation independently of thromboxane A(2) and integrin alpha 2 beta 1. Poly(PHG)-induced tyrosine phosphorylation of glycoprotein VI (GPVI)-related molecules and failed to activate GPVI/FcR gamma-deficient platelets. Binding of GPVI to poly(PHG) was confirmed by a surface plasmon resonance spectroscopy, suggesting that poly(PHG) activates platelets through GPVI. Poly(PHG) is an useful research tool to investigate GPVI-mediated signals and a substitute for collagen in platelet functional assays. (c) 2008 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

Patients admitted to the hospital with heart failure (HF) include those with new-onset of acute HF and those with acute exacerbation of chronic HF (CHF). In therapy for new-onset acute HF associated with acute myocardial infarction, therapy to inhibit left ventricular (LV) remodeling in the convalescent phase is required in addition to that needed to overcome the acute phase. Hitherto, CHF therapy was aimed at improving LV contractability, whereas more recently the aim has shifted to resting the heart. Most patients with HF should be routinely managed with a combination of 3 types of drugs: a diuretic an angiotensin converting enzyme inhibitor and/or an angiotensin II receptor blocker and a β-blocker. The administration of β-blockers is of particular importance. For HF unresponsive to medical therapy, non-pharmacological therapies are considered. When a HF patient fails to respond to all available therapies, heart transplantation becomes necessary. Of the 1,000 HF patients admitted to our hospital, two cases received heart transplants. 11 cases were indicated for heart transplantation but died before registration. It should be remembered that although in Japan the possibility of receiving a heart transplant is very low, it is by no means entirely impossible.

Lilium formosanum Wallace has remarkable traits such as 'precocious flowering' ability, i.e., it reaches anthesis within 12 months from seed germination, with multiple shooting of flower stalks. To verify the possibility of the usefulness of these traits in lily breeding, nine combinations of interspecific crosses (L. formosanum as the seed parent; L. auratum, L. speciosum, L. regale,'Lollypop','Pink Tiger','Zaza','Le Reve','Marco Polo', and 'African Queen' as pollen parents) were carried out with cut-style pollination and ovary-slice culture. Germination was observed in all nine interspecific crosses and 53 hybrids were obtained. Thirty (56.6%) of the 53 hybrids and two self-pollinated progenies of L. formosanum reached anthesis within 24 months from germination through ovary-slice culture. Multiple shooting of flower stalks was recognized in 11 (36.7%) of those flowered hybrids. Four hybrids, the pollen parents of which were Asiatic hybrid lilies with colored flowers, expressed 'precocious flowering' ability., multiple shooting of flower stalks, and entirely colored flowers simultaneously. These results suggest the possibility of breeding new types of cultivars with triple favorable traits from the cross between L. formosanum and Asiatic hybrid lilies with colored flowers.