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In VitroBioassay of Allelopathy in Four Bamboo Species;Bambusa multiplex, Phyllostachys bambusoides, P. nigra, Sasa kurilensis, Using Sandwich Method and Protoplast Co-Culture Method with Digital Image Analysis

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American Journal of Plant Sciences 2017年第7期8卷 1699-1710页

作者： Shinjiro ogita Hamako Sasamoto Biotechnology Research Center and Department of Biotechnology Toyama Prefectural University Imizu Japan Department of Life Sciences Faculty of Life and Environmental Sciences Prefectural University of Hiroshima Shoubara Japan Faculty of Environment and Information Sciences Yokohama National University Yokohama Japan Research Institute for Integrated Science Kanagawa University Hiratsuka Japan

Moderately strong allelopathic activities were found in four bamboo species, Bambusa multiplex cv. Houraichiku;Phyllostachys bambusoides cv. Madake;P. nigra cv. Hachiku;Sasa kurilensis cv. Chishimazasa, which are of different classification or of different ecological distributions, using the “Sandwich Method”, which assays the dried leaves on growth of lettuce seedlings. Only small difference of activity was found among the four bamboo species. In addition, “Protoplast Co-culture Method” for assay of allelopathy in a 50 μL liquid medium using a 96 well culture plate, was applied to the suspension cultures of the four bamboo species. Protoplasts were isolated from two-week cultured suspension cells of four bamboo species using Cellulase RS and Pectolyase Y-23 in 0.6 M mannitol. At low protoplast densities of bamboo, B. multiplex and P. bambusoides stimulated the recipient lettuce growth, i.e., non-spherically cell enlargement and cell divisions observed under an inverted microscope, while protoplasts of P. nigra and S. kurilensis were less stimulatory or inhibitory. Inhibitory effect of S. kurilensis was the strongest among four bamboo species. Furthermore, highly inhibitory effects of S. kurilensis protoplasts on yellow color accumulation of lettuce protoplasts were clearly observed by analysis of a scanned digital image of a 96-well culture plate. Differences and causes of the allelopathic activities were discussed comparing with other plant species studied using the same assay methods.

关键词： Allelopathy Bamboo Bioassay Digital Image Analysis Protoplast Culture

Tremor Associated with Muscle Fatigue Is Temporary and Beneficial: A Case Report

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Journal of Biomedical Science and Engineering 2022年第3期15卷 103-107页

作者： Hiroyuki Tamaki Futoshi ogita Department of Sports and Life Science National Institute of Fitness and Sports Kanoya Japan

Enhanced physiological tremor (EPT) associated with fatiguing static muscle contractions is generally thought to limit continuous muscle work. However, the EPT dies away, allowing the continuous muscle contraction to be maintained at low-level workload. Here, we report a case study of a 22-year-old male volunteer who performed static plantar flexion at 30% maximal voluntary contraction (MVC) for 66 min with repeated EPT events. Comparative analysis of 1 min pre- and post-EPT phases revealed lower electromyographic (EMG) activity, heart rate, and blood pressure levels during the post-EPT phases, while the popliteal artery blood-flow volume and vascular conductance were raised. EPT may therefore be a mechanism for removing factors that inhibit continuous muscle contraction. This unique case report provides new insight into the functional significance of EPT during prolonged static muscle contractions.

关键词： Enhanced Physiological Tremor Electromyography Blood Flow

Cellular Growth Dynamics Affects Allelopathic Activity in Coffee Cell Culture

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American Journal of Plant Sciences 2023年第8期14卷 857-868页

作者： Muchamad Imam Asrori Shinjiro ogita Graduate School of Comprehensive Scientific Research Prefectural University of Hiroshima Hiroshima Japan

Cellular growth dynamics and allelopathic activity in coffee cell cultures were examined as follows: First, we compared allelopathic activity of seven woody plant calli, Coffea canephora, Derris indica, Ficus carica L., Juniperus conferta, Prunus persica, Punica granatum, and Sonneratia ovata, using a modified “sandwich method bioassay” and found that coffee callus showed the strongest growth inhibition to lettuce seedling nearly 90% of hypocotyl and 96% of root. This coffee callus actively proliferated, with a 21-fold increase during five weeks of subculture, with a growth curve comprising two typical phases: a lag phase of 0 - 2 weeks of culture and an exponential phase of 3 - 5 weeks of culture. Allelopathic activity varied depending on the growth phase of the coffee callus. The strongest allelopathic activity was detected in 1 - 2-week-old callus showing nearly 100% inhibitory effect on lettuce seedling growth. As the allelopathic activity of coffee calli is extremely high, beyond the natural level in coffee leaves and green beans, we focused on analyzing the allelopathic activity of its aqueous extracts using high-performance liquid chromatography. Several prominent peaks, including two reference alkaloids, theobromine and caffeine, which are known allelochemicals in coffee plants, and three distinct unknown peaks were identified at 270 nm in coffee calli during the lag phase (1 - 2 weeks of culture). The higher value of the total phenolic content in the lag phase also suggested a key biosynthetic pathway in relation to the allelopathic activity of coffee callus will be activated in the lag phase.

关键词： Allelopathic Activity Coffea canephora Callus Growth Phase Sandwich Method Bioassay

A β-Glucosidase Activity Potentially Involved in Cell Division and Wall Development of PhyllostachysBamboo Suspension Cells

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American Journal of Plant Sciences 2012年第8期3卷 1066-1072页

作者： Shinjiro ogita Shinya Ohki Taiji Nomura Yasuo Kato Laboratory of Plant and Cell Engineering Department of Biotechnology Toyama Prefectural University Toyama Japan.

We propose a novel Madake (Phyllostachys bambusoides) bamboo suspension culture model for investigation of key enzyme(s) activity involved in growth/differentiation. Sedimented Cell Volume (SCV) and fresh weight (FW) of the suspension cultured cells reached 34% (v/v) and 8.7 g in 10 μM 2,4-dichlorophenoxyacetic acid (2,4-D)-containing medium while only 7% (v/v) SCV and 1.9 g FW of the cells in 10 μM gibberellic acid (GA3)-containing medium in 14 days. Proportion of mitotically active cells (S to G2/M phases) at a log phase was identified as 29.5% in the former cells with tiny cytoplasmic features while 5.4% in the latter cells with elongation, wall thickening, and lignification by using flow cytometry and laser scanning microscopic analysis. The total β-glucosidase (BGL) activity under the 2,4-D condition increased from 4.8 U in day 2 to 26.2 U in day 14 (ca. 5.5-fold) while a slight reduction, from 4.4 U in day 2 to 2.1 U in day 14 (ca. 0.5-fold), occurred when cell division was suppressed under the GA3 condition. Ratio of the BGL activity of the soluble fractions to the membrane-associated fractions varied depending of the culture condition. The ratio was stable (2 to 8) during the culture period under the 2,4-D condition. Interestingly, the activity of the soluble enzyme fractions increased up to ca. 65% under the GA3 condition in inverse proportion to the membrane-associated enzymes. All together, it was strongly suggested that the detected specificity/variability of BGL activity is potentially involved in cell division and lignification in Madake bamboo cells.

关键词： Bamboo β-Glucosidase Cell Wall Development Phyllostachys bambusoides Suspension Culture

Knowledge and Tube Feeding Practices for Older Adult Patients among Japanese Registered Nurses

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International Journal of Clinical Medicine 2013年第4期4卷 208-216页

作者： Mihoko ogita Hiroko Utsunomiya Kazuko Nin Hidenori Arai Deparment of Community Network and Collaborative Medicine Kyoto University Hospital Kyoto Japan Department of Human Health Sciences Kyoto University Graduate School of Medicine Kyoto Japan

The aim of this study was to examine nurses’ thinking regarding the indications for tube feeding in older adults with eating difficulties and to determine whether the team approach can improve their knowledge and tube feeding practices. We sent self-administered questionnaires to 436 nurses and collected them from November to December 2010 (response rate, 70.0%). The results indicated that approximately 30% of the Japanese nurses thought that older patients with dementia should be on tube feeding and more than half of the nurses answered that tube feeding is indicated for aspiration-prone, frail, older adults. Moreover, our findings showed that nurses who organize multidisciplinary teams performed more interventions for dysphagia before introducing tube feeding than the reference group as analyzed by multivariate adjustment (odds ratio, 2.1-6.6). In conclusion, a multidisciplinary team approach is expected to make better decisions on the treatment and care of older patients with difficulty eating, including the need for tube

关键词： Tube Feeding Percutaneous Endoscopic Gastrostomy Older Adults Knowledge Practice Nursing Multidisciplinary Team

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采用新的制造方法来改善铝铅轴瓦性能

汽车与新动力 1995年第6期 22-30页

作者： ogita Y Ido Y 梁炬仁

介绍一种新的制造轴瓦材料的热挤压成形工艺,用这种方法可以改善铝铅轴瓦性能。疲劳强度比常规烧结法生产的铝铅轴承合金提高30%,抗咬合性比其它合金提高10%。

关键词：轴承工艺

A Versatile Liquid Culture Method to Control the in VitroDevelopment of Shoot and Root Apical Meristems of Bamboo Plants

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American Journal of Plant Sciences 2020年第2期11卷 262-275页

作者： Most Tanziman Ara Taiji Nomura Yasuo Kato Shinjiro ogita Graduate School of Comprehensive Scientific Research Prefectural University of Hiroshima Nanatsukacho Shobara Hiroshima Japan Biotechnology Research Center and Department of Biotechnology Toyama Prefectural University Kurokawa Imizu Toyama Japan

We focus on controlling morphological and histochemical responses of the shoot apical meristem (SAM) and root apical meristem (RAM) of bamboo node by using a simple and versatile liquid culture system. First, nodes of 11 different bamboo species that belong to seven major bamboo genera (Bambusa, Dendrocalamus, Phyllostachys, Tetragonocalamus, Chimonobambusa, Pleioblastus, and Sasa) were cultured using 2 mL per well of a liquid medium in a 6-well microplate to form a small-scale liquid culture environment (SLCE). The dormant lateral buds of all bamboo nodes resumed expanding and elongating within 7 days in the SLCE. The dormant and active lateral buds were sectioned longitudinally and stained with Sytox green (SG) to monitor mitotic activity and counterstained with safranin (SF) to detect the inward region of the SAM region. Further, mitotic activity was calculated using a digital imaging analysis, which showed an increase of up to 1.2- to 3.8-fold in terms of the SG/SF ratio after 7 days in the culture. Moreover, we used in vitro node cultures of two typical bamboo species, the sympodial clump-forming type (Bambusa multiplex Raeush, Bm) and the monopodial single culm-forming type (Phyllostachys meyeri McClure, Pm), and noted the following: 1) since gradual white-to-green tinge shoots were observed, we investigated the relation between color variation in the outer regions of culm and node tissues and their suitability as explants. By checking the autofluorescence property of whole shoots under LED 365 nm illumination with an RGB (red, green, and blue) digital imaging analysis using ImageJ software, we specified the color variation of explants as the relative intensity of the blue value. 2) Since the obtained shoots of a 1-month-old culture box showed growth variation, we distinguished shoot types based on plant height, i.e., short (less than 5 cm), medium (ca. 5 - 10 cm), and tall (more than 10 cm). Tall shoots that have ca. 5 nodes on average were suitable for expla

关键词： Bamboo Digital Imaging Node Culture RAM SAM SLCE

The Mutated Acetolactate Synthase Gene from Rice as a Non-Antibiotic Selection Marker for Transformation of Bamboo Cells

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American Journal of Plant Sciences 2012年第3期3卷 368-372页

作者： Shinjiro ogita Nanaka Kikuchi Taiji Nomura Yasuo Kato Laboratory of Plant and Cell Engineering Department of Biotechnology Toyama Prefectural University Toyama Japan

Previously, we developed a particle bombardment-mediated transformation protocol in Phyllostachys nigra bamboo by expressing hygromycin phosphotransferase gene (HPT) and neomycin phosphotransferase II gene (NPT II). Although these marker genes could introduce to several tissue cultured organs (e.g. leaves, buds, and calli) of Phyllostachs bamboo species, some organs showed a high susceptibility and/or a low selectivity to hygromycin and kanamycin. In this report, therefore, we describe advantages and technical details for generating stable transgenic bamboo cells using the particle bombardment method with the mutated-acetolactate synthase gene (mALS) from rice (W548L/S627IOsALS) as a non-antibiotic selection marker. A facile and efficient transformation was achieved with the mALS gene and enhanced fluorescent protein gene (mCherry). Approximately 490 and 1400 mCherry-expressing cells/dish/shot in average were observed in both P. bambusoides and P. nigra under fluorescent stereo-microscope. Stable transgenic bamboo cell lines were generated in a selection medium supplemented with 0.1 μM of bispyribac-sodium (BS) as ALS inhibitor. The integration of mALS gene was identified by in vivo ALS enzyme assay and a PCR-restriction fragment length polymerphism (RFLP) based detection procedures.

关键词： Bamboo Mutated Acetolactate Synthase Gene Particle Bombardment Suspension Culture

Relation between Amino Acids Profiles and Recalcitrancy of Cell Growth or Salt Tolerance in Tissue and Protoplast Cultures of Three Mangrove Species, Avicennia alba, Bruguiera sexangula, and Sonneratia alba

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American Journal of Plant Sciences 2013年第7期4卷 1366-1374页

作者： Shinpei Tsuchiya Shinjiro ogita Yoshifumi Kawana Tomoya Oyanagi Ai Hasegawa Hamako Sasamoto Biotechnology Research Center and Department of Biotechnology Toyama Prefectural University Toyama Japan Graduate School of Environment and Information Sciences Yokohama National University Yokohama Japan

Amino acids profiles were investigated in tissues, cultured cells, i.e. callus or suspension cells, and their protoplasts of three mangrove species, Avicennia alba, Bruguiera sexangula, and Sonneratia alba. Original tissues of cultured cells of three mangrove species were cotyledons and hypocotyls, leaves, and cotyledons, respectively. In protoplasts isolated from cultured cells, glutamine and alanine were the major amino acids. Different contents of glycine, proline and serine were observed among protoplasts of three mangrove species. Large differences in the major amino acids were found among cultured cells and their protoplasts while no difference was found between callus and suspension cells independent of additional salt in culture medium. Protoplasts of original tissues, young leaves and cotyledons, contained alanine and glutamine and/or asparagine. In suspension cells of B. sexangula, total contents of amino acids were low while their protoplasts showed similar value as of other samples. Protoplasts of leaf and cotyledons of A. alba and cotyledons of A. lanata, A. marina and S. alba were also investigated. The total contents of amino acids and their profiles might be related to the recalcitrance for the growth and salt tolerance or halophilic nature of cells and basal media used for the maintenance of cell cultures or protoplast cultures of the mangrove species. This is the first report on callus induction from hypocotyls of A. alba.

关键词： Amino Acid Avicenniaceae Callus Culture Mangrove Plants Protoplast Culture Rhizophoraceae Sonneratiaceae Suspension Culture