Tropical Technology Center Ltd

Uruma, Japan

Tropical Technology Center Ltd

Uruma, Japan

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Zheng H.-J.,Shanghai Institute of Planned Parenthood Research | Zheng H.-J.,Chinese National Human Genome Center at Shanghai | Tsukahara M.,Tropical Technology Center Ltd. | Liu E.,Shanghai Institute of Planned Parenthood Research | And 6 more authors.
Stem Cells and Development | Year: 2015

The gene trap method for embryonic stem cells is an efficient method for identifying new genes that are involved in development. Using this method, we identified a novel gene called helicase family gene related to gastrulation (helG). Helicase family proteins regulate many systems in the body that are related to cell survival. HelG encodes a protein of 137kDa, which contains a DExH helicase motif that is now named DHX30. HelG is strongly expressed in neural cells (ie, in the headfold, neural plate, neural tube, and brain) and somites during embryogenesis. Growing homozygous mutant embryos have neither differentiated somites nor brains. In these mutants, development was retarded by embryonic day 7.5 (E7.5), and the mutants died at E9.5. After the purification of HelG, an untwisting experiment was performed to confirm the helicase activity of HelG for DNA in vitro. We report for the first time that a helicase family gene is required for differentiation during embryogenesis; this gene might interact with polynucleotides to regulate some genes that are important for early development and has a structure similar to that of a human DExH box helicase. © Copyright 2015, Mary Ann Liebert, Inc.


Yasumoto K.,Kitasato University | Yasumoto-Hirose M.,Tropical Technology Center Ltd | Yasumoto J.,University of Ryukyus | Murata R.,Kitasato University | And 7 more authors.
Marine Biotechnology | Year: 2014

Bacteria, including cyanobacteria, as well as some fungi, are known to deposit calcium carbonate (CaCO3) extracellularly in calcium-containing artificial medium. Despite extensive investigation, the mechanisms involved in extracellular formation of CaCO3 by bacteria have remained unclear. The ability of synthetic amines to remove carbon dioxide (CO2) from natural gas led us to examine the role of biogenic polyamines in CaCO3 deposition by bacteria. Here, we demonstrated that biogenic polyamines such as putrescine, spermidine, and spermine were able to react with atmospheric CO2 and the resultant carbamate anion was characterized by using nuclear magnetic resonance (NMR) analysis. Biogenic polyamines accelerated the formation of CaCO3, and we artificially synthesized the dumbbell-shaped calcites, which had the same form as observed with bacterial CaCO3 precipitates, under nonbacterial conditions by using polyamines. The reaction rate of calcification increased with temperature with an optimum of around 40 °C. Our observation suggests a novel scheme for CO2 dissipation that could be a potential tool in reducing atmospheric CO2 levels and, therefore, global warming. © 2014 Springer Science+Business Media New York.


Umemura M.,Japan National Institute of Advanced Industrial Science and Technology | Koike H.,Japan National Institute of Advanced Industrial Science and Technology | Yamane N.,Japan National Institute of Advanced Industrial Science and Technology | Koyama Y.,Japan National Institute of Advanced Industrial Science and Technology | And 13 more authors.
DNA Research | Year: 2012

Aspergillus oryzae has been utilized for over 1000 years in Japan for the production of various traditional foods, and a large number of A. oryzae strains have been isolated and/or selected for the effective fermentation of food ingredients. Characteristics of genetic alterations among the strains used are of particular interest in studies of A. oryzae. Here, we have sequenced the whole genome of an industrial fungal isolate, A. oryzae RIB326, by using a next-generation sequencing system and compared the data with those of A. oryzae RIB40, a wild-type strain sequenced in 2005. The aim of this study was to evaluate the mutation pressure on the non-syntenic blocks (NSBs) of the genome, which were previously identified through comparative genomic analysis of A. oryzae, Aspergillus fumigatus, and Aspergillus nidulans. We found that genes within the NSBs of RIB326 accumulate mutations more frequently than those within the SBs, regardless of their distance from the telomeres or of their expression level. Our findings suggest that the high mutation frequency of NSBs might contribute to maintaining the diversity of the A. oryzae genome. © 2012 The Author.


Kuba-Miyara M.,University of Ryukyus | Agarie K.,University of Ryukyus | Sakima R.,Tropical Technology Center Ltd | Imamura S.,Tropical Technology Center Ltd | And 5 more authors.
International Immunopharmacology | Year: 2012

Degranulation inhibitors in plants are widely used for prevention and treatment of immediate-type allergy. We previously isolated a new ellagic acid glucoside, okicamelliaside (OCS), from Camellia japonica leaves for use as a potent degranulation inhibitor. Crude extracts from leaves also suppressed allergic conjunctivitis in rats. In this study, we evaluated the in vivo effect of OCS using a pure sample and performed in vitro experiments to elucidate the mechanism underlying the extraordinary high potency of OCS and its aglycon. The IC 50 values for degranulation of rat basophilic leukemia cells (RBL-2H3) were 14 nM for OCS and 3 μM for aglycon, indicating that the two compounds were approximately 2 to 3 orders of magnitude more potent than the anti-allergic drugs ketotifen fumarate, DSCG, and tranilast (0.17, 3, and > 0.3 mM, respectively). Antigen-induced calcium ion (Ca 2+) elevation was significantly inhibited by OCS and aglycon at all concentrations tested (p < 0.05). Upstream of the Ca 2+ elevation in the principle signaling pathway, phosphorylation of Syk (Tyr525/526) and PLCγ-1 (Tyr783 and Ser1248) were inhibited by OCS and aglycon. In DNA microarray-screening test, OCS inhibited expression of proinflammatory cytokines [interleukin (IL)-4 and IL-13], cytokine-producing signaling factors, and prostaglandin-endoperoxidase 2, indicating that OCS broadly inhibits allergic inflammation. During passive cutaneous anaphylaxis in mice, OCS significantly inhibited vascular hyperpermeability by two administration routes: a single intraperitoneal injection at 10 mg/kg and per os at 5 mg/kg for 7 days (p < 0.05). These results suggest the potential for OCS to alleviate symptoms of immediate-type allergy. © 2012 Elsevier B.V. All rights reserved.


Patent
TROPICAL TECHNOLOGY CENTER Ltd. and University of Ryukyus | Date: 2010-04-06

A therapeutic agent comprising fucoxanthin or fucoxanthinol as an active component is disclosed. The therapeutic agent is effective and high clinical utility for medical treatment and prevention of virus-associated malignancy such as adult T-cell leukemia and Burkitt lymphoma.


Yamada O.,Japanese National Research Institute of Brewing | Takara R.,Tropical Technology Center Co. | Hamada R.,Japanese National Research Institute of Brewing | Hayashi R.,Japanese National Research Institute of Brewing | And 2 more authors.
Journal of Bioscience and Bioengineering | Year: 2011

To assess the position of Kuro-Koji molds in black Aspergillus, we performed sequence analysis of approximately 2500 nucleotides of partial gene fragments, such as histone 3, on a total of 57 Aspergillus strains, including Aspergillus kawachii NBRC 4308, 12 Kuro-Koji molds isolated from awamori breweries in Japan, Aspergillus niger ATCC 1015, and A. tubingensis ATCC10550. Sequence results showed that all black Aspergillus strains could be classified into 3 types, type N which includes A. niger ATCC 1015, type T which includes A. tubingensis ATCC 10550, and type L which includes A. kawachii NBRC 4308. Phylogenetic analysis showed these three types belong to different clusters. All 12 Kuro-Koji molds isolated from awamori breweries were classified as type L, thus we concluded type L represents the industrial Kuro-Koji molds. We found all type L strains lack the An15g07920 gene which is required for ochratoxin A biosynthesis in black Aspergillus. This sequence is present in the genome of A. niger CBS 513.88 and has homology to the polyketide synthase fragment of A. ochraceus which is involved in ochratoxin A biosynthesis. Based on the industrial importance and the safety of Kuro-Koji molds, we propose to classify the type L strains as Aspergillus luchuensis, as initially reported by Dr. Inui. © 2011 The Society for Biotechnology, Japan.


Ikehara T.,Tropical Technology Center Ltd | Imamura S.,Tropical Technology Center Ltd | Yoshino A.,Tropical Technology Center Ltd | Yasumoto T.,Okinawa Institute of Science and Technology
Toxins | Year: 2010

Okadaic acid and its analogs (OAs) responsible for diarrhetic shellfish poisoning (DSP) strongly inhibit protein phosphatase 2A (PP2A) and thus are quantifiable by measuring the extent of the enzyme inhibition. In this study, we evaluated the suitability of the catalytic subunit of recombinant human PP2A (rhPP2Ac) for use in a microplate OA assay. OA, dinophysistoxin-1(DTX1), and hydrolyzate of 7-O-palmitoyl-OA strongly inhibited rhPP2Ac activity with IC 50 values of 0.095, 0.104, and 0.135 nM, respectively. The limits of detection and quantitation for OA in the digestive gland of scallops and mussels were 0.0348 μg/g and 0.0611 μg/g respectively, and, when converted to the whole meat basis, are well below the regulation level proposed by EU (0.16 μg/g whole meat). A good correlation with LC-MS data was demonstrated, the correlation coefficient being 0.996 with the regression slope of 1.097. © 2010 by the authors; licensee Molecular Diversity Preservation International, Basel, Switzerland.


PubMed | Tropical Technology Center Ltd.
Type: Journal Article | Journal: Toxins | Year: 2011

Okadaic acid and its analogs (OAs) responsible for diarrhetic shellfish poisoning (DSP) strongly inhibit protein phosphatase 2A (PP2A) and thus are quantifiable by measuring the extent of the enzyme inhibition. In this study, we evaluated the suitability of the catalytic subunit of recombinant human PP2A (rhPP2Ac) for use in a microplate OA assay. OA, dinophysistoxin-1(DTX1), and hydrolyzate of 7-O-palmitoyl-OA strongly inhibited rhPP2Ac activity with IC(50) values of 0.095, 0.104, and 0.135 nM, respectively. The limits of detection and quantitation for OA in the digestive gland of scallops and mussels were 0.0348 g/g and 0.0611 g/g respectively, and, when converted to the whole meat basis, are well below the regulation level proposed by EU (0.16 g/g whole meat). A good correlation with LC-MS data was demonstrated, the correlation coefficient being 0.996 with the regression slope of 1.097.

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