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  1. Article ; Online: Deep learning-based classification of adequate sonographic images for self-diagnosing deep vein thrombosis.

    Yusuke Nakayama / Mitsuru Sato / Masashi Okamoto / Yohan Kondo / Manami Tamura / Yasuko Minagawa / Mieko Uchiyama / Yosuke Horii

    PLoS ONE, Vol 18, Iss 3, p e

    2023  Volume 0282747

    Abstract: Background Pulmonary thromboembolism is a serious disease that often occurs in disaster victims evacuated to shelters. Deep vein thrombosis is the most common reason for pulmonary thromboembolism, and early prevention is important. Medical technicians ... ...

    Abstract Background Pulmonary thromboembolism is a serious disease that often occurs in disaster victims evacuated to shelters. Deep vein thrombosis is the most common reason for pulmonary thromboembolism, and early prevention is important. Medical technicians often perform ultrasonography as part of mobile medical screenings of disaster victims but reaching all isolated and scattered shelters is difficult. Therefore, deep vein thrombosis medical screening methods that can be easily performed by anyone are needed. The purpose of this study was to develop a method to automatically identify cross-sectional images suitable for deep vein thrombosis diagnosis so disaster victims can self-assess their risk of deep vein thrombosis. Methods Ultrasonographic images of the popliteal vein were acquired in 20 subjects using stationary and portable ultrasound diagnostic equipment. Images were obtained by frame split from video. Images were classified as "Satisfactory," "Moderately satisfactory," and "Unsatisfactory" according to the level of popliteal vein visualization. Fine-tuning and classification were performed using ResNet101, a deep learning model. Results Acquiring images with portable ultrasound diagnostic equipment resulted in a classification accuracy of 0.76 and an area under the receiver operating characteristic curve of 0.89. Acquiring images with stationary ultrasound diagnostic equipment resulted in a classification accuracy of 0.73 and an area under the receiver operating characteristic curve of 0.88. Conclusion A method for automatically identifying appropriate diagnostic cross-sectional ultrasonographic images of the popliteal vein was developed. This elemental technology is sufficiently accurate to automatically self-assess the risk of deep vein thrombosis by disaster victims.
    Keywords Medicine ; R ; Science ; Q
    Subject code 571
    Language English
    Publishing date 2023-01-01T00:00:00Z
    Publisher Public Library of Science (PLoS)
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  2. Article ; Online: Development of a filter device for the prevention of aquatic bacterial disease using a single-chain variable fragment (scFv)-conjugated affinity silk

    Harsha Prakash / Mitsuru Sato / Katsura Kojima / Atsushi Sato / Shinpei Maruyama / Takahiro Nagasawa / Miki Nakao / Tomonori Somamoto

    Scientific Reports, Vol 12, Iss 1, Pp 1-

    2022  Volume 8

    Abstract: Abstract Infectious disease is one of the most serious problems in the aquaculture industry for ornamental or edible fish. This study attempted to develop a new device for preventing an aquatic bacterial disease, ulcer disease, caused by Aeromonas ... ...

    Abstract Abstract Infectious disease is one of the most serious problems in the aquaculture industry for ornamental or edible fish. This study attempted to develop a new device for preventing an aquatic bacterial disease, ulcer disease, caused by Aeromonas salmonicida (As), using “affinity silk”. Affinity silk is a silk protein-containing fibroin L-chain (FibL) fused to the single-chain variable fragment (scFv). It can be easily processed into different formats such as fibers, gels, sponges, or films. A transgenic silkworm that could express a cDNA construct containing FibL fused to an scFv derived from a monoclonal antibody (MAb) against As was successfully generated. An enzyme-linked immunosorbent assay was used to detect As by employing 96-well plates coated with scFv-conjugated affinity silk. As could be captured efficiently by glass wool coated with affinity silk in the column. Furthermore, the air-lift water filter equipped with the affinity silk-coated wool could considerably reduce the concentration of As in water and was estimated to have sufficient ability to trap a lethal dose of As. These findings show that the “affinity silk filter” is a potential device for the prophylaxis of aquatic animal diseases.
    Keywords Medicine ; R ; Science ; Q
    Subject code 572
    Language English
    Publishing date 2022-06-01T00:00:00Z
    Publisher Nature Portfolio
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  3. Article ; Online: Development and validation of scFv-conjugated affinity silk protein for specific detection of carcinoembryonic antigen

    Mitsuru Sato / Hiroshi Kitani / Katsura Kojima

    Scientific Reports, Vol 7, Iss 1, Pp 1-

    2017  Volume 8

    Abstract: Abstract The production costs for monoclonal antibodies (MAbs) utilized in medical diagnostic kits are inevitably high because the MAbs are mostly obtained from hybridoma cell culture. Here, we report the development and validation of a novel affinity ... ...

    Abstract Abstract The production costs for monoclonal antibodies (MAbs) utilized in medical diagnostic kits are inevitably high because the MAbs are mostly obtained from hybridoma cell culture. Here, we report the development and validation of a novel affinity silk protein produced by transgenic silkworm technology as a possible alternative diagnostic tool for cancers. We generated a transgenic silkworm expressing a cDNA construct containing fibroin L-chain fused to a single-chain variable fragment (scFv) derived from a MAb against carcinoembryonic antigen (CEA). The transgenic cocoons were dissolved in aqueous lithium bromide solution, applied to 96-well plates, and analysed by enzyme-linked immunosorbent assay. The scFv-conjugated affinity silk protein specifically recognized CEA as well as the parental MAb. The binding activity was retained after several months of storage in coated plates or concentrated solution. Thus, the scFv-conjugated affinity silk protein provides a potentially useful alternative to conventional MAbs in medical diagnostic kits.
    Keywords Medicine ; R ; Science ; Q
    Subject code 572
    Language English
    Publishing date 2017-11-01T00:00:00Z
    Publisher Nature Publishing Group
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  4. Article ; Online: Far rapid synthesis of giant DNA in the Bacillus subtilis genome by a conjugation transfer system

    Mitsuhiro Itaya / Mitsuru Sato / Miki Hasegawa / Nobuaki Kono / Masaru Tomita / Shinya Kaneko

    Scientific Reports, Vol 8, Iss 1, Pp 1-

    2018  Volume 6

    Abstract: Abstract Bacillus subtilis offers a platform for giant DNA synthesis, which is mediated by the connection of overlapping DNA segments called domino DNA, in the cloning locus of the host. The domino method was successfully used to produce DNA fragments as ...

    Abstract Abstract Bacillus subtilis offers a platform for giant DNA synthesis, which is mediated by the connection of overlapping DNA segments called domino DNA, in the cloning locus of the host. The domino method was successfully used to produce DNA fragments as large as 3500 kbp. However, domino DNA is limited to <100 kbp because of size restrictions regarding the transformation (TF) of B. subtilis competent cells. A novel conjugal transfer (CT) method was designed to eliminate the TF size limit. The CT method enables rapid and efficient domino reactions in addition to the transfer of giant DNA molecules of up to 875 kbp to another B. subtilis genome within 4 hours. The combined use of the TF and CT should enable significantly rapid giant DNA production.
    Keywords Medicine ; R ; Science ; Q
    Language English
    Publishing date 2018-06-01T00:00:00Z
    Publisher Nature Publishing Group
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  5. Article: Specific binding of the WASP N-terminal domain to Btk is critical for TLR2 signaling in macrophages

    Sakuma, Chisato / Hiroshi Kitani / Mitsuru Sato / Takato Takenouchi

    Molecular Immunology. 2015 Feb., v. 63

    2015  

    Abstract: Wiskott–Aldrich syndrome protein (WASP) is an adaptor molecule in immune cells. Recently, we revealed that WASP is involved in lipopolysaccharide-TLR4 signaling in macrophages by association of Bruton's tyrosine kinase (Btk) with the WASP N-terminal ... ...

    Abstract Wiskott–Aldrich syndrome protein (WASP) is an adaptor molecule in immune cells. Recently, we revealed that WASP is involved in lipopolysaccharide-TLR4 signaling in macrophages by association of Bruton's tyrosine kinase (Btk) with the WASP N-terminal domain. Btk has been shown to play important roles in the signaling of several TLRs and to modulate the inflammatory response in macrophages. In this study, we evaluated the importance of the interaction between Btk and WASP in TLR2 signaling by using bone marrow-derived macrophage cell lines from transgenic (Tg) mice expressing anti-WASP N-terminal domain single-chain variable fragment (scFv) or VL single-domain intrabodies. In this Tg bone marrow-derived macrophages, specific interaction between WASP and Btk were strongly inhibited by masking of the binding site in the WASP N-terminal domain. There was impairment of gene expression of TNF-α, IL-6, and IL-1β and phosphorylation of inhibitor of κB α/β (IKKα/β) and nuclear factor (NF)-κB upon stimulation with TLR2 ligands. Furthermore, tyrosine phosphorylation of WASP following TLR2-ligand stimulation was severely inhibited in the Tg bone marrow-derived macrophages, as shown by the impairment in WASP tyrosine phosphorylation following lipopolysaccharide stimulation. These results strongly suggest that the association between the WASP N-terminal domain and Btk plays an important role in the TLR2-signaling pathway in macrophages.
    Keywords binding sites ; gene expression ; genetically modified organisms ; inflammation ; interleukin-1beta ; interleukin-6 ; ligands ; lipopolysaccharides ; macrophages ; mice ; phosphorylation ; Toll-like receptor 2 ; tumor necrosis factor-alpha ; tyrosine
    Language English
    Dates of publication 2015-02
    Size p. 328-336.
    Publishing place Elsevier Ltd
    Document type Article
    ZDB-ID 424427-8
    ISSN 1872-9142 ; 0161-5890
    ISSN (online) 1872-9142
    ISSN 0161-5890
    DOI 10.1016/j.molimm.2014.08.004
    Database NAL-Catalogue (AGRICOLA)

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  6. Article ; Online: Human recombinant erythropoietin improves motor function in rats with spinal cord compression-induced cervical myelopathy.

    Takahiro Tanaka / Hidetoshi Murata / Ryohei Miyazaki / Tetsuya Yoshizumi / Mitsuru Sato / Makoto Ohtake / Kensuke Tateishi / Phyo Kim / Tetsuya Yamamoto

    PLoS ONE, Vol 14, Iss 12, p e

    2019  Volume 0214351

    Abstract: OBJECTIVE:Erythropoietin (EPO) is a clinically available hematopoietic cytokine. EPO has shown beneficial effects in the context of spinal cord injury and other neurological conditions. The aim of this study was to evaluate the effect of EPO on a rat ... ...

    Abstract OBJECTIVE:Erythropoietin (EPO) is a clinically available hematopoietic cytokine. EPO has shown beneficial effects in the context of spinal cord injury and other neurological conditions. The aim of this study was to evaluate the effect of EPO on a rat model of spinal cord compression-induced cervical myelopathy and to explore the possibility of its use as a pharmacological treatment. METHODS:To develop the compression-induced cervical myelopathy model, an expandable polymer was implanted under the C5-C6 laminae of rats. EPO administration was started 8 weeks after implantation of a polymer. Motor function of rotarod performance and grip strength was measured after surgery, and motor neurons were evaluated with H-E, NeuN and choline acetyltransferase staining. Apoptotic cell death was assessed with TUNEL and Caspase-3 staining. The 5HT, GAP-43 and synaptophysin were evaluated to investigate the protection and plasticity of axons. Amyloid beta precursor protein (APP) was assessed to evaluate axonal injury. To assess transfer of EPO into spinal cord tissue, the EPO levels in spinal cord tissue were measured with an ELISA for each group after subcutaneous injection of EPO. RESULTS:High-dose EPO maintained motor function in the compression groups. EPO significantly prevented the loss of motor neurons and significantly decreased neuronal apoptotic cells. Expression of 5HT and synaptophysin was significantly preserved in the EPO group. APP expression was partly reduced in the EPO group. The EPO levels in spinal cord tissue were significantly higher in the high-dose EPO group than other groups. CONCLUSION:EPO improved motor function in rats with compression-induced cervical myelopathy. EPO suppressed neuronal cell apoptosis, protected motor neurons, and induced axonal protection and plasticity. The neuroprotective effects were produced following transfer of EPO into the spinal cord tissue. These findings suggest that EPO has high potential as a treatment for degenerative cervical myelopathy.
    Keywords Medicine ; R ; Science ; Q
    Subject code 616
    Language English
    Publishing date 2019-01-01T00:00:00Z
    Publisher Public Library of Science (PLoS)
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  7. Article ; Online: Somaclonal variation is induced de novo via the tissue culture process

    Mitsuru Sato / Munetaka Hosokawa / Motoaki Doi

    PLoS ONE, Vol 6, Iss 8, p e

    a study quantifying mutated cells in Saintpaulia.

    2011  Volume 23541

    Abstract: Background The origin of somaclonal variation has not been questioned previously, i.e., "pre-existing mutations" in explants and "newly induced mutations" arising from the tissue culture process have not been distinguished. This is primarily because ... ...

    Abstract Background The origin of somaclonal variation has not been questioned previously, i.e., "pre-existing mutations" in explants and "newly induced mutations" arising from the tissue culture process have not been distinguished. This is primarily because there has been no reliable molecular method for estimating or quantifying variation. Methodology/principal findings We adopted a petal-variegated cultivar of Saintpaulia 'Thamires' (Saintpaulia sp.) as the model plant. Based on the difference between the pre- and post-transposon excision sequence of the promoter region of flavonoid 3', 5'-hydoroxylase (F3'5'H), we estimated mutated (transposon-excised) cell percentages using a quantitative real-time PCR. Mutated cell percentages in leaf laminae used as explants was 4.6 and 2.4% in highly or low variegation flower plants, respectively, although the occurrences of blue color mutants in their regenerants were more than 40%. Preexisting mutated cell percentages in cultured explants were considerably lower than the mutated plant percentage among total regenerants via tissue culture. Conclusions/significance The estimation of mutated cell percentages became possible using the quantitative real-time PCR. The origins of mutations were successfully distinguished; it was confirmed that somaclonal variations are mainly caused by newly generated mutations arising from tissue culture process.
    Keywords Medicine ; R ; Science ; Q
    Subject code 571
    Language English
    Publishing date 2011-01-01T00:00:00Z
    Publisher Public Library of Science (PLoS)
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  8. Article ; Online: Immortalization and Characterization of Porcine Macrophages That Had Been Transduced with Lentiviral Vectors Encoding the SV40 Large T Antigen and Porcine Telomerase Reverse Transcriptase

    Takato Takenouchi / Hiroshi Kitani / Shunichi Suzuki / Michiko Nakai / Dai-ichiro Fuchimoto / Mitsutoshi Tsukimoto / Hiroki Shinkai / Mitsuru Sato / Hirohide Uenishi

    Frontiers in Veterinary Science, Vol

    2017  Volume 4

    Abstract: The domestic pig is an important agricultural animal, and thus, infectious diseases that affect pigs can cause severe economic losses in the global swine industry. Various porcine pathogens target macrophages, which are classical innate immune cells. ... ...

    Abstract The domestic pig is an important agricultural animal, and thus, infectious diseases that affect pigs can cause severe economic losses in the global swine industry. Various porcine pathogens target macrophages, which are classical innate immune cells. Although macrophages basically protect the host from pathogens, they also seem to contribute to infectious processes. Therefore, cultured macrophages can be used to develop in vitro models for studying not only genes associated with porcine innate immunity but also the infectious processes of porcine pathogens. However, the availability of porcine macrophage cell lines is limited. In this study, we describe a novel immortalized porcine kidney-derived macrophage (IPKM) cell line, which was generated by transferring the SV40 large T antigen (SV40LT) and porcine telomerase reverse transcriptase (pTERT) genes into primary porcine kidney-derived macrophages using lentiviral vectors. The IPKM displayed a typical macrophage morphology and was routinely passaged (doubling time: about 4 days). These cells were immunostained for macrophage markers. In addition, they exhibited substantial phagocytosis of polystyrene microbeads and released inflammatory cytokines upon lipopolysaccharide (LPS) stimulation. Furthermore, the maturation and secretion of interleukin-1β were observed after nigericin-induced inflammasome activation in LPS-primed IPKM. These findings suggest that IPKM exhibit the typical inflammatory characteristics of macrophages. By transferring the SV40LT and pTERT genes using lentiviral vectors, we also successfully immortalized macrophages derived from the peripheral blood of a low-density lipoprotein receptor-deficient pig. These results suggest that the co-expression of SV40LT and pTERT is an effective way of immortalizing porcine macrophages.
    Keywords porcine macrophages ; immortalization ; SV40 large T antigen ; porcine telomerase reverse transcriptase ; lentiviral vector ; Veterinary medicine ; SF600-1100
    Subject code 630 ; 616
    Language English
    Publishing date 2017-08-01T00:00:00Z
    Publisher Frontiers Media S.A.
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  9. Article ; Online: Critical roles of the WASP N-terminal domain and Btk in LPS-induced inflammatory response in macrophages.

    Chisato Sakuma / Mitsuru Sato / Takato Takenouchi / Joe Chiba / Hiroshi Kitani

    PLoS ONE, Vol 7, Iss 1, p e

    2012  Volume 30351

    Abstract: While Wiskott-Aldrich syndrome protein (WASP) plays critical roles in TCR signaling as an adaptor molecule, how it transduces innate immune signals remains to be elucidated. To investigate the roles of WASP in innate immune cells, we established bone ... ...

    Abstract While Wiskott-Aldrich syndrome protein (WASP) plays critical roles in TCR signaling as an adaptor molecule, how it transduces innate immune signals remains to be elucidated. To investigate the roles of WASP in innate immune cells, we established bone marrow-derived macrophage (BMDM) cell lines from WASP15 transgenic (Tg) mice overexpressing the WASP N-terminal region (exons 1-5). Upon LPS stimulation, WASP15 Tg BMDM cell lines produce lower levels of inflammatory cytokines, such as TNF-α, IL-6, and IL-12p40 than the wild-type BMDM cell line. In addition, the production of nitric oxide by WASP15 Tg BMDM cells in response to LPS and IFN-γ was significantly impaired. Furthermore, we uncovered that the WASP N-terminal domain associates with the Src homology (SH) 3 domain of Bruton's tyrosine kinase (Btk). Overexpression of the WASP N-terminal domain diminishes the extent of tyrosine phosphorylation of endogenous WASP in WASP15 Tg BMDM cells, possibly by interfering with the specific binding between endogenous WASP and Btk during LPS signaling. These observations strongly suggest that the interaction between WASP N-terminal domain and Btk plays important roles in the LPS signaling cascade in innate immunity.
    Keywords Medicine ; R ; Science ; Q
    Language English
    Publishing date 2012-01-01T00:00:00Z
    Publisher Public Library of Science (PLoS)
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  10. Article ; Online: A System Utilizing Metal Hydride Actuators to Achieve Passive Motion of Toe Joints for Prevention of Pressure Ulcers

    Minako Hosono / Shuichi Ino / Mitsuru Sato / Kazuhiko Yamashita / Takashi Izumi

    Rehabilitation Research and Practice, Vol

    A Pilot Study

    2012  Volume 2012

    Abstract: This paper describes the influence of human toe movement on blood flow and the design of a toe joint passive motion system for preventing pressure ulcers. First, we measured lower extremity blood flow in the foot during active and passive motion of the ... ...

    Abstract This paper describes the influence of human toe movement on blood flow and the design of a toe joint passive motion system for preventing pressure ulcers. First, we measured lower extremity blood flow in the foot during active and passive motion of the toe to facilitate the design of new rehabilitation equipment. Also, the flexion and extension angles and the force of the toe joints were measured to determine appropriate specifications for the system. Increases in blood flow were observed at the external malleolus during movement. Flexion and extension angles and the force of the toe joints were found to differ significantly among participants. It is shown that a toe joint passive motion system can be effective in preventing pressure ulcers. On the basis of these results, a device using alloys of metal hydride (MH) as an actuator that is suitable for the system to initiate toe motion, was developed.
    Keywords Medicine (General) ; R5-920
    Subject code 629
    Language English
    Publishing date 2012-01-01T00:00:00Z
    Publisher Hindawi Limited
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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