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Background:Recently, immunometabolism has gathered attention of many immunologists. It has been widely recognized that metabolic reprogramming in each immune cell brings different effects on different cells and is important for regulating their functions. Along with the progress of statistical genetics, serum metabolites were shown to be under genetic regulations1). Metabolic changes are now considered not only to be mere phenotypes of cells but also to be key factors for controlling immune cell differentiation, proliferation and function through regulating gene expressions eventually. Although genome-wide association studies have brought deep insights into SLE pathogenesis, the precise pathway from genome to metabolome has been largely unknown, and vice versa.Objectives:The aim of this study is to investigate metabolomic regulation in SLE in relation to gene expressions by integrating plasma metabolome data and transcriptome data.Methods:We collected plasma samples from patients with SLE (n=57) who met the 1997 American College of Rheumatology criteria for SLE. Gender- and age-matched healthy controls (HCs) (n=56) were recruited. Metabolic profiles focusing on 39 amino acids were analyzed with liquid chromatography (LC)-mass spectrometry. Transcriptome data of SLE patients were obtained from our RNA-sequencing data of each immune cell subset (total 19 subsets). Whole-genome sequencing was also performed.Results:Our previous experiment showed that about 160 peaks were detected from comprehensive LC-TOFMS and amino acids were useful for distinguishing SLE patients from HCs. Both partial least squares discriminant analysis (PLS-DA) and random forest, a machine learning algorithm, revealed the importance of histidine (His), one of the essential amino acids, to classify SLE patients from HCs, whose plasma level was lower in SLE patients. In addition, inverse correlation between His level and titer of ds-DNA as well as damage index (SDI) was detected. His level was correlated neither with PSL dosage nor with type I interferon (IFN) signature. Receiver operating characteristic (ROC) analysis showed the best predictability for SLE with the combination of specific amino acids including His. Our transcriptome analysis has revealed the significance of oxidative phosphorylation (OXPHOS) in B cells for SLE pathogenesis. Interestingly, OXPHOS signature was inversely correlated with His level in SLE B cells.Conclusion:His may be an important factor for SLE pathogenesis especially in B cells independently from IFN signal. SLC15A4, a transporter of His on lysosome, is one of the SLE GWAS SNPs and has been reported to play an important role in IFN production in B cells through regulation of TLR7/9 activation 2). We also identified that SLE patients with risk allele of SLC15A4 had tendency to show higher plasma His level, indicating His homeostasis could become a novel treatment target for SLE. Moreover, the inverse correlation of His level to SDI as well as OXPHOS signature suggests that His might play a key role for promoting organ damages in SLE.References:[1]Nat Genet.2017;49:568. 2)Immunity. 2014;41:375. 3)Semin Arthritis Rheum.2019;48:1142Disclosure of Interests: :Yukiko Iwasaki: None declared, Yusuke Takeshima: None declared, Masahiro Nakano: None declared, Mineto Ota: None declared, Yasuo Nagafuchi: None declared, Akari Suzuki: None declared, Yuta Kochi: None declared, Tomohisa Okamura: None declared, Takaho Endo: None declared, Ichiro Miki: None declared, Kazuhiro Sakurada: None declared, Kazuhiko Yamamoto Grant/research support from: Astellas, BMS, MitsubishiTanabe, Pfizer, Ayumi, Takeda, Chugai, Eisai, Taisho Toyama, UCB, and ImmunoFuture, Keishi Fujio Grant/research support from: Astellas, BMS, MitsubishiTanabe, Pfizer, Ayumi, Takeda, Chugai, Eisai, Taisho Toyama, Eli Lilly, Sanofi, and UCB

Background A novel approach to differentiate somatic vs. germline BRCA1/2 mutations in cfDNA using a beta binomial model (AACR 2018, abst #4272) may enable identification of pts with advanced GI tumors who are likely to benefit from a PARP inhibitor. The results from the GOZILA study, the Nationwide Cancer Genome Screening Project utilizing Guardant360, are presented here. Methods Pts with advanced GI tumors who were appropriate for any systemic therapy and had progressed following at least one prior treatment were eligible. We investigated the prevalence of somatic vs. germline BRCA1/2 mutations and the association between BRCA1/2 mutations, TMB and MSI. Results From Jan 2018 to Mar 2019, 850 pts were prospectively enrolled among 26 major cancer centers in Japan. Of 40 (4.7%) actionable BRCA1/2 mutations, 14 were germline: esophagus (squamous cell carcinoma only), 2 pts (N = 62, 3.2%); stomach, 2 pts (N = 131, 1.5%); colorectum, 3 pts (N = 377, 0.8%); pancreas, 3 pts (N = 157, 1.9%); biliary tract, 4 pts (N = 77, 5.2%); and others, none (N = 31, 0%). Notably, the majority of BRCA1/2 mutations (75%) in pts with esophageal, pancreas and biliary tract cancers were germline, while 81% of BRCA1/2 mutations in pts with stomach and colorectal cancers were somatic (p = 0.001). Median TMB of pts with germline BRCA1/2 mutations was significantly lower than those with somatic BRCA1/2 mutations (p = 0.042). In addition, all pts with germline BRCA1/2 mutations were microsatellite stable, while 19% of pts with somatic BRCA1/2 mutations were MSI-high. Conclusions Analysis of cfDNA identified a unique subset of pts with germline BRCA1/2 mutations in advanced GI cancers. Given the association of these findings with efficacy of PARP inhibitor and immune-based therapies, these findings suggest that the identification of BRCA1/2 mutations and their germline-somatic origin may have future implications for therapy selection. Clinical trial identification UMIN000029315. Legal entity responsible for the study SCRUM-Japan GI-SCREEN. Funding Guardant Health, Ono Pharmaceutical. Disclosure Y. Kawamoto: Honoraria (self): Taiho Pharmaceutical Co., Ltd.; Honoraria (self): Daiichi Sankyo Co., Ltd.; Honoraria (self): Takeda Pharmaceutical Co., Ltd.; Honoraria (self): Chugai Pharmaceutical Co., Ltd.; Honoraria (self): Merck Biopharma Co., Ltd.; Honoraria (self): Eli Lilly Japan K.K.. Y. Nakamura: Research grant / Funding (institution): Ono Pharmaceutical; Research grant / Funding (institution): Taiho Pharmaceutical. M. Ikeda: Honoraria (self), Advisory / Consultancy, Research grant / Funding (institution): Novartis Pharma; Honoraria (self), Advisory / Consultancy, Research grant / Funding (institution): Bayer Yakuhin; Research grant / Funding (institution): Bristol-Myers Squibb; Honoraria (self), Advisory / Consultancy, Research grant / Funding (institution): Eisai; Honoraria (self): Taiho Pharmaceutical; Honoraria (self), Advisory / Consultancy, Research grant / Funding (institution): Eli Lilly Japan; Research grant / Funding (institution): Yakult; Advisory / Consultancy: Otsuka Pharmaceutical; Advisory / Consultancy: Daiichi-Sankyo; Honoraria (self): Sumitomo Dainippon Pharma; Honoraria (self), Advisory / Consultancy: Teijin Pharma; Honoraria (self): EA Pharma; Honoraria (self): MSD; Advisory / Consultancy, Research grant / Funding (institution): ASLAN Pharmaceuticals; Advisory / Consultancy, Research grant / Funding (institution): Chugai Pharmaceutical; Research grant / Funding (institution): Ono Pharmaceutical; Research grant / Funding (institution): AstraZeneca; Research grant / Funding (institution): Merck Serono; Research grant / Funding (institution): Nano Carrier; Honoraria (self): Gilead. H. Bando: Honoraria (self): Taiho Pharmaceutical company; Honoraria (self): Eli Lilly; Research grant / Funding (self): Sysmex; Research grant / Funding (self): AstraZeneca. T. Esaki: Honoraria (self), Research grant / Funding (institution): Taiho; Honoraria (self): Chugai; Honoraria (self), Research grant / Funding (institution): Ono; Honoraria (self): Takeda; Honoraria (self): Bayer; Honoraria (self): Eli Lilly; Research grant / Funding (institution): Astellas; Research grant / Funding (institution): MSD; Research grant / Funding (institution): Daiichi-Sankyo; Research grant / Funding (institution): Merck Serono; Research grant / Funding (institution): Dainippon Sumitomo; Research grant / Funding (institution): Novartis. M. Ueno: Honoraria (self), Research grant / Funding (institution): Taiho Pharmaceutical; Honoraria (self), Research grant / Funding (institution): Yakult Honsha; Honoraria (self), Research grant / Funding (institution): AstraZeneca; Honoraria (self): Novartis; Honoraria (self): Lilly; Honoraria (self): Teijin Pharma; Honoraria (self), Advisory / Consultancy, Research grant / Funding (institution): Shire; Honoraria (self), Research grant / Funding (institution): Ono Pharmaceutical; Honoraria (self), Research grant / Funding (institution): Merck Serono; Research grant / Funding (institution): Daiichi Sankyo; Research grant / Funding (institution): Eisai; Research grant / Funding (institution): MSD; Research grant / Funding (institution): NanoCarrier; Research grant / Funding (institution): Dainippon Sumitomo Pharma; Research grant / Funding (institution): Incyte; Research grant / Funding (institution): ASLAN Pharmaceuticals. T. Nishina: Honoraria (self), Research grant / Funding (institution): Taiho pharmaceutinal; Honoraria (self), Research grant / Funding (institution): Chugai Pharma; Honoraria (self), Research grant / Funding (self): Merck Serono; Honoraria (self), Research grant / Funding (institution): Bristol-Myers Suibb; Honoraria (self): Nihonkayaku; Honoraria (self), Research grant / Funding (institution): Lilly Japan; Honoraria (self), Research grant / Funding (institution): Ono Pharmaceutical; Research grant / Funding (institution): Dainippon Sumitomo Pharma; Research grant / Funding (institution): MSD; Research grant / Funding (institution): Diichi Sankyo. E. Oki: Speaker Bureau / Expert testimony: Taiho Pham; Speaker Bureau / Expert testimony: Yakult Honsha; Speaker Bureau / Expert testimony: Merck; Speaker Bureau / Expert testimony: Chugai Pham; Speaker Bureau / Expert testimony: Takeda Pharm; Speaker Bureau / Expert testimony: Eli Lilly; Speaker Bureau / Expert testimony: Bayer. T. Denda: Speaker Bureau / Expert testimony: DAIICHI SANKYO COMPANY, LIMITED. T. Mizukami: Advisory / Consultancy, Speaker Bureau / Expert testimony, Research grant / Funding (institution): Eli Lilly Japan; Advisory / Consultancy: Bristol-Myers Squibb Company; Speaker Bureau / Expert testimony: Takeda Pharmaceutical; Research grant / Funding (institution): TAIHO Pharmaceutical. N. Okano: Honoraria (self): Taiho Pharmaceutical; Honoraria (self): Merck Serono; Honoraria (self): Eisai; Honoraria (self): Ono Pharmaceutical; Honoraria (self): Yakult Honsha; Honoraria (self): Takeda; Honoraria (self): J-Pharma; Honoraria (self): Kyowa Hakko Kirin. M.I. Lefterova: Shareholder / Stockholder / Stock options, Full / Part-time employment: Guardant Health. J.I. Odegaard: Shareholder / Stockholder / Stock options, Full / Part-time employment: Guardant Health. H. Taniguchi: Honoraria (self): Chugai; Honoraria (self), Research grant / Funding (self): Takeda; Honoraria (self): Taiho; Honoraria (self): Eli Lilly; Research grant / Funding (self): Daiichi-Sankyo; Research grant / Funding (self): Sysmex. C. Morizane: Honoraria (self), Research grant / Funding (institution): Pfizer; Honoraria (self), Advisory / Consultancy: Novartis; Honoraria (self), Advisory / Consultancy, Research grant / Funding (institution): Yakult Honsha; Honoraria (self): Lilly; Honoraria (self), Research grant / Funding (institution): Nobelpharma; Honoraria (self): Fujifilm; Honoraria (self): Teijin Pharma; Honoraria (self), Advisory / Consultancy, Research grant / Funding (institution): Taiho Pharmaceutical; Honoraria (self), Research grant / Funding (institution): Eisai; Advisory / Consultancy: Abbvie; Research grant / Funding (institution): ONO PHARMACEUTICAL; Research grant / Funding (institution): J-Pharma. T. Yoshino: Research grant / Funding (self): Novartis Pharma K.K.; Research grant / Funding (self): MSD.K.K.; Research grant / Funding (self): Sumitomo Dainippon Pharma Co., Ltd.; Research grant / Funding (self): CHUGAI PHARMACEUTICAL Co., Ltd.; Research grant / Funding (self): Sanofi K.K.; Research grant / Funding (self): DAIICHI SANKYO Co., Ltd.; Research grant / Funding (self): PAREXEL International Inc.; Research grant / Funding (self): ONO PHARMACEUTICAL Co., Ltd. All other authors have declared no conflicts of interest.

Objective and design: P-selectin is a cell adhesion molecule of the selectin family. This study evaluated the effects of novel, low molecular weight P-selectin inhibitors in a cell adhesion assay and a murine model of peritonitis.¶Materials: U937 or HL60 was used for cell adhesion assay. Human polymorphonuclear cells were studied for the production of superoxide. BALB/c mice were used for the in vivo study.¶Treatment: The thioglycollate (TG)-induced accumulation of leukocytes in mice was measured 6 h after the treatment. KF38789 or antibody (1 mg/kg) was injected intravenously prior to TG injection and at 3 h following initial injection.¶Results: Low molecular weight, non-carbohydrate inhibitors against P-selectin- mediated cell adhesion were tested. One of the most potent inhibitors, KF38789, inhibited the bind-ing of U937 cells to immobilized P-selectin immunoglobu-lin G chimeric protein (P-selectin-Ig) with an IC50 value of 1.97 μM. Cell adhesion to both E-selectin-Ig and L-selectin-Ig were not affected even by 100 μM of KF38789. Moreover, KF38789 inhibited P-selectin-induced superoxide production from human polymorphonuclear cells. Intravenously injected KF38789 significantly inhibited the TG-induced accumulation of leukocytes in the mouse peritoneal cavity (p < 0.01).¶Conclusion: A novel low molecular weight compound, KF38789, specifically inhibited P-selectin-dependent cell adhesion and the leukocyte recruitment in mouse peritonitis.

This paper presents a method of estimating the initial rotor position of a surface permanent-magnet synchronous motor without a position sensor. The estimation is performed by using the nonlinear magnetization characteristics of the stator core caused by the magnet of the rotor. This method is based on the principle that the d-axis current value for the voltage vector applied to the motor under some conditions increases as the voltage vector generated from the inverter approaches the N pole of the rotor. During the estimation process, the rotor is practically at standstill. The experimental results show that the average of the estimation error is /spl plusmn/3.8 electrical degrees.

A novel pyridobenzazepinone derivative (Z)-11-(5-carboxypentylidene)-6-methyl-5, 11-dihydropyrido +ad4,3-C] [1] benzazepin-5 (6H)-one (CAS 127654-03-9, KF 13218), inhibited human and bovine platelet thromboxane synthase with IC50 values of 27 +/- 5.8 nmol/l (mean +/- S.E.M.) and 36 +/- 6.9 nmol/l, respectively. The compound did not inhibit cyclooxygenase or 5-lipoxygenase up to a dose of 100 mumol/l and did not antagonize thromboxane A2/prostaglandin H2 receptors. KF13218 inhibited arachidonic acid-induced thromboxane B2 production by human intact platelets with an IC50 value of 5.3 +/- 1.3 nmol/l. The IC50 value of KF13218 for the intact platelets was about 5 times lower than that for the microsomal enzyme. The inhibition of thromboxane synthase in platelets by KF13218 was sustained after removal of the extracellular compound. After oral dosing in rat from 0.03 mg/kg to 3 mg/kg, KF13218 dose-dependently inhibited the thromboxane B2 production in serum, and the inhibition was retained for 72 h. KF13218, at a dose of 0.1 mg/kg p.o. prevented mortality induced by sodium arachidonate in rabbit. It is concluded that KF13218 is a potent, selective and long lasting thromboxane synthase inhibitor.

A series of 11-[[2-[(arylsulfonyl)amino]ethyl]thio]-6,11- dihydrodibenz[b,e]oxepin-2-carboxylic acids and related derivatives were synthesized. The compounds were tested for their antagonizing effects on guinea pig platelet TXA2/PGH2 receptors. Structure-activity relationships are discussed. (+/-)-11-[[2-[(Styrylsulfonyl)amino]ethyl]-thio]-6,11- dihydrodibenz[b,e]oxepin-2-carboxylic acid (41) and (+/-)-11-[[2-[(phenylsulfonyl)amino]ethyl]thio]-6,11- dihydrodibenz[b,e]thiepin-2-carboxylic acid (4af) were the most promising compounds with K(i) values of 6.5 +/- 0.29 and 3.7 +/- 0.31 nM, respectively, for the TXA2/PGH2 receptor. These compounds also significantly inhibited U-46619-induced guinea pig platelet aggregation ex vivo (10 mg/kg po). Compound 41 was resolved into its optically active form. The (-)-isomer was 60-fold more potent than the (+)-isomer in the TXA2/PGH2 receptor binding assay. Some compounds tested in this study showed both TXA2/PGH2 receptor antagonizing and TXA2 synthase inhibitory effects.

The initial rotor position estimation is one of some serious problems about sensorless control of permanent magnet synchronous motors. This paper describes an estimation method of the initial rotor position for a slotless permanent magnet motor. The method is based on the variation of the current response when the voltage vector is applied to the motor. To estimate the polarity of the rotor magnetic pole, the rotor is slightly moved. As a result, the average of the estimation error is 3.42 electrical degrees.

This paper describes the better shape design of the permanent magnet to reduce the torque ripple in interior permanent magnet synchronous motors (IPMSMs) without deteriorating other performances. We make various motor models to analyze the cogging torque and the torque ripple. Using the two-dimensional finite element method, these motor models are discussed under the same condition. As a result, we have found the better shape of the permanent magnet.

The role of macrophage migration inhibitory factor (MIF), a proinflammatory cytokine, was tested using a mouse asthma model. One hundred and four male BALB/c mice were used in this study. Mice were actively sensitized with an intraperitoneal injection of ovalbumin (OVA) and challenged with repeated nebulization of 1 w/v% OVA. Polyclonal anti-MIF antibody was intraperitoneally injected at 10 mg/kg during the antigen challenge period. Bronchoalveolar lavage (BAL) was performed 8 h after the last challenge. Airway hyperresponsiveness to inhaled methacholine was measured 24 h after the last challenge. Antigen challenge to immunized mice induced increase in inflammatory cells and concentration of Th2 cytokines in BAL fluid (BALF), and caused the development of airway hyperresponsiveness. Anti-MIF antibody significantly decreased the numbers of inflammatory cells including macrophages, eosinophils, lymphocytes and neutrophils in BALF from OVA-challenged mice. Prednisolone decreased the numbers of eosinophils, lymphocytes and neutrophils but not macrophages. Anti-MIF antibody reduced airway hyperresponsiveness. Anti-MIF antibody affected neither the cytokine levels in BALF nor the IgE levels in serum. MIF was involved in the antigen-induced inflammatory cell accumulation in the lung and airway hyperresponsiveness without affecting immune responses.

SKG mice have a point mutation of the zeta-associated protein of 70 kD (ZAP-70) and spontaneously develop a severe polyarthritis in the conventional condition, whereas they are healthy under the specific pathogen free (SPF) condition. The purpose of this study was to investigate the cytokine production from splenocytes in SKG mice developing arthritis under the SPF condition.SKG and BALB/c mice were intraperitoneally injected with zymosan A under the SPF condition. Spleen was isolated 1, 2 or 8 weeks after the intraperitoneal injection of saline or zymosan A. Splenocytes were cultured with concanavalin A. Cytokine production and proliferation were measured 48 and 72 h after the culture.An intraperitoneal injection of zymosan A induced severe polyarthritis with increased levels of rheumatoid factor and interleukin 6 (IL-6) only in SKG mice. Splenocytes from SKG mice did not proliferate well maybe because of less productivity of IL-2. The IL-4 production from splenocytes of SKG mice was higher, while interferon-gamma production was lower than those of BALB/c mice. An injection of zymosan A reduced the IL-4 production only in SKG mice.SKG mice do not develop arthritis under the SPF condition possibly because of a low proliferative activity of T cells and Th2-predominance.

The ideal second-line treatment regimens for Helicobacter pylori infection may differ between the areas, countries and races.The aim was to confirm which was the better regimen for second-line therapy after treatment failure with a standard triple therapy in Japan, a high dosage of levofloxacin- or metronidazole-based therapy.Sixty outpatients with persistent H. pylori infection after a standard triple therapy were enrolled in this prospective, open-label and randomised trial.The subjects were randomly administered levofloxacin (300 mg b.d.)- or metronidazole (500 mg b.d.)-based therapy with lansoprazole (30 mg b.d.) and amoxicillin (1000 mg b.d.) for 7 days, and the cure rates and side effects were analysed. Antimicrobial susceptibility was also examined before second-line therapy using the E-test.Good compliance was obtained without severe side effects in both the groups except for two patients. The cure rates, expressed as intention-to-treat and per-protocol analyses, respectively, were 70.0 and 72.4% in the levofloxacin group, and 96.7 and 100% in the metronidazole group. Each regimen often overcame even clarithromycin-resistant strains.Metronidazole-based triple therapy is recommended as second-line therapy in Japan, and levofloxacin-based therapy can be an alternative treatment option.

This paper describes an initial rotor position estimation of surface permanent magnet synchronous motor without sensor. The estimation is performed by using the nonlinear magnetization characteristics of the stator core caused by the magnet of the rotor. The validity of the proposed estimation method is verified by the experiment.

The Engineering Department of the University of Tokushima Graduate School conduct this study to investigate traffic signals with a localizing function (that indicates the location of the traffic signal device) for visually-impaired persons, as the study project leader. We at the Traffic Management and Control Division, Traffic Department, Tokushima Prefectural Police, intend to actively support this study, including providing expertise for operation of traffic signals and a field for experimental operation, as a member of the project team. The objective of this study is to examine traffic signal devices with a localizing function to help visually-impaired persons cross streets safely, with a view towards the spread of these devices in Japan and international standardization of audible traffic signal devices. These investigational devices, imported from Germany by Tokushima University, feature various functions, such as auditory localization, acoustic signals (mechanical sounds), tactile pulse signals, and adjustment of sound levels according to the level of ambient noise. Since the existing Japanese audible traffic signal devices do not have such functions, this study is intended to investigate the convenience of the German devices and the problems they may cause pedestrians and residents of adjacent properties. The devices will be attached to audible traffic signal devices presently in use in Tokushima Prefecture, for a one-year experimental operation, launched in April 1999, to investigate their suitability to actual traffic conditions in Japan.

The single neuron used has three input connections that have associated weights. Each gain parameter corresponding with each weight of the single neuron is determined in accordance with the learning method based on the delta rule. The structure of this system is very simple and practical. The motor speed control system with 100 W DC servomotor has been simulated and experimented upon. These results show the good tuning performance of this controller. >

This paper presents a fuzzy reasoning-based new auto-tuning method for the speed controller (PI controller) in vector-controlled induction motor drive systems. A fuzzy rule plane based on the phase plane is introduced, and the trajectory of the motor speed step response is described in this plane. At characteristic points defined on this trajectory, a fuzzy inference is implemented to obtain appropriate gains of the controller. This system has been applied to the model drive system with 0.75 kW induction motor, and the performance of this system has been substantiated by simulations. >

This paper presents an auto-tuning method for the indirect vector controlled induction motor drives for the start up. The tuning algorithm is based on the rotor flux behavior of the induction motor for a stepwise torque command. An effect of the stator core loss of the induction motor, which appears in the transient motor terminal voltage caused by the undesirable rotor flux variation, is taken into account when the tuning is implemented. This method can tune the accurate inverse of the rotor time constant. Simulation results of the laboratory model system verify the validity of the proposed auto-tuning method.

The purpose of this work is the development of an estimation method of initial rotor position for surface permanent magnet (PM) synchronous motor without a position sensor. The principle of the estimation is based on the variation of the current response caused by the magnetic saturation of the stator core when the voltage vector is applied to the motor. The initial rotor position is identified by observing the maximum d-axis current caused by the magnetic saturation. This method can be performed without motor parameters and the additional hardware. A decision method of the optimal voltage vector applied to the motor is also proposed to accurately implement the estimation. Experimental results show that the initial rotor position can be estimated without rotating the rotor by using the optimal voltage vector.

The purpose of this work is development of an estimation method of initial rotor position for a surface permanent magnet synchronous motor. The principle of the estimation is based on the variation of the current response caused by the magnetic saturation of the stator core when the voltage vector is applied to the motor. This method can be performed without motor parameters and the additional hardware. The decision method of the optimum voltage vector applied to the motor is also proposed to accurately implement the estimation. Experimental results show that the initial rotor position can be estimated without rotating the rotor by using the optimum voltage vector.

Objective and Design: A newly synthesized inhibitor of pyrimidine de novo biosynthesis, KF20444 (6,7-dihydro-10-fluoro-3-(2-fluorophenyl)-5H-benzo [6,7] cyclohepta [1,2-b] quinoline-8-carboxylic acid), was evaluated as an inhibitor of dihydroorotate dehydrogenase (DHO-DHase) and tested in the rat collagen-induced arthritis (CIA) model.¶Material and Methods: Female Sprague Dawley rats, 5 weeks-old, were used for evaluation of KF20444 in the CIA model. Arthritis was evaluated by arthritis score, serum anti-type II collagen antibody titer, body weight loss, radiographical and histological changes.¶Treatment: KF20444 was orally administered 5 times per week (0.3, 1, 3 mg/kg/day).¶Results: KF20444 inhibited rat liver dihydroorotate dehydrogenase in vitro with Ki = 8.5 ± 3.2 nM, which was a comparable effect to that of brequinar sodium (Ki = 25.3 ± 5.3 nM). The anti-proliferative effect of KF20444 was caused by cell cycle arrest at the S-phase. Treatment with 3 mg/kg/day of KF20444 completely prevented the development of CIA based on reduction of the arthritis score. The 50% effective dose (ED50) of KF20444 on arthritis score was 0.64 mg/kg. KF20444 ameliorated body weight loss associated with disease onset. The compound also inhibited the increase in serum anti-type II collagen antibody level, and reduced both pannus formation and bone erosion. Importantly, KF20444 suppressed the development of arthritis, even when it was administered after booster immunization of collagen.¶Conclusions: KF20444 is a novel immunosuppressant which inhibits DHO-DHase and its effects in CIA suggest that it could be useful in the treatment of rheumatoid arthritis.¶

A novel pyridobenzazepinone derivative (Z)-11-(5-carboxypentylidene)-6-methyl-5, 11-dihydropyrido +ad4,3-C] [1] benzazepin-5 (6H)-one (CAS 127654-03-9, KF 13218), inhibited human and bovine platelet thromboxane synthase with IC50 values of 27 +/- 5.8 nmol/l (mean +/- S.E.M.) and 36 +/- 6.9 nmol/l, respectively. The compound did not inhibit cyclooxygenase or 5-lipoxygenase up to a dose of 100 mumol/l and did not antagonize thromboxane A2/prostaglandin H2 receptors. KF13218 inhibited arachidonic acid-induced thromboxane B2 production by human intact platelets with an IC50 value of 5.3 +/- 1.3 nmol/l. The IC50 value of KF13218 for the intact platelets was about 5 times lower than that for the microsomal enzyme. The inhibition of thromboxane synthase in platelets by KF13218 was sustained after removal of the extracellular compound. After oral dosing in rat from 0.03 mg/kg to 3 mg/kg, KF13218 dose-dependently inhibited the thromboxane B2 production in serum, and the inhibition was retained for 72 h. KF13218, at a dose of 0.1 mg/kg p.o. prevented mortality induced by sodium arachidonate in rabbit. It is concluded that KF13218 is a potent, selective and long lasting thromboxane synthase inhibitor.

SRPX2 (Sushi repeat-containing protein, X-linked 2) has recently emerged as a multifunctional protein that is involved in seizure disorders, angiogenesis and cellular adhesion. Here, we analyzed this protein biochemically. SRPX2 protein was secreted with a highly post-translational modification. Chondroitinase ABC treatment completely decreased the molecular mass of purified SRPX2 protein to its predicted size, whereas heparitinase, keratanase and hyaluroinidase did not. Secreted SRPX2 protein was also detected using an anti-chondroitin sulfate antibody. These results indicate that SRPX2 is a novel chondroitin sulfate proteoglycan (CSPG). Furthermore, a binding assay revealed that hepatocyte growth factor dose-dependently binds to SRPX2 protein, and a ligand–glycosaminoglycans interaction was speculated to be likely in proteoglycans. Regarding its molecular architecture, SRPX2 has sushi repeat modules similar to four other CSPGs/lecticans; however, the molecular architecture of SRPX2 seems to be quite different from that of the lecticans. Taken together, we found that SRPX2 is a novel CSPG that is overexpressed in gastrointestinal cancer cells. Our findings provide key glycobiological insight into SRPX2 in cancer cells and demonstrate that SRPX2 is a new member of the cancer-related proteoglycan family.

e19557 Background: Informing the participants about study result is increased demand in recent years. Some studies reported that almost all participants want to receive study result. However, how much information do participants want and how do researcher offer were not mentioned. Moreover there were concerned about resource for providing study result and the attitude regarding informing result to participants in Japan was not studied. The purpose of this study was aimed to discuss and propose the way of informing study results to Japanese participants. Methods: Summarizing study result and a self-administrated questionnaire survey were conducted. Summary was created from abstract of academic paper with lay language and illustration using color. Objective study was Oral Uracil and Tegafur Compared With Classic Cyclophosphamide, Methotrexate, Fluorouracil As Postoperative Chemotherapy in Patients With Node-Negative, High-Risk Breast Cancer: National Surgical Adjuvant Study for Breast Cancer 01 Trial “N-SAS-BC01 trial”(J Clin Oncol 2009). Objective subject was participants of N-SAS-BC01 trial in National Cancer Center Hospital. A cross table and inductive analyze were performed. Results: Summary of result was made A4 (letter size) form 4sheet. 58 patients responded to questionnaire of 116 patients (50%). In 58 responders, 92% of them preferred to be informed of the result. About 80% of responders answered that summary was clear to understand and 30% of them reported that the document should contain more information. The way of receive result preferred by the mail. When the result included bad news, 67% of responders wanted to receive result; on the other hand, three patients answered that they would not to receive because of psychological harmful influence. Conclusions: These answers suggested that participants in Japan desired to inform study result. Therefore, Clinical study result should be provided participants whenever it takes time to get results, including bad news in a careful way. Summary of result should be created using lay language and illustration. Planning the cost, keeping participant contact list, and noticing participants that study result would be informed were needed at the start of the study.

We examined whether antirheumatic drugs alter cytokine- or lipopolysaccharide-induced expression of adhesion molecules on vascular endothelial cells. Human umbilical cord vein endothelial cells were co-cultured with various antirheumatic drugs in the presence of inflammatory cytokines, and adhesion molecule expression was measured by cell enzyme-linked immunosorbent assay and Northern blot analysis. Among these antirheumatic drugs, gold sodium thiomalate significantly inhibited intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 expression on vascular endothelial cells and suppressed cellular binding between human monocytic cell lines, including U937 and HL-60 cells, and interleukin-1 beta-stimulated vascular endothelial cells. It is speculated that down-regulation of adhesion molecules might be one of the novel mechanisms of action of gold sodium thiomalate.

A series of 4-hydroxy-3-quinolinecarboxylic acid derivatives (6) and 4-hydroxy-2-oxo-1,2-dihydro-3-quinolinecarboxylic acid derivatives (7) were designed and synthesized as 5-HT3 receptor antagonists. Molecular modeling studies suggested that the 3-carbonyl moiety in 6 was almost coplanar to the plane of an aromatic ring, but in 7 there was a 30 degrees deviation. 4-Hydroxy substitution in quinoline derivatives enhanced affinity for the 5-HT3 receptors, and endo-N-(8-methyl-8-azabicyclo[3.2.1]oct-3-yl)-4-hydroxy-3- quinolinecarboxamide (6f) exhibited the most potent activity in the Bezold-Jarisch (B-J) reflex test (ED50 = 0.1 micrograms/kg, iv) among quinoline derivatives 6. Although 4-hydroxy-2-oxo-1,2-dihydro-3-quinolinecarboxamide derivatives (7a) exhibited higher affinity (e.g., 7d: Ki = 0.48 nM) for the 5-HT3 receptors than ondansetron (Ki = 7.6 nM) or granisetron (Ki = 2.1 nM), these amides showed less potent activity in the B-J reflex test than the reference compounds. Interestingly, the ester derivatives 7c, 7f, and 7h eliminated affinity for the 5-HT3 receptors. These unusual structure-activity relationships and the deviation of the 3-carbonyl moiety from the plane of an aromatic ring suggest that the active conformation of 7a might be different from the proposed one for the preceding 5-HT3 antagonists. Thus, 6f was chosen for further studies. No receptor binding for a variety of ligands was significantly antagonized by 6f. Comparing the ratios of the ED50 value in the B-J reflex test (rat, iv) with the LD50 value in acute lethal toxicity (mouse, iv), 6f was proved to have a 600-fold wider margin of safety than ondansetron. Compound 6f dose-dependently attenuated both the incidence and frequency of emetic episodes induced by cisplatin in the dog (ED50 = 14 micrograms/kg, iv) more potently than ondansetron (ED50 = 210 micrograms/kg, iv). Compound 6f (KF-20170) is now under further investigation as a drug for treating gastrointestinal disorder.

We characterized the thromboxane A2/prostaglandin H2 receptors in porcine coronary artery. The binding of [3H]SQ 29,548, a thromboxane A2 antagonist, to coronary arterial membranes was saturable and displaceable. Scatchard analysis of equilibrium binding showed a single class of high affinity binding sites with a dissociation constant of 18.5 +/- 1.0 nM and the maximum binding of 80.7 +/- 5.2 fmol/mg protein. [3H]SQ 29,548 binding was concentration-dependently inhibited by thromboxane A2 antagonists such as SQ 29,548, BM13505 and BM13177 or the thromboxane A2 agonists such as U46619 and U44069. KW-3635, a novel dibenzoxepin derivative, concentration-dependently inhibited the [3H]SQ 29,548 binding to thromboxane A2/prostaglandin H2 receptors in coronary artery with an inhibition constant of 6.0 +/- 0.69 nM (mean +/- S.E.M.).

The effect of KW-3635 (sodium (E)-11-[2-(5,6-dimethyl-1-benzimidazolyl) ethylidene]-6,11-dihydrodibenz[b,e] oxepine-2-carboxylate monohydrate, CAS 127166-41-0), a novel thromboxane A2 (TxA2) receptor antagonist, on collagen-induced coronary ischemia was studied in guinea-pigs. Under pentobarbital anaesthesia, intravenous injection (i.v.) of collagen (1 mg/kg) induced abnormal ECG changes such as ST-T changes, elevation of T-wave arrhythmia and cardiac arrest in severe cases. The changes of ECG (leads I, II and III) were recorded for 10 min following collagen injection. KW-3635 (25-50 mg/kg p.o.) remarkably improved the collagen-induced ischemic ECG changes. The effect of KW-3635 was more potent than those of daltroban, isbogrel and ticlopidine. Neither nifedipine nor propranolol had any effect. The plasma thromboxane B2 level in the KW-3635-treated animals was lower in comparison with those in both the control and daltroban-treated animals. These results suggest that TxA2 may play a role in this model of coronary ischemia and that KW-3635 is effective in the treatment of ischemic heart disease.

MEG-01s, an established human megakaryoblastic leukemia cell line, exhibited specific high-affinity binding sites for [3H]iloprost, a stable prostaglandin (PG) I2 analogue, for [3H]SQ-29548, a stable thromboxane (TX) A2 antagonist and, for [3H]PGE2/PGE1, but not for [3H]PGD2. In the MEG-01s cells, iloprost/PGI2, or PGE1 stimulated cAMP production with ED50 values practically identical to the IC50 values for the [3H] iloprost binding. STA2 and U46619, TXA2/PGH2 agonists, PGE2/PGE1, iloprost/PGI2, and thrombin elevated the intracellular concentrations of Ca2+ ([Ca2+]i), as determined by Fura-2 fluorescence signals. Elevation of [Ca2+]i by PGE2/PGE1 and iloprost, but not that by TX-agonists or thrombin, was totally dependent on the presence of extracellular Ca2+. This effect by PGE2/PGE1 was partially inhibited by prior treatment of the cells with islet-activating protein (IAP), while that by TX-agonists or by PGI2/iloprost was not affected. We tentatively conclude from these results that: (1) MEG-01s cells express (a) PGI2/PGE1 receptor(s) coupled to adenylate cyclase and Ca2+ influx, a TXA2/PGH2 receptor coupled to the phosphatidylinositol-turnover-Ca2+ system, and the PGE2/PGE1 receptor coupled to Ca2+ influx; (2) the receptors for TXA2/PGH2 and iloprost and those for PGE2/PGE1 and thrombin are coupled to IAP-insensitive and IAP-sensitive GTP-binding proteins, respectively, and function in a different manner to elevate [Ca2+]i. Thus, the MEG-01s cell line is a pertinent model for studying eicosanoid receptor-mediated signal transduction in platelet/megakaryocyte systems.

The fecundity and egg retention of anadromous female sockeye salmon (Oncorhynchus nerka) collected during 1971–82 from several stocks in British Columbia undergoing controlled fertilization to enhance adult sockeye production were examined. The relationship between egg number and postorbital–hypural length based on 863 females representing 14 stocks was not consistent between all age-types, stocks, and years, probably because of inadequate sample size in some instances. Combined samples, however, revealed a significant positive relationship between postorbital–hypural length and egg number for age 1.2, 1.3, and 2.2 females. Mean absolute fecundity for the respective age-types was 3218, 4125, and 3544 eggs. For samples of 10 or more females, significant stock and annual differences were detected when individual mean absolute fecundity was adjusted to a postorbital–hypural length of 447 mm, but not for females of different age. A comparison of mean fecundities for coastal stocks with historical data for interior British Columbia stocks suggests that coastal stocks are 18% more fecund than interior stocks. Possible causal mechanisms for this regional difference are hypothesized. Examination of 796 carcasses (representing five stocks) for egg retention revealed a range from totally spawned to totally unspawned females, with 56% of the carcasses containing 20 eggs or less and 68% containing 50 eggs or less. The mean egg retention based on all samples combined was estimated to be 6.5% of the mean individual fecundity. This value was reduced to 3.9% when stock means were averaged.

The specific deposited heat or the specific Let transfer number, i e. the ratio of heat atsorbed by the base metal to the weight of deposited metal is measured for various kinds of electrodes, the core diameter being 4 mm The value varies from 12 to 25 Ka' per gram for ordinary arc length.The value is independent of the current employed but increases as the arc length increases, because the melting speed of electrod metal is proportional to the current while the thermal efficiency is independent of the current, and the total heat input increases as the arc length increases (3), (4)It will be higher for those electrodes and polarity which have lower melting speeds.Fig. 3, 4 shov those relations.

Heat transferred to the base metal in arc welding was measured by using two plate widths, namely 35 mm and 11 mm, and the writers attempted to separate the heat into two component parts, one being transferred to a comparatively wide surface of the plate, and the other being delivered to the crater.As both the melting speed of welding rod, and the heat absorbed by the base metal are proportional to the welding current, the heat absorbed can be expressed by equivalent voltage.For example, in case of Shinko B-17 welding rod, when the arc voltage is kept 30 V, total heat absorbed by the base metal is 23.5 V (78%) and 18.5 V (62%), width of plate being 35 mm and 11 mm respectively, while heat delivered to the crater was estimated to be about 16.3 V (54%) (See Fig. 11, Fig. 12, Fig. 17.)Some discussions are given on the papers of Rosenthal (1-2) and also Naka (15-17) on the thermal efficiency of welding arc.

Leukotriene A4 hydrolase was purified to apparent homogeneity from the guinea pig lung. The molecular weight was determined to be 70 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The enzyme exhibited two active forms with different pI values (5.7 and 5.4) depending on the presence or absence of SH-reducing reagents during purification procedures. No significant differences were observed between both forms of the enzyme as regards the catalytic properties. The N-terminal 20 amino acid sequence (PEVVDTXSLASPATVXRTKH) showed a 90% identity to the human enzyme with a constitutive substitution of Ile-3 and Ser-14 (human) by Val-3 and Thr-14 (guinea pig), respectively.

11(S),12(S)-oxido-5Z,7E,9E,14Z-eicosatetraenoic acid (11,12-LTA4) was chemically synthesized from 12-hydroperoxy-eicosatetraenoic acid (12-HPETE). 11,12-LTA4 methyl ester was nonenzymically hydrolyzed to at least four products. These products were identified to be epimers of 5,12(S)-dihydroxy-eicosatetraenoic acid methyl ester (5,12(S)-diHETE-Me) and epimers of 11,12-diHETE-Me. In addition to the above nonenzymic products, 11,12-LTA4 was converted to 11,12-LTC4 by the rat liver glutathione S-transferase, and to an isomer of 11,12-diHETE by homogenates of the guinea pig adrenal gland and liver.