1-4) 1) Fuscoporia obliqua 5,6) 2,8,9)

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1 ANTI-AGING MEDICINE Received: Jan 29, 2007 Accepted: May 19, 2007 Published online: June 1, 2007 Original Article Double Blind Study of Health Claims for Food Containing Extract of Kabanoanatake (Charga: Fuscoporia obliqua) (RCT: randomized controlled trial). Yoshikazu YONEI 1), Yoko TAKAHASHI 1), Koji MATSUSHITA 2), Miwako WATANABE 3), Toshito YOSHIOKA 4) 1) Anti-Aging Medical Research Center, Doshisha University 2) Carna Medsalon 3) Maressia Garden Clinic 4) Graduate School of Commerce, Waseda University Anti-Aging Medicine 4 (1) : 1-10, 2007 Char ga Fuscoporia obliqua Anti-Aging Medicine 4 (1) : 1-10, 2007 (c) Japanese Society of Anti-Aging Medicine

2 1-4) 1) Fuscoporia obliqua 5,6) 2,8,9)

3 10) 11) 12,13) 14,15) Table 1-3 Table 1 Height Weight Total body water Total body muscle Fat-free mass Total skeletal muscle Total body fat BMI Body fat percentage Waist-to-hip ratio Basal metabolism Systolic blood pressure Diastolic blood pressure Table 2 Height Weight Total body water Total body muscle Fat-free mass Total skeletal muscle Total body fat BMI Body fat percentage Waist-to-hip ratio Basal metabolism Systolic blood pressure Diastolic blood pressure Table 3 Height Weight Total body water Total body muscle Fat-free mass Total skeletal muscle Total body fat BMI Body fat percentage Waist-to-hip ratio Basal metabolism Systolic blood pressure Diastolic blood pressure Physical Measurements (Control Group) cm L /m 2 kcal ± ± ± ± ± ± ± ± ± ±0.05 1,312.40± ± ± ± ± ± ± ± ± ± ± ± ±0.05 1,314.27± ± ± Physical Measurements (Charga 5ml Group) cm L /m 2 kcal Physical Measurements (Charga 15ml Group) cm L /m 2 kcal ± ± ± ± ± ± ± ± ± ±0.05 1,269.71± ± ± ± ± ± ± ± ± ± ± ± ±0.08 1,303.33± ± ± ± ± ± ± ± ± ± ± ± ±0.05 1,261.79± ± ± ± ± ± ± ± ± ± ± ± ±0.08 1,302.83± ± ±

4 Table 4 Table 4 Weight Total body water Total body muscle Fat-free mass Total skeletal muscle Total body fat BMI Body fat percentage Waist-to-hip ratio Basal metabolism Systolic blood pressure Diastolic blood pressure Physical Measurements Control ± ± ± ± ±0.06 5ml 0.99±0.02** 0.99±0.01** 0.99±0.01** 0.99±0.01** 0.99±0.02** 1.00±0.06** 0.99±0.02** 1.01±0.05** ** 0.99±0.01** 0.96±0.08** 0.97±0.07** 15ml 1.00± ± ± ± ±0.10 (n=20, mean±sd, ANOVA with contrast, *p<0.05, **p<0.01; significantly different from the other groups. ) Table 5 White blood cell count Red blood cell count Hemoglobin Hematocrit Platelet count GOT GPT γ-gtp AL_P CPK Urea nitrogen Creatinine Uric acid Total cholesterol HDL-Chol LDL-Chol Triglyceride Na K Cl Fasting blood glucose CRP HbA1c IGF-I DHEA-s Cortisol Insulin Homocysteine NK cell activity Clinical Laboratory Test (Control Group) /mm 3 005,450.00±1, ,850.00±1, x/mm 3 g/dl 10 4 x/mm 3 µg/dl µu/ml ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ,432.50± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ,494.00± ± ± ± ± Table 5-7 Table 6 White blood cell count Red blood cell count Hemoglobin Hematocrit Platelet count GOT GPT γ-gtp AL_P CPK Urea nitrogen Creatinine Uric acid Total cholesterol HDL-Chol LDL-Chol Triglyceride Na K Cl Fasting blood glucose CRP HbA1c IGF-I DHEA-s Cortisol Insulin Homocysteine NK cell activity Clinical Laboratory Test (Charga 5ml Group) /mm x/mm 3 g/dl 10 4 x/mm 3 µg/dl µu/ml 05,490.00±1, ,505.00±1, ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ,701.50± ,646.50± ± ± ± ± ± ± ± ±

5 ANTI-AGING MEDICINE Table 7 Clinical Laboratory Test (Charga 15ml Group) Table 8 Clinical Laboratory Test Control 5ml 15ml White blood cell count Red blood cell count Hemoglobin Hematocrit Platelet count GOT GPT γ-gtp AL_P CPK Urea nitrogen Creatinine Uric acid Total cholesterol HDL-Chol LDL-Chol Triglyceride Na K Cl Fasting blood glucose CRP HbA1c IGF-I DHEA-s Cortisol Insulin Homocysteine NK cell activity /mm x/mm 3 g/dl 10 4 x/mm3 µg/dl µu/ml 05,695.00±1, ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ,472.50± ± ± ± ± ,515.00±1, ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ,575.50± ± ± ± ± White blood cell count Red blood cell count Hemoglobin Hematocrit Platelet count GOT GPT γ-gtp AL_P CPK Urea nitrogen Creatinine Uric acid Total cholesterol HDL-Chol LDL-Chol Triglyceride Na K Cl Fasting blood glucose CRP HbA1c IGF-I DHEA-s Cortisol Insulin Homocysteine NK cell activity 1.08± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±0.10* 1.00±0.19* 0.98±0.24* 1.06±0.25** 0.99± ±0.35* 0.93± ± ± ± ± ± ±0.62* 1.00± ±0.02* 1.01± ± ± ± ± ± ± ± ± ± ± ± ± ± ±0.10* 1.04± ± ± ± ± ± ± ± ± ± ± ±0.37* 1.00± ± ± ± ± ± ± ± ± ± ± ±0.17 (n=20, mean±sd, ANOVA with contrast, *p<0.05, **p<0.01; significantly different from the other groups.) Table 8 Table 9-11 Table 9 8-OHdG generation rate Isoprostane generation rate 8-OHdG/ creatinine Isoprostane/ creatinine (n=20, mean ± SD, paired t-test) Oxidization Stress Markers (Control Group) ng//hr ng//hr 1.35± ± ± ± ± weeks later 1.07± ± ± ± ± Table 10 Oxidization Stress Markers (Charga 5ml Group) 8-OHdG generation rate Isoprostane generation rate 8-OHdG/ creatinine Isoprostane/ creatinine (n=20, mean ± SD, paired t-test) ng//hr ng//hr 0.97± ± ± ± ± weeks later 0.89± ± ± ± ± Table 11 Oxidization Stress Markers (Charga 15ml Group) 8-OHdG generation rate Isoprostane generation rate 8-OHdG/ creatinine Isoprostane/ creatinine (n=20, mean ± SD, paired t-test) ng//hr ng//hr 1.15± ± ± ± ± weeks later 0.96± ± ± ± ±

6 Table 12 Table 12 Oxidization Stress Markers 8-OHdG generation rate Isoprostane generation rate 8-OHdG/ creatinine Isoprostane/ creatinine (n=20, mean±sd, ANOVA with contrast) Table Control 0.91± ± ± ± ±0.67 5ml 0.92± ± ± ± ± ml 0.89± ± ± ± ±0.54 Table 14 Physical Symptoms (Charga 5ml Group) Tired eyes Blurry eyes Eye pain Stiff shoulders Muscular pain/stiffness Palpitations Dyspnea Tendency to gain weight Weight loss; thin Lethargy No feeling of good health Thirst Skin problems Anorexia Early satiety Epigastralgia Liable to catch colds Coughing and sputum Diarrhea Constipation Headache Dizziness Tinnitus Lumbago Arthralgia Edematous Easily breaking into a sweat Frequent urination Hot flash Cold skin 3.0± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±1.0 Table 15 Physical Symptoms (Charga 15ml Group) Table 13 Physical Symptoms (Control Group) Tired eyes Blurry eyes Eye pain Stiff shoulders Muscular pain/stiffness Palpitations Dyspnea Tendency to gain weight Weight loss; thin Lethargy No feeling of good health Thirst Skin problems Anorexia Early satiety Epigastralgia Liable to catch colds Coughing and sputum Diarrhea Constipation Headache Dizziness Tinnitus Lumbago Arthralgia Edematous Easily breaking into a sweat Frequent urination Hot flash Cold skin 2.9± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± Tired eyes Blurry eyes Eye pain Stiff shoulders Muscular pain/stiffness Palpitations Dyspnea Tendency to gain weight Weight loss; thin Lethargy No feeling of good health Thirst Skin problems Anorexia Early satiety Epigastralgia Liable to catch colds Coughing and sputum Diarrhea Constipation Headache Dizziness Tinnitus Lumbago Arthralgia Edematous Easily breaking into a sweat Frequent urination Hot flash Cold skin 2.7± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±

7 ANTI-AGING MEDICINE Table 16 Table 16 Physical Symptoms Tired eyes Blurry eyes Eye pain Stiff shoulders Muscular pain/stiffness Palpitations Dyspnea Tendency to gain weight Weight loss; thin Lethargy No feeling of good health Thirst Skin problems Anorexia Early satiety Epigastralgia Liable to catch colds Coughing and sputum Diarrhea Constipation Headache Dizziness Tinnitus Lumbago Arthralgia Edematous Easily breaking into a sweat Frequent urination Hot flash Cold skin Control 1.07± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±0.60 5ml 0.95± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ml 1.09± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±0.47* 1.01± ±0.37 (n=20, mean±sd, Kriskal-Wallis ANOVA with contrast, *p<0.05; significantly different from the other groups.) Table Table 20 Table 17 Mental Symptoms (Control Group) Irritability Easily angered Loss of motivation No feeling of happiness Nothing to look forward in life Daily life is not enjoyable Loss of confidence Reluctance to talk with others Depressed A sense of uselessness Shallow sleep Difficulty falling asleep Pessimism Lapse of memory Inability to concentrate Inability to solve problems Inability to make judgments readily Inability to sleep because of worries A sense of tension Feeling of anxiety for no special reason A vague feeling of fear 2.6± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±0.4 Table 18 Mental Symptoms (Charga 5ml Group) Irritability Easily angered Loss of motivation No feeling of happiness Nothing to look forward in life Daily life is not enjoyable Loss of confidence Reluctance to talk with others Depressed A sense of uselessness Shallow sleep Difficulty falling asleep Pessimism Lapse of memory Inability to concentrate Inability to solve problems Inability to make judgments readily Inability to sleep because of worries A sense of tension Feeling of anxiety for no special reason A vague feeling of fear 2.3± ± ± ± ± ± ± ± ± ± ± ± ± ±0.5 Table 19 Mental Symptoms (Charga 15ml Group) Irritability Easily angered Loss of motivation No feeling of happiness Nothing to look forward in life Daily life is not enjoyable Loss of confidence Reluctance to talk with others Depressed A sense of uselessness Shallow sleep Difficulty falling asleep Pessimism Lapse of memory Inability to concentrate Inability to solve problems Inability to make judgments readily Inability to sleep because of worries A sense of tension Feeling of anxiety for no special reason A vague feeling of fear 1.6± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±

8 Table 20 Mental Symptoms Irritability Easily angered Loss of motivation No feeling of happiness Nothing to look forward in life Daily life is not enjoyable Loss of confidence Reluctance to talk with others Depressed A sense of uselessness Shallow sleep Difficulty falling asleep Pessimism Lapse of memory Inability to concentrate Inability to solve problems Inability to make judgments readily Inability to sleep because of worries A sense of tension Feeling of anxiety for no special reason A vague feeling of fear Control 1.11± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±0.44 5ml 1.01± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±0.34** 1.03± ± ± ml 1.03± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±0.53 (n=20, mean±sd, Kriskal-Wallis ANOVA with contrast, **p<0.01; significantly different from the other groups.)

10 Fuscoporia obliqua 16) 16) 16) 5) 6) 17) 18)

11 18) 19) 01) 02) 0 3) 0 4) 5) 6) 0 7) 8) 9) 10) 11) Yonei Y, Mizuno Y. The human dock of tomorrow -Annual health checkup for anti-aging-. Ningen Dock 19:5-8, Yonei Y, Iwaita Y, Muramatsu K, Mizuno Y. The anti-aging secrets of Japanese executives. Anti-Aging Medical Research 2:61-69, Yonei Y, Mizuno Y, Kido M, Kaku L, Yasuda H. Research on toxic metal levels in scalp hair of the Japanese. Anti-Aging Medical Research 2:11-20, Yonei Y, Takahashi Y, Sakura S, Yoshikawa T. The effects of walking on quality of life and various symptoms and issues relating to aging. Anti-Aging Medical Research 3, (in press) Ichimura T, Watanabe O, Maruyama S. Inhibition of HIV-1 protease by water-soluble lignin-like substance from an edible mushroom, Fuscoporia obliqua. Biosci Biotechnol Biochem. 62: , Okada J.Cystine and Fuscoporia oblique powder interfere with influenza virus infection in mice. The Japanese Journal of Veterinary Research 51:32-33, Miura T, Tsuchiya M, Itoh Y, Okamoto H, Noji M, Suzuki I. Antidiabetic activity of Fuscoporia oblique in KK-Ay mice. The Journal of Traditional Medicines 21: 39-41, Yonei Y, Mizuno Y, Togari H, Sato Y. Muscular resistance training using applied pressure and its effects on the promotion of growth hormone secretion. Anti-Aging Medical Research 1:13-27, Yonei Y, Mizuno Y, Katagiri E. Effects of cosmetics therapy using isoflavone and pine bark extract on the skin and QOL: A doubleblind placebo-controlled trial.anti-aging Medical Research 1:48-58, McCord JM, Fridovich I. Superoxide dismutase. An enzymic function for erythrocuprein (hemocuprein). J Biol Chem 244: , Bedogni G, Malavolti M, Severi S, Poli M, Mussi C, Fantuzzi AL, Battistini N. Accuracy of an eight-point tactile-electrode impedance method in the assessment of total body water. Eur J Clin Nutr 56: , ) 13) 14) 15) 16) 17) 18) 19) Floyd RA, Watson JJ, Wong PK, Altmiller DH, Rickard RC. Hydroxyl free radical adduct of deoxyguanosine: sensitive detection and mechanisms of formation. Free Radic Res Commun 1: , Toyokuni S, Tanaka T, Hattori Y, Nishiyama Y, Yoshida A, Uchida K, Hiai H, Ochi H, Osawa T. Quantitative immunohistochemical determination of 8-hydroxy-2'-deoxyguanosine by a monoclonal antibody N45.1: its application to ferric nitrilotriacetate-induced renal carcinogenesis model. Lab Invest 76: , Morrow JD, Awad JA, Kato T, Takahashi K, Badr KF, Roberts LJ 2nd, Burk RF. Formation of novel non-cyclooxygenase-derived prostanoids (F2-isoprostanes) in carbon tetrachloride hepatotoxicity. An animal model of lipid peroxidation. J Clin Invest 90: , Milne GL, Morrow JD. Isoprostanes and related compounds: update Antioxid Redox Signal 8: , He J, Feng XZ, Lu Y, Zhao B. Three new triterpenoids from Fuscoporia obliqua. J Asian Nat Prod Res 3:55-61, Munzel T, Heitzer T, Harrison DG. The physiology and pathophysiology of the nitric oxide/superoxide system. Herz 22: , Koyama T, Taka A, Togashi H. Cardiovascular effects produced by a traditional fungal medicine, Fuscoporia obliqua extract, and microvessels in the left ventricular wall of stroke-prone spontaneously hypertensive rat (SHRSP). Clin Hemorheol Microcirc 35: , Jankovic BD. From immunoneurology to immunopsychiatry: neuromodulating activity of anti-brain antibodies. Int Rev Neurobiol 26: , 1985.

1,2) 3-5) 6) 7) Table 1 Table 1 Number of subjects Age (y.o.) Men / Women Weight (kg) Height (cm) Body mass index Systolic BP (mmhg) Diastolic BP (mmh

1,2) 3-5) 6) 7) Table 1 Table 1 Number of subjects Age (y.o.) Men / Women Weight (kg) Height (cm) Body mass index Systolic BP (mmhg) Diastolic BP (mmh Received: Jun 19, 2007 Accepted: Dec 6, 2007 Published online: Jan 29, 2008 Original Article The effects of walking with pedometers on quality of life and various symptoms and issues relating to aging