ORALS

SESSION: OxidativeMonAM-R1	Yoshikawa International Symposium (2nd Intl. Symp. on Oxidative Stress for Sustainable Development of Human Beings)
Mon. 28 Nov. 2022 / Room: Ballroom B
Session Chairs: Toshikazu Yoshikawa; Haruhiko Inufusa; Session Monitor: TBA

11:55: [OxidativeMonAM02] OS Plenary
Antioxidant treatment for Sleep Apnea
Haruhiko Inufusa¹ ; Fuhua Yang² ; Naomi Okada³ ; Toshikazu Yoshikawa⁴ ; ¹TIMA, Gifu, Japan; ²Division of Anti-Oxidant Research, Gifu University, Gifu, Japan; ³Gifu University, Gifu, Japan; ⁴Louis Pasteur Center, Kyoto, Japan;
Paper Id: 424 [Abstract]

The number of severely ill patients requiring treatment for sleep apnea syndrome (SAS) is estimated to be over 9 million in Japan and 400 million worldwide. The Space Shuttle Challenger tragedy and the Exxon Valdez maritime accident were caused by SAS, resulting in significant global economic losses. The only effective treatment currently available is CPAP, a forced-air ventilator worn during sleep, which is often discontinued by patients due to its complexity and noise.<br />About 70% of SAS patients have elevated mCRP and oxidative stress in their blood, indicating inflammation somewhere in the body.　Twendee X (TWX) is a very strong antioxidant composition and reduces inflammation which contains L-glutamine, ascorbic acid (VC), L-cystine, coenzyme Q10, succinic acid, fumaric acid, vitamin B2, and niacin. Clinical research of SAS using TWX for SAS was conducted at Takashima Clinic, Utsunomiya Japan. A survey of SAS patients was also conducted at Eyez Inc.The number of snoring as a major symptom of SAS was monitored. 2 weeks of treatment with TWX significantly reduced the number of snoring in 3 out of 5 patients, and 4 weeks of treatment significantly reduced the number of snoring in all 5 patients. 4 weeks of treatment with TWX showed the following effects in 30 SAS patients. TWX reduced snoring by 85%, nighttime waking phenomenon by 84%, deep sleep by 91%, daytime sleepiness by 84%, apnea by 85%, and headache upon awakening by 96%.<br />TWX is a supplement with no side effects and can be safely taken by children, suggesting that it may be effective for about 80% of SAS patients.

References:
1). Novel compositions and uses thereof WO 2008071790 A3\n2). Antioxidant composition and its use in diabetes EP 1932542 A1\n3). Pretreatment Twendee X® reduced acute ischemic brain damage. Kusaki M, Inufusa H, Abe K, JSCVD 2017; 26: 1191-1196\n4). Clinical and Pathological Benefit of Twendee X in Alzheimer's Disease Transgenic Mice with Chronic Cerebral Hypoperfusion. Liu Xia, Yamashita T, Abe K, et al. J Stroke Cerebrovasc Dis. 2019 Jul;28(7):1993-2002.

SESSION: OxidativeMonAM-R1	Yoshikawa International Symposium (2nd Intl. Symp. on Oxidative Stress for Sustainable Development of Human Beings)
Mon. 28 Nov. 2022 / Room: Ballroom B
Session Chairs: Toshikazu Yoshikawa; Haruhiko Inufusa; Session Monitor: TBA

12:20: [OxidativeMonAM03] OS
The antioxidant Twendee X and cancer treatment
Naomi Okada¹ ; Fuhua Yang² ; Taku Nakashima³ ; Haruhiko Inufusa⁴ ; Toshikazu Yoshikawa⁵ ; ¹Gifu University, Gifu, Japan; ²Division of Anti-Oxidant Research, Gifu University, Gifu, Japan; ³Department of Molecular and Internal Medicine, Graduate school of Biomedical & Health sciences, Hiroshima University, Hiroshima, Japan; ⁴TIMA, Gifu, Japan; ⁵Louis Pasteur Center, Kyoto, Japan;
Paper Id: 124 [Abstract]

As examples of cancer treatment, immunotherapy is now widely recognized as a powerful cancer therapy in addition to conventional therapies such as operation, chemotherapy and radiotherapy. Although they are all effective treatments, chemotherapy and radiotherapy, which take advantage of small differences between cancer cell (tumor cell) and normal cell, damage normal cells as well as cancer cells. As a result, adverse events are almost inevitable and are often difficult to continue treatment depending on the degree. 　As these treatments are administered over months or years, physical and mental damage accumulate, leading to a decline in physical strength and immunity. These adverse events are largely due to oxidative stress. That is, the tumor itself, and reactive oxygen species (ROS) generated by treatment for it, can cause damage to various cells due to oxidative stress. And oxidative stress can cause a decline in physical strength and immunity, which leads to tumor growth and a negative spiral. Thus, suppression of oxidative stress in tumor and cancer treatment may be very effective in not only alleviating symptoms but also prolonging prognosis. Since oxidative stress-mediated mechanisms of carcinogenesis also exist, suppression of oxidative stress may help to prevent cancer. An antioxidant compound Twendee X (TWX) which composed of vitamins, amino acids, and CoQ 10 is rated as the strongest and safe antioxidant in the world by ICDD (France). Cytoprotective effects of TWX against radiation exposure were examined using murine alveolar epithelial cells (LA-4). After allowing LA-4 cells to adhere, 25ug/ml of TWX was added in the medium. The cells were irradiated with 30Gy of radiation 2 hours after the addition, and after 72 hours, the effect on cell proliferative capacity was examined. The result showed that TWX significantly inhibited the radiation-induced decrease in cell proliferative capacity. In addition, TWX can be used in single and continuous doses in both healthy and pregnant mice and also increased NK activity, which is tumor immunity. In summary, TWX not only reduced adverse events during cancer treatment but also increased tumor immunity. We believe TWX can be an aid in the treatment and prevention of cancer.

References:
1. Novel compositions and uses thereof WO 2008071790 A3
2. Antioxidant composition and its use in diabetes EP 1932542 A1
3. Suplatast tosilate reduces radiation-induced lung injury in mice through suppression of oxidative stress. Yusuke Izumi, Taku Nakashima, Noboru Hattori, Free Radic Biol Med.2019;136:52-59
4. Balancing efficacy of and host immune responses to cancer therapy:the yin and yang effects. Shbaked Y, Nat Rev Clin Oncol.2016 Oct;13(10):611-26.
5. Systematic Review: Generating Evidence-Based Guidelines on the Concurrent Use of Dietary Antioxidants and Chemotherapy or Radiotherapy. Akiko Nakayama, Karen P, Obianuju Lgbokwe, and Leffrey D. white. Cancer Invest. 2011 Dec:29(10): 655-667.

SESSION: OxidativeMonAM-R1	Yoshikawa International Symposium (2nd Intl. Symp. on Oxidative Stress for Sustainable Development of Human Beings)
Mon. 28 Nov. 2022 / Room: Ballroom B
Session Chairs: Toshikazu Yoshikawa; Haruhiko Inufusa; Session Monitor: TBA

12:45: [OxidativeMonAM04] OS Keynote
Oxidative stress is major cause of allergic reaction in COVID-19 infection
Haruhiko Inufusa¹ ; Fuhua Yang² ; Naomi Okada³ ; Toshikazu Yoshikawa⁴ ; ¹TIMA, Gifu, Japan; ²Division of Anti-Oxidant Research, Gifu University, Gifu, Japan; ³Gifu University, Gifu, Japan; ⁴Louis Pasteur Center, Kyoto, Japan;
Paper Id: 123 [Abstract]

Oxidative stress and immune activity are closely related with disease or symptoms. Allergic reaction depending on imbalance of immune system which is easy to be damaged by oxidative stress. COVID-19 pneumonia starts at the lower and back side of lung, and CT scan shows typical interstitial pneumonia. Interstitial pneumonia is kind of allergic reaction, thus pneumonia start high blood flow area of lung and very rapidly spread to the whole lung. It is well known that interstitial pneumonia shows extremely high oxidative stress, damages lung tissue, and some patients show irreversible damage of the lung. High risks of COVID-19 infection are high age with underlying disease, especially diabetes mellitus, obesity, heart disease, hypertension, after stroke, cancer, neurogenic disorder, dementia and so on. All these risk factors show extremely high oxidative stress. Oxidative stress damages vascular endothelium and inflammation of vascular wall produces thrombosis, which are reasons of Kawasaki disease like vasculitis and cerebral infraction. Oxidative stress plays a major role in these symptoms.

References:
1. Novel compositions and uses thereof WO 2008071790 A3
2. Antioxidant composition and its use in diabetes EP 1932542 A1

SESSION: OxidativeMonPM1-R1	Yoshikawa International Symposium (2nd Intl. Symp. on Oxidative Stress for Sustainable Development of Human Beings)
Mon. 28 Nov. 2022 / Room: Ballroom B
Session Chairs: Fuhua YANG; Yoshiaki Harakawa; Session Monitor: TBA

14:00: [OxidativeMonPM105] OS Plenary
Antioxidant treatment for infertility
Haruhiko Inufusa¹ ; Fuhua Yang² ; Naomi Okada³ ; Toshikazu Yoshikawa⁴ ; ¹TIMA, Gifu, Japan; ²Division of Anti-Oxidant Research, Gifu University, Gifu, Japan; ³Gifu University, Gifu, Japan; ⁴Louis Pasteur Center, Kyoto, Japan;
Paper Id: 122 [Abstract]

The decline in fertility is a major problem in many countries, especially in Japan, where the total fertility rate in 2018 is 1.42. Half of infertility couples have male causes, and the other half have female causes. Oxidative stress can be a major cause of infertility. There are many reports that the number of spermatozoa is decreased by mobile phone that emits electromagnetic waves that cause oxidative stress in the pocket near the testicle, resulting in a decrease in motility. The decline in pregnancy rates with age in women is partly due to the firmness of the egg membrane due to oxidative stress.
Administration of an antioxidant composition would increase the pregnancy rate was examined. The monitor questionnaire was conducted by EYEZ,INC., which is a third-party institution, and the permission for secondary use of the results listed on the company's website (public data) was obtained. Twendee Mtcontrol (TwM) was used as antioxidant composition, which contains L-glutamine, ascorbic acid (VC), L-cystine, coenzyme Q10, succinic acid, fumaric acid, vitamin B2, niacin, Pantothenic acid, thiamine (VB1), pyridoxamine (VB6), folic acid, biotin, vitamin B12, lactoferrin. Twenty-eight couples who had been infertile for more than 2 years and wanted to raise their children joined the monitor questionnaire. For 1 year, both couples took TwM and their pregnancy was monitored. As a result, 9 pairs became pregnant, and the rate was 32% (9/28). Pregnancy rate is about 16% for a couple of more than two years of infertility when they receive infertility treatment at hospital. TwM showed almost 3 times higher pregnancy rate than infertility treatment at hospital. The possibility of antioxidant treatment for infertility was suggested.

References:
1.Novel compositions and uses thereof WO 2008071790 A3\n2.Antioxidant composition and its use in diabetes EP 1932542 A1\n3.Pretreatment Twendee X® reduced acute ischemic brain damage. Kusaki M, Inufusa H, Abe K, JSCVD 2017; 26: 1191-1196\n4.Clinical and Pathological Benefit of Twendee X in Alzheimer's Disease Transgenic Mice with Chronic Cerebral Hypoperfusion. XiaLiu, Abe K https://doi.org/10.1016/j.jstrokecerebrovasdis.2019.03.029

SESSION: OxidativeMonPM1-R1	Yoshikawa International Symposium (2nd Intl. Symp. on Oxidative Stress for Sustainable Development of Human Beings)
Mon. 28 Nov. 2022 / Room: Ballroom B
Session Chairs: Fuhua YANG; Yoshiaki Harakawa; Session Monitor: TBA

15:15: [OxidativeMonPM108] OS
Effect of antioxidant Twendee X on radiation-induced oxidative stress
Yoshiaki Harakawa¹ ; Fuhua Yang² ; Toshikazu Yoshikawa³ ; Haruhiko Inufusa⁴ ; ¹Division of Anti-oxidant Research, Gifu University, Gifu, Japan; ²Division of Anti-Oxidant Research, Gifu University, Gifu, Japan; ³Louis Pasteur Center, Kyoto, Japan; ⁴TIMA, Gifu, Japan;
Paper Id: 418 [Abstract]

It is known that when a living organism is exposed to radiation, water molecules in the body ionize, producing reactive oxygen species, which in turn cause inflammation.<br />Twendee X (TwX) is an antioxidant supplement consisting of eight ingredients (coenzyme Q10, vitamin C, vitamin B2, niacin, L-cystine, succinic acid, fumaric acid, and L-glutamine). We investigated whether TwX and other substances can inhibit oxidative stress generated by radiation.<br />C57BL/6J mice were orally administered the test substance TwX (15 mg/kg) for 3 days at 24-hour intervals, irradiated with 6 Gy of radiation on the fourth day, and blood samples were taken 72 hours later, and survival was observed as is. Vitamin C (15 and 150 mg/kg), Resveratrol, Astaxanthin, Protandim, and Pycnogenol were used as antioxidants for comparison to TwX. Plasma hydroperoxide level was measured as blood oxidative stress (d-ROMs test).<br />Increased d-ROMs test value by radiation was reduced by one third in the group treated with TwX (15 mg/kg) before irradiation for 3 days (t-test, p<0.05), and the oxidative stress level was lower in the group treated for 7 days than in normal mice. In addition, the administration of TWX suppressed radiation-induced deaths.<br />Vitamin C (150 mg/kg) and Astaxanthin treatment groups reduced d-ROMs test values, but the difference was not statistically significant. The low concentrations of vitamin C (15 mg/kg), Pycnogenol, and Protandim did not change the d-ROMs test values compared to the control group, and for the Resveratrol group, the d-ROMs test values increased compared to the control group. Furthermore, Twendee Mtcontrol (TwM), which added seven active ingredients to TwX, reduced radiation-related deaths with fewer doses than TwX.<br />TwX significantly reduced radiation-induced oxidative stress at a small dose of 15 mg/kg, while other single antioxidants with normal amount have no significant improvement in animal radiation model. High dose antioxidants may cause side effects. TwX, TwM are composed only of substances that exist in the human body, and a small dose of them showed high antioxidant activity. This may have reduced inflammation caused by oxidative stress, ultimately leading to an increase in survival rate. The antioxidant potential of Twendee series in this experiment suggests the possibility of future applications for various oxidative stress-related diseases.

References:
[1] Tadokoro K, et al. Clinical Benefits of Antioxidative Supplement Twendee X for Mild Cognitive Impairment: A Multicenter, Randomized, Double-Blind, and Placebo-Controlled Prospective Interventional Study. J Alzheimers Dis. 2019;71(3):1063-1069.\n[2] Yamamoto T, Kinoshita M, Shinomiya N, Hiroi S, Sugasawa H, Matsushita Y, Majima T, Saitoh D, Seki S. Pretreatment with ascorbic acid prevents lethal gastrointestinal syndrome in mice receiving a massive amount of radiation. J. Radiat. Res., 2010. 51, 145-156．\n[3] Yoshikawa T, et al. [All about Free Radicals – from Health to Environmental Pollution -] Kasseisanso , Free Radical no subete – Kennkou kara kannkyouosen made – (in Japanese), Maruzen Publishing, 2000, 78-79.\n[4] Seki Y. [Evaluation of total oxidative stress by d-ROMs testing] d-ROMs test niyoru sankasutoresu sougouhyouka (in Japanese). Seibutsu siryou bunseki (Journal of analytical bio-science), 2010. 32(4), 301-306.

SESSION: OxidativeTuePM1-R1	Yoshikawa International Symposium (2nd Intl. Symp. on Oxidative Stress for Sustainable Development of Human Beings)
Tue. 29 Nov. 2022 / Room: Ballroom B
Session Chairs: Shigeru Hirano; Session Monitor: TBA

14:00: [OxidativeTuePM105] OS Plenary
Antioxidant treatment for Long- COVID and mRNA vaccine side effects
Haruhiko Inufusa¹ ; Fuhua Yang² ; Naomi Okada³ ; Toshikazu Yoshikawa⁴ ; ¹TIMA, Gifu, Japan; ²Division of Anti-Oxidant Research, Gifu University, Gifu, Japan; ³Gifu University, Gifu, Japan; ⁴Louis Pasteur Center, Kyoto, Japan;
Paper Id: 425 [Abstract]

COVID-19 infection produces large amounts of Reactive Oxygen Species (ROS) in the human body, and the attack on the immune system by ROS causes a cytokine storm. Therefore, COVID19 infection generally does not cause severe disease in children with low ROS levels, but often causes severe disease and death in the elderly or those with underlying disease already ROS levels are high. The major problem with after COVID-19 pandemic is not only Long-COVID, but also the large number of patients who suffer from the tremendous side effects caused by mRNA vaccines. Pfizer and other genetic vaccines produce spike proteins and antibodies against them. Not only the spike protein itself causes ROS, but also pseudouridine and Lipid Nanoparticles (LNP) contained in the vaccine induce inflammation and increase ROS. In other words, both diseases are oxidative stress diseases. Twendee X (TwX) is a very strong antioxidant composition and reduces inflammation which L-glutamine, ascorbic acid (VC), L-cystine, coenzyme Q10, succinic acid, fumaric acid, vitamin B2, and niacin was used for patients. Patients took 3 tablets of TwX once a day for 4 weeks and answered a web-based questionnaire about their symptoms before and after taking TwX. A score of 0 on the questionnaire indicated no symptoms, while a score of 5 indicated severe symptoms. A total of 149 patients with mRNA vaccine sequelae completed the monitoring questionnaire completely. The monitored symptoms were Fatigue, Breathing difficulty, Chest pain, Smell or taste disorder, Headache, Brain fog, Joint pain, and Dizziness. All monitored symptoms showed significant post-medication score reductions and symptom improvement. Especially in patients with moderate to severe disease with a score of 3 or higher, the mean score dropped to less than half of the pre-medication level for all items. TwX were thought to be effective in relieving symptoms in these patients, many of whom suffer from mRNA vaccine sequelae worldwide.

References:
1). Novel compositions and uses thereof WO 2008071790 A3\n2). Antioxidant composition and its use in diabetes EP 1932542 A1\n3). Pretreatment Twendee X® reduced acute ischemic brain damage. Kusaki M, Inufusa H, Abe K, JSCVD 2017; 26: 1191-1196\n4). Clinical and Pathological Benefit of Twendee X in Alzheimer's Disease Transgenic Mice with Chronic Cerebral Hypoperfusion. Liu Xia, Yamashita T, Abe K, et al. J Stroke Cerebrovasc Dis. 2019 Jul;28(7):1993-2002.\n5). Antioxidant composition Twendee X may improve long COVID symptoms. You F, Tanaka S, Inufusa H, et al.　Brain Supplement. 2022; 4: 7-12.\n6). Effects of Antioxidant composition Twendee X on side effects of SARS -COV-2 mRNA vaccine. You F, Tanaka S, Inufusa H, et al. Brain Supplement. 2022; 4: 1-6.

SESSION: OxidativeTuePM1-R1	Yoshikawa International Symposium (2nd Intl. Symp. on Oxidative Stress for Sustainable Development of Human Beings)
Tue. 29 Nov. 2022 / Room: Ballroom B
Session Chairs: Shigeru Hirano; Session Monitor: TBA

14:25: [OxidativeTuePM106] OS
Role of Antioxidants in Oxidative Stress Induced by High Intake of Alcohol
Fuhua Yang¹ ; Yoshiaki Harakawa² ; Toshikazu Yoshikawa³ ; Haruhiko Inufusa⁴ ; ¹Division of Anti-Oxidant Research, Gifu University, Gifu, Japan; ²Division of Anti-oxidant Research, Gifu University, Gifu, Japan; ³Louis Pasteur Center, Kyoto, Japan; ⁴TIMA, Gifu, Japan;
Paper Id: 429 [Abstract]

Although there are genetic differences in the metabolism of alcohol, large amounts of alcohol intake can cause hangovers and, in severe cases, death due to acute alcohol poisoning. Once alcohol enters the body, it is metabolized mainly in the liver, where it is broken down into the highly hepatotoxic acetaldehyde. It is thought that excessive alcohol intake causes insufficient metabolism in the body, and the acetaldehyde induces oxidative stress, resulting in hangovers and organ damage. Therefore, we examined the effects of large amounts of alcohol on the body using Twendee S (TwS), an antioxidant formula consisting of coenzyme Q10, vitamin C, vitamin B2, niacin, L-cystine, succinic acid, fumaric acid, and L-glutamine. Mice were intraperitoneally administered ethanol, and blood acetaldehyde and oxidative stress were measured. The results showed that acetaldehyde in the blood increased at 2 hours after ethanol administration and was hardly detectable at 24 hours. However, oxidative stress remained high even after 72 hours of ethanol administration, and antioxidant capacity was low. In contrast, TwS-treated group suppressed blood acetaldehyde and the rising oxidative stress without reducing antioxidant capacity. In the mouse acute alcohol intoxication experiment, all mice in the Control group, to which nothing was administered, died after 12 hours. However, in the group that received 4 doses of TwS at 12-hour intervals the survival rate was 100%. These results suggest that heavy alcohol intake increases oxidative stress in the body, and that this oxidative stress may be a factor in acute poisoning deaths. In contrast, the antioxidant compound TwS suppressed acute poisoning deaths by speeding up the metabolism of alcohol and suppressing elevated oxidative stress. In conclusion, heavy and prolonged alcohol intake causes significant oxidative stress in the body, which may lead to aging and disease. Therefore, countermeasures against oxidative stress caused by alcohol intake are important, and TwS was shown to have the potential to reduce this risk.

References:
1). Markus Graf V. Matuschka-Greinffenclau, Hans Peter Jander. Alcohol metabolism moderating composition. WO 2005077464 A1. 2005-08-25.\n2). Markus Graf Matuschka Von Greiffenclau, Haruhiko Inufusa. Composition for accelerating alcohol metabolism and for reducing the risk of alcohol induced diseases. WO 2012095509 A8. 2012-10-04.\n3). Petrella C, Carito V Ceccanti M, et al. Oxidative stress inhibition by resveratrol in alcohol-dependent mice. Nutrition. 2020;79-80:110783.

SESSION: OxidativeTuePM1-R1	Yoshikawa International Symposium (2nd Intl. Symp. on Oxidative Stress for Sustainable Development of Human Beings)
Tue. 29 Nov. 2022 / Room: Ballroom B
Session Chairs: Shigeru Hirano; Session Monitor: TBA

14:50: [OxidativeTuePM107] OS
Potential for Antioxidant Therapy for Diabetes and Diabetic Complications
Yoshiaki Harakawa¹ ; Fuhua Yang² ; Toshikazu Yoshikawa³ ; Haruhiko Inufusa⁴ ; ¹Division of Anti-oxidant Research, Gifu University, Gifu, Japan; ²Division of Anti-Oxidant Research, Gifu University, Gifu, Japan; ³Louis Pasteur Center, Kyoto, Japan; ⁴TIMA, Gifu, Japan;
Paper Id: 426 [Abstract]

Diabetes is known to be a disease caused by oxidative stress, and also known to generates its own oxidative stress, resulting in numerous complications. Twendee X (TwX) is an antioxidant combination supplement with international and Japanese patents, and has been certified as effective in preventing dementia from a randomized, double-blind clinical trial led by the Japan Society for Dementia Prevention in 2019. In recent years, we have also successfully developed Twendee Mtcontrol (TwM), an antioxidant with seven more ingredients added to TwX to increase its antioxidant effect. The antioxidants TwX and TwM were tested in animal models to see if they can treat diabetes and diabetic complications. To examine the effect of antioxidants on diabetes, a diabetic disease model was developed. TwX or TwM was orally administered daily for 2 months. Afterwards, a glucose tolerance test was performed, and blood glucose levels were measured over time. In addition, insulin levels were measured under the same conditions. The results showed that the peak blood glucose level was significantly improved in the TwX group, and the peak level was also improved in the TwM group compared to the control group. Insulin levels also tended to be lower than controls in both groups. In addition, to verify the effectiveness of the supplements in preventing actual diabetic complications, we tested it on SDT fatty rats, which develop diabetic complications early in the course of the disease. These rats develop elevated blood pressure and cataracts at a much earlier stage due to the progression of diabetes mellitus. SDT fatty rats (5-week-old males) were divided into three groups (n=8): control (non-treated), TwX 40 mg/kg/Day, and TwM 40 mg/kg/Day. At 17 weeks of age, most rats in the non-treated group showed severe cataracts, whereas those in the TwM-treated group had relatively mild cataracts. Systolic blood pressure also tended to increase with age in the non-treated group, but the TwX and TwM groups significantly suppressed the age-related increase in blood pressure. These results suggest that TwX and TwM reduced blood glucose levels in an insulin-independent manner, thereby reducing diabetic complications such as hypertension and cataracts. TwX reduced blood glucose levels more, and TwM had a pronounced cataract-preventive effect. TwM may have a broader range of disease-preventive effects. however, further studies are required for the detailed mechanism of action. These results suggest the possibility of antioxidant therapy for diabetes mellitus and its complications.

References:
[1] Yamagishi S. [Diabetes and Oxidative Stress] Tounyoubyou to Sanka Sutoresu (in Japanese), Medical Review Co., Ltd., 2011, 3.\n[2] Masuyama T, Katsuda Y, and Shinohara M. A novel model of obesity-related diabetes: introgression of the Lepr^fa allele of the Zucker fatty rat into nonobese Spontaneously Diabetic Torii (SDT) rats. Exp. Anim. 54(1): 13-20, 2005.\n[3] Matsui K, Ota T, Matsushita M, et al. Diabetes-associated complications in Spontaneously Diabetic Torii Fatty Rats. Exp. Anim. 57(2): 111-121, 2008.

SESSION: OxidativeWedPM1-R1	Yoshikawa International Symposium (2nd Intl. Symp. on Oxidative Stress for Sustainable Development of Human Beings)
Wed. 30 Nov. 2022 / Room: Ballroom B
Session Chairs: Koji Fukui; Koji Abe; Session Monitor: TBA

14:00: [OxidativeWedPM105] OS Plenary
Twendee X improves cognitive function in vitamin E-deficient mice
Koji Fukui¹ ; Fuhua Yang² ; Yugo Kato¹ ; Yoshiaki Harakawa³ ; Toshikazu Yoshikawa⁴ ; Haruhiko Inufusa⁵ ; ¹Shibaura Institute of Technology, Saitama, Japan; ²Division of Anti-Oxidant Research, Gifu University, Gifu, Japan; ³Division of Anti-oxidant Research, Gifu University, Gifu, Japan; ⁴Louis Pasteur Medical Research Center, Kyoto, Japan; ⁵TIMA, Gifu, Japan;
Paper Id: 419 [Abstract]

Reactive oxygen species may attack several living organs and gradually accumulate oxidative products such as lipid hydroperoxides. Recently, several kinds of evidence have been demonstrated that accumulation of oxidative products induces onset and progression of several severe diseases such as inflammations, cardiovascular events and more [1]. One severe disease is dementia, and it also deeply relates to senescence. To prevent age-related neurodegenerative disorders, treatment with antioxidant is effective. Twendee X, a mixed antioxidant supplement contains 8 substances [2] and anyone can buy it. However, the detailed scientific evidence is not enough. In this study, we fed Twendee X to vitamin E-deficient mice and check cognitive function.<br />We created vitamin E-deficient mice and Twendee X treated for one month. Cognitive function was measured by the Morris Water maze task [3].<br />Vitamin E deficient mice were significantly decrease learning ability. Treatment with Twendee X was significantly improved it. Brain neurotrophic factors in Twendee X-treated mouse were significantly increase compared to the untreated groups.<br />Twendee X has a potential as a powerful antioxidant supplement through protection against neuronal dysfunction.

References:
[1] Yoshida, N, Kato, Y, Takatsu, H, et al., Biomedicines 10 (2022) 28.\n[2] Tadokoro, K, Morihara, R, Ohta, Y, et al., J Alzheimer’s Dis, 71 (2019) 1063-1069.\n[3] Fukui, K, Onodera, K, Shinkai, T, et al., Ann NY Acad Sci, 928 (2001) 169-176.

SESSION: OxidativeWedPM1-R1	Yoshikawa International Symposium (2nd Intl. Symp. on Oxidative Stress for Sustainable Development of Human Beings)
Wed. 30 Nov. 2022 / Room: Ballroom B
Session Chairs: Koji Fukui; Koji Abe; Session Monitor: TBA

14:50: [OxidativeWedPM107] OS
Anti-oxidative approach to prevent Alzheimer's type of dementia
Koji Abe¹ ; Haruhiko Inufusa² ; Fuhua Yang³ ; Toshikazu Yoshikawa⁴ ; ¹National Center of Neurology and Psychiatry, Tokyo, Japan; ²TIMA, Gifu, Japan; ³Division of Anti-Oxidant Research, Gifu University, Gifu, Japan; ⁴Louis Pasteur Center, Kyoto, Japan;
Paper Id: 423 [Abstract]

Alzheimer's disease (AD) and chronic cerebral hypoperfusion (CCH) often coexist in dementia patients in aging societies. The hallmarks of AD including amyloid-β (Aβ)/phosphorylated tau (pTau) and pathology-related events such as neural oxidative stress and neuroinflammation play critical roles in pathogenesis of AD with CCH. A large number of lessons from failures of drugs targeting a single target or pathway on this so complicated disease indicate that disease-modifying therapies targeting multiple key pathways hold potent potential in therapy of the disease. In the present study, we used a novel mouse model of AD with CCH to investigate a potential therapeutic effect of a free radical scavenger, Edaravone (EDA) on AD with CCH via examining motor and cognitive capacity, AD hallmarks, neural oxidative stress, and neuroinflammation. Compared with AD with CCH mice at 12 months of age, EDA significantly improved motor and cognitive deficits, attenuated neuronal loss, reduced Aβ/pTau accumulation, and alleviated neural oxidative stress and neuroinflammation. These findings suggest that EDA possesses clinical and pathological benefits for AD with CCH in the present mouse model and has a potential as a therapeutic agent for AD with CCH via targeting multiple key pathways of the disease pathogenesis. In a clinical setting, anti-oxidative dietary supplement TwendeeX prevented cognitive function or even improved it with MMSE (minimental score examination) in mild cognitive impairment population. These results strongly suggest a promising function of anti-oxidative approach to prevent dementia.

References:
1). Tadokoro K, Morihara R, Abe K, et al. Clinical Benefits of Antioxidative Supplement Twendee X for Mild Cognitive Impairment: A Multicenter, Randomized, Double-Blind, and Placebo-Controlled Prospective Interventional Study. J Alzheimers Dis. 2019; 71(3): 1063-1069.\n2.) Feng T, Yamashita T, Abe K, et al. Clinical and Pathological Benefits of Edaravon for Alzheimar”s Disease with Chronic Cerebral Hypoperfusion in a Novel Mouse Model. J Alzheimars Dis. 2019; 71(1);327-339. \n3). Feng T, Yamashita T, Abe K, et al. In Vitro Free Radical Scavenging Activities of Dietary Supplements by Electron Spin Resonance. Brain Supplements. 2020; 2: 1-12.\n4). Tadokoro K, Ohta Y, Abe K, et al. Prevention of Cognitive Decline in Alzheimer’s Disease by Novel Sntioxidative Supplements. Int J Mol Sci. 2020 Mar 13:21(6)

SESSION: OxidativeWedPM1-R1	Yoshikawa International Symposium (2nd Intl. Symp. on Oxidative Stress for Sustainable Development of Human Beings)
Wed. 30 Nov. 2022 / Room: Ballroom B
Session Chairs: Koji Fukui; Koji Abe; Session Monitor: TBA

15:15: [OxidativeWedPM108] OS
Involvement of anti-oxidant Twendee X in immunity.
Fuhua Yang¹ ; Naomi Okada² ; Haruhiko Inufusa³ ; Toshikazu Yoshikawa⁴ ; ¹Division of Anti-Oxidant Research, Gifu University, Gifu, Japan; ²Gifu University, Gifu, Japan; ³TIMA, Gifu, Japan; ⁴Louis Pasteur Center, Kyoto, Japan;
Paper Id: 103 [Abstract]

Many diseases and physical disorders are caused by both external and internal factors, but the common denominator is inflammation and oxidative stress. Suppressing oxidative stress on a daily basis and maintaining a high level of self-immunity can help prevent and treat disease and maintain good health.
Intestines are involved in about 60% of our immune system. There are over 100 species of, or 100 trillions of bacteria in our intestines, and their ideal balance is 20% of good, 10% of bad, and 70% of opportunistic bacteria. This balance is lost by disturbed life rhythm and various diseases, in other words “oxidative stress”, being said to decrease bacterial diversity. Reducing oxidative stress and increasing diversity of intestinal bacteria is important for maintaining healthy life. Composed of vitamins, CoQ10, and amino acids, Twendee X (TwX) is an antioxidant which is confirmed to have a dementia-prevention effect. Effects of oxidative stress reduction by taking antioxidants on the bacterial diversity of the intestinal microflora and the balance of the intestinal microflora are examined.
Rats were given an antifungal agent OPP for 2 weeks, then blood and feces samples were collected to analyze blood oxidative stress and intestinal bacterial diversity. TwX was administrated from 1 week before OPP initiation. Although OPP rat group increased blood oxidative stress by 5%, no significant change was observed in microbial flora diversity. A rat group given TwX since 1 week before decreased oxidative stress by 13.3% and increased bacterial diversity 1.5 times higher, compared to rats which wasn’t given, and the number of Akkermansia, anti-inflammatory bacteria, was increased 3.5 times more. In the questionnaire survey for healthy participants who were given TwX for 1 month, the number of good bacteria and intestinal bacterial diversity were increased. Their defecation status and QOL were improved with that.
TwX tends to reduce oxidative stress, to increase the number of bacteria and diversity in intestinal microflora. It was suggested that this enables to protect human body from harmful substances invaded inside, improve immune system, and improve daily defecation status and QOL.

References:
1. Novel compositions and uses thereof WO 2008071790 A3\n2. Antioxidant composition and its use in diabetes EP 1932542 A1\n3. Composition for accelerating alcohol metabolism and for reducing the risk of alcohol induced diseases WO 2012095509 A8\n4. Composition for moderating triglyceride and cholesterol levels WO 2009062910 A1

SESSION: OxidativeWedPM2-R1	Yoshikawa International Symposium (2nd Intl. Symp. on Oxidative Stress for Sustainable Development of Human Beings)
Wed. 30 Nov. 2022 / Room: Ballroom B
Session Chairs: Hironori Nakagami; Session Monitor: TBA

15:55: [OxidativeWedPM209] OS
Usefulness of Antioxidant Combination Products for Chronic Fatigue Syndrome
Fuhua Yang¹ ; Yoshiaki Harakawa² ; Toshikazu Yoshikawa³ ; Haruhiko Inufusa⁴ ; ¹Division of Anti-Oxidant Research, Gifu University, Gifu, Japan; ²Division of Anti-oxidant Research, Gifu University, Gifu, Japan; ³Louis Pasteur Center, Kyoto, Japan; ⁴TIMA, Gifu, Japan;
Paper Id: 427 [Abstract]

In Chronic Fatigue syndrome (CFS), a complex combination of factors such as infections and excessive stress in daily life causes abnormalities in the immune system, endocrine system, brain, and neurological functions. The pathogenesis of the syndrome is not yet clear, and no treatment has been established. Recently, it has been reported that CFS-like symptoms are also observed in COVID-19 sequelae and SARS-CoV-2 vaccine sequelae, and there is an urgent need to clarify the pathogenesis mechanism and establish prevention and treatment methods. Abnormal energy metabolism due to mitochondrial dysfunction and inflammation in the brain have been reported as one of the causes of CFS. Since oxidative stress is highly related to both of these factors, it was thought that suppressing oxidative stress may lead to improvement of various symptoms of CFS. Therefore, we analyzed the symptom changes before and after the use of an antioxidant combination supplement, Twendee Mtcontrol (TwM), taken by people who had been diagnosed with chronic fatigue syndrome. 2 months of TwM intake resulted not only in an anti-fatigue effect, but also in improvement of pains and QOL, compared to before intake. Although the effects of various antioxidants have been reported for CFS, most of them are mainly for fatigue, and there are no reports showing significant effects on various symptoms. This may be due to differences in the combination of antioxidants and their antioxidant capacity. The results of this study may be useful for future treatment and prevention of CFS.

References:
1). Feng T, Yamashita T, Abe K, et al. In Vitro Free Radical Scavenging Activities of Dietary Supplements by Electron Spin Resonance. Brain Supplements. 2020; 2: 1-12.\n2). You F, Tanaka S, Inufusa H, et al. Effects of Antioxidant composition Twendee X on side effects of SARS -COV-2 mRNA vaccine. Brain Supplement. 2022; 4: 1-6.\n3). You F, Tanaka S, Inufusa H, et al. Antioxidant composition Twendee X may improve long COVID symptoms. Brain Supplement. 2022; 4: 7-12.

SESSION: OxidativeWedPM2-R1	Yoshikawa International Symposium (2nd Intl. Symp. on Oxidative Stress for Sustainable Development of Human Beings)
Wed. 30 Nov. 2022 / Room: Ballroom B
Session Chairs: Hironori Nakagami; Session Monitor: TBA

16:20: [OxidativeWedPM210] OS
Antioxidants and Cancer Cells
Fuhua Yang¹ ; Yoshiaki Harakawa² ; Naomi Okada³ ; Toshikazu Yoshikawa⁴ ; Haruhiko Inufusa⁵ ; ¹Division of Anti-Oxidant Research, Gifu University, Gifu, Japan; ²Division of Anti-oxidant Research, Gifu University, Gifu, Japan; ³Gifu University, Gifu, Japan; ⁴Louis Pasteur Center, Kyoto, Japan; ⁵TIMA, Gifu, Japan;
Paper Id: 428 [Abstract]

Oxidative stress is involved in over 150 different diseases, and cancer is one of the most common. High oxidative stress can damage proteins and DNA that are essential to the body, leading to the formation of cancer over time. Oxidative stress in the body is also elevated in carcinoma carriers. When the body's oxidative stress is elevated, inflammatory substances increase, and the body's cancer immunity is reduced, which is thought to promote cancer cell growth and metastasis. Since antioxidants are thought to play an effective role in breaking this vicious cycle, we used the antioxidant compound Twendee X (TwX) to verify the effect of TwX on cancer cells through in vivo and in vitro experiments. Human colon cancer RPMI4788 cells are used as cancer cells. In in vitro experiments, the addition of 60 μg/ml of TwX to RPMI4788 cell culture medium slowed cell proliferation from 1 hour after addition, whereas the same concentration of the antioxidant Vitamin C did not inhibit cell proliferation. The inhibition of cell proliferation by the addition of TwX was not observed in normal cells. A mouse model of lung metastasis was created by transplanting RPMI cells (2 x 104 cells/ml) into nude mice via intravenous tail injection. After 6 weeks of treatment with TwX (40 mg/kg/day), blood samples were taken, and oxidative stress in the plasma, lung　metastasis, and NK activity was measured. The results showed that the oxidative stress level of the tumor-bearing mice was twice as high as that of normal mice, but TwX suppressed the increase from the normal level to less than half. Lung weights were also significantly lighter than controls. NK activity was significantly lower NK activity than normal mice, while the TWX group had twice higher NK activity as the carriers and 1.5 times higher than the normal mice. These results suggest that TwX may suppress cancer metastasis by suppressing elevated oxidative stress, thereby increasing NK activity, which is cancer immunity. However, similar results may not be obtained in the case of single-component antioxidants. We believe that the results of this study will help antioxidants in cancer treatment and prevention.

References:
1. Composition for protection against cell-damaging effects. WO/2013/072441 23.05.2013 (patent)\n2. Novel compositions and uses thereof WO 2008071790 A3\n3. Suplatast tosilate reduces radiation-induced lung injury in mice through suppression of oxidative stress. Yusuke Izumi, Taku Nakashima, Noboru Hattori, Free Radic Biol Med.2019;136:52-59