Jacobs Journal of Anatomy and Physiology

Fish is Best Food to Get Anti-Aging and Long Life. NOx Elimination Should be stopped to Produce Much Fish and To Protect Global Warming

*Shoichiro Ozaki
Department Of Physiology, Institute Of Physical And Chemical Research, Hirosawa Wokoshi Saitama , Japan

*Corresponding Author:
Shoichiro Ozaki
Department Of Physiology, Institute Of Physical And Chemical Research, Hirosawa Wokoshi Saitama , Japan
Email:ozaki-0991@jcom.zaq.ne.jp

Published on: 2019-03-21

Abstract

I found the reason why glucosamine, hyaluronic acid are used as much as health food. I found anti-aging reagent: sulfo disaccharides which co-working with Klotho. The disaccharides have glucosamine structure and similar structure with hyaluronic acid and chondroitin. Relations of the disaccharide, Klotho, hyaluronic acid, chondroitin, glucosamine with health and anti-aging were studied. Klotho makes disaccharide from glucosamine and glucuronic acid and co-works on site with produced disaccharide and gives stable Ca homeostasis and consequent health; anti-aging and long life. Fish is best food to get anti-aging and long life. The earth is warmed by burning of fossil and large amount of CO2 and NOx are emitted. Many developed countries eliminating NOx as pollution gas. Then CO2 assimilationist and plankton growth are blocked and fish production is blocked. I am insisting NOx elimination should be stopped to produce enough fish to get anti-aging and long life and to protect global warming.

Keywords

Anti-aging reagent, Klotho, Glucosamine, hyaluronic acid, Anti-aging food NOx, Protection of global warming.

Introduction

Anti-aging and long life is dream of persons for thousand years.Average life of Japanese is men 80.50 (third), women 86.83 (top in the world). I wonder why Japanese live longer than other country. I believe that Japanese food based on fish is fit and good for long life. Fish contain glucosamine, hyaluronic acid and chondroitin. Glucosamine, hyaluronic acid, chondroitin are now used as health food by many persons in Japan. Suntory sold more than 20 million bottles of glucosamine and chondroitin as nutrition supporting food. Setagaya shizenshokuhin sold 200 million bags of glucosamine, hyaluronic acid and chondroitin as health food for 14 years. Taishoseiyaku are selling glucosamine and chondroitin. Zeria Shinyaku is selling chondroitin as medicine for 60 years. Wada calcium pharmaceutical sold 10 million bags containing glucosamine, chondroitin sulfate and collagen as nutrition supporting food. About 7 million persons are drinking and eating these materials and enjoying health and long life. Japanese eat fish contains glucosamine, hyaluronic acid and chondroitin. Food will be concerned with long life. I tried to find the reason way glucosamine; hyaluronic acid and chondroitin are so effective by the studies of glucosamine derivatives. I found one reason why these compounds are connected with health and long life.  I found anti-aging reagent. This anti-aging gene is produced by Klotho (anti-aging gene) from glucosamine, chondroitin.

Glucosamine, hyaluronic acid, chondroitin are now used as health food by many persons in Japan. Suntory sold more than 20 million bottles of glucosamine and chondroitin as nutrition supporting food. Setagaya shizenshokuhin sold 200 million bags of glucosamine, hyaluronic acid and chondroitin as health food for 14 years. Taishoseiyaku are selling glucosamine and chondroitin. Zeria Shinyaku is selling chondroitin as medicine for 60 years. Wada calcium pharmaceutical sold 10 million bags containing glucosamine, chondroitin sulfate and collagen as nutrition supporting food. About 7 million persons are drinking and eating these materials and enjoying health and long life. Japanese eat fish contains glucosamine, hyaluronic acid and chondroitin. Food will be concerned with long life. I tried to find the reason way glucosamine; hyaluronic acid and chondroitin are so effective by the studies of glucosamine derivatives. I found one reason why these compounds are connected with health and long life. [1-7] I found anti-aging reagent. This anti-aging gene is produced by Klotho (anti-aging gene) from glucosamine, chondroitin. Anti-aging reagent co-work with Klotho and contribute for anti-aging and long life. Therefore glucosamine and chondroitin are effective food for anti-aging and long life. [8, 9].

I wish to recommend eating fish. But I found fish production of Japan decreased rapidly to 0-10 % of most fish production year 1970. I investigated the reason why fish production of Japan decreased so rapidly. I found that NOx and NP in waste water are eliminated as pollution substance almost completely. Plankton growth is inhibited and fish and sea weed cannot grow at such NP low concentration of sea. Fish price increased 5 times and we cannot buy fish easily and we cannot live longer. We may lose long life record of Japan. I am now insisting that NOx, NP elimination should be eliminated to increase NP concentration of sea to promote plankton CO2 assimilation to increase fish production. [10-14]

Klotho and sulfo- disaccharide

Nabeshima found that mutation of mouth Klotho gene leads to a syndrome resembling aging [15].The mouse having this gene can alive 30% longer. Since then many reports [16-45] are published. Petotou et al [30] reported that saccharides are involved in the binding of Klotho and FGF. Martin -Lumas report the activation of fibroblast growth glucuronidase capable factors by heparin [41] Klotho is a nobel β-of hydrolyzing β glycuronides [38] Nabeshima found a sulfo disaccharide from mouse liver cell. This saccharide binded with αKlotho and regulated the binding of αKlotho and FGF23. Mass analysis of this compound showed Mass number 843.28.C30H49N7O19S. Corresponding Thr-His-Gln-O-(β-D-3-sulfo-glucuronosyl) (1-4)-2-acetoamid-2- deoxy-D-glucopyranoside 9205 I planned to identify the real structure of this natural disaccharide by organic synthesis starting from known starting materials. Several disaccharides having similar structures were synthesized and relation of chemical structure and binding activity with Klotho and FGF 23 were studied [1-9]

Synthesis of sulfo disaccharide and measurements of binding activity

I have synthesized following compounds and measured their binding activity with Klotho and FGF23 [1] Synthesis of 9294; p-methoxyphenyl-3-sulfo-p-beta-D-glucronosyl-(1-4) - acetoamino-3-deoxy-glucopyranosyside 9294 is synthesized from 2 easily and this is most easily obtainable ant-aging reagent [3].

Structure activity relationship

9188,9583,9279,9244 were measured. The activities of these 5 compounds were almost same activity irrespective of (1-4) or (1-3) bond, glucopyranoside or galactopyranoside, p-methoxyphenyl, or esteron, or serine [1].

Structure of sulfo disaccharides

I planned to determine chemical structure of natural disaccharide by synthesis. But I cannot determine (1-3) bond or (1-4) bond. Because both bond compounds showed activity. Disacharides have close relation with hyaluronic acid and chondroitin. Hyaluronic acid and chodroitin have (1-3) bond. Therefore I wish to propose (1-3) bonded two structures: Sulfo-Glucuronosyl (1-3)-Glucoside , Sulfo-Glucuronosyl(1-3)-Galactoside as natural disacharides

Relation of Klotho, sulfo-disaccharide, glucosamin, hyaruronic acid and chondroitin

Klotho is an enzyme that in humans is encoded by the KL gene [10] This gene encodes protein that is related to β-glucuronidases. Klotho is a novelβ-glucuronidases [38]. β-Glucuronidase is an enzyme that can cleave glucuronosyl bond and this enzyme must do reverse reaction that is this enzyme can bind glucuronosyl group. Therefore Klotho can make disaccharide from glucuronoic acid and glucosamine.

                                                                                                   Hyalunoic acid is poly(glucuronosyl(1-3)glucoside)

                                                                                                  and chondroitin is polt(glucuronosyl(1-3)galactoside).

Hyalunoic acid gives gulcronic acid by Klotho

Chondroitin gives glucronyic acid by Kloth

Depolymerization of chitin gives N-acetylglucosamine. Depolymarization of chitosan gives glucosamine

Klotho combine glucronic acid and glucosamine to afford disaccharide

Klotho co-work with produced disaccharide on site to afford stable Ca homeostasis and subsequent Health and Anti-aging

These schemes indicate that Klotho is playing multiple roles.

1) Production of glucuronic acid

2) Production of disaccharide

3) Production of stable Ca homeostasis co-working 

with disaccharide These schemes show that disaccharides, Klotho, hyalunoicacid, chondroitin, glucosamine are working closely related all together and giving us health and anti-aging. Klotho is a regulator of Calcium homeostasis [26] working with produced disaccharide[3] Klotho makes disaccharide from glucosamine and glucuronic acid and co-works with produced disaccharide on site and gives stable Ca homeostasis and consequent health and anti-aging.

Glucosamine derivatives

When we look at glucosamine derivative from natural products, we can find many compounds like hyalunoic acid, chondroitin, chondroitinsulfate, chitin, chitosan, keratan sulfate, proteochonroitinsulfate (chondromuco protein). Hyalunoic acid is poly (glucuronosyl (1-3)glucoside) and chondroitin is polt(glucuronosyl(1-3)galactoside). Soft bone, cartilage: Proteochonroitinsulfate is proteio-glycan aggregate composed of hyarunoic acid and core-protein.

Anti-aging foods [7-9]

Following story exists for long time, never throw away umbilical cord, naval string and keep it at poulownia chest of drawers for long time, and when you get incurable sickness, boil it in water for long hours, then drink the solution , then heavy sickness will be cured. I wish to explain the reason why umbilical cord (main component is hyaluronic acid) is so effective Hyaluronic acid is a main umbilical cord (connecting constitutional substance of naval string, an tubes between placenta of mother and unborn-baby for the supply of nutrition). Hyaluronic acid was first isolated from the bovine vitreous body in 1934 by Karl Meyer, a professor at Colombia University. The name hyaluronic acid is derived from hyaloids which means vitreous body in Greek. Hyaluronic acid is a linear polysaccharide. It has a repetitive structural unit composed of the disaccharides N-Acetyl D-glucosamine and D-glucuronic acid. Hyaluronic acid in our bodies generally exists as a high molecular weight of several million times. Hyaluronic acid, chondroitin, glucosamine are now sold from several companies as health food and several million persons are drinking and eating these materials and enjoying health and long life. By the studies of disaccharide, I found the reason why these compounds are consumed as health food. For good health, anti-aging and long life, eating of food containing hyaluronic acid, chondroitin, glucosamine and derivative is essential. 

Eating of fish, whole body or head of fish or eye of fish, like sea bream(tai), house-macckerel (aji), mackerel pike (sanma), sardine (iwashi), shirasu (small sardine), iriko (boiled and dried small sardine),mezashi (dried sardine),shishamo, ayu, gold eye sea bream(kinme dai), loach(dojou), eel(unagi),carp(koi), curucian carp(funa), flat fish(hirame,karei),sand lance (ikanago), silver stripe rouna heming(kibinago),stone fish (okoje),blach rockfish(mebaru),herring(nishin), tsumire(crashed mixture of small fish), shark fin(fukahire), tsukudani of small fish, Among fish, clammy (nurunuru) fish like eel, anglerfish (ankou),loach, cat fish(namazu),sea cucumber(namako),octopus(tako),moray eel(utsubo), pike congera sea eel(hamo),clammy turtle (suppon) are good as anti-aging food, because main component of clammy substance at skin is chondroitin. Eating of eel (unagi no kabayaki) at hottest day of the year(doyono ushinohi) is popular as health food in Japan. Eel is served without head. But if it served with head, the efficiency will be increased. Anglerfish (ankou) is delicious and whole body is eaten. Octopus is clammy and dried under sun and eaten as health food. But octopus is washed with washing machine and clammy chondroitin is washed away. This octopus is not as effective as unwashed one.Turtle (suppon) is clammy and famous as delicious anti-aging food. Husk of shrimp and crab is composed with chitin and chitosan. Depolymerization of chitin gives N-acetyl glucosamine. Depolarization of chitosan gives glucosamine. Eating of shrimp and crab with head and husk is recommended. Eating of nebaneba (stringy and sticky) vegetable like sea tangle, kelp, mozuku, yam? nattou(fermented soybean)is recommended. Eating of cartilaginous cow, pig and chicken tissues is recommended. Hyaluronic acid was isolated from eyeball of cow and was named from Greek name glassily hyaloid. Hyaluronic acid is found in the highest concentrations at eye and joints. Most chondroitin appears to be made from extracts of cartilaginous cow and pig tissues (cow trachea and pig ear and nose, pig pitreous gel, but other sources such as bird cartilage are also used. The people who do not like to eat fish are advised to eat tablet of glucosamine, hyaluronic acid and chondroitin. Nonfood benefit of hyaluronic acid Hyaluronic acid is used for anti-aging skin handling [46], wound Healing [47, 48] cosmetic use [49]. Hyaluronic acid has been used to treat osteoarthritis of the knee via injecting it into the joint [50, 51].

Relation of plankton CO2 assimilation, fish production and NOx [10-14]

360 Billion tone CO2 was produced in the world in 2016 by the burning of 140 billion tone fossil. To protect global warming, we must fix CO2 same amount of CO2 as emission. We are emitting 360 billion tones CO2. We are fixing 283 billion tone .Therefore 142 billion tone CO2 is in creasing. We must fix 142 billion tones CO2 by proportion of emission of each country. NOx: Amount of NOx is estimated from the fact that NOx is produced about 1/25 of produced CO2. The ratio C/N = 25/ 1 is same as plant composition ratio C/N = 25/1. Plant eat CO2 and nutrient N by the ratio C/N =25/1. 14.4 billion tone NOx is estimated to be produced when 360 billion tone CO2 is produced. Many governments such as United State, Japan , Germany ,United Kingdom ,France consider NOx as pollution gas and eliminated NOx by the reaction with ammonia. They ignored the importance of NOx as fertilizer. Amount of NOx is so much, CO2 assimilation is retarded very much. CO2 fixing is retarded very much. And global warming is accelerated very much. For the production of ammonia, much fossil is used and much CO2 is produced and global warming is accelerated so fast. World fish production in 2016 increased to 200 million tone, about double of 93 million tone in1997. China produced 79.38 million tone, Indonesia 22.21, India 10.11,Vietnam 6.21increasing fish production very much. Japan produced 16 million tone fish, top in the world in 1960 but fish production decreased to 4.64 million tone 7th place in 2016. This is huge decrease. Japan is eliminating 3 million tone N and P since 1980. Therefore NP concentration of sea decreased remarkably. Plankton cannot grow at this low concentration. Therefore fish decreased. 12 million tone fish was not produced in recent years. Japanese can alive longest by eating fish as main protein source. Because fish contain hyaluronic acid, glucosamine and chondroitin which precursor of anti-aging reagents which co-working with anti-aging gene Klotho [1- 9] Japanese may lose long life record. (Men 80.50 (third), women 86.63 (top) from the fact that fish production was reduced remarkably by NOx elimination law.

Summary

We can live longer by eating food containing glucosamine, chondroitin. I recommend you to eat fish and cartilaginous caw, chicken tissue. Eating of nebaneba (stringy and sticky) vegetable like sea tangle, kelp, mozuku, yam nattou (fermented soybean) is recommended. Because anti-aging gene Klotho makes anti-aging disaccharide from glucosamine and glucuronic acid and co-working with produced disaccharide and contribute to Ca homeostasis and subsequent health and anti-aging. Glucosamine, chondroitin and hyarunoic acid are starting material to make anti-aging reagents : sulfo disaccharide. NOx elimination should be stopped to get much fish and protect global warming.

Reference

1. Ozaki Shoichiro. Sulfo disaccharides co-working with Klotho. Studies on structure, structure activity relation and function. World J of Pharmacy and Pharmaceutical Sci 2015 4(8):152-175

2. Ozaki Shoichiro Glucosamine Derivatives. Sulfo disaccharide co-working with Klotho. Nutrition and Food Science 2015 5.416

3. Ozaki Shoichiro. Synthesis of anti-aging reagent: Sulfo disaccharide co-working with anti-aging gene. Archeves of Medicines 2015 6:17

4. Ozaki Shoichiro. Nutrition for good health, anti-aging and long life. Hyaluronic acid,glucosamine and chodroitin. Maternal and Paediatric Nutrition Journal 2015 1: e102.

5. Ozaki Shoichiro Food containing hyaluronic acid and chondroitin is essential for anti-aging (2016) International J of Aging & Clinical Research 1, 101.

6. Ozaki Shoichiro. Toward anti-aging and long life. Jacobs Journal of Physiology. 2016 2(1) 12

7. Ozaki Shoichiro. Secret of anti-aging: Anti-aging food containing glucosamine, hyaluronic acid and chondroitin. JJ Physiology 2016 2(1): 13-17

8. Ozaki Shoichiro. Food effective for anti-aging and long life. New Food Industry 2017 59(2) 91-94

9. Ozaki Shoichiro. Method to anti-aging and long life Rikuryo Science (2017) Vol 59 10-12

10.Ozaki Shoichiro. NOx is Best Compound to Reduce CO2. Eur J Exp Biol 2017 7:12.

11. Ozaki Shoichiro. Method to fit Paris agreement for protection of global warming . Int J Waste Res 2017 7(4)318

12. Ozaki Shoichiro. Effect of NOx elimination on electricity price, fish production, GDP and protection of global warming. Int J of Waste Res 2018 8 (1) 1000328

13. Ozaki Shoichiro. Promotion of Plankton CO2 assimilation by effective use of NOx and NP is best method to produce much fish and protect global warming 2018 J Marine Sci Res Oceanography Volume 1 issue (1). 1

14. Ozaki Shoichiro.NOx and NP in wast water fix CO2 and control global warming and climate Int J Biochem Physiol 2018 3(4): 000140

15. Kuro-o M, Matsumura Y, Aizawa H, Kawaguchi H, Nabeshima YI et al (November 1997). “Mutation of the mouse klotho gene leads to a syndrome resembling aging “. Nature 390 (6655): 45-51.

16. Arking DE, Krebsova A, Macek M Sr, Macek M Jr, Arking A, et al 2002. “Association of human aging with a functional variant of klotho”. Proc. Natl. Acad. Sci. U.S.A.99 (2): 856-61. Bibcode:2002 PNAS;99-856A.

17. Xiao NM, Zhang YM, Zheng Q, Gu J May 2004 “Klotho is a serum factor related to human aging”. Chin. Med. J. 117 (5): 742-747: May 2004 .

18. Takayama Y, Herz J, Kahn CR, Rosenblatt KP, Kuro-o M Dena B. Dubal et al. Life Extension Factor Klotho   Enhances Cognition, Cell Reports, 2014.

19.Nabeshima Y (2000)]”Challenge of overcoming aging-related disorders.”J. Dermatol. Sci. 24(1): 15 21.

20. Kuro-O, “Klotho and aging”. Biochimica et Biophysica Acta M 2000 1790 (10): 1049-1058.

21. Tsujikawa H, Kurotaki Y, Fujimori T, Fukuda K, Nabeshima Y.”Klotho, a gene related to a syndrome resembling human premature aging, functions in a negative regulatory circuit of vitamin D endocrine system”. Mol. Endocrinol. December 2003-17 (12): 2393-403.

22. Xiao NM, Zhang YM, Zheng Q, Gu J May 2004. “Klotho is a serum factor related to human aging”. Chin. Med. J. May 2004 117 (5): 742-7.

23. Zhang R, Zheng F 2008. “PPAR-gamma and aging: one link through klotho?”. Kidney Int. 2008 74 (6): 702-4

24. Imura A, Tsuji Y, Murata M, Maeda R, Nabeshima Y et al. 2007. “alpha-Klotho as a regulator of calcium homeostasis” Science 316 (5831): 1615-8. Bibcode: 2007Sci...316.1615I

25. Fukumoto S. “Chronic kidney disease (CKD) and bone. Regulation of calcium and phosphate metabolism by FGF23/Klotho”. 2009 Clin Calcium 19 (4): 523-8.

26. Nabeshima Y. “[Discovery of alpha-Klotho and FGF23 unveiled new insight into calcium and phosphate homeostasis]”. 2008 Clin Calcium 18 (7): 923-34.

27. Nabeshima Y. (2007) Molecular function of α-Klotho in calcium homeostasis. Igakunoayumi 222(3) 225-230

28. Prié D, Ureña Torres P, Friedlander G 2009. “Fibroblast Growth Factor 23-Klotho: a new axis of phosphate balance control”. Med Sci (Paris) 25 (5) 489-95.

29. Razzaque MS.. “FGF23-mediated regulation of systemic phosphate homeostasis: is Klotho an essential player?”. Am. J. Physiol. 2009 Renal Physiol. 296(3): F470-6.

30. Prié D, Ureña Torres P, Friedlander G. “[Fibroblast Growth Factor 23-Klotho: a new axis of phosphate balance control]”. Med Sci 2009 25 (5): 489-95.

31. Torres PU, Prié D, Beck L, et al. “Klotho gene, phosphocalcic metabolism, and survival in dialysis.”. J Ren Nutr 2009 19 (1): 50-6.

32. Medici, D.; Razzaque, M. S.; Deluca, S.; Rector, T. L.; Hou, B.; Kang, K.; Goetz, R.; Mohammadi, M.; Kuro-o, M.; Olsen, B. R.; Lanske, B.. “FGF-23-Klotho signaling stimulates proliferation and prevents vitamin D-induced apoptosis”. The Journal of Cell Biology 2008 182 (3): 459-465.

33. Wolf I, Laitman Y, Rubinek T et al. “Functional variant of KLOTHO: a breast cancer risk modifier among BRCA1 mutation carriers of Ashkenazi origin.”Oncogene 2010 29 (1): 26-33.

34. Torres PU, Prié D, Molina-Blétry V et al.”Klotho: an antiaging protein involved in mineral and vitamin D metabolism.” Kidney Int. 2007 71 (8): 730-7.

35. Halaschek-Wiener J, Amirabbasi-Beik M, Monfared N, et al. Mary Bridger, Joanna, ed. «Genetic variation in healthy oldest-old.». PLoS ONE 2009 4 (8): e6641. Bibcode:2009PLoSO...4.6641H.

36. Shimoyama Y, Nishio K, Hamajima N, Niwa T.»KLOTHO gene polymorphisms G-395A and C1818T are associated with lipid and glucose metabolism, bone mineral density and systolic blood pressure in Japanese healthy subjects.» Clin Chim Acta 2009 406 (1-2): 134-138.

37. Kurosu H, Kuro-o M.»The Klotho gene family and the endocrine fibroblast growth factors.». Curr Opin Nephrol Hypertens 2008 17 (4): 368-72.

38. Choi BH, Kim CG, Lim Y et al. «Transcriptional activation of the human Klotho gene by epidermal growth factor in HEK293 cells; role of Egr-1.» Gene 2010 450 (1-2): 121- 127.

39. Menon R, Pearce B, Velez DR, et al. «Racial disparity in pathophysiologic pathways of preterm birth based on genetic variants.» Reprod Biol Endocrinol 2009 7: 62.

40. Halaschek-Wiener J, Amirabbasi-Beik M, Monfared N, et alMary Bridger, Joanna, ed. «Genetic variation in healthy oldest-old.»PLoS ONE 2009 4 (8): e6641. Bibcode:2009PLoSO...4.6641H.

41. Chen SN, Cilingiroglu M, Todd J, et al. «Candidate genetic analysis of plasma high-density lipoprotein-cholesterol and severity of coronary atherosclerosis.» BMC Med. Genet. 2009 10: 111.

42. J. L. dePaz, M. Martin-Lomus (2005)Eur J. Org.Chem., 1849-1859

43. Rafael Ojeda, Jesus Angulo,Pedro M, Manuel Martin-Lumas .The activation of fibloblast grouth factors by heparin: Synthesis and structureal study of rationally modified heparin-like oligosaccharides. Can.J Chem 2002 80:917-936

44. Tubeur C, Jean-Maurice Mullet, Bono F, H,Maurice J-M Petitou,P. Sinay. Bioorganic and Medicinal Chem1999 2003- 2012

45. Tohyama O, Imura A, Iwano A,Jean-Noel Freund, Henrissat R, et al. Nabeshima YKlotho is a novelβ-glucuronidases capable of hydrolyzing steroidβ-glucuronides J.Biological Chemistry 2004 279(11): 9777-9764

46. Tammi R, Ripellino JA, Margolis RU, Maibach HI, Tammi M “Hyaluronate accumulation in human epidermis treated with retinoic acid in skin organ culture”. J Invest. Dermatol 1989 92 (3): 326-32.

47. Chen WY, Abatangelo G “Functions of hyaluronan in wound repair”. Wound Repair Regen 1999 7 (2): 79-89.

48. Longaker MT, Chiu ES, Adzick NS, Stern M, Harrison MR, et al.”Studies in fetal wound healing. V. A prolonged presence of hyaluronic acid characterizes fetal wound fluid”. Ann Surg 1991 213 (4): 292-296

49. Edwards, PC, Fantasia. “Review of long-term adverse effects associated with the use of chemically-modified animal and nonanimal source hyaluronic acid dermal fillers.” Clinical interventions in aging JE 2007 2 (4): 509-519.

50. Rutjes AW, Jüni P, da Costa BR, Trelle S, Nüesch E, et al.”Viscosupplementation for osteoarthritis of the knee: a systematic review and meta-analysis”. Ann. Intern. Med 2012 157 (3): 180-191.

51. Frampton JE “Hylan G-F 20 single-injection formula”. Drugs Aging 2010; 27(1) 77-85.