| 1. |
Toshihiro Ona,Junko Johzuka, Rapid and synchronized dormancy-breaking jumbo leek scale powder supplementation promotes liver glutathione production to contribute to biological defense, The 31st International Congress on Nutrition and Integrative Medicine (ICNIM 2023), 2023.07, The number of diseases that derive from a lack of ability to eliminate oxidative stress in the body has increased dramatically in recent years, probably due to aging and the entry of nanoparticles, such as plastics, into the body. In medicine, glutathione injections are given as a treatment method. On the other hand, detoxification with glutathione has also been reported as a method of removing nanoparticles. Therefore, methods to support the increase and maintenance of glutathione in the body to optimal levels are attracting attention. In particular, the intake of foods and nutrients containing cysteine, the rate-limiting step in glutathione production in the body, has been proposed. In this study, we investigated the efficacy of jumbo leek scale powder to enhance glutathione production in the liver as a dietary supplement by using the sample that was heat-treated after promoting phase transition of its contents by rapid and synchronized dormancy-breaking.. |
| 2. |
Toshihiro Ona,Junko Johzuka, Rapid and synchronized dormancy-breaking jumbo leek bulb inhibits postprandial carbohydrate degradation and absorption to be assumed preprandial ingestion, The 8th International Electronic Conference on Medicinal Chemistry, 2022.11, In order to prevent obesity and diabetes, it is important to avoid overeating, especially carbohydrates. Therefore, we evaluated the effect of taking jumbo leek bulb before a meal on the decomposition and absorption inhibition of carbohydrates after a meal. In this case, we used a product that was heat-treated after promoting phase transition of its content by rapid and synchronized dormancy-breaking (RSDB), and investigated by decreasing two enzyme activities, namely pancreatic α-amylase and α-glucosidase, and report the results.. |
| 3. |
Toshihiro Ona,Junko Johzuka, Rapid and synchronized dormancy-breaking to enhance medicinal properties of plant seeds and bulbs, The 3rd International Electric Conference on Food: Food, Microbiome, and Health, 2022.10, In the development of functional foods for health, real food is favorable which is prepared without extraction or concentration considered from long human eating habits. Achieving this target, we have examined the possibility of dormancy-breaking of plant seeds and bulbs by the following phenomena.
Plant seeds and bulbs hydrolyze the ingredients into water soluble forms to use and break toxicity substances to inhibit germination. For them to sprout and grow, a lot of nutrients as well as life itself reside, including sugar, amino acids, minerals, hormones, and even substances both boosting immunity and restoring body to prevent diseases and defects after sprouting.
However, the ingredients become uneven if they are not awakened synchronizedly. Therefore, such a rapid method was newly developed as proprietary Grandir recipe™ with maximum 5 days of dormancy-breaking.
Here, the in vitro study was performed to determine the enhanced efficacy and decreased toxicity by rapid and synchronized dormancy-breaking in case of oral administration as foods in grape seeds, ginger rhyzomes and jumbo leek scales.. |
| 4. |
Toshihiro Ona, Junko Johzuka, Inhibition of postprandial carbohydrate degradation and absorption assuming preprandial ingestion of rapid and synchronized dormancy-breaking jumbo leek bulb powder, The 30th International Congress on Nutrition and Integrative Medicine (ICNIM 2022), 2022.07, To prevent obesity and diabetes, it is important not to overeat, especially limiting carbohydrates. Therefore, we report here on the evaluation of the inhibitory effect on the decomposition and absorption of carbohydrates after a meal. When jumbo leek bulb powder was heat-treated after promoting the phase transition of its content by rapid and synchronized dormancy-breaking and ingested before a meal, it decreased the activity of two enzymes, pancreatic α-amylase and α-glucosidase.. |
| 5. |
Toshihiro Ona,Junko Johzuka, Rapid and synchronized dormancy breaking to enhance medicinal properties of seeds and bulbs, 2nd Edition of Food Science and Technology Virtual, 2022.04, In the development of functional foods, real food is favorable to take without extraction or concentration considered from long human eating habits. To achieve this, we have examined the possibility of dormancy-breaking by the following phenomena.
Plant seeds and bulbs change the ingredients into water soluble to use and break toxicity substances to inhibit germination. For them to sprout and grow, a lot of nutrients as well as life itself reside, including sugar, amino acids, minerals, hormones, and even substances boosting immunity to prevent diseases after sprouting.
However, the ingredients become uneven if they are not awakened at the same time. Therefore, a rapid method of awaking the plant seeds and bulbs synchronizedly was developed as proprietary Grandir recipe™ with maximum 5 days of dormancy breaking.
Here, the in vitro study was conducted to determine the enhanced efficacy and decreased toxicity by rapid and synchronized dormancy breaking (RSDB) in case of oral administration as food as follows.
1. The anti-cancer (cytotoxicity) and pro-immunity of natural killer cell were confirmed using RSDB grape seed endosperms of Kyoho (Vitis labruscana) grown in Japan and Isabella (Vitis labruscana) grown in India. They are suitable for fight and prevention against cancer diseases and against infection.
2. The liver fat reduction and liver activation were confirmed using RSDB ginger rhizome (Zingiber officinale) grown in Japan.
3. The inhibition of the carbohydrates breakdown was confirmed using RSDB jumbo leek rhizome (Allium ampeloprasum) grown in Japan, suitable for fight and prevention against obesity and diabetes.
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| 6. |
Toshihiro Ona,Teruyuki Seino,Junko Johzuka, Liver mitochondrial activation and fat-lowering effects of rapid and synchronized dormancy-breaking ginger rhizome assumed to be administered orally, 7th International Electronic Conference on Medicinal Chemistry, 2021.11, [Background/Aims] In ginger rhizome, the percentage of 6-gingerol (GING) is high before heating and the percentage of 6-shogaol (SHOG) increases after heating, and SHOG has been reported to activate liver metabolism. We have studied the method of rapid and synchronized dormancy-breaking treatment with Awakening liquidTM to promote phase transition of content components in seeds, resulting in improved functionality. In this study, we applied this method to ginger rhizome. After the treatment and heating, we evaluated liver activation and final fat-reduction before and after metabolism in the liver, assuming oral administration, including comparison between GING and SHOG.
[Results] At 100nM, apoptotic toxicity was observed in GING both before and after metabolism, and promotion of metabolic activity was observed in SHOG, about 2 times greater after metabolism than before. In SHOG 1000nM, apoptotic toxicity was observed before metabolism, and promotion of metabolic activity was observed after metabolism by detoxification. The SHOG 100 nM decreased liver fat by about 20%. In dormancy-breaking ginger rhizome endosperm, SHOG was found to increase, containing about 8 times more than untreated and about 1.5 times more than non-dormancy-breaking one. There was also a concentration-dependent decrease in liver fat, which was about 27% at 2.0 mg/ml. However, apoptotic toxicity was observed at 3.0 mg/ml and above. Similarly, untreated sample showed little effect at 1.0 mg/ml, and apoptotic toxicity was observed at 2.0 mg/ml and above. In the model experiment assuming oral administration, rapid and synchronized dormancy-breaking ginger rhizome showed high effects on liver mitochondrial activation and fat lowering.. |
| 7. |
Root vegetables, bulbs and fruits that are not fully ripe need to be ripened by ethylene gas treatment. Using a proprietary method, we have developed an edible liquid that can rapidly ripen them in 1-2 days. The use of the chemical ethylene can be avoided and an eco-friendly life is realized! Kiwis, melons, mangoes, dekopons, avocados, bananas, sweet potatoes, potatoes, pumpkins, etc., with the taste of full ripeness! Kiwis soaked in the liquid for two days ripened without becoming soft, while the sugar content of steamed Anno potatoes ripened from 21 to 29. Avocados ripened while still green. Commercialization of citrus fruits picked from agricultural waste. Ripening period↓, sugar content↑, greenish taste↓, bitter taste↓, nutritional value (composition)↑, appearance↑, unit price↑, cost↓
Breaking the dormancy of seeds and bulbs of dormant plants requires a long period of time and is difficult to synchronize. We have developed an edible liquid that can be used to wake up seeds and bulbs quickly and synchronizedly within 5 days. It can also be used for germination. The intrinsic functional ingredients are bioconverted to water soluble form. Harmful ingredients are reduced, umami is increased, and health consciousness is realized. Garlic, ginger, peanuts, walnuts, sesame seeds, chestnuts, etc. can be awakened to become highly functional with the life force of a baby! Commercialization of seeds from agricultural waste. Product differentiation↑, Health-consciousness↑, Deliciousness↑, Active ingredients↑, Harmful ingredients↓, Unit price↑
Examples of commercialization using the liquid, "Kyoho no Mezame (Awakened Kyoho grape seed nuts)" (grape seed endosperm that breaks dormancy quickly and synchronizedly) and "Ginger no Mezame (Awakened ginger rhizome)" (ginger rhizome that breaks dormancy quickly and syncronizedly) are also exhibited. The ingredients are not concentrated or extracted, but are used as they are in foods that have been eaten for a long time, making them safe and reliable and highly functional.. |
| 8. |
Root vegetables, bulbs and fruits that are not fully ripe need to be ripened by ethylene gas treatment. We have developed an edible liquid that can be rapidly ripened in 1-2 days by our original method. On the other hand, it takes a long time to break the dormancy of seeds and bulbs of dormant plants, and synchronization is difficult. Using a proprietary method, we have developed an edible liquid that can be used to wake up seeds and bulbs quickly and synchronizedly within five days. It can also be used for germination and biotransformation of intrinsic functional ingredients into water-soluble form, making them tasty, effective, and harmful. It will contribute to the functionalization of foods and the development of a sixth industry.
The world's first rapid ripening "Post-ripening Liquid" and the world's first rapid and synchronized dormancy-breaking "Awakening Liquid". The exhibition will feature the world's first "Post-ripening liquid" for rapid ripening and the world's first "Awakening liquid" for rapid and synchronized dormancy breaking. Furthermore, as an example of commercialization using the liquid, "Kyoho no Mezame (Awakened Kyoho grape seed nuts," which is a rapid and synchronized dormancy-breaking Kyoho grape seed embryo milk, was also exhibited. The ingredients are not concentrated or extracted, and the food, which has been consumed for a long time, is successfully made safe and secure with high functionality.. |
| 9. |
Junko Johzuka,Toshihiro Ona , One hour in vivo-like phenotypic screening system for anti-cancer drugs using a high precision surface Plasmon resonance device, 6th International Electronic Conference on Medicinal Chemistry, 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 |
| 10. |
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, 6th International Electronic Conference on Medicinal Chemistry, 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.. |
| 11. |
Root vegetables, bulbs and fruits that are not fully ripe need to be ripened by ethylene gas treatment. We have developed an edible liquid that can rapidly ripen them in 1-2 days by our original method. On the other hand, it takes a long time to break the dormancy of seeds and bulbs of dormant plants, and synchronization is difficult. Using a proprietary method, we have developed an edible liquid that can be used to wake up seeds and bulbs quickly and synchronizedly within five days. It can also be used for germination and biotransformation of intrinsic functional ingredients into water-soluble form, making them tasty, effective, and harmful. It will contribute to the functionalization of foods and the development of a sixth industry.. |
| 12. |
Toshihiro - Ona, Junko - Johzuka, Killing efficacy of rapid and synchronized dormancy-broken Kyoho grape seed endosperm against human breast and liver cancers, The 26th International Congress on Nutrition and Integrative Medicine (ICNIM 2017), 2019.07, In ICNIM2017, we reported that rapid and synchronized dormancy-broken domestic Kyoho grape seed endosperm (RSDB-GSE) possessed direct killing efficacy against pancreatic cancer cell with low toxicity in case of oral administration. It was achieved by phase transition of endosperm ingredients by the Grandir recipe for RSDB. Furthermore, in ICNIM 2018, we showed activation of natural killer (NK) cell by RSDB-GSE for pro-immune to fight against cancers. From these, RSDB-GSE was expected to express synergistic effect by these two types of efficacy against anti-cancer at similar physiological concentration. Here, we examined direct killing efficacy against breast and liver cancer cells.. |
| 13. |
Toshihiro - Ona, Junko - Johzuka, Pro-immune efficacy by rapid and simultaneous dormancy-broken grape seed endosperm, The 26th International Congress on Nutrition and Integrative Medicine (ICNIM 2017), 2018.07, Pro-immune efficacy by rapid and simultaneous dormancy-broken grape seed endosperm. |
| 14. |
Toshihiro - Ona, Junko - Shibata, Evaluation of efficacy and toxicity for dormancy-breaking grape seed supplement against pancreatic cancer, The 25th International Congress on Nutrition and Integrative Medicine (ICNIM 2017), 2017.07, Evaluation of efficacy and toxicity for dormancy-breaking grape seed supplement against pancreatic cancer. |
| 15. |
Toshihiro - Ona, Junko - Shibata, Development of universal method for skin stimulus test using epidermal keratinocyte mitochondria as a forefront of sensory system, The 24th International Congress on Nutrition and Integrative Medicine (ICNIM 2016), 2016.07, Development of universal method for skin stimulus test using epidermal keratinocyte mitochondria as a forefront of sensory system. |
| 16. |
Toshihiro - Ona, Junko - Shibata, Akio - Hayakawa, Shinya - Sakamoto, Rapid in vivo-like phenotypic assay of epidermis pro-turnover efficacy with toxicity by gold kiwi fruit grown in Saga prefecture, Japan, The 23rd International Congress on Nutrition and Integrative Medicine (ICNIM 2014), 2015.07, Rapid in vivo-like phenotypic assay of epidermis pro-turnover efficacy with toxicity by gold kiwi fruit grown in Saga prefecture, Japan. |
| 17. |
Jyunichi - Ohshima, Kazuya - Iizuka, Futoshi - Ishiguri, Shinso - Yokota, Toshihiro - Ona, Relationship between various extracted basic densities and cell morphology in Eucalyptus, International Symposium on Wood Science and Technology 2015 (IAWPS 2015), 2015.03, Wood density is an important index for mechanical and pulp properties of wood. However, it is affected by the existence of extraneous compounds. Here, the relationship between various extracted basic densities and cell morphology were investigated by their within-tree variations in Eucalyptus camaldulensis and E. globulus grown at the same site of Western Australia with the age of 14. For various extracted basic densities, examined were for basic density (BD), extractives-free basic density (EF-BD), alkali-extractives-free basic density (AF-BD), total-extractives-free basic density (TF-BD), and extraneous compounds-free basic density (ECF-BD). For cell morphology, investigated were fiber, vessel, apotracheal and paratracheal axial parenchyma cells, proportion of cell types, and derived wood properties. In E. camaldulensis, BD had significant negative relationships with fiber length, and tangential diameter and wall thickness in paratracheal axial parenchyma only (example of BD vs wall thickness of paratracheal axial parenchyma in Fig. 1, r=-0.530**), while EF-BD, TF-BD, and ECF-BD had the relationships with many cell morphology. In E. camaldulensis, BD was influenced by the presence of extraneous compounds and did not clearly reflect the cell morphology. In E. globulus, almost all fiber and vessel morphology had significant correlations with all basic densities (example of BD vs fiber wall thickness in Fig. 2, r=0.585**), whereas apotracheal and paratracheal axial parenchyma cell morphology had low or no correlations. Vessel percentage did not relate to all basic densities, although other proportion of cell types significantly related to them. Highly significant correlations were obtained between derived wood properties and all basic densities. In contrast to E. camaldulensis, E. globulus showed distinctly high correlations between almost all cell morphology and all basic densities. From these, in E. globulus, BD expresses cell morphology well without the disturbance by extraneous compounds. Based on the obtained results, between-species difference was clearly observed for two eucalypt species grown at the same site with the same age in the relationship between BD and cell morphology. Our results will help the usage of BD and extracted BDs for wood and pulp & paper industries.. |
| 18. |
Junko - Shibata, Toshihiro - Ona, Development of in vivo-like cell-based screening system for rapid and quantitative toxicity and efficacy predictions against epidermis activation, Drug Discovery 2014, 2014.09, Animal test is widely used for compound toxicity and evaluation although the results obtained are different from human clinical test even using a monkey.
For in vitro compound screening against epidermis activation, cocultivation of keratinocyte with a target compound is often hired since animal test is inhibited for cosmetic development in EU. The 3D cell culture is performed using extracellular matrix and additionally fibroblast cells from dermis to reconstitute skin structure. However, this requires long time culture duration (ca. 1 month) because the method is based on endpoint. Furthermore, it will not allow simultaneous determination of toxicity of apoptosis and necrosis, and efficacy. Moreover, the cell activity decreases during long term culture causing low experimental repeatability.
We have developed a custom-made human cell-based assay device, High-Precision Surface Plasmon Resonance (HP-SPR), and in vivo-like method for anti-cancer drug screening. This monitors early stage of mitochondrial response to stimuli within live cells giving endpoint toxicity and efficacy predictions rapidly (ca. 1 hour) without cell culture.
We proved HP-SPR validity for prediction of target compound toxicity and efficacy at physiological concentration for epidermis activation, and successfully established new in vivo-like test.. |
| 19. |
Toshihiro - Ona, Junko - Shibata, New in vivo-like chemosensitivity test for physiological concentration of anti-cancer drug: Rapid and reliable prediction within 1 h, Drug Discovery 2014, 2014.09, Conventional technology of in vivo-like test for anti-cancer drug screening is based on expensive 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 previous device methods depend on the pharmaceutical mode of action, and give no relation to the conventional method. And a separate set of experiment is required to obtain both efficacy and toxicity results. Our new technology overcomes these all problems sensing dynamic cellular reaction against target compound(s) by laser.
The technological break points are as follows.
1. New device is developed for real-time monitoring of mitochondrial polarization status within live cells without both labeling and invasion.
2. Early mitochondrial polarization change rate after addition of target compound (s) is revealed as a key to quantitatively predict the final compound efficacy and toxicity against live cells.
3. 3D cell activity is obtained by conditioning 2D attached cells covered by extracellular matrix for short time with no cell division.. |
| 20. |
Toshihiro - Ona, Junko - Shibata, Cell-based rapid and quantitative toxicity and efficacy monitoring in consecutive before and after the compound metabolism within liver for two ginger compounds at physiological concentration, 9th World Congress on Alternatives and Animal Use in the Life Sciences, 2014.08, [Objectives] Animal test is often used for compound toxicity and efficacy evaluation although the results obtained are different from clinical test and show total metabolism [1]. We have developed an original human cell-based assay device, High-Precision Surface Plasmon Resonance (HP-SPR), and method [2,3]. Here we examined the applicability of HP-SPR to elucidate toxicity and efficacy including metabolism of two ginger compounds as examples working against liver before and after their metabolism.
[Methods] Human liver cell Hep G2 was used for two ginger compounds of 6-shogaol (SHOG) and 6-gingerol (GING). The 2D cultured cells were self-attached to an HP-SPR sensor chip surface and covered by collagen to obtain in vivo like cell status [3], and monitored.
[Results] In SHOG (heated GING), the efficacy of liver activation was observed at 100nM. This activation was twice higher in after metabolism than in before metabolism. Apoptotic effect as toxicity was observed at 1000nM to inhibit liver activation before the metabolism. However, after the metabolism, its effect was revered and detoxification was processed. In GING, 100nM showed apoptotic effect as toxicity before and after the metabolism, and no effect was observed below 100nM. The different mode of action was successfully monitored between two ginger compounds.
1. Damia G, D'Incalci M. Eur J Cancer, 2009; 45:2768-2781.
2. Ona T, Shibata J. Anal Bioanal Chem, 2010; 398:2505-2533.
3. Ona T. WO 2013/039112 A1.. |
| 21. |
Junko - Shibata, Shouhei - Yano, Teruyuki - Seino, Toshihiro - Ona, Anti-liver cancer effect of Chinese chive using in vivo-like test, The 22st International Congress on Nutrition and Integrative Medicine (ICNIM 2014), 2014.07, 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.. |
| 22. |
Toshihiro - Ona, Junko - Shibata, Activation of skin by AHCC: comparison between whole and low molecular weight fraction, The 22st International Congress on Nutrition and Integrative Medicine (ICNIM 2014), 2014.07, 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.
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| 23. |
Do you have problems in evaluation of foods and food compounds for anti-metabolic syndrome and anti-ageing together with toxicity? You may feel the test cost high, the result acquisition slow, and want to get results equivalent to clinical test, synergistic combination, consulting… We will overcome these for you with our sample screening service; that is, a rapid, high sensitive, label-free and non-invasive cell-based sensing technology for reliable efficacy prediction of bioactive foods and food compounds at physiological concentration within 1 h regardless to the mode of action.. |
| 24. |
Junko - Shibata, Toshihiro - Ona, Development of label-free, rapid and reliable efficacy prediction of anti-cancer drug at physiological concentration using high-precision surface plasmon resonance, The Twelfth Asian Conference on Analytical Sciences (ASIANALYSIS XII), 2013.08, Conventional chemosensitivity test is designed for efficacy prediction of target drug(s) using cell culture technique supplemented with the drug(s). In case of drug evaluation at physiological concentration to achieve in vivo like condition, it is based on three dimesion (3D) cell culture and endpoint assay necessitating time-consuming experimental procedure for about 2-4 weeks and labeling agents, sometimes causing low reliability of drug efficacy prediction.
We previously developed label-free cell-based sensing technology of High-Presision Surface Plasmon Resonance (HP-SPR) for new chemosensitivity test using second dimension (2D) cultured cells within 1 h after the addition of the drug regardless to the mode of action [1]. We achieved this by a custom-made instrument and method to monitor early stage of mitochondrial response to stimuli within live cells utilizing HP-SPR as a measurement principle. Furthermore, we have found mitochondrial polarization status change rate quantitatively implied endpoint cell status.
In the present study, we examined validity of HP-SPR for evaluation of anti-cancer drugs at physiological concentration to perform in vivo like test, using pancreatic cancer cell lines. To accomplish this, self-attached 2D cultured cells on a sensor chip were covered by extracellular matrix to activate cells into in vivo like status and subsequently monitored HP-SPR signal change after the drug addition for 1 h. The obtained HP-SPR signal change rate correlated to cell viability by conventional 3D cell culture chemosensitivity test of collagen gel droplet embedded culture drug sensitivity test [2].
Consequently, we proved HP-SPR validity as a label-free, rapid and reliable new chemosensitivity test for prediction of drug efficacy at physiological concentration regardless to the mode of action and successfully established new in vivo like test.
References
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| 25. |
Toshihiro - Ona, Junko - Shibata, Activation of skin by AHCC, The 21st International Congress on Nutrition and Integrative Medicine (ICNIM 2013), 2013.07, 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. This will not allow the simultaneous prediction of efficacy and toxicity. Alternative method using specialized device such as impedance monitoring depends on the pharmaceutical mode of action, and gives no relation to the conventional method.
New technology is brought by a custom-made instrument and method to monitor early stage of mitochondrial response to stimuli utilizing high-precision surface plasmon resonance as a measurement principle. And we have found mitochondrial polarization status change rate quantitatively implied endpoint cell status. From these, we overcome all problems described above and achieved prediction of endpoint cell status as efficacy and toxicity against target compound(s) correlating to conventional cell-based chemosensitivity test.
Expected applications for compound screening are anti-cancer, -Alzheimer disease and -aging (skin care), diabetes, pro-fat burning, -metabolism, -hair restoration and -body temp, even suitable in case of DDS.
We report the application of our new technology for skin activation by AHCC.
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| 26. |
toshihiro - ona, Shibata, J., Rapid and reliable cell-based sensing technology for efficacy prediction of anti-cancer and anti-metabolic syndrome compounds, 2012 Bio International Convention, 2012.06, We successfully developed a rapid, high sensitive, label-free and non-invasive cell-based sensing technology for reliable efficacy prediction of anti-cancer and anti-metabolic syndrome compounds in their physiological concentrations. This technology senses dynamic cellular reaction against target compounds (including combined use) and determines their efficacy within 1 h after the compound addition regardless to the pharmaceutical mode of action.. |
| 27. |
Development of rapid and reliable efficacy evaluation method for anti-cancer drugs and anti-metabolic syndrome in physiological concentration. |
| 28. |
Development of rapid and reliable efficacy evaluation method for cancer drugs in physiological concentration. |
| 29. |
Development of efficacy evaluation system for cancer drugs by optical monitoring mitochondrial activity in living cells . |
| 30. |
Rapid reliable cell-based sensing technology for efficacy prediction of anti-cancer and anti-metabolic syndrome compounds. |
| 31. |
Developement of rapid and reliable cell-based evaluation technology for efficacy of anti-cancer and anti-metabolic syndrome compounds. |
| 32. |
Nano-bio-device for health related market. |
| 33. |
Nano-bio-device for real-time-monitoring of live cells . |
| 34. |
Efficacy evaluation device of anti-cancer and anti-metabolic syndrome compounds with their combined use and its application. |
| 35. |
Quantitative evaluation of anti-cancer drug efficacy derived from trees with drug mechanism independent manner by high precision SPR-development of signaling echo method-. |
| 36. |
Quantitative evaluation of anti-breast cancer drug efficacy with drug mechanism independent manner by high precision SPR. |
| 37. |
Quantitative evaluation of anti-cancer drug efficacy with drug mechanism independent manner by high precision SPR-development of signaling echo method-. |
| 38. |
Developement of forest resources possessing high bioactivity. |
| 39. |
Evaluation of cell response of mitochondrial inner membrane polarization by real-time monitoring without labelling. |
| 40. |
Nanodevice for quick and quantitative evaluation in therapeutic potential of cancer drug cocktail, and ppt level ultra high sensitivity sensor quantifying environmental abnormal multi components . |
| 41. |
最先端ラマン分光法の高感度高速性の紹介と応用展開例
Introduction of high relationship between molecular and lattice information obtained by Raman spectroscopy and dynamic physical properties indices such as in polymer solution. Intoroduction of most advanced Raman spectroscopy in high sensitivity and high time-resolution with their applications.. |
| 42. |
Development of high sensitivity device system aiming practical cancer therapy. |
| 43. |
Application of custom-made spectroscopic device for polymer analysis and biophotonics. |
| 44. |
Rapid and quantitative method for evaluating the personal therapeutic potential of pancreatic cancer drugs. |
| 45. |
Rapid and quantitative method for evaluating the therapeutic potential of liver cancer drugs. |
| 46. |
Rapid and quantitative method for evaluating the personal therapeutic potential of pancreatic cancer drugs. |
| 47. |
Rapid and quantitative method for evaluating the therapeutic potential of liver cancer drugs. |
| 48. |
Cell-based automatic device for evaluation of cancer drug efficacy. |
| 49. |
Development of cell-based quantitative evaluation method and instrument on personal therapeutic potential of cancer drugs for apoptosis by high precision surface plasmon resonance sensor. |
| 50. |
Development of cavity mirror enhanced Raman spectroscopy (CMERS). |
| 51. |
Development of forest products possessing high functionality. |
| 52. |
Rapid and quantitative method and instrument for evaluating the personal therapeutic potential of cancer drugs. |
| 53. |
Development of wave-guided surface plasmon spectrometer for cancer drug efficacy evaluation aiming personal therapy. |
| 54. |
Development of cell-based quantitative evaluation method for cell cycle-arrest type cancer drugs for apoptosis by high precision surface plasmon resonance sensor. |
| 55. |
Quantitative prediction of therapeutic potential of cancer drugs including pharmacokinetic interactions for apoptosis by monitoring mitochondrial polarity. |
| 56. |
Quantitative Screening Method and Instrumentation of polyphenol for apoptotic efficacy against cancer including pharmacodynamic interactions. |
| 57. |
Quantitative prediction of anti-cancer efficacy of polyphenols by AKT and ERK1/2 down regulation. |
| 58. |
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. |
| 59. |
Quantitative prediction of cancer drug efficacy by AKT and ERK1/2 down regulation in pancreatic cancer. |
| 60. |
Quantitative prediction of therapeutic potential of cancer drugs including pharmacokinetic interactions for apoptosis in 5 min. |
| 61. |
Developement of rapid and quantitative screening system for anti-cancer compound derived from tree. |
| 62. |
Developement of method and instrumentation for linking health, environment and anaysis. |
