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Structural characteristics and refolding of in vivo aggregated hyperthermophilic archaeon proteins

Mitsuo Umetsu, Kouhei Tsumoto, Kumar Ashish, Shigeki Nitta, Yoshikazu Tanaka, Tadafumi Adschiri, Izumi Kumagai
2003 FEBS Letters  
Several recombinant proteins in inclusion bodies expressed in Escherichia coli have been measured by Fourier transform infrared and solid-state nuclear magnetic resonance spectra to provide the secondary structural characteristics of the proteins from hyperthermophilic archaeon Pyrococcus horikoshii OT3 (hyperthermophilic proteins) in inclusion bodies. The L L-strand-rich single chain Fv fragment (scFv) and K K-helix-rich interleukin (IL)-4 lost part of the native-like secondary structure in
more » ... lusion bodies, while the inclusion bodies composed of the hyperthermophilic proteins of which the native form is K K-helix rich, are predominated by K K-helix structure. Further, the secondary structure of the recombinant proteins solubilized from inclusion bodies by detergent or denaturant was observed by circular dichroism (CD) spectra. The solubilization induced the denaturation of the secondary structure for scFv and IL-4, whereas the solubilized hyperthermophilic proteins have retained the K K-helix structure with the CD properties resembling those of their native forms. This indicates that the hyperthermophilic proteins form native-like secondary structure in inclusion bodies. Refolding of several hyperthermophilic proteins from in vivo aggregated form without complete denaturation could be accomplished by solubilization with lower concentration (e.g. 2 M) of guanidine hydrochloride and removal of the denaturant via stepwise dialysis. This supports the existence of proteins with nativelike structure in inclusion bodies and suggests that non-native association between the secondary structure elements leads to in vivo aggregation. We propose a refolding procedure on the basis of the structural properties of the aggregated archaeon proteins.
doi:10.1016/s0014-5793(03)01441-8 pmid:14741340 fatcat:ofsws5hfvndpldtaix5ima7eoe

IAN Family Critically Regulates Survival and Development of T Lymphocytes

Takeshi Nitta, Mariam Nasreen, Takafumi Seike, Atsushi Goji, Izumi Ohigashi, Tadaaki Miyazaki, Tsutomu Ohta, Masamoto Kanno, Yousuke Takahama, Philippa Marrack
2006 PLoS Biology  
Citation: Nitta T, Nasreen M, Seike T, Goji A, Ohigashi I, et al. (2006) IAN family critically regulates survival and development of t lymphocytes. PLoS Biol 4(4): e103.  ... 
doi:10.1371/journal.pbio.0040103 pmid:16509771 pmcid:PMC1393758 fatcat:byxh2kijrnd7zgqnmbjjhdnkda

Development of Cast Reinforced Aluminum Alloy Matrix Composite Belt Conveyor Roller

Hiroyuki Omura, Shin Nitta, Izumi Murashima, Yosuke Takahashi, Naomi Nishi
1998 Journal of Japan Foundry Engineering Society  
doi:10.11279/jfes.70.213 fatcat:kragw3oqcjfo3bl2pcd7e3v6ru

A volume diagnosis method for identifying systematic faults in lower-yield wafer occurring during mass production

Tsutomu Ishida, Izumi Nitta, Koji Banno, Yuzi Kanazawa
2014 2014 19th Asia and South Pacific Design Automation Conference (ASP-DAC)  
doi:10.1109/aspdac.2014.6742968 dblp:conf/aspdac/IshidaNBK14 fatcat:p4tb2amhvzhnvbmbnkoj3bse7m

Non-Gaussian Statistical Timing models of die-to-die and within-die parameter variations for full chip analysis

Katsumi Homma, Izumi Nitta, Toshiyuki Shibuya
2008 2008 Asia and South Pacific Design Automation Conference  
Statistical Timing Analysis (SSTA) is a method that calculates circuit delay statistically with process parameter variations, die-to-die (D2D) and within-die (WID) variations. In this paper, we model that WID parameter variations are independent for each cell and line in a chip and D2D variations are governed by one variation on a chip. We propose a new method of computing a full chip delay distribution considering both D2D and WID parameter variations. Experimental results show that the
more » ... d method is more accurate than previous methods on actual chip designs.
doi:10.1109/aspdac.2008.4483961 dblp:conf/aspdac/HommaNS08 fatcat:m6ud35fpyrbjzcv6fhslvjxs3i

Expression of a synthetic gene encoding canine milk lysozyme in Escherichia coli and characterization of the expressed protein

Takumi Koshiba, Tomohiro Hayashi, Ishido Miwako, Izumi Kumagai, Teikichi Ikura, Keiichi Kawano, Katsutoshi Nitta, Kunihiro Kuwajima
1999 Protein Engineering Design & Selection  
Our previous studies have also revealed, however, that there is a group of calcium-binding lysozymes, although only several such examples have been identified at present (Nitta et al., 1987; Nitta and  ...  Among the known examples of calcium-binding lysozymes, only canine and equine milk lysozymes are known to show a stable equilibrium molten globule state (Nitta et al., 1993; Van Dael et al., 1993; Griko  ... 
doi:10.1093/protein/12.5.429 pmid:10360984 fatcat:yhwjsqyuabgejgu2bl52x4unh4

Evaluation of Trace Elements in Augmentation of Statin-Induced Cytotoxicity in Uremic Serum-Exposed Human Rhabdomyosarcoma Cells

Hitoshi Uchiyama, Masayuki Tsujimoto, Naomi Shimada, Koji Tsutsui, Ayaka Nitta, Takuya Yoshida, Taku Furukubo, Satoshi Izumi, Tomoyuki Yamakawa, Hidehisa Tachiki, Tetsuya Minegaki, Kohshi Nishiguchi
2018 Toxins  
Patients with end-stage kidney disease (ESKD) are at higher risk for rhabdomyolysis induced by statin than patients with normal kidney function. Previously, we showed that this increase in the severity of statin-induced rhabdomyolysis was partly due to uremic toxins. However, changes in the quantity of various trace elements in ESKD patients likely contribute as well. The purpose of this study is to determine the effect of trace elements on statin-induced toxicity in rhabdomyosarcoma cells
more » ... ed to uremic serum (US cells) for a long time. Cell viability, apoptosis, mRNA expression, and intracellular trace elements were assessed by viability assays, flow cytometry, real-time RT-PCR, and ICP-MS, respectively. US cells exhibited greater simvastatin-induced cytotoxicity than cells long-time exposed with normal serum (NS cells) (non-overlapping 95% confidence intervals). Intracellular levels of Mg, Mn, Cu, and Zn were significantly less in US cells compared to that in NS cells (p < 0.05 or 0.01). Pre-treatment with TPEN increased simvastatin-induced cytotoxicity and eliminated the distinction between both cells of simvastatin-induced cytotoxicity. These results suggest that Zn deficiencies may be involved in the increased risk for muscle complaints in ESKD patients. In conclusion, the increased severity of statin-induced rhabdomyolysis in ESKD patients may be partly due to trace elements deficiencies. Key contribution: Our results highlighted deficiencies of various trace elements such Mg, Mn, Cu, and Zn are potential cause of the increased statin-induced cytotoxicity in ESKD patients.
doi:10.3390/toxins10020053 pmid:29370118 pmcid:PMC5848154 fatcat:gvzuqmqkkrfc3kkeibylzr4qp4

C5a receptor (CD88) promotes motility and invasiveness of gastric cancer by activating RhoA

Takayoshi Kaida, Hidetoshi Nitta, Yuki Kitano, Kensuke Yamamura, Kota Arima, Daisuke Izumi, Takaaki Higashi, Junji Kurashige, Katsunori Imai, Hiromitsu Hayashi, Masaaki Iwatsuki, Takatsugu Ishimoto (+7 others)
2016 OncoTarget  
Purpose: Anaphylatoxin C5a is a strong chemoattractant of the complement system that binds the C5a receptor (C5aR). The expression of C5aR is associated with poor prognosis in several cancers. However, the role of C5aR in gastric cancer (GC) is unknown. The aim of this study was to examine the role of C5aR on GC cell motility and invasion. Experimental Design: The mechanism of invasion via C5aR was assessed by analyzing cytoskeletal rearrangement and RhoA activity after C5a treatment. Moreover,
more » ... we investigated the relationship between C5aR expression and the prognosis of GC patients. Results: Two human GC cell lines (MKN1 and MKN7) had high C5aR expression. An invasion assay revealed that C5a stimulation promoted the invasive ability of MKN1 and MKN7 cells and that this was suppressed by knockdown of C5aR using siRNA or a C5aR-antagonist. Moreover, overexpression of C5aR in GC cells enhanced the conversion of RhoA-guanosine diphosphate (RhoA-GDP) to RhoA-guanosine triphosphate (RhoA-GTP) after C5a stimulation and caused morphological changes, including increased expression of stress fibers and filopodia. Examination of tumor specimens from 100 patients with GC revealed that high C5aR expression (35 of 100 samples, 35.0%) was associated with increased invasion depth, vascular invasion and advanced stage. The 5-year overall survival of patients with high or low C5aR expression was 58.2% and 88.5%, respectively (p=0.008). Conclusions: This study is the first to demonstrate that C5aR promotes GC cell invasion by activating RhoA and is associated with a poor prognosis in GC patients. Therefore, this study provides a biomarker for GC patients who require an advanced therapeutic strategy.
doi:10.18632/oncotarget.12656 pmid:27756879 pmcid:PMC5356699 fatcat:za2u36hsavbjjiqjfaqarf6hvy


Yuki Tsuruta, Yuko Sasaki, Makiko Izumi, Misako Miyashita, Yukio Tsuruta, Mitsuyo Itabashi, Takashi Takei, Ken Tsuchiya, Kousaku Nitta
, Misako Miyashita, Yukio Tsuruta 2 )東京都健康長寿医療センター 腎臓内科 Yuko Sasaki, Mitsuyo Itabashi, Takashi Takei 3 )東京女子医科大学 第4内科 Kousaku Nitta 4 )東京女子医科大学 血液浄化療法科 Ken Tsuchiya  ...  たのに対し,ベンズブロマロン投与で同様に尿酸 値を低下させても血管内皮機能は改善しなかった ことが報告されている 5) .XO 阻害薬が尿酸値を 介さずに血管内皮機能を改善させる可能性が示唆 されており,今後リスク集団である血液透析患者 において,XO 阻害薬が心血管疾患などに与える 1 )医療法人社団敬天会 鶴田板橋クリニック Yuki Tsuruta, Makiko Izumi  ... 
doi:10.6032/gnam.41.33 fatcat:i7pxifjabraohla25lm4sh53nq

Ultrasound-guided central venous catheterization via distal femoral vein (a case report)

Joho Tokumine, Kenichi Nitta, Koji Teruya, Tatsuya Higa, Ayako Haga, Junichi Okubo, Shunsuke Izumi, Kazuhiro Sugahara
2008 Journal of the Japanese Society of Intensive Care Medicine  
Ultrasound-guided central venous catheterization via distal femoral vein (a case report
doi:10.3918/jsicm.15.79 fatcat:r2bgj5zcfzfstnozftjzyfmagu

Association of gene expression involving innate immunity and genetic variation in interleukin 28B with antiviral response

Yasuhiro Asahina, Kaoru Tsuchiya, Masaru Muraoka, Keisuke Tanaka, Yuichiro Suzuki, Nobuharu Tamaki, Yoshihide Hoshioka, Yutaka Yasui, Tomoji Katoh, Takanori Hosokawa, Ken Ueda, Hiroyuki Nakanishi (+7 others)
2011 Hepatology  
Innate immunity plays an important role in host antiviral response to hepatitis C viral (HCV) infection. Recently, single nucleotide polymorphisms (SNPs) of IL28B and host response to peginterferon a (PEG-IFNa) and ribavirin (RBV) were shown to be strongly associated. We aimed to determine the gene expression involving innate immunity in IL28B genotypes and elucidate its relation to response to antiviral treatment. We genotyped IL28B SNPs (rs8099917 and rs12979860) in 88 chronic hepatitis C
more » ... ents treated with PEG-IFNa-2b/RBV and quantified expressions of viral sensors (RIG-I, MDA5, and LGP2), adaptor molecule (IPS-1), related ubiquitin E3-ligase (RNF125), modulators (ISG15 and USP18), and IL28 (IFNk). Both IL28B SNPs were 100% identical; 54 patients possessed rs8099917 TT/rs12979860 CC (IL28B major patients) and 34 possessed rs8099917 TG/rs12979860 CT (IL28B minor patients). Hepatic expressions of viral sensors and modulators in IL28B minor patients were significantly up-regulated compared with that in IL28B major patients (%3.3-fold, P < 0.001). However, expression of IPS-1 was significantly lower in IL28B minor patients (1.2-fold, P 5 0.028). Expressions of viral sensors and modulators were significantly higher in nonvirological responders (NVR) than that in others despite stratification by IL28B genotype (%2.6-fold, P < 0.001). Multivariate and ROC analyses indicated that higher RIG-I and ISG15 expressions and RIG-I/IPS-1 expression ratio were independent factors for NVR. IPS-1 down-regulation in IL28B minor patients was confirmed by western blotting, and the extent of IPS-1 protein cleavage was associated with the variable treatment response. Conclusion: Gene expression involving innate immunity is strongly associated with IL28B genotype and response to PEG-IFNa/ RBV. Both IL28B minor allele and higher RIG-I and ISG15 expressions and RIG-I/IPS-1 ratio are independent factors for NVR. (HEPATOLOGY 2012;55:20-29) Abbreviations: CH-C, chronic hepatitis C; c-GTP, c-glutamyl transpeptidase; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; HCV, hepatitis C virus; HMBS, hydroxymethylbilane synthase; IL28, interleukin 28; IPS-1, IFNb promoter stimulator 1; ISG15, interferon-stimulated gene 15; MDA5, melanoma differentiation associated gene 5; NVR; nonvirological responders; PEG-IFNa, pegylated interferona; SNP, single nucleotide polymorphism; RIG-I, retinoic acidinducible gene I; RBV, ribavirin; RNF125, ring-finger protein 125; ROC, receiver operator characteristic; SVR, sustained viral responder; TVR, transient virological responder; USP18, ubiquitin-specific protease 18; VR, virological responder. From the
doi:10.1002/hep.24623 pmid:21898478 fatcat:cs73b5do4jfijlsyjs3s2pdwge

The Cytokine RANKL Produced by Positively Selected Thymocytes Fosters Medullary Thymic Epithelial Cells that Express Autoimmune Regulator

Yu Hikosaka, Takeshi Nitta, Izumi Ohigashi, Kouta Yano, Naozumi Ishimaru, Yoshio Hayashi, Mitsuru Matsumoto, Koichi Matsuo, Josef M. Penninger, Hiroshi Takayanagi, Yoshifumi Yokota, Hisakata Yamada (+4 others)
2008 Immunity  
Nitta is acknowledged. This study was supported by MEXT Grant-in-Aid for Scientific Research on Priority Areas ''Immunological Self'' and the Mitsubishi Foundation.  ...  Retrovirus Infection PCR-cloned cDNA fragments encoding an open-reading frame of mouse RANKL, OX40L, CD30L, CD40L, or FasL were cloned in the retrovirus vector pMSCV-IRES-EGFP (Nitta et al., 2006) .  ...  selection, we surveyed oligonucleotide microarray data from our previous study where we compared gene-expression profiles between positively selected TCR-transgenic thymocytes and wild-type thymocytes (Nitta  ... 
doi:10.1016/j.immuni.2008.06.018 pmid:18799150 fatcat:ffsz6l6pdfaijakk45nqqi5uta

Short-Term Effects of Acupuncture on Open-Angle Glaucoma in Retrobulbar Circulation: Additional Therapy to Standard Medication

Shin Takayama, Takashi Seki, Toru Nakazawa, Naoko Aizawa, Seri Takahashi, Masashi Watanabe, Masayuki Izumi, Soichiro Kaneko, Tetsuharu Kamiya, Ayane Matsuda, Akiko Kikuchi, Tomoyuki Yambe (+3 others)
2011 Evidence-Based Complementary and Alternative Medicine  
Background. The relation between glaucoma and retrobulbar circulation in the prognosis has been indicated.Purpose. To investigate the effects of acupuncture on retrobulbar circulation in open-angle glaucoma (OAG) patients.Methods. Eleven OAG patients (20 eyes with OAG) who were treated by topical antiglaucoma medications for at least 3 months were enrolled. Acupuncture was performed once at acupoints BL2, M-HN9, ST2, ST36, SP6, KI3, LR3, GB20, BL18, and BL23 bilaterally. Retrobulbar circulation
more » ... was measured with color Doppler imaging, and intraocular pressure (IOP) was also measured at rest and one hour after rest or before and after acupuncture.Results. TheΔvalue of the resistive index in the short posterior ciliary artery (P<.01) and theΔvalue of IOP (P<.01) were decreased significantly by acupuncture compared with no acupuncture treatment.Conclusions. Acupuncture can improve the retrobulbar circulation and IOP, which may indicate the efficacy of acupuncture for OAG.
doi:10.1155/2011/157090 pmid:21437193 pmcid:PMC3062145 fatcat:r4vktmpslfev7b3gdhsr6xxo4u

Aire-dependent production of XCL1 mediates medullary accumulation of thymic dendritic cells and contributes to regulatory T cell development

Yu Lei, Adiratna Mat Ripen, Naozumi Ishimaru, Izumi Ohigashi, Takashi Nagasawa, Lukas T. Jeker, Michael R. Bösl, Georg A. Holländer, Yoshio Hayashi, Rene de Waal Malefyt, Takeshi Nitta, Yousuke Takahama
2011 Journal of Experimental Medicine  
., 2006; Nitta et al., 2009) . Thus, instead of CCR7 and CXCR4, the XCL1-XCR1 chemokine axis may play a major role in regulating the medullary accumulation of tDCs.  ...  negative selection, particularly in the intrathymic presentation of peripheral antigens, by cooperating with promiscuous gene expression in mTECs (Gallegos and Bevan, 2004; Koble and Kyewski, 2009; Nitta  ... 
doi:10.1084/jem.20102327 pmid:21300913 pmcid:PMC3039864 fatcat:vrgp54retfdzzeco3lou755wgu

Experimental study of sympathetic nerve activity in animal with total artificial heart

1991 Jinko Zoki  
Yambe T, Nitta S, Katahira Y, Sonobe T, Naganuma S, Akiho H, Kakinuma Y, Izutsu K, Sugawara S, Tanaka M, Miura M,* Sato N,* Mohri H,* Yoshizawa M,** Izumi K,*** Takeda H*** Experimental study of sympathetic  ... 
doi:10.11392/jsao1972.20.1434 fatcat:4mkkskw355g2jojtau4jfuyqge
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