Phytochemical Indole-3-carbinol

Indole-3-carbinol is a phytochemical in the class of Indoles, by the breaking down of the glucosinolate glucobrassicin,  found abundantly in cabbage, kale, Brussels sprouts, rutabaga, mustard greens, broccoli, etc.

1, Breast cancer
Breast cancer (malignant breast neoplasm) is a cancer started in the tissues of the breast either from the inner lining of milk ducts (Ductal carcinoma) or the lobules (Lobular carcinoma) that supply the ducts with milk. There is also rare cases that breast cancer starts in other areas of the breast. In 2010, over 250,000 new cases of breast cancer were expected to be diagnosed in women in the U.S. alone and the risk of getting invasive breast cancer during life time of a women is 1/8.
Indole-3-carbinol may act NAG-1 modulators involved in natural products-induced NAG-1 transcriptional signaling pathway used in prevention and treatment of cancer(1). According to the University of Urbino "Carlo Bo",, The indole-3-carbinol cyclic tetrameric derivative (CTet) inhibited breast cancer cell proliferation by endoplasmic reticulum stress and autophagy-related cell death induction, AKT(2). In human breast cancer cell lines MCF-7 and MDA-MB-231, the derivative also enhances ER stress response, as the main upstream molecular mechanism through which CTet acts in both hormone-responsive and triple-negative breast cancer cells(3). In support to the above, the study of University of Urbino Carlo Bo, Urbino (PU),  showed that the derivative may also be a potential anticancer combination agent for both hormone-responsive and triple-negative breast tumors(4).

References

(1) Nonsteroidal anti-inflammatory drug activated gene-1 (NAG-1) modulators from natural products as anti-cancer agents by Yang MH¹, Kim J², Khan IA³, Walker LA⁴, Khan SI⁵.(PubMed)
(2) The Indole-3-carbinol cyclic tetrameric derivative CTet synergizes with cisplatin and doxorubicin in triple-negative breast cancer cell lines by De Santi M¹, Galluzzi L, Duranti A, Magnani M, Brandi G.(PubMed)
(3) Induction of endoplasmic reticulum stress response by the indole-3-carbinol cyclic tetrameric derivative CTet in human breast cancer cell lines by Galluzzi L¹, De Santi M, Crinelli R, De Marco C, Zaffaroni N, Duranti A, Brandi G, Magnani M(PubMed)
(4) Antitumoral activity of indole-3-carbinol cyclic tri- and tetrameric derivatives mixture in human breast cancer cells: in vitro and in vivo studies by Brandi G¹, Fraternale A, Lucarini S, Paiardini M, De Santi M, Cervasi B, Paoletti MF, Galluzzi L, Duranti A, Magnani M.(PubMed).

2.  Lung cancer
Lung cancer is defined as a condition of the abnormal growth of the cells in the lung's tissue. Most common form of primary lung cancers are derived from epithelial cells. In Us, Lung cancer is the leading cause of cancer deaths, causing 158,683 people deaths, including 88,329 men and 70,354 women, according to 2007 statistic.
In mice study, Indole-3-carbinol showed to inhibit ling cancer via modulation of the receptor tyrosine kinase/PI3K/Akt signaling pathway(1), in dose dependent manner(2), In support of the above, the study by Medical College of Soochow University, indicated that I3C reduces radiosensitivity of lung cancer cells by mediating EGFR expression, indicating that EGFR may be an important target for I3C-mediated radioresistance in lung cancer. In deed, the individual of indole-3-carbinol or silibinin, showed to inhibit 43 and 36% reductions, respectively, in the multiplicity of tumors on the surface of the lung and 83 and 50% reductions, respectively, and mixtures of I3C and silibinin are more potent than the individual compounds for the chemoprevention of lung cancer in A/J mice, according to University of Minnesota(4).


References
(1) Indole-3-carbinol inhibited tobacco smoke carcinogen-induced lung adenocarcinoma in A/J mice when administered during the post-initiation or progression phase of lung tumorigenesis by Qian X¹, Melkamu T, Upadhyaya P, Kassie F.(PubMed)
(2) Dose-dependent inhibition of tobacco smoke carcinogen-induced lung tumorigenesis in A/J mice by indole-3-carbinol by Kassie F¹, Matise I, Negia M, Upadhyaya P, Hecht SS.(PubMed)
(3) [EGFR-dependent impact of indol-3-carbinol on radiosensitivity of lung cancer cells].

[Article in Chinese] by Xiao X¹, Meng Q, Xu J, Jiao Y, Rosen EM, Fan S.(PubMed)

(4) Enhanced inhibition of lung adenocarcinoma by combinatorial treatment with indole-3-carbinol and silibinin in A/J mice by Dagne A¹, Melkamu T, Schutten MM, Qian X, Upadhyaya P, Luo X, Kassie F.(PubMed)

 

 

3. Pancreatic cancer

Cancer of pancreas or pancreatic cancer is defined as condition of a malignant neoplasm of the pancreas, as the cells have become cancerous and divided uncontrollably. Most pancreatic cancers start in the ducts (exocrine component of the pancreas) that carry pancreatic juices with cancer of an epithelium that originates in glandular tissue accounts for over 75 - 90% of all cases of exocrine cancers.

I3C would be effective for enhancing sensitivity of pancreatic cancer cells to gemcitabine via down-regulation of miR-21, according to the Seoul National University College of Medicine(1). Indole-3-carbinol (I3C) also showed to lower the LD(50) of gemcitabine (Gemzar) in decreasing growth of both male (MiaPaca2) and female (SU86.86) pancreatic cancer cells, according to ¹FDA/National Center for Toxicological Research(2). In pancreatic cell lines, ther phytochemical showed to induced gemcitabine cytotoxicity, through up-regulation of hENT1 expression involved in the additive effect of I3C and gemcitabine(3).

 

 

References

(1) Chemosensitivity induced by down-regulation of microRNA-21 in gemcitabine-resistant pancreatic cancer cells by indole-3-carbinol by Paik WH¹, Kim HR, Park JK, Song BJ, Lee SH, Hwang JH.(PubMed)

(2) Gender differences in gemcitabine (Gemzar) efficacy in cancer cells: effect of indole-3-carbinol. by Lyn-Cook BD¹, Mohammed SI, Davis C, Word B, Haefele A, Wang H, Hammons G.(PubMed)

(3) Enhanced efficacy of gemcitabine by indole-3-carbinol in pancreatic cell lines: the role of human equilibrative nucleoside transporter 1 by Wang H¹, Word BR, Lyn-Cook BD.(PubMed)

 

 

4. Prostate cancer

Prostate cancer is defined as a condition in which the cells of prostate has become cancerous, causing abnormal cell growth which spread to the distant parts of the body. Most prostate cancers are slow growing and enlarged prostate and prostate cancer may be detected during the Physical (rectum) exams.

Indole-3-carbinol, has showed a strong evidence in supporting the use of  phytochemicals as epigenetic modulators for prostate cancer chemoprevention(1). Indole-3-carbinol (I3C), a common phytochemical in cruciferous vegetables, inhibited prostate cancer through induced G1 cell-cycle arrest and apoptosis in prostate cancer cells(2). In human PC-3 prostate cancer cell lines, the phytochemical also induced G1 cell cycle arrest of that leading to apoptosis, and regulates the expression of apoptosis-related genes(3). According to the Division of Cancer Prevention, National Cancer Institute, broccoli-derived compounds indole-3-carbinol (I3C) and 3,3'-diindolylmethane (DIM), showed to inhibit prostate cancer through modulation of the effect of androgen on CCL2-mediated pathways(4).

 

 

references

(1) Phytochemicals from cruciferous vegetables, epigenetics, and prostate cancer prevention. by W Watson G¹, M Beaver L, E Williams D, H Dashwood R, Ho E.(PubMed)

(2) Indole-3-carbinol and prostate cancer by Sarkar FH¹, Li Y.(PubMed)

(3) Indole-3-carbinol (I3C) induced cell growth inhibition, G1 cell cycle arrest and apoptosis in prostate cancer cells by Chinni SR¹, Li Y, Upadhyay S, Koppolu PK, Sarkar FH.(PubMed)

(4) Indole-3-carbinol and 3',3'-diindolylmethane modulate androgen's effect on C-C chemokine ligand 2 and monocyte attraction to prostate cancer cells by Kim EK¹, Kim YS, Milner JA, Wang TT.(PubMed)

4.
Dietary indole-3-carbinol (I3C) prevents the development of estrogen-enhanced cancers including breast, endometrial and cervical cancers(1) due to it antitumor activities associated not only with its regulation of estrogen activity and metabolism, but also its modulation of ER transcription activity(2) through both antiestrogenic and estrogenic activity(3). In human cervical cancer cells, in comparison to diindolylmethane (DIM), a major in vivo product of indole-3-carbinol (I3C), both I3C and DIM caused accumulation of DNA strand breaks in three cervical cancer cell lines.  I3C  increased apoptosis in target tissues in vivo.(4). In mice study, according to the The Long Island Campus of Albert Einstein College of Medicine, showed to prevent the cervical-vaginal cancer promoted high doses of estrogen in these mices(5) and increased PTEN expression in the cervical epithelium of the transgenic mouse in vivo(6)

References
(1) Indole-3-carbinol is a negative regulator of estrogen by Auborn KJ¹, Fan S, Rosen EM, Goodwin L, Chandraskaren A, Williams DE, Chen D, Carter TH.(PubMed)
(2) Indole-3-carbinol is a negative regulator of estrogen receptor-alpha signaling in human tumor cells. by Meng Q¹, Yuan F, Goldberg ID, Rosen EM, Auborn K, Fan S.(PubMed)
(3) Indolo[3,2-b]carbazole: a dietary-derived factor that exhibits both antiestrogenic and estrogenic activity by Liu H¹, Wormke M, Safe SH, Bjeldanes LF.(PubMed)
(4) Indole-3-carbinol and diindolylmethane induce apoptosis of human cervical cancer cells and in murine HPV16-transgenic preneoplastic cervical epithelium by Chen DZ¹, Qi M, Auborn KJ, Carter TH.(PubMed)
(5) Indole-3-carbinol prevents cervical cancer in human papilloma virus type 16 (HPV16) transgenic mice by Jin L¹, Qi M, Chen DZ, Anderson A, Yang GY, Arbeit JM, Auborn KJ.(PubMed)
(6) Indole-3-carbinol prevents PTEN loss in cervical cancer in vivo by Qi M¹, Anderson AE, Chen DZ, Sun S, Auborn KJ.(PubMed)

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Herbal Therapy - Chickweed (Stellaria media)

Chickweed is a cool-season annual plant often eaten by chickens., genus S. media, belonging to the family Caryophyllaceae, native to Europe. The herb has been used in herbal medicine to treat skin conditions, indigestion, eczema, cough, rashes, etc. and as astringent, carminative, demulcent, diuretic, expectorant, laxative, refrigerant, vulnerary agent.

Chemical constituents(1)(2)
1. Igenin 6-C-beta-D-galactopyranosyl-8-C-alpha-L-arabinopyranoside
2. Apigenin 6-C-alpha-L-arabinopyranosyl-8-C-beta-D-galactopyranoside
3. Apigenin 6-C-beta-D-galactopyranosyl-8-C-beta-L-arabinopyranoside
4. Apigenin 6-C-beta-D-glucopyranosyl-8-C-beta-D-galactopyranoside
5. Apigenin 6, 8-di-C-alpha-L-arabinopyranoside
6. Alkaloidal glycoside
7. Glucodichotomine B
8. Neolignan glycosides
9. Dichotomosides A, B, C, and D
10. Phenylpropanoid glycoside
11. Dichotomoside E
12. Saponins
13. Mucilage
14. PABA
15. Coumarin,
16. Flavonoids (rutin)
17. Hydroxycoumarin


References
(1) [Studies on chemical constituents from stellaria media. I].[Article in Chinese] by Dong Q¹, Huang Y, Qiao SY.(PubMed)
(2) Bioactive constituents from Chinese natural medicines. XIV. New glycosides of beta-carboline-type alkaloid, neolignan, and phenylpropanoid from Stellaria dichotoma L. var. lanceolata and their antiallergic activities by Morikawa T¹, Sun B, Matsuda H, Wu LJ, Harima S, Yoshikawa M.(PubMed)


1. Chickweed and Obesity
Obesity is defined as a medical condition of excess body fat has accumulated overtime, while overweight is a condition of excess body weight relatively to the height. According to the Body Mass Index(BMI), a BMI between 25 to 29.9 is considered over weight, while a BMI of over 30 is an indication of obesity. According to the statistic, 68% of American population are either overweight or obese.
In progesterone-induced obesity model in female albino mice, conducted by the CMR College of Pharmacy, showed that Methanolic extract of S. media (MESM) showed to inhibit the increase in body weight, adipose tissue weight and size, and upturned obesity and associated complications and may bebeneficial in suppression of obesity induced by progesterone(3). Other study suggested that the anti-obesity effects of Llyophilized juice extracted from the plant in high-fat-diet fed mice may be partly mediated through delaying the intestinal absorption of dietary fat and carbohydrate by inhibiting digestive enzymes(4).

References
(1) [Studies on chemical constituents from stellaria media. I].[Article in Chinese] by Dong Q¹, Huang Y, Qiao SY.(PubMed)
(2) Bioactive constituents from Chinese natural medicines. XIV. New glycosides of beta-carboline-type alkaloid, neolignan, and phenylpropanoid from Stellaria dichotoma L. var. lanceolata and their antiallergic activities by Morikawa T¹, Sun B, Matsuda H, Wu LJ, Harima S, Yoshikawa M.(PubMed)
(3)Antiobesity effect of Stellaria media against drug induced obesity in Swiss albino mice by Chidrawar VR¹, Patel KN, Sheth NR, Shiromwar SS, Trivedi P.(PubMed)
(4) Quality assessment and anti-obesity activity of Stellaria media (Linn.) Vill by Rani N¹, Vasudeva N, Sharma SK.(PubMed)

2. Chickweed and Hepatitis
Chickweed (Stellaria media (Linn.) Villars) has been used in traditional Chinese medicine over 200 years, mainly for the treatment of dermatitis and other skin diseases. In the study to evaluated the  anti-hepatitis B virus (HBV),  using the human HBV-transfected liver cell line HepG2.2.15, researched at the Jiangsu Provincial Academy of Chinese Medicine indicated that the herbal  30 μg/mL SM-3 effectively suppressed the secretion of HBsAg and HBeAg after 6 days of treatment,  Consistent with the reduction in HBV antigens, SM-3 also reduced the level of HBV DNA in a dose-dependent manner(1). In support of the result, the ¹Siberian State Medical University, indicated a hepaprotective effect of the herb water-soluble polysaccharide fraction at a dose of 100 mg/kg(2). In the evaluation of anti-hepatoma activity on five human liver-cancer cell lines, i.e. HepG2/C3A, SK-HEP-1, HA22T/VGH, Hep3B and PLC/PRF/5, with 15 most common herbs used in Canada folk medicine, chickweed may be effective in portect the liver against the growth of all five cell lines, depending to further study validation(3).

References
(1) Anti-hepatitis B virus activity of chickweed [Stellaria media (L.) Vill.] extracts in HepG2.2.15 cells by Ma L¹, Song J, Shi Y, Wang C, Chen B, Xie D, Jia X.(PubMed)
(2) Evaluation of hepatoprotective activity of water-soluble polysaccharide fraction of Stellaria media L. by Gorina YV¹, Saprykina EV, Gereng EA, Perevozchikova TV, Krasnov EA, Ivanova EV, Fait EA, Baranova OV.(PubMed)
(3) In vitro anti-hepatoma activity of fifteen natural medicines from Canada by Lin LT¹, Liu LT, Chiang LC, Lin CC.(PubMed)

3. Chickweed and Antimicrobial effects
HSV-2 is a members of the herpesvirus family and can be spread through contact with saliva,
 Stellarmedin A, a novel antiviral protein purified from Stellaria media, showed to inhibit the an proliferative and peroxidase activities of HSV-2 protein, according to the study by the Jiangsu Province and Chinese Academy of Sciences(1). Homologous defensins, Sm-AMP-D1 and Sm-AMP-D2, were isolated from seeds of common chickweed Stellaria media L, according to Russian Academy of Sciences, showed a strong  inhibitory activity against phytopathogenic fungi and oomycetes and may be a potent promising for engineering pathogen resistance in crops(2). α-hairpinin, a novel antifungal peptide isolated from Stellaria media seeds, also exhibited high broad-spectrum activity against fungal phytopathogens(3).

References
(1) Purification and characterization of a novel anti-HSV-2 protein with antiproliferative and peroxidase activities from Stellaria media by Shan Y¹, Zheng Y, Guan F, Zhou J, Zhao H, Xia B, Feng X.(PubMed)
(2) Isolation, molecular cloning and antimicrobial activity of novel defensins from common chickweed (Stellaria media L.) seeds by Slavokhotova AA¹, Odintsova TI, Rogozhin EA, Musolyamov AK, Andreev YA, Grishin EV, Egorov TA.(PubMed)
(3) Novel antifungal α-hairpinin peptide from Stellaria media seeds: structure, biosynthesis, gene structure and evolution by Slavokhotova AA¹, Rogozhin EA, Musolyamov AK, Andreev YA, Oparin PB, Berkut AA, Vassilevski AA, Egorov TA, Grishin EV, Odintsova TI.(PubMed)









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Food therapy - Fennel

Fennel is a species of Foeniculum Vulgare, belong to the family Apiaceae (Umbelliferae), and native to the shores of the Mediterranean. It is now widely cultivated all around the globe to use as food and herb.
Nutritients
1. Carbohydrates
2. Fiber
3. Protein
4. Thiamine (Vitamin B1)
5. Riboflavin (Vitamin B2)
6. Niacin (Vitamin B3)
7. Pantothenic acid (B5)
8. Vitamin B6
9. Folate (Vitamin B9)
10. Vitamin C
11. Calcium
12. Copper
13. Iron
14. Magnesium
15. Molybdenum
16. Phosphorus
17. Potassium
18. Zinc

Chemical constituents(1)(2)
1. 1,3-benzenediol
2. 1-methoxycyclohexene
3. O-cymene
4. Sorbic acid,
5. 2-hydroxy-3-methyl-2-cyclopenten-1-one
6. Estragole
7. Limonene-10-ol
8. 3-methyl-2-cyclopenten-1-one
9. Linoleic acid
10. Oleic acid
11. Undecanal
12. 2,4-undecadienal
13. trans-Anethole
14. Fenchone
15. Limonene

1. Fennel anti microbial activities
The immune system is the set of cells and their activity against antigens or infectious agents that comprises of the body's defense system against diseases. The immune system does a great job of keeping people healthy and preventing infections. Beside foods and nutritional supplements, herbs also play a important role in helping the immune system defend against viruses and bacteria attacks.
According to the study by the Ataturk University, chemical constituents from the inflorescence, leaf stems, and whole aerial parts of fennel, including anethole, and hexane extract showed effectively against most of the foodborne pathogenic, saprophytic, probiotic, and mycotoxigenic microorganisms tested(2). In comparison of essential oil from 3 different species of fennels, such as Essential oils of the fruits of three organically grown cultivars of Egyptian fennel (Foeniculum vulgare var. azoricum, Foeniculum vulgare var. dulce and Foeniculum vulgare var. vulgare, researched indicated that all three cultivars showed similar antimicrobial activity(3). Unfortunately, the ¹Universidade do Algarve study, suggested that essential oils of fennel showed a very low antimicrobial activity, but exerted a pro-oxidant activity(4).

References
(1) Antimycobacterial activity of constituents from Foeniculum vulgare var. dulce grown in Mexico.by Esquivel-Ferriño PC¹, Favela-Hernández JM, Garza-González E, Waksman N, Ríos MY, del Rayo Camacho-Corona M.(PubMed)
(2) Chemical composition, antimicrobial and antioxidant activities of essential oils from organically cultivated fennel cultivars by Shahat AA¹, Ibrahim AY, Hendawy SF, Omer EA, Hammouda FM, Abdel-Rahman FH, Saleh MA.(PubMed)
(3) Chemical composition, antimicrobial and antioxidant activities of essential oils from organically cultivated fennel cultivars by Shahat AA¹, Ibrahim AY, Hendawy SF, Omer EA, Hammouda FM, Abdel-Rahman FH, Saleh MA.(PubMed)
(4) Foeniculum vulgare essential oils: chemical composition, antioxidant and antimicrobial activities by Miguel MG¹, Cruz C, Faleiro L, Simões MT, Figueiredo AC, Barroso JG, Pedro LG.(PubMed)

2. Fennel as antioxidants
Free radicals are atoms, molecules, or ions with unpaired electrons through chemical bonds with other atoms or molecules during a chemical reaction. They may have positive, negative or zero charge. The unpaired electrons cause radicals to be highly chemically reactive in the human body, leading to aging and cancers.
The comparison of commercial essential oils (EO) containing Citrus aurantium, C. limon, Cupressus sempervirens, Eucalyptus globulus, Foeniculum vulgare and Thymus vulgaris showed that  F. vulgare EOs is effective against lipid peroxidation(3). In support of the above, the University of Wyoming suggested that Fennel oils might have higher antioxidant capacity in distillation time (DT) depend-manner. DT can be used to obtain fennel essential oil with differential composition(4).
Some researchers suggested that the potent antioxidant properties of medicinal plants may depend on variety conditions such as environmental conditions, climatic and seasonal variations, geographical regions of growth, degree of ripeness, growing practices, and many other factors such as postharvest treatment and processing(5).

References
(1) Antimycobacterial activity of constituents from Foeniculum vulgare var. dulce grown in Mexico.by Esquivel-Ferriño PC¹, Favela-Hernández JM, Garza-González E, Waksman N, Ríos MY, del Rayo Camacho-Corona M.(PubMed)
(2) Chemical composition, antimicrobial and antioxidant activities of essential oils from organically cultivated fennel cultivars by Shahat AA¹, Ibrahim AY, Hendawy SF, Omer EA, Hammouda FM, Abdel-Rahman FH, Saleh MA.(PubMed)
(3) Anti-oxidant, anti-inflammatory and anti-proliferative activities of Moroccan commercial essential oils by Aazza S, Lyoussi B, Megías C, Cortés-Giraldo I, Vioque J, Figueiredo AC, Miguel MG.(PubMed)(4) Distillation time modifies essential oil yield, composition, and antioxidant capacity of fennel (Foeniculum vulgare Mill) by Zheljazkov VD¹, Horgan T, Astatkie T, Schlegel V.(PubMed)
(5) Antioxidant activity and protecting health effects of common medicinal plants.

Škrovánková S¹, Mišurcová L, Machů L.(PubMed)

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3.

The Science of Soy - The East Viewpoints: Part A6 - Soy and Cancers in Japanese

By Kyle J. Norton(Draft article)

Soy foods, including tofu have been in traditional Chinese diet over thousands of year, according to Chinese literature. The reduced risk of chronic disease, including metabolic syndrome such as cardiovascular diseases, obesity and diabetes and lesser menopause symptoms in advanced age, may be aided by eating a lot of soy food accompanied with large portion of vegetables and fruits. Indeed, according to the study, only 10% of women in the East are experience symptoms of menopause in advanced age compared to over 70% of their Western counterparts.
According to Dr. Mark Messina, Ph.D., Soy foods contributed from 6.5%8 to 12.8%7 of total protein intake in older adult in Japan.(b)

The approval of cardiovascular benefit of soy by FDA in 1999 accompanied with the discovery of health benefits in clinical studies over past decade, prompted the promotion and advertisement of soy's health benefits in every aspect in Western society. Evidences could be seen by walking through the supermarkets and drug stores. Soy supplements and products such as tofu, soy milk, soy-based infant formula, and meatless “texturized vegetable protein” burgers were widely available. According to the United Soybean Board’s 2004–2005, 25% of Americans consume soy foods or beverages at least once per week, and 74% view soy products as healthy.

Today, the promotion of soy are no longer existed, it may be results of discovery of adverse effect in single ingredient and animal studies, as intake of soy is associated to induce risk certain mammary cancers and infertility. The publication of the result have drawn many criticisms. According to Thomas Badger, director and senior investigator at the Arkansas Children’s Nutrition Center in Little Rock, these effects are seen only under certain experimental conditions that are not likely to occur in humans—and therein lies the crux of the debate(a). Equol (4',7-isoflavandiol), an isoflavandiol metabolized from daidzein may be the causes, as 90% of Eastern population are equol producers but only 30% in the West.
The explanation of the positive effect of soy isoflavones in reduced risk of mammary cancers by University of Goettingen may be interesting, as researchers said" Most importantly, there is dispute as to whether isoflavones derived from soy or red clover have negative, positive or any effect at all on the mammary gland or endometrium. It is beyond any doubt that soy products may have cancer preventing properties in a variety of organs including the mammary gland. However, these properties may only be exerted if the developing organ was under the influence of isoflavones during childhood and puberty.

Soybean is the genus Glycine, belonging to the family Fabaceae, one of the legumes that contains twice as much protein per acre as any other major vegetable or grain crop, native to Southeast Asia. Now, it is grown worldwide with suitable climate for commercial profits.
Nutrients
1. Carbohydrates
2. Dietary fiber
3. Fat
4. Protein
5. Essential amino acid
6. Vitamin A
7. Vitamin B6
8. Vitamin B12
9. Vitamin C
10. Vitamin K
11. Calcium
12. Iron
13. Magnesium
14. Phosphorus
15. Potassium
16. Sodium
17. Zinc
18. Etc.
Phytochemicals
1. Isoflavones
2. Genistein
3. Saponins
4. Beta-sitosterol
5. Daidzein

I. Soy in Eastern population
A. The Japanese population
Japan, an island nation in the Pacific Ocean, lies to the east of the Sea of Japan, China, North Korea, South Korea and Russia, stretching from the Sea of Okhotsk in the north to the East China Sea and Taiwan in the south(1c). According to Moriyama, Japanese women and men live longer and healthier than everyone else on Earth, it may be result of healthier Japanese diet and lifestyle. According to the World Health Organization, the Japanese have an average of 75 years healthy living with disability-free, it may be due to average soy intake 10 to 70 times higher than in Western people(1a)(1b).

A6. Soy and cancers in Japan population
Cancer is a class of diseases in which a group of cells growing and multiplying disordered and uncontrollable way in our body, have become progressively worse and damaged other healthy tissues, sometimes spreads to other organs in the body via lymph or blood and results may be in death.

Epidemiological studies, linking soy and soy products in reduced risk and treatment have been contradictive(1)(2)(3)(4), but in Japanese population, soy and its products have been found effectively in reduced risks of and treatment of cancers, it may be result of equol producer status or long term exposure since childhoods. Some researchers suggested that it may be result of traditional Japanese diet with high in soy, vegetables, fish.

1. Breast cancer
Breast cancer (malignant breast neoplasm) is a cancer that starts in the tissues of the breast either from the inner lining of milk ducts (Ductal carcinoma) or the lobules (Lobular carcinoma) that supply the ducts with milk. there is also rare cases that breast cancer starts in other areas of the breast.
According to the evaluation based on a systematic review of epidemiologic evidence among the Japanese population, there is no associated of intakes of individual soy foods with the risk of breast cancer but evidence supported the biological plausibility of a protective effect of isoflavones on breast cancer risk(5). The study in the testing of oral administration of  IF-rich tablet (20 and 40 mg/day) on climacteric women,  showed  the product not only reduced risk of breast cancer but also improved of bone density, hypertension and climacteric symptoms, cardiovascular diseases, gynecological problems and possible immune potentiation(6)
Japanese women have lower incidence of and mortality from breast cancer, compared to Caucasians, it may be result of large amount intake soy protein and isoflavones, as high dietary intake of phytoestrogens, mainly in the form of soy products, can produce circulating levels of phytoestrogens that are known experimentally to have oestrogenic effects(7).

2. Lung cancer
Lung cancer is defined as a condition of the abnormal growth of the cells in the lung's tissue. Most common form of primary lung cancers are derived from epithelial cells. In Us, Lung cancer is the leading cause of cancer deaths, causing 158,683 people deaths, including 88,329 men and 70,354 women, according to 2007 statistic.
Soy intakes are associated to reduced risk of lung cancer with epidermal growth factor receptor (EGFR) mutations which play substantial roles in genesis and proliferation of non-small-cell lung cancers (NSCLCs), according to the Aichi Cancer Center Research Institute.(8). In Asian populations, not only Japanese, in the review of total of 11 epidemiologic studies that consisted of 8 case-control and 3 prospective cohort studies, although consumption of soy food is associated with lower lung cancer risk, intervention studies that use unified measures of soy intake are needed to fully characterize such an association due to different methods used to assess soy consumption across reviewed studies(9).
In deed, isoflavone intake was associated with a decreased risk of lung cancer in never smokers, but not in current or past smokers, according to a population-based prospective cohort study in 36,177 men and 40,484 women aged 45-74 y with no history of cancer at baseline in 1995-1999, conducted by the National Cancer Center Tokyo Japan(10).

3.  Bowel (Colorectal ) Cancer
Bowel cancer also known as colorectal cancer, is defined as a condition of the abnormal proliferation of cells in the colon, rectum, or vermiform appendix. Bowl is divided in 2 parts, the first part of the bowel, the small bowl, is involved with the digestion and absorption of food. The 2nd part, the large bowel which consist the the colon and rectum, is involved in absorption of water from the small bowel contents and broken down of certain materials in the feces into substances of which some of them to be re absorbed and reused by the body. Bowel cancer is relatively very common and slowly growing and progress cancer and in predictable way.
Bowel cancer is the third most commonly diagnosed cancer in developed countries, including US and Canada.
According to the Research Center for Cancer Prevention and Screening, National Cancer Center in Japan, the investigated the association between dietary soy and isoflavone intake and incidence of colorectal cancer in a prospective cohort study of 83,063 Japanese men and women, ages 45 to 74 years, intake of isoflavones, miso soup, and soy food was not associated with colorectal cancer in either men or women. By colorectal cancer subsite, the risk of proximal colon cancer in men decreased with increasing consumption of isoflavones, miso soup, and soy food  in Compared with men in the lowest quartiles of isoflavones(11) and High isoflavone intake was associated with reduced risk of CI and MI in Japanese women, especially in postmenopausal women(12).
But for patients with Acromegaly, a syndrome that results when the anterior pituitary gland produces excess growth hormone (GH), the risk colorectal cancer is increased substantially(13)(14).

4. Gastric cancer
Stomach cancer is defined as condition of abnormal growth of the mucus-producing cells of the inside lining of the stomach. Adenocarcinoma is the most common type of stomach cancer.
According to the National Cancer Center, Goyang, Korea, high level of consumption of non-fermented soy foods, rather than fermented soy foods, is associated to reducing GC risk, in the reviews of 20 studies assessing the effect of the consumption of fermented soy food on GC risk(15).
4-hydroxy-2 (or 5)-ethyl-5 (or 2)-methyl-3(2H)-furanone (HEMF), a chemical constituent from of Japanese-style fermented soy sauce, reduced hydrogen peroxide concentration in human polymorphonuclear leucocytes stimulated by arachidonic acid or 12-O-tetradecanoylphorbol-13-acetate,through inhibition of carcinogenesis in this system by acting at the post-initiation stage, in female rats(16). Long-term effect of shoyu (Japanese soy sauce), in rat study did not appear to be a carcinogen in the rat; its prolonged use impaired neither health nor longevity but with a mild gastritis(17).

5. Cancer of Endometrium/Endometrial Cancer
Endometrium is the inner lining of the mammalian uterus and very susceptible hormone change, particular to menstrual cycle. Endometrial cancer is a late adulthood cancer defined as a condition of which the cells of the endometrial lining of uterus have growth uncontrollable or become cancerous as a result of the alternation of cells DNA. It's the fourth most common cancer among women overall, after breast cancer, lung cancer, and bowel cancer.
Greater consumption of isoflavone-containing foods is associated with a reduced risk of endometrial cancer in this population of nonhysterectomized postmenopausal women(18).
 In support to the above, the University of Hawaii, in the study of Endometrial cancer cases (n = 332) diagnosed between 1985 and 1993 which were identified from the five main ethnic groups in the state (Japanese, Caucasian, Native Hawaiian, Filipino, and Chinese) through the rapid-reporting system of the Hawaii Tumor Registry, showed that plant-based diets low in calories from fat, high in fiber, and rich in legumes (especially soybeans), whole grain foods, vegetables, and fruits reduce the risk of endometrial cancer(20). According to the National Cancer Center, Tokyo, Japan, in the study of Forty nine thousand one hundred and twenty-one women of age 45-74 years who responded to a 5-year follow-up survey, researchers found that there was no evidence of a protective association between soy food or isoflavone intake and endometrial cancer risk(19).

6. Cervical Cancer
Cervix is the lower part of uterus that opens at the top of the vagina. Cervix acts an transition area for vaginal lining (squamous epithelium) change to uterus type (columnar epithelium) through the transitional area (squamous columnar epithelium) to host the development of the fetus. Cervical cancer is malignant neoplasm of the cervix uteri or cervical area caused by abnormal cells growth with alternation of cells DNA.
According to the study by, there was a the marginally significant inverse association between CIN3 and tofu and green leafy vegetables among Japanese women(21).

7. Prostate cancer
See The Science of Soy - The East Viewpoints: Part A3 - Soy and Prostate Cancer in Japanese Men

8. Thyroid Cancer
Thyroid is one of the largest endocrine glands found in the neck, below the Adam's Apple with the function of regulating the body use of energy, make of proteins by producing its hormones as a result of the stimulation of thyroid-stimulating hormone (TSH) produced by the anterior pituitary.
Thyroid cancer is defined as condition in which the cells in the thyroid gland have become cancerous.
According to the National Institute of Health Sciences, Tokyo, Japan, in the evaluation of the proliferating cell nuclear antigen labeling indices elevated in the ID diet(iodine-deficient diet) Female F344 rat groups suggest that isoflavones may not be involved in the mechanisms underlying the synergistic goitrogenic effect of soybean with iodine deficiency(22) but the study by showed the positive effect of dietary defatted soybean  in synergistically stimulated the growth of rat thyroid with iodine deficiency, partly through a pituitary-dependent pathway(23). No study was found in the search of the key word - soy and thyroid cancer in Japanese population" in  PubMed).

Taken altogether, Soy and its by products may be association to reduce risk and  for treatment certain cancers in Japanese population, including breast, lung, bowel, gastric, prostate, endometrial, cervical cancers, etc.  Due to lack of qualities of the studies found in PubMed, further study may be necessary to improve its validation even in Japanese population. As always, all articles written by Kyle J. Norton are for information & education only, please consult your Doctor & Related field specialist before applying.

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References
(1) Long-term exposure to dietary sources of genistein induces estrogen-independence in the human breast cancer (MCF-7) xenograft model by Andrade JE¹, Ju YH, Baker C, Doerge DR, HelferichWG.(PubMed)
(2) Genistein stimulates growth of human breast cancer cells in a novel, postmenopausal animal model, with low plasma estradiol concentrations by Ju YH¹, Allred KF, Allred CD, Helferich WG.(PubMed)
(3) Daidzein-estrogen interaction in the rat uterus and its effect on human breast cancer cell growth by Gaete L¹, Tchernitchin AN, Bustamante R, Villena J, Lemus I, Gidekel M, Cabrera G, Astorga P.(PubMed)
(4) The soybean peptide lunasin promotes apoptosis of mammary epithelial cells via induction of tumor suppressor PTEN: similarities and distinct actions from soy isoflavone genistein by Pabona JM¹, Dave B, Su Y, Montales MT, de Lumen BO, de Mejia EG, Rahal OM, Simmen RC.(PubMed)
(5) Soy intake and breast cancer risk: an evaluation based on a systematic review of epidemiologic evidence among the Japanese population by Nagata C¹, Mizoue T, Tanaka K, Tsuji I, Tamakoshi A, Matsuo K, Wakai K, Inoue M, Tsugane S, Sasazuki S; Research Group for the Development and Evaluation of Cancer Prevention Strategies in Japan(PubMed)
(6) Isoflavones for prevention of cancer, cardiovascular diseases, gynecological problems and possible immune potentiation by Watanabe S¹, Uesugi S, Kikuchi Y.(PubMed)
(7) Phytoestrogens and breast cancer--promoters or protectors? by Rice S¹, Whitehead SA.(PubMed)
(8) Soy consumption reduces the risk of non-small-cell lung cancers with epidermal growth factor receptor mutations among Japanese by Matsuo K¹, Hiraki A, Ito H, Kosaka T, Suzuki T, Hirose K, Wakai K, Yatabe Y, Mitsudomi T, Tajima K.(PubMed)
(9) Soy intake is associated with lower lung cancer risk: results from a meta-analysis of epidemiologic studies by Yang WS¹, Va P, Wong MY, Zhang HL, Xiang YB.(PubMed)
(10) Isoflavone intake and risk of lung cancer: a prospective cohort study in Japan by Shimazu T¹, Inoue M, Sasazuki S, Iwasaki M, Sawada N, Yamaji T, Tsugane S; Japan Public Health Center-based Prospective Study Group.(PubMed)
(11) Dietary soy and isoflavone intake and risk of colorectal cancer in the Japan public health center-based prospective study by Akhter M¹, Inoue M, Kurahashi N, Iwasaki M, Sasazuki S, Tsugane S; Japan Public Health Center-Based Prospective Study Group(PubMed)
(12) Association of dietary intake of soy, beans, and isoflavones with risk of cerebral and myocardial infarctions in Japanese populations: the Japan Public Health Center-based (JPHC) study cohort I by Kokubo Y¹, Iso H, Ishihara J, Okada K, Inoue M, Tsugane S; JPHC Study Group.(PubMed)
(13) The prevalence and associated factors of colorectal neoplasms in acromegaly: a single center based study by Yamamoto M¹, Fukuoka H, Iguchi G, Matsumoto R, Takahashi M, Nishizawa H, Suda K, Bando H, Takahashi Y.(PubMed)
(14) Risk of colorectal neoplasm in patients with acromegaly and its relationship with serum growth hormone levels by Matano Y¹, Okada T, Suzuki A, Yoneda T, Takeda Y, Mabuchi H(PubMed)
(15) Fermented and non-fermented soy food consumption and gastric cancer in Japanese and Korean populations: a meta-analysis of observational studies by Kim J¹, Kang M, Lee JS, Inoue M, Sasazuki S, Tsugane S.(PubMed)
(16) Inhibition of benzo[a]pyrene-induced mouse forestomach neoplasia and reduction of H2O2 concentration in human polymorphonuclear leucocytes by flavour components of Japanese-style fermented soy sauce by Kataoka S¹, Liu W, Albright K, Storkson J, Pariza M.(PubMed)
(17) Long-term effect of shoyu (Japanese soy sauce) on the gastric mucosa of the rat by MacDonald WC, Dueck JW.(PubMed)
(18) Legume, soy, tofu, and isoflavone intake and endometrial cancer risk in postmenopausal women in the multiethnic cohort study by Ollberding NJ¹, Lim U, Wilkens LR, Setiawan VW, Shvetsov YB, Henderson BE, Kolonel LN, Goodman MT.(PubMed)
(19) Soy food and isoflavone intake and endometrial cancer risk: the Japan Public Health Center-based prospective study by Budhathoki S¹, Iwasaki M, Sawada N, Yamaji T, Shimazu T, Sasazuki S, Inoue M, Tsugane S; JPHC Study Group(PubMed)
(20) Association of soy and fiber consumption with the risk of endometrial cancer by Goodman MT¹, Wilkens LR, Hankin JH, Lyu LC, Wu AH, Kolonel LN.(PubMed)
(21) Association between dietary calcium and vitamin D intake and cervical carcinogenesis among Japanese women by Hosono S¹, Matsuo K, Kajiyama H, Hirose K, Suzuki T, Kawase T, Kidokoro K, Nakanishi T, Hamajima N, Kikkawa F, Tajima K, Tanaka H.(PubMed)
(22) Lack of effect of soy isoflavone on thyroid hyperplasia in rats receiving an iodine-deficient diet by Son HY¹, Nishikawa A, Ikeda T, Imazawa T, Kimura S, Hirose M.(PubMed)
(23) Dramatic synergism between excess soybean intake and iodine deficiency on the development of rat thyroid hyperplasia by Ikeda T¹, Nishikawa A, Imazawa T, Kimura S, Hirose M.(PubMed)

Dehydroascorbic acid (DHA)(*) 12% concentration regained the youthfulness of Forearm between the Wrist and Elbow in stimulated reproduction of Collagen against Sagging skin

Kyle J. Norton

I have truly believed that scientists have found a natural ingredient for define-aging, but it was shelved as it offers no commercial value. If one is published with effectiveness more superior than the currently sold products, it may be a death spell to many companies which rely totally in selling cosmetic products.
Anti-aging and maintaining our youthful appearance has been researched throughout human civilization. Today, these researchers have become more aggressive as finding a workable agent may produce billions of dollars of profits to the found company. According to statistics, Global Anti-Aging Products Market will reach $291.9 Billion by 2015.

Dehydroascorbic acid (DHA)(*) 12% concentration may be a potent solution in regained the youthfulness of forearm between the wrist and elbow in stimulated reproduction of collagen against Skin sagging due to aging progression.

Abstract
Introduction
Skin aging is one of the most visible ageing processes that occur constantly in our skin organ. Many ingredients from synthetic and natural sources have been proven in studies to have certain effects on skin aging. In our previous study, Dehydroascorbic acid (DHA) at 4% showed a significant improvement in smoothing of the skin, skin elasticity, firming, disappearance of black and white heads, liver spots, pigmentation, and healing acne and acne scars. In this study, Dehydroascorbic acid (DHA) at 12% is tested for its effectiveness in stimulated reproduction of collagen in the forearm between the wrist and elbow.

Method of Experimentation
10 Patients at age 59 -60 (5 females and 5 males; 5 Caucasians, 3 Asians and 2 Africans) were recruited to test for aging process due to loss of collagen causes of forearm between the wrist and elbows.

Patients must also agree during the study that they will not to use any other topical products (such as including moisturizers, sunscreens, and fragrances, make-up and any facial procedures such as peels, facials, microdermabrasion, and injection of botulinum toxin type A or dermal fillers) but the testing solution. The study also examines the patient with any cosmetic surgery and/or medication use of which may interfere with the study, as well as history of facial skin diseases. All patients are also required to sign an informed consent. Solutions of Dehydroascorbic acid (DHA)(*)were remixed every 3 days in order to maintain their refreshness and concentration levels.
Patients were instructed to stir the solution well with wooden chopstick or ice cream stick and wear glove when apply the solution to prevent oxidation causes of brownness in the palm and figure nails due to high concentration of the solution, as well as to cover the applied areas before going out to extreme sunlight.
 Patients were also instructed to apply the provided solution on their forearm between the wrist and elbow for a duration of 12 weeks. Forearm up to the wrist examinations were conducted and recorded every 4 weeks.

Results
All patients completed the trial. Topical administration of Dehydroascorbic acid (DHA)(*) 12% showed  to stimulate the reproduction of collagen in enhancing the youthfulness of the applied area up to 95% in 4 patients

Conclusion
Treatment of concentrations of Dehydroascorbic acid (DHA)(*) at 12 %  exhibited a significant efficacy in rejuvenation of  the youthfulness of forearm between the wrist and elbow through reproduction of collagen  skin smoothing, elasticity, firming, probably through anti-inflammatory and skin absorption pathways, in doses, skin aging progression and durative depending manner. It may be used to rejuvenate the skin in other parts of the body. Since the experimental group is small in nature, further studies with a larger sample size are necessary to improve the ingredient’s validation.

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Introduction
Skin aging is one of the most visible aging processes that occur constantly in our skin. According to the Clinical Centre of Nis, certain plant extracts may have the ability to scavenge free radicals to protect the skin matrix through the inhibition of enzymatic degradation, or to promote collagen synthesis in the skin, and affect skin elasticity and tightness (a). Another study suggested that free radicals induced domino effects in production of reactive oxygen species and can react with DNA, proteins, and fatty acids causing oxidative damage and impairment of the antioxidant system, leading to injuries, damage of the regulation pathways of skin causing wrinkles, roughness, appearance of fine lines, lack of elasticity, and de-pigmentation or hyperpigmentation marks (b).
Walking through the drug and commercial cosmetic stores, one can see hundreds of anti-aging creams and products displayed. Many of them have also gone through certain studies for its efficacy with advertisement through media and celebrities. In fact, according to statistics, Global Anti-Aging Products Market will be worth $291.9 Billion by 2015.
Searching through the key word of Dehydroascorbic Acid (Oxidized Vitamin C, DHAA) and skin has yielded no related studies or clinical trials. It may be the result of the solution having no commercial value as it can be made by any cosmetic purchaser, or the studies indicate that L-ascorbic acid must be formulated at pH levels less than 3.5 to enter the skin with maximal concentration of 20% for optimal percutaneous absorption. But according to Dr. Douglas Q. Kitt, Dehydroascorbic Acid (DHAA) permeates stratum corneum at a rate up to 12 times faster than AA. This supports the concept that lower concentrations of DHAA in topical preparations can enhance skin vitamin C levels with less potential side effects (c).
Ingredient 1: Dehydroascorbic Acid (DHA) (vitamin C supplement oxidized form)
Vitamin C, also known as L-ascorbic acid, is a water-soluble vitamin found in fresh fruits, berries and green vegetables. It is best known for its free radical scavenging activity and regenerating oxidized vitamin E for immune system support. In skin aging, the vitamin may improve solar radiation protection and epidermal aging (1) through production of collagen due to its antioxidant activity (2)(3).
Epidemiological studies linking vitamin C in prevention of skin damage and aging have produced some certain results (3a)(3b)(3c), but the large sample size and multi-centred studies are necessary to validate its effectiveness. Vitamin C oxidized form or dehydroascorbic acid (DHA) processed antiviral and virucidal effects (23) showed to prevent H2O2-induced cell death by increasing the GSH levels mediated by the GPx and GR activities and PPP (17), and regulated neuronal energy metabolism, through facilitating the utilization of glucose via the PPP for antioxidant purposes (18) by increasing antioxidant potential in the central nervous system (19). Dehydroascorbic acid (DHA) may be a potential anti-cancer agent to treat aggressive cancers (20)(21). Dr. Toohey at the Cytoregulation Research said "rapidly-dividing tumour cells make unusually large amounts of homocysteine thiolactone and that administered dehydroascorbic acid enters the cells and converts the thiolactone to mercaptopropionaldehyde which kills the cancer cells" (22).
1. Plasma levels of vitamin C L-ascorbic acid is known for its effects on skin-whitening and against the anti-oxidation causes of skin aging. During the aging process, levels of vitamin C was found to slowly deplete according to the Tokyo Metropolitan Institute of Gerontology in the measurement of plasma and urine of C57BL/6 male mice during 3 to 30 months of aging(4). Therefore, restoration of physiological levels of vitamin C inside the cells might improve the lysosomal degradation (protection of cell from the degradative enzymes through protection of the cytosol) in the outflow pathway cells and prevent the pathogenesis of glaucomadegrade proteins(5).
Unfortunately, some researchers suggested that higher levels of circulating vitamin C may not provide protection against incident radiographic knee OA, and be associated with an increased risk of knee OA(6) and the ¹Panorama Research Institute and Regenerative Sciences Institute, insisted "careful attention to individual and family medical history and personal genomic data may prove essential to make wise dietary and supplement choices to be combined with exercise”(7).
2. The effects
According to the Minghsin University of Science and Technology, in doses of a dependent manner, the concentration of L-ascorbic acid induced absorption of the collagen solution in exhibition of smoothing wrinkles and clear up spots(8). Ascorbic acid (AA) is essential in stimulating collagen gene expression. In type 1 and type 4 collagen and SVCT2, the vitamin was found to enhance the expression of type 1 and type 4 collagens and SVCT2 mRNA in cultured human skin fibroblasts at 100 μM AA placed every 24h for 5 days to prevent depletion(9). The Chiang Mai University study also supported the effects of ascorbic acid in the anti-aging process through exhibition pro and active MMP-2 inhibitory(10). Other studies suggested the combined vitamins, including vitamin C in a single formulation had a slightly lower degradation rate and more stable formulations as compared to different preparations containing only one of the vitamins(11)(12). Amazingly, in vivo, application of vitamin C showed a significant reduction of oxidative stress in the skin, an improvement of the epidermal-dermal microstructure and a reduction of fine lines and wrinkles in aged skin within a relatively short period of time of product application(13). The Bruce and Associates study also insisted the effectiveness of vitamin C application over a 12 week period as the vitamin enhanced the overall intensity of pigmentation, fine lines and wrinkles, tactile roughness, and laxity with a 100% satisfaction of overall appearance of the tested subjects’ skin(14).
1. Skin roughness and Scaliness
Skin roughness due to aging is a result of dead skin cells shedding more slowly, and building up in the upper layer of skin causing skin complexion to become rough and dull. According to the University of California at San Francisco, they are the result of aging and anatomic site of which demonstrated a significant influence on skin roughness and scaliness(25). Another study suggested that frictional properties of skin are dependent on more than water content or non-apparent sweating and the role of sebum secretion may be one possible factor(26), or it may be result of depletion of surface lipid content(27).
2. Wrinkles
Wrinkles are also associated with aging, hormonal status, smoking, and intercurrent disease according to the study by the University of California-San Francisco(28). Some researchers suggested that vitamin C may play an important role in collagen production due to its antioxidant properties(28). Another study indicated a positive anti-wrinkle effect through consumption of a mixture of soy isoflavones, lycopene, vitamin C, vitamin E, and fish oil(29).
3. Skin elasticity and firming
Skin sagging is a result of a loss of collagen and elastin of the skin due to aging in combination with the dreaded pull of gravity. The study of a novel dietary supplement (Imedeen Prime Renewal) including soy extract, fish protein polysaccharides, extracts from white tea, grape seed and tomato, vitamins C and E as well as zinc and chamomile extract, showed to inhibit forehead, periocular and perioral wrinkles, mottled pigmentation, laxity, sagging, under eye dark circles, and overall appearance. After 6 months treatment, it was suggested that the formula may provide improved condition, structure and firmness of the skin in post-menopausal women(30). Other studies insisted that since vitamin C (Vit. C) in the form of L-ascorbic acid (Asc) can accelerate wound healing(33)(32) and protect fatty tissues from oxidation damage(33), it may play an integral role in collagen synthesis of reducing wrinkles and skin sagging(34)(35)
5. Liver and aging spots
Liver spot is a condition of brown or black spots appearing on the surface of the skin due to aging and exposure to ultraviolet radiation from the sun. According to skin care guides, high concentration of Vitamin C may improve the skin and lighten the spots(36). Other articles insisted that liver spots may be the result of a deficiency of vitamin C(37). According to the Duke University Medical Center, the application of topical solutions containing vitamins C and E provided protection for human skin against damage caused by ultraviolet radiation(38)(39)(40)(41).
6. Pigmentation
Skin pigmentation disorder is a result of damage most likely caused by UV sunlight, or unhealthy cells due to aging affecting the production of melanin. Vitamin C topical and by iontophoresis are found effectively in aiding post-laser hyperpigmentation  or short and long term treatment (42) for melism (43). In an experiment where 29 females with melisma enrolled for iontophoresis, a vitamin C solution was applied to one side of the face, while distilled water was applied to the other side as a control, treatment with colorimeter site showed a significant decrease in the luminance value, (a photometric measure of the luminous intensity per unit area of light) compared to that of the control site (44).
8. Acne and Acne scars
According to Dr. GEORGE E. MORRIS, M.D., in a study of 60 patients with acne being given 8 oz of citrus juice twice daily and vitamin C in 3 gm a day, after 4 months 43 showed improvement, 10 failed to show improvement and 7 did not return for follow up(45). Some articles showed that vitamin C may be effective in the reduction of the formation of acne scars through it’s anti-inflammatory and free radical scavenger activities(46), but no study has been found through searching on PubMed.

9. Back and White heads
Black and white heads are the result of a skin pore becoming clogged with sebum from the body's natural oil and becoming black heads if oxidized. No study has been found specifically for the key word vitamin C and back and white heads on PubMed.

10. Fine Lines
No study and been found.

Method
A total of 10p atients were recruited with ages ranging from 59 - 60 (5 Caucasians, 3 Asians and 2 Africans, and 5 females and 5 males) to test for the effectiveness in the difference of concentrations of Dehydroascorbic acid (DHA) at 12% ( 24000mg diluted into 100 mL of hot water which is let cool before refrigerating).

Patients are eligible to enter this study
Selection was based on those who had moderate to severe forearm wrinkles. Patients must also agree during the study that they will not to use any other topical products but the testing solution, such as moisturizers, sunscreens, fragrances, make-up, and any facial procedures such as peels, facials, microdermabrasion, and injection of botulinum toxin type A or dermal fillers. The study also examines the patient with any cosmetic surgery and medication use which may interfere with the study, as well as history of facial skin diseases. All patients are also required to sign an informed consent.
Patients were instructed to stir the solution well with wooden chopstick or ice cream stick and wear glove when apply the solution to prevent oxidation causes of brownness in the palm and figure nails due to high concentration of the solution, as well as to cover the applied areas before going out to extreme sunlight.
 Patients were also instructed to apply the provided solution on their forearm up to the wrist for a duration of 12 weeks. Facial skin examinations were conducted and recorded every 4 weeks.

Total patients study       Genders             Ages
Ethnicity                         5 Females,          59 - 60
5 Caucasians,                  5 Males,
3 Asians,
2 Africans

Initial examination
Based on scale of 0 - 4 (0 - Not applicable, 1 - Minimum, 2 - Mild, 3 - Moderated, 4 - Severe)
6 patients with wrinkle on the forearm between the wrist and elbow range from mild to moderated
4 patients with wrinkle on the forearm up to the wrist range from moderated to severe.

Treatment solution
1. Equipment's contained a solution provided or mixed by patients as instructed. All solutions were refrigerated to protect its effectiveness.
2. Solution was applied twice per day, once in the morning and once in the evening.
Patients applied the solution first to the affected areas using the sponge path containing Dehydroascorbic acid (DHA)(*) at 12% with little pressure to enhance the absorption for about 1 minute. The sponge path would then be cleared and the solution would be returned to the refrigerator.

Patients were also advised not to let the solution get into their eyes. If this did happen, patients were advised to use cold water to clean them out.

Outcome measure
The results will be measured 3 times:
1. The end of week 4
2. The end of week 8
3. The end of week 12
Patients were measured for the intensity of the applicable areas and to check for efficacy of the solution by comparing to the base line.

Results
Observation report
At the end of weeks 4
Dehydroascorbic acid (DHA)(*) at 12% concentration showed to exhibit the reproduction of collagen as follow
6 Patient with reduction of skin sagging of forearm between the wrist and elbow of 75%
4 Patient with reduction of forearm between the wrist and elbow of 80%

At the end of weeks 4
Dehydroascorbic acid (DHA)(*) at 12% concentration showed to exhibit the reproduction of collagen as follow
6 Patient with reduction of skin sagging of forearm between the wrist and elbow of 85%
4 Patient with reduction of skin sagging of forearm between the wrist and elbow of 90%

At the end of weeks 4
Dehydroascorbic acid (DHA)(*) at 12% concentration showed to exhibit the reproduction of collagen as follow
6 Patient with reduction of skin sagging of forearm between the wrist and elbow of 90%
4 Patient with reduction of skin sagging of forearm between the wrist and elbow of 95%

As the data indicates, patient satisfaction with the solution and the progression of skin sagging reduction.

Tolerability
Through over 12 weeks of application with the designed solution, no adverse effect has been reported in the group.

Discussion
Dehydroascorbic acid (DHA)(*) showed the most efficacy in enhanced smoothness, firmness, and elasticity of the skin through stimulation the production of collagen on forearm between the wrist and elbow, probably through its antioxidant activities via inflammatory pathways, and skin absorption property. It may be used in other parts of the body including facial skin.
The efficacy of the solutions indicated a logical thinking through maintaining healthy skin by preventing the loss of collagen due to aging is the most effective way to slow down the aging progression. Dehydroascorbic acid (DHA)(*) at 12%  concentration may provide us with more insight with this astonished result, but further study is necessary to rule out any adverse effect and improve its validation, due to small sample size.


Please make sure that you discuss the use of any topical solution in the study with your doctor or related field specialist before applying. Please Donate for funding the larger sample size study to improve the validation of this discovery.                            

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References
 (*) http://authors.library.caltech.edu/11677/1/BORjbc37a.pdf (The oxidation of ascorbic acid and its reduction in vitro and vivo)
(a) Skin ageing: natural weapons and strategies by Binic I¹, Lazarevic V, Ljubenovic M, Mojsa J, Sokolovic D.(PubMed)
(b) Phytoconstituents as photoprotective novel cosmetic formulations by Saraf S¹, Kaur CD.(PubMed)
(c) Topical Dehydroascorbic Acid (Oxidized Vitamin C) Permeates Stratum Corneum More Rapidly Than Ascorbic Acid by Douglas Q Kitt
(1) Active ingredients against human epidermal aging by Lorencini M¹, Brohem CA², Dieamant GC², Zanchin NI³, Maibach HI(PubMed)
(2) CAM use in dermatology. Is there a potential role for honey, green tea, and vitamin C? by Barbosa NS¹, Kalaaji AN².(PubMed)
(3) The science behind vitamins by Linder J.(PubMed)
(3a) Split-face study of topical 23.8% L-ascorbic acid serum in treating photo-aged skin by Xu TH¹, Chen JZ, Li YH, Wu Y, Luo YJ, Gao XH, Chen HD(PubMed)
(3b) Use of topical ascorbic acid and its effects on photodamaged skin topography by Traikovich SS.(PubMed)
(3c) Formulation and in-vivo evaluation of a cosmetic multiple emulsion containing vitamin C and wheat protein by Akhtar N¹, Yazan Y(PubMed)
(4) Ascorbic acid levels in various tissues, plasma and urine of mice during aging by Iwama M¹, Amano A, Shimokado K, Maruyama N, Ishigami A.(PubMed)
(5) Ascorbic Acid Modulation of Iron Homeostasis and Lysosomal Function in Trabecular Meshwork Cells by Xu P¹, Lin Y, Porter K, Liton PB(PubMed)
(6) High plasma levels of vitamin C and E are associated with incident radiographic knee osteoarthritis by Chaganti RK¹, Tolstykh I², Javaid MK³, Neogi T⁴, Torner J⁵, Curtis J⁶, Jacques P⁷, Felson D⁴, Lane NE⁸, Nevitt MC⁹; Multicenter Osteoarthritis Study Group (MOST)(PubMed)
(7) Trade-offs between anti-aging dietary supplementation and exercise by Mendelsohn AR¹, Larrick JW(PubMed)
(8) [The study of absorption efficiency and restoring effects of collagen and ascorbic acid on aged skin by fluorescence and reflection spectroscopy].[Article in Chinese] by Yang BW¹, Lin YM, Wang SY, Yeh DC.(PubMed)
(9) Ascorbic acid enhances the expression of type 1 and type 4 collagen and SVCT2 in cultured human skin fibroblasts by Kishimoto Y¹, Saito N, Kurita K, Shimokado K, Maruyama N, Ishigami A.(PubMed)
(10) Nanoscale gelatinase A (MMP-2) inhibition on human skin fibroblasts of Longkong (Lansium domesticum Correa) leaf extracts for anti-aging by Manosroi A¹, Kumguan K, Chankhampan C, Manosroi W, Manosroi J.(PubMed)
(11) Benefits of combinations of vitamin A, C and E derivatives in the stability of cosmetic formulations by Gianeti MD¹, Gaspar LR, Camargo FB Jr, Campos PM.(PubMed)
(12) Stability of vitamin C derivatives in topical formulations containing lipoic acid, vitamins A and E. by Segall AI¹, Moyano MA.(PubMed)
(13) Topical activity of ascorbic acid: from in vitro optimization to in vivo efficacy by Raschke T¹, Koop U, Düsing HJ, Filbry A, Sauermann K, Jaspers S, Wenck H, Wittern KP.(PubMed)
(14) Evaluation of a prescription strength 4% hydroquinone/10% L-ascorbic acid treatment system for normal to oily skin by Bruce S¹, Watson J(PubMed)
(15) Fatal vitamin C-associated acute renal failure by McHugh GJ, Graber ML, Freebairn RC.(PubMed)
(16) Ascorbic acid overdosing: a risk factor for calcium oxalate nephrolithiasis by Urivetzky M, Kessaris D, Smith AD.(PubMed)
(17) Dehydroascorbic acid prevents oxidative cell death through a glutathione pathway in primary astrocytes by Kim EJ¹, Park YG, Baik EJ, Jung SJ, Won R, Nahm TS, Lee BH.(PubMed)
(18) The oxidized form of vitamin C, dehydroascorbic acid, regulates neuronal energy metabolism by Cisternas P¹, Silva-Alvarez C, Martínez F, Fernandez E, Ferrada L, Oyarce K, Salazar K, Bolaños JP, Nualart F.(PubMed)
(19) Vitamin C crosses the blood-brain barrier in the oxidized form through the glucose transporters by Agus DB¹, Gambhir SS, Pardridge WM, Spielholz C, Baselga J, Vera JC, Golde DW.(PubMed)
(20) Expression and/or activity of the SVCT2 ascorbate transporter may be decreased in many aggressive cancers, suggesting potential utility for sodium bicarbonate and dehydroascorbic acid in cancer therapy by McCarty MF.(PubMed)
(21) Gelatin crosslinked with dehydroascorbic acid as a novel scaffold for tissue regeneration with simultaneous antitumor activity by Falconi M¹, Salvatore V, Teti G, Focaroli S, Durante S, Nicolini B, Mazzotti A, Orienti I.(PubMed)
(22) Dehydroascorbic acid as an anti-cancer agent by Toohey JI.(PubMed)
(23) Antiviral and virucidal activities of natural products by Arakawa T¹, Yamasaki H, Ikeda K, Ejima D, Naito T, Koyama AH.(PubMed)
(24) Topical L-ascorbic acid: percutaneous absorption studies, by Pinnell SR¹, Yang H, Omar M, Monteiro-Riviere N, DeBuys HV, Walker LC, Wang Y, Levine M.(PubMed)
(25) Use of topical ascorbic acid and its effects on photodamaged skin topography by Traikovich SS.(PubMed)
(26) Frictional properties of human skin: relation to age, sex and anatomical region, stratum corneum hydration and transepidermal water loss by Cua AB¹, Wilhelm KP, Maibach HI.(PubMed)
(27) Skin surface lipid and skin friction: relation to age, sex and anatomical region by Cua AB¹, Wilhelm KP, Maibach HI.(PubMed)
(28) Skin aging. Effect on transepidermal water loss, stratum corneum hydration, skin surface pH, and casual sebum content by Wilhelm KP¹, Cua AB, Maibach HI.(PubMed)
(29) Wrinkle reduction in post-menopausal women consuming a novel oral supplement: a double-blind placebo-controlled randomized study by Jenkins G¹, Wainwright LJ, Holland R, Barrett KE, Casey J.(PubMed)
(30) Effect of a novel dietary supplement on skin aging in post-menopausal women by Skovgaard GR¹, Jensen AS, Sigler ML.(PubMed)
(31) Vitamin C: a wound healing perspective, by Moores J.(PubMed)
(32) Nutrition 411: revisiting vitamin C and wound healing by Collins N.(PubMed)
(33) Ef[The study of absorption efficiency and restoring effects of collagen and ascorbic acid on aged skin by fluorescence and reflection spectroscopy].[Article in Chinese] by Yang BW¹, Lin YM, Wang SY, Yeh DC.(PubMed)
(34) Regulation of collagen synthesis in human dermal fibroblasts in contracted collagen gels by ascorbic acid, growth factors, and inhibitors of lipid peroxidation by Gessin JC¹, Brown LJ, Gordon JS, Berg RA(PubMed)
(35) Electroporation-mediated topical delivery of vitamin C for cosmetic applications by Zhang L¹, Lerner S, Rustrum WV, Hofmann GA.(PubMed)
(36) Liver Spot Removal: Vitamin C Products(Skin care guide)
(37) Lack of vitamin leads to brown spot on hand(livestrong)
(38) A topical antioxidant solution containing vitamins C and E stabilized by ferulic acid provides protection for human skin against damage caused by ultraviolet irradiation by Murray JC¹, Burch JA, Streilein RD, Iannacchione MA, Hall RP, Pinnell SR.(PubMed)
(39) UV photoprotection by combination topical antioxidants vitamin C and vitamin E by Lin JY¹, Selim MA, Shea CR, Grichnik JM, Omar MM, Monteiro-Riviere NA, Pinnell SR.(PubMed)
(40) Protective effects of topical antioxidants in humans by Dreher F¹, Maibach H.(PubMed)
(41) Effectiveness of antioxidants (vitamin C and E) with and without sunscreens as topical photoprotectants by Darr D¹, Dunston S, Faust H, Pinnell S.(PubMed)
(42) Successful short-term and long-term treatment of melasma and postinflammatory hyperpigmentation using vitamin C with a full-face iontophoresis mask and a mandelic/malic acid skin care regimen by Taylor MB¹, Yanaki JS, Draper DO, Shurtz JC, Coglianese M.(PubMed)
(43) Intravenous vitamin C in the treatment of post-laser hyperpigmentation for melasma: a short report by Lee GS.(PubMed)
(44) A randomized, double-blind, placebo-controlled trial of vitamin C iontophoresis in melisma by Huh CH¹, Seo KI, Park JY, Lim JG, Eun HC, Park KC.(PubMed)
(45) USE OF VITAMIN C IN ACNE VULGARIS  by GEORGE E. MORRIS, M.D.(Jama dermatology)