Hokkaido System Science Ltd. Co.

Hokkaido, Japan

Hokkaido System Science Ltd. Co.

Hokkaido, Japan
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Kozuka M.,Hokkaido University | Murao S.,Hokkaido University | Yamane T.,Hokkaido University | Inoue T.,Hokkaido System Science Co. | And 2 more authors.
Journal of Nutritional Science and Vitaminology | Year: 2015

Lysozyme (EC 3.2.1. 17) is a hydrolytic enzyme that cleaves the β-(1, 4)-glycosidic bond between N-acetylmuramic acid and N-acetylglucosamine in peptidoglycan, a major bacterial cell wall polymer. In the food industry, lysozyme is used as an additive mainly in the production of wine and beer. Lysozyme was found to be localized in the egg shell membrane. In this study, we found that lysozyme was easily purified from the egg shell membrane and that the enzyme also had antibacterial activity. Furthermore, we found that the antibacterial activity of purified lysozyme from the egg shell membrane was lower than that of purified lysozyme from the egg white at alkaline pH. The method for rapid purification of lysozyme developed in this study should contribute to the food industry. © 2015, Center for Academic Publications Japan. All rights reserved.


Yamane T.,Hokkaido University | Yamane T.,Shiga University of Medical Science | Murao S.,Hokkaido University | Kato-Ose I.,Hokkaido University | And 8 more authors.
Biochemical and Biophysical Research Communications | Year: 2013

Legumain (EC 3.4.22.34) is an asparaginyl endopeptidase. Strong legumain activity was observed in the mouse kidney, and legumain was found to be highly expressed in tumors. We previously reported that bovine kidney annexin A2 was co-purified with legumain and that legumain cleaved the N-terminal region of annexin A2 at an Asn residue in vitro and in vivo. In this study, we found a p53-binding site in intron 1 of the human legumain gene using computational analysis. To determine whether transcription of the legumain gene is regulated by p53, HCT116 cells were transfected with p53 siRNA and the effect of knockdown of p53 expression on legumain expression was examined. The results showed that expression levels of both legumain mRNA and protein were decreased in the siRNA-treated cells. Furthermore, enzyme activity of legumain was also increased by doxorubicin and its activity was reduced by knockdown of p53 in HCT116 cells. These results suggest that legumain expression and its enzyme activity are regulated by p53. © 2013 Elsevier Inc.


Patent
Daiichi Fine Chemical Co. and Hokkaido System Science Co. | Date: 2011-09-29

The invention provides a nucleic-acid-transfecting composition which exhibits low cytotoxicity, which facilitates an effective nucleic acid transfection into a cell, and which improves expression of the nucleic acid in the cell. The composition for transfecting a nucleic acid into a cell, contains a di(C_(12-16 )alkyl)dimethylammonium halide and a phospholipid.


Imamura S.,Chiyoda Corporation | Haruna M.,Chiyoda Corporation | Goshima T.,Chiyoda Corporation | Kanezashi H.,Chiyoda Corporation | And 2 more authors.
Foodborne Pathogens and Disease | Year: 2016

The development of procedures for the efficient removal or inactivation of noroviruses from contaminated oysters is of great interest in oyster production. However, there is a critical limitation for evaluating the depuration efficacy of presently available procedures, as no suitable cell culture system currently exists to cultivate noroviruses. Thus, we applied a next-generation sequencing (NGS) technique to characterize norovirus genotypes in pre- and post-depurated oysters. As a result, we revealed the diversity of noroviruses in pre- and post-depurated oysters. Although the applied depuration procedure could reduce the number of bacterial agents to the level recommended by the Japanese Ministry of Health, Labour and Welfare, no significant changes were observed in the detection rate and the proportion of norovirus group (G) I and GII genotypes. To our knowledge, this is the first report to evaluate the profile of noroviruses in pre- and post-depurated oysters, specifically with respect to norovirus removal, using NGS; the findings imply that the removal of noroviruses from oysters through depuration is not presently sufficient. Further studies are needed to develop a more suitable depuration procedure for removing and/or inactivating noroviruses from contaminated oysters. © Mary Ann Liebert, Inc. 2016.


Watanabe Y.,RIKEN | Watanabe Y.,University of Tsukuba | Numata K.,RIKEN | Murata S.,Keio University | And 10 more authors.
Genomics | Year: 2010

The functionality of sense-antisense transcripts (SATs), although widespread throughout the mammalian genome, is largely unknown. Here, we analyzed the SATs expression and its associated promoter DNA methylation status by surveying 12 tissues of mice to gain insights into the relationship between expression and DNA methylation of SATs. We have found that sense and antisense expression positively correlate in most tissues. However, in some SATs with tissue-specific expression, the expression level of a transcript from a CpG island-bearing promoter is low when the promoter DNA methylation is present. In these circumstances, the expression level of its opposite-strand transcript, especially when it is poly(A)-negative was coincidentally higher. These observations suggest that, albeit the general tendency of sense-antisense simultaneous expression, some antisense transcripts have coordinated expression with its counterpart sense gene promoter methylation. This cross-strand relationship is not a privilege of imprinted genes but seems to occur widely in SATs. © 2010 Elsevier Inc.


Matsuura H.,Aichi University of Technology | Hokonohara H.,Osaka University | Sugita T.,Hokkaido System Science Co. | Takagi A.,Osaka University | And 3 more authors.
Journal of Applied Physics | Year: 2011

This paper investigates the roles of a solution (decane) in deoxyribonucleic acid (DNA) observation with a scanning tunneling microscope. Our study indicates that decane prevents continuous water adsorption from air and subsequent ionization of the water to create specific conditions for DNA observation. Analysis of the tunneling current reveals that the current with decane became twice as stable in deviation and the current is sustained 1 nm further in the z-direction than without decane. The apparent barrier height with decane is also decreased by a factor of 0.18. These properties enable us to measure bulky DNA (4 nm) at the highest success ratio ever attained. © 2011 American Institute of Physics.


Kim S.,Tokyo University of Agriculture and Technology | Matsumoto M.,Hokkaido System Science Co. | Chiba K.,Tokyo University of Agriculture and Technology
Chemistry - A European Journal | Year: 2013

Recent progress in the RNA therapeutics has increased demand for the synthesis of large quantities of oligoribonucleotides. The assembly of RNA oligomers relies mainly on solid-phase approaches. These allow rapid product purification and the ability to drive a target reaction to completion through the use of excess reagents. Despite the known advantages of solid-phase synthesis, some issues in the process remain to be addressed, such as low and limited scale, reagent accessibility, and the use of a very large excess of reagents. Herein, we report a highly efficient and practical method of liquid-phase synthesis of RNA oligomers by using alkyl-chain-soluble support. We demonstrate the utility of the liquid-phase method through 21-mer RNA synthesis on a gram scale. The assembly of RNA oligomers relies principally on solid-phase approaches, although some alternative methods have been developed to date. A highly efficient and practical method of liquid-phase synthesis for RNA oligomers by using an alkyl-chain-type soluble support is reported. The utility of the liquid-phase method through 21-mer RNA synthesis on a gram scale is described (see scheme). Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.


Yamane T.,Hokkaido University | Hachisu R.,Hokkaido System Science Co. | Yuguchi M.,Hokkaido System Science Co. | Takeuchi K.,Shiga University of Medical Science | And 6 more authors.
Biochemical and Biophysical Research Communications | Year: 2013

Legumain (EC 3.4.22.34) is an asparaginyl endopeptidase. Strong legumain activity was observed in the mouse kidney, and legumain was highly expressed in tumors. We previously reported that bovine kidney annexin A2 was co-purified with legumain and that legumain cleaved the N-terminal region of annexin A2 at an Asn residue in vitro. In this study, to determine whether annexin A2 is cleaved by legumain in vivo, siRNA-lipoplex targeting mouse legumain was injected into mouse tail veins. Mouse kidneys were then isolated and the effect of knockdown of legumain expression on annexin A2 cleavage was examined. The results showed that both legumain mRNA and protein expression levels were decreased in the siRNA-treated mouse kidneys and that legumain activity toward a synthetic substrate, Z-Ala-Ala-Asn-MCA, was decreased by about 40% in the kidney but not in the liver or spleen. Furthermore, cleavage of annexin A2 at the N-terminal region was decreased in the mouse kidney that had been treated with the legumain siRNA-lipoplex. These results suggest that legumain siRNA was delivered to the kidney by using LipoTrust and that the reduced legumain expression inhibited legumain-induced degradation of annexin A2 in vivo. © 2012 Elsevier Inc.


Patent
Hokkaido System Science Co. | Date: 2015-04-08

A nucleic acid synthesis method enabling a reaction in a fluid (flow) with a highly dispersible liquid-phase support to improve coupling efficiency is provided. The method for synthesizing an oligonucleotide comprising: sequentially condensing and oxidizing a nucleoside phosphoramidite compound in the presence of an acid/azole complex compound using a starting raw material, i.e., hydrophobic group-bonded nucleoside represented by Formula (1):where R^(1): an alkylene group having 1 to 12 carbon atoms,R^(2): an alkylene group having 1 to 22 carbon atoms, R^(3) andR^(4) each independently represent an alkyl group having 1 to 22 carbon atoms or the like, R^(5): a single bond or an alkylene group having 1 to 22 carbon atoms, R^(6): each independently an alkyl group having 6 to 30 carbon atoms, n represents an integer of 2 to 6, X represents a hydrogen atom, hydroxyl group, or the like, Y: a protecting group deprotectable under an acidic condition, and Z: an adenyl group, a guanyl group, or the like having a polar group optionally protected by a protecting group,wherein a condensation reaction is performed by preliminarily dissolving the hydrophobic group-bonded nucleoside or hydrophobic group-bonded oligonucleotide and the nucleoside phosphoramidite compound in a nonpolar solvent, and contacting the resulting solution with the acid/azole complex compound or a solution containing the complex compound.


Patent
Hokkaido System Science Co. | Date: 2012-05-30

A nucleic acid synthesis method enabling a reaction in a fluid (flow) with a highly dispersible liquid-phase support to improve coupling efficiency is provided. The method for synthesizing an oligonucleotide comprising: sequentially condensing and oxidizing a nucleoside phosphoramidite compound in the presence of an acid/azole complex compound using a starting raw material, i.e., hydrophobic group-bonded nucleoside represented by Formula (1): where R^(1): an alkylene group having 1 to 12 carbon atoms, R^(2): an alkylene group having 1 to 22 carbon atoms, R^(3 )and R^(4 )each independently represent an alkyl group having 1 to 22 carbon atoms or the like, R^(5): a single bond or an alkylene group having 1 to 22 carbon atoms, R^(6): each independently an alkyl group having 6 to 30 carbon atoms, n represents an integer of 2 to 6, X represents a hydrogen atom, hydroxyl group, or the like, Y: a protecting group deprotectable under an acidic condition, and Z: an adenyl group, a guanyl group, or the like having a polar group optionally protected by a protecting group,

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