|toshihiro - ona||Last modified date：2021.05.28|
Associate Professor / Sustainable Resources Science / Department of Agro-environmental Sciences / Faculty of Agriculture
|toshihiro - ona||Last modified date：2021.05.28|
|1.||Junko Johzuka,Toshihiro Ona,, One hour in vivo-like phenotypic screening system for anti-cancer drugs using a high precision surface plasmon resonance device, MDPI, 10.3390/ECMC2020-07366, 6th International Electronic Conference on Medicinal Chemistry session Round Table on Natural Products, 2020.11, [Background/Aims] In anti-cancer drug (candidate) screening, the demand exists for evaluation at physiological concentrations similar to in vivo. This is often performed by three-dimensionally (3D) cultured cells. It necessitates a long culture period of 2-4 weeks with tedious experimental procedures based on endpoint assay and labeling agents causing low reliability. The previous device methods depend on the pharmaceutical mode of action, little related to the conventional method. Furthermore, a separate set of experiment is required to obtain both efficacy and toxicity. Here, we report on a high precision surface Plasmon resonance-3D system to overcome these all problems sensing dynamic cellular reaction against target compound(s) by laser.
[Results] We developed the system with average fluctuation of 50 ndeg/s combined with two-dimensionally cultured cells attached onto a sensor chip by applying collagen on the top. The 3D cell activity was shortly obtained by this without cell division. New system gave real-time monitoring of mitochondrial membrane potential (MMP) within live cells without both labeling and invasion. It allowed in vivo-like phenotypic screening for anti-cancer drugs within 1h of drug addition. The data were collected as the stable linear signal change parts for at least 5min after 25min following drug addition. The results provided compatibility to clinically related chemosensitivity test (P<0.001) using two cell lines of pancreatic cancer and three anti-cancer drugs to represent differences in individual gene expression and drug mode of action. Early MMP change rate is concluded as a key to quantitatively predict the efficacy and toxicity..
|2.||Toshihiro Ona,Junko Johzuka, Rapid and synchronized dormancy-broken Kyoho grape seed endosperm without extraction or concentration shows significant effect on both anti-cancer and pro-immunity, MDPI, 10.3390/ECMC2020-07249, 6th International Electronic Conference on Medicinal Chemistry session Round Table on Natural Products, 2020.11, [Background/Aims] Anti-cancer effect has been reported in oral administration of grape seed extract (GSE) although the clinical trial failed because of its toxicity. Grape seeds should be toxic to prevent them to be eaten by animals. However, once they wake up, they change the ingredients into a form which dissolves easily in water and break toxicity substances inhibiting germination. In order for seeds to sprout and grow, a lot of nutrients as well as life itself reside. The objective of this study is to determine the in vivo-like efficacy and toxicity on anti-cancer and pro-immunity of rapid and synchronized dormancy-broken grape seed endosperm (RSDB-GSE) by our proprietary Grandir recipe. It was performed in case of oral administration using digested and absorbed RSDB-GSE.
[Results] Direct killing efficacy against all cancer cells was observed in a dose-dependent manner at 2.5 mg/ml or more, and it was the maximum at 5.0 mg/ml for pancreas and liver and at 7.5 mg/ml for breast. The cell viability was less than 20%. At concentrations higher than these, it showed toxicity which was thought to be cell functional shutdown by suspension of mitochondrial polarization. No effect was observed for normal cells. The RSDB-GSE showed general direct cancer killing efficacy for organs. T lymphocytes cytotoxic activity by NK cells was observed in a dose-dependent manner at 3.75 mg/ml or more, and it was kept at least until 5.0 mg/ml. From this, pro-immunity was shown by RSDB-GSE. Consequently, it is expected to express synergistic anti-cancer effect, even orally administered..
|3.||Junko - Shibata, Shouhei Yano, Teruyuki Seino, Toshihiro - Ona, Anti-liver cancer effect of Chinese chive using in vivo-like test, Organizing Committee of ICNIM2014, 227-229, 2014.08, Chinese chive is a perennial herb belonging to Allium genus cultivated BC era in China. In Japan, it is introduced in old book Manyoshu in 8th century. In herbal medicine, dried leaves of Chinese chive are used as called Kyusai. The efficacy spectrum is widely reported in tonicity, recover from exhaustion, improvements in vascular flow and cholesterol, preventions in arteriosclerosis and myocardial infarct, pro-body temperature, prevention and suppression in tumors, anti-microbial and so on.
As active compounds, Chinese chive contains sulphuric amino compounds of methin and aliin, vitamins C, A and E, minerals of Ca、K、Mg、Se、Fe、P, chlorophylls and dietary fibers.
In Japan, Chinese chive is cultivated from Hokkaido to Okinawa. In Hokkaido, Shiriuchi town in Oshima district is top for the production using a brand name of “Kitano Hana” with 1,700t/y. The first picking leaves are thick and sweat and attract customers with increased handling volume. Shiriuchi town is eager to add more values to “KitanoHana”and has started various projects.
On the other hand, in a liver, absorbed food compounds are directly works against a liver first. After this, metabolized compounds within a liver with chemical modification work against a liver secondary.
In active compound evaluation, physiological concentration results are demanded equally as in vivo. For this, in vivo-like chemosensitivity test is often hired using 3D cell culture with extracellular matrix. However, this requires time-consuming experimental process with 2-4 weeks and labeling agents causing misjudge, and has difficulty in simultaneous efficacy and toxicity measurement. We conquered all these problems with development of High Precision-Surface Plasmon Resonance (HP-SPR) device and applications.
Here we examined and conformed the nature of extract from Chinese chive for anti-cancer efficacy and toxicity against liver before and after their metabolism within liver as real-time manner by HP-SPR..
|4.||Toshihiro - Ona, Junko - Shibata, Activation of skin by AHCC: comparison between whole and low molecular weight fraction, Organizing Committee of ICNIM2014, 173-175, 2014.08, Skin aging has intrinsic and extrinsic processes. Intrinsic process is derived from genetic factors such as chronicle age and reduction in estrogen. Extrinsic one is from environmental factors such as exposure to sunlight and pollutants, and diet. These generate free radicals (ROS). When ROS amount exceeds cell scavenging capacity, the oxidative stress occurs.
The oxidative stress at epidermis accelerates the formation of Advanced Glycation End Products (AGE) by protein glycation of plasma membrane. AGE inhibits SOD activity, reduces DNA repair ability and stability, mitochondrial function, cell division and delays cell cycle and migration, and induces apoptosis. This implies the decrease in cell metabolism.
As a result, epidermis becomes thin with 10-50% reduction by atrophy of stratum spinosum. Furthermore, the skin glycation reduces skin elasticity. These increase skin vulnerability and fragility and decrease in desquamation to delay wound healing and in replacement speed of lipids to disturb barrier function.
The compound screening for epidermis activation often hires cocultivation of keratinocyte with a target compound. On the other hand, 3D cell culture is reported with the use of extracellular matrix and even with fibroblast cells from dermis to reconstitute skin. However, this necessitates long time culture duration (ca. 1 month) because of endpoint method, and has difficulty in simultaneous determination of efficacy and toxicity. Moreover, the reduction in cell activity causes low experimental repeatability by long term culture.
We developed a custom-made instrument and method to monitor early stage of mitochondrial response to stimuli within live cells utilizing High-Precision Surface Plasmon Resonance (HP-SPR) as a measurement principle. This brings endpoint efficacy and toxicity predictions extremely before endpoint without cell culture.
At last ICNIM meeting, we elucidated the validity of AHCC for pro-skin turnover. This time, we report on the validity difference by comparison between whole and low molecular weight fraction of AHCC..
|5.||Toshihiro - Ona, Activation of skin by AHCC, ICNIM2013実行委員会, 7-10, 2013.08, We successfully developed a rapid, high sensitive, label-free and non-invasive cell-based sensing technology for reliable efficacy prediction of bioactive compounds at physiological concentration within 1 h regardless to the mode of action. Conventional technology is based on endpoint assay of 3D cell culture with extracellular matrix necessitating time-consuming experimental process and labeling agents causing interference with a target compound that provide low reliability. The conventional device method depends on the pharmaceutical mode of action, and gives no relation to the conventional method. New technology overcomes these all problems. Expected applications for compound screening are; 1. Anti-cancer, -Alzheimer disease, -aging (skin care), diabetes2. Pro-fat burning, -metabolism, -hair restoration, -body temp.|
|6.||Rapid efficacy evaluation of cancer drugs using light sensing technology.|
|7.||Reliable evaluation of functional compounds using light sensing technology for mitochondria in live cells .|
|8.||Nano-bio-device for real-time-monitoring of live cells.|
|9.||Nano-bio-device for real-time-monitoring of live cells.|
|10.||Nano-bio-device for real-time-monitoring of live cells.|
|11.||Ona, T., Nishijima, H., Kosaihira, A. and Shibata, J., Development of cell-based quantitative evaluation method for cell cycle-arrest type cancer drugs for apoptosis by high precision surface plasmon resonance sensor, SPIE, Biophotonics: Photonic Solutions for Better Health Care-Progress in Biomedical Optics and Imaging, Vol. 9 (38), Eds. Popp, J., Drexler, W., Tuchin, V.V. and Matthews, D.L., 69910R, 2009.04.|
|12.||Are your mitochondria healthy?.|
|13.||Rapid and quantitative evaluation method and instrumentation using surface plasmon resonance for personal therapeutic potential of cancer drugs for apoptosis by sensing mitochondrial membrane potential.|
|14.||Kosaihira, A., Fukumori, T., Sakai, K. and Ona, T., Improvement of forest resources for recyclable forest products, Springer-Verlag, 170-172, A new combination device comprised of surface plasmon resonance and fluorescence microscopy for a rapid screening of anticancer phenolic compounds., 2004.03.|
|15.||Ibuki, T., Tokida, Y., Matsuda, N., Czempinski, K., Muller-Roeber, B., Ona T. and Uozumi, N., Improvement of forest resources for recyclable forest products, Springer-Verlag, 167-169, Characterization of potassium channel from Arabidopsis thaliana., 2004.03.|
|16.||Yokota, S., Nakayama, K., Sagawa, A., Urabe, F., Ona, T., Asada, T. and Yoshizawa, N., Improvement of forest resources for recyclable forest products, Springer-Verlag, 161-162, Possible effects of properties in polyphenol oxidases on rooting ability of Eucalyptus camaldulensis cutting shoots., 2004.03.|
|17.||Yamamoto, H., Kojima, Y., Okuyama, T., Ona, T. and Okayama, T., Improvement of forest resources for recyclable forest products, Springer-Verlag, 139-143, An essay on the fine structure of the wood cell wall related to the physical properties of the recycled paper., 2004.03.|
|18.||Murakami, S., Ona, T., Sakai, K., Saito, K. and Fukushima, K., Improvement of forest resources for recyclable forest products, Springer-Verlag, 111-116, Effect of deuterium exchange in lignin on its structural analysis using FT-Raman spectroscopy., 2004.03.|
|19.||Mizumoto, M., Seino, T., Sakai K., Ona, T., Ishida, Y. and Ohtani, H., Improvement of forest resources for recyclable forest products, Springer-Verlag, 107-110, Rapid characterization of total fatty acids in wood by reactive thermal desorption-gas chromatography with tetrabutylammonium hydroxide., 2004.03.|
|20.||Haisaki, H., Tateishi, M., Seino, T., Sakai, K., Ona, T., Ohshima, J., Adachi, K., Yokota S. and Yoshizawa, N., Improvement of forest resources for recyclable forest products, Springer-Verlag, 100-104, Assessment of vessel anatomical feature in Eucalyptus camaldulensis by pyrolysis-gas chromatography., 2004.03.|
|21.||Tateishi, M., Seino, T., Sakai, K., Ona, T., Ohshima, J., Adachi, K., Yokota, S. and Yoshizawa, N., Improvement of forest resources for recyclable forest products, Springer-Verlag, 95-99, Rapid assessment of vessel morphology by pyrolysis-gas chromatography., 2004.03.|
|22.||Ohshima, J., Yokota, S., Yoshizawa, N. and Ona, T., Improvement of forest resources for recyclable forest products, Springer-Verlag, 89-94, Representative heights assessing whole-tree values and the within-tree variations of derived wood properties in Eucalyptus camaldulensis and E. globulus., 2004.03.|
|23.||Ohshima, J., Adachi, K., Yokota, S., Yoshizawa, N. and Ona T., Improvement of forest resources for recyclable forest products, Springer-Verlag, 83-88, Within-tree variation of vessel morphology and frequency and representative heights for estimating the whole-tree values in Eucalyptus camaldulensis and E. globulus., 2004.03.|
|24.||Ohshima, J., Yokota, S., Yoshizawa, N. and Ona, T., Improvement of forest resources for recyclable forest products, Springer-Verlag, 77-82, Within-tree variation of detailed fiber morphology and its position representing the whole-tree value in Eucalyptus camaldulensis and E. globulus., 2004.03.|
|25.||Ishiguri, F., Yokota, S., Yoshizawa, N. and Ona, T., Improvement of forest resources for recyclable forest products, Springer-Verlag, 74-76, Radial variation of cell morphology in three acacia species., 2004.03.|
|26.||Yoshizawa, N., Ishiguri, F., Yokota, S. and Ona, T., Improvement of forest resources for recyclable forest products, Springer-Verlag, 71-73, Formation and structure of reaction wood fibers forming no G-layer in some hardwood species., 2004.03.|
|27.||Okayama, T., Kibatani, Y., Nakayama, K., Kojima Y. and Ona, T., Improvement of forest resources for recyclable forest products, Springer-Verlag, 52-59, Fiber properties and papermaking potential of recycled Acacia pulp., 2004.03.|
|28.||Seino, T., Yoshioka, A., Takai, M., Kojima, Y., Ishikura, Y., Ona, T., Ishida, Y. Ohtani, H. and Tsuge S., Improvement of forest resources for recyclable forest products, Springer-Verlag, 47-51, Characterization of photo-yellowing trigger compounds repressing paper recyclability of Eucalyptus globulus by Pyrolysis-GC/MS., 2004.03.|
|29.||Kojima, Y., Isaji, S. and Ona T., Improvement of forest resources for recyclable forest products, Springer-Verlag, 40-46, Production of high brightness CTMP from Eucalyptus globulus and their light-induced color reversion., 2004.03.|
|30.||Ona, T., Kawana, J., Kibatani, Y., Ishikura, Y., Kojima, Y. and Okayama, T., Improvement of forest resources for recyclable forest products, Springer-Verlag, 29-32, Toward the construction of efficient link between forest recycling and paper recycling using trees with high performance for paper recycling., 2004.03.|
|31.||Ona, T., Tateishi, M., Nozaki, H., Seino, T., Yoon, S-L., Isaji, S. and Kojima, Y., Improvement of forest resources for recyclable forest products, Springer-Verlag, 24-28, Feasibility Study of Tree Selection for High Pulp Yield, Brightness and Recyclable Chemithermomehanical Paper Production using Eucalyptus globulus.
|32.||Ona, T., Improvement of forest resources for recyclable forest products, Springer-Verlag, 3-7, Overview of the project “Development of forest resources with high performance for paper recycling.
|33.||Kosaihira, A., Fukumori, T., Sakai, K. and Ona, T. , 10th European Conference on the Spectroscopy of Biological Molecules, JATEPress , 71, Rapid screen of antiproliferative phenolic compounds with simultaneous analysis of cancer cell reaction by a new combination device of surface plasmon resonance and fluorescence microscopy., 2003.08.|
|34.||Ona, T., Ohshima, J., Adachi, K., Yokota, S. and Yoshizawa, N. , 10th European Conference on the Spectroscopy of Biological Molecules, JATEPress , 200a, In situ determination of vessel element length in tree by Fourier transform Raman spectroscopy. , 2003.08.|
|35.||T. Ona, T. Sonoda, K. Ito, M. Shibata, Y. Ootake, J. Ohshima, S. Yokota and N. Yoshizawa, Spectroscopy of Biological Molecules: New Directions, Kluwer Academic Publishers, 545-546 In situ determination of proportion of cell types in wood by FT-Raman spectroscopy, 1999.08.|