A story about oxygen nanobubbles: introduction

The document attached to this blog is a Freedom of Information Act Appeal submitted to the US  Department of Health And Human Services on July 2013.

I can certify that the document is original, except for erased information related to my home address and personal IDs. More than 1 year has passed since my request without receiving any responses, that’s why I decided to start today the disclosure of information about RNS60.

My story related to oxygen nanobubbles (charge-stabilized nanobubbles or RNS60) started in a rainy Thursday on May 2013.

More information will be disclosed in the future, however I do not know in which date I’ll post more information about RNS60, as well as the level of disclosure or for how long I’ll engage on scientific & ethical discussions about RNS60.



News Division
Office of the assistant Secretary for public Affairs
US   Department of Health And Human Services.
7700 Wisconsin Avenue, Suite 920,
Bethesda, Md 20814, U.S.A

Jorge Hernan Ramirez, US and Colombian citizen, resident in Cali, Colombia, through this paper, present my appeal to the answer of my request (request number 20133730) for information about RNS60 received by myself at my personal address on June 16, 2013.


RNS60 as a novel therapeutic compound consisting of physically modified saline: fact or a fairy tale?

RNS60 is a product developed by Revalesio Corporation consisting of physically modified saline containing charge-stabilized nano-structures (CSN) with high concentrations of oxygen. In principle, RNS60 could not be cataloged as a drug because it has no active pharmacological principle, it consist of oxygen nanobubbles in saline, presumably stable in solution, without adverse effects and tremendous potential applications in several inflammatory, neurological, and cardiovascular diseases.

Disclosure FOI Request Log Date range from 05/13/2013 to 05/17/2013 Report

Revalesio is a biotechnology company founded in Tacoma, Washington in 2000. The original name of the company was Microdiffusion Inc. and changed its name to Revalesio in 2007. Goods and services offered by Revalesio Corporation registered in the United States and Australia includes eye topical solutions, drinking water, agricultural products, and solutions for cleansing wounds and burns (Table 1).

In May 2013 the news media from Colombia and other Latin American countries announced a multicentric phase II clinical trial on five Colombian cities (Cali, Bogotá, Barranquilla, Medellín, and Bucaramanga) for the prevention of neurological sequelae in patients with stroke. This study is on the design stage by several neurologists, leaded by the famous Colombian neuroscientist  Rodolfo Llinas and sponsored by the colombian pharmaceutical company Tecnoquimicas through an alliance with Revalesio Corporation. Tecnoquimicas is an important pharmaceutical company, specialized in generic medicines, with their headquarters in Cali, Colombia; Tecnoquimicas also have several manufacturing plants in other Latin American countries.

No preclinical studies has been published evaluating the effects of RNS60 on animal models of stroke. Several active pharmaceutical principles (e.g., tilirazad) with positive results on animal models showed no benefits on preventing the neurological sequelae of stroke in human clinical trials.

Revalesio Corporation claims the existence of these nanobubbles without any convincing scientific data published in the scientific literature. In the following appeal several pharmacological and ethical questions regarding the research with RNS60 are presented. In addition, the RNS60 story is plagued by misleading advertising, several potential of conflict of interests, and the possibility of scientific fraud.

Trademark serial number Goods and services
77123216 Eye drops; eye wetting solutions; saline solution for the eyes; topical eye medications; contact lens wetting solutions; saline solutions for contact lens use; saline solutions for medical use
77123314 Agricultural services, namely hydroponics, aquaculture and gardening services relating to the growing of plants
77123285 Agricultural products and forest products, namely raw fruit and vegetables, natural plants and flowers
77123285 Drinking water; mineral water; flavored waters; soft drinks; sports drinks; oxygenated drinking water; oxygenated flavored waters; oxygenated soft drinks; oxygenated sports drinks.
77123234 Surgical and medical apparatus, namely aqueous solution for the cleansing, irrigating and debriding of wounds and burns

Table 1. Revalesio Corporation goods and services

Reliant water is a sport drinking beverage owned by Revalesio Corporation. This drinking water has been promoted to improve exercise performance in athletes through misleading advertisement. The price of 32 water bottles of Reliant is $100 USD. The effectiveness of Reliant Water on exercise has not been evaluated by scientific studies. Revalesio Corporation have no drug available in the pharmaceutical market and their pipeline only includes RNS60.

Four clinical trials (phase I and phase II) evaluating the effects of RNS60 on acute myocardial infarction, asthma and multiple sclerosis have been registered at clinicaltrials.gov (completion dates September 2011 and October 2011 for two of these clinical trials). Another clinicaltrials.gov record report the existence of a phase I study to determine the effect of the topical application of Revera Wound Care on venous leg ulcers, the reported completion date of this study was Oct 2010 but no results have been published to this day.

Box 1. Clinical trials registered at clinicaltrials.gov evaluating the effects of RNS60 on humans

Eleven short scientific abstracts have reported the effects of RNS60 on animal models of diseases, in vitro pharmacological studies (tissues and cell cultures), and humans (phase I clinical trial on asthma) have been published in several scientific journals (Table 2). Of these scientific abstracts only two complete manuscripts evaluating the effects of RNS60 have been published and some authors were not included in the previously published scientific abstracts. In addition, the studies conducted on ischemia (Circulation 2011 / J Amer Coll Cardiol 2013) and alzheimer (Neuroscience 2012 / Journal Neurochemistry 2013) are duplicated abstracts .

Abstract Authors Year Journal Complete publication in scientific journal
P05 Multiple Sclerosis: Treatment Mechanism of Action of RNS60, a Novel Therapeutic, Suppresses Relapsing-Remitting EAE Via Tregs: Implications for MS Therapy (P05.181) Susanta Mondal, Supurna Ghosh, Richard Watson and Kalipada Pahan 2013 Neurology No
Abstract 17400: Charge-Stabilized Nanostructures Reduce Ischemia-Reperfusion Injury in A Pig Model in vivo Andreas Kalmes; Supruna Ghosh; Richard Watson 2011 Circulation No
rns60, a novel therapeutic, inhibits glial activation and memory loss in a mouse model of Alzheimer’s disease K. K. MODI, A. JANA, S. GHOSH, R. WATSON, K. PAHAN; 2012 Neuroscience No
RNS60, a novel anti-inflammatory therapeutic, protects neurons and memory in a transgenic mouse model of Alzheimer’s disease K. K. MODI, A. JANA, S. GHOSH, R. WATSON, K. PAHAN; 2013 Journal Neurochemistry No
RNS60 DEMONSTRATES EARLY PROMISE IN FIRST HUMAN ASTHMA TRIAL Watson, R; Ghosh, S; Kalmes, A; Nadeau, K 2011 Annals of asthma, allergy and immunology No
RNS60, a Novel Therapeutic, Modifies T-Helper Cells and Inhibits Experimental Allergic Encephalomyelitis: Implications for Treating Multiple Sclerosis Mondal, S; Ghosh, S; Watson, R; Pahan, K 2012 Neurology Yes(J Biol Chem)
Suppression of NF-kappa B Activation and Inflammation in Glial Cells by RNS60, a Novel Therapeutic: Implications for Neurodegenerative Disorders Khasnavis, S; Jana, A; Roy, A; Wood, T; Ghosh, S; Watson, R; Pahan, K 2012 Neurology Yes (Plos One)
Isotonic Saline Subjected to Taylor-Couette-Poiseuille Flow Demonstrates Anti-Inflammatory Activity in a Rat Model of Allergic Asthma Ghosh, S; Mega, TL; German, S; Burke, LM; Diegel, ML; Wood, AB; Watson, RL 2011 Journal of Allergy and Clinical Immunology No
Isotonic Saline Subjected to Taylor-Couette-Poiseuille Flow Demonstrates Anti-Inflammatory Activity In Vitro Ghosh, S; Mega, TL; German, S; Burke, LM; Diegel, ML; Wood, AB; Watson, RL 2011 Journal of Allergy and Clinical Immunology No
Electrokinetically Altered Normal Saline Modulates Ion Channel Activity James C. Choi, Tony L. Mega, Sean German, Anthony B. Wood, Richard L. Watson. 2012 Biophysics Journal No

One scientific paper (Bhushan et al. 2013) probing the existence of nanobubbles on sample of RNS60 have been recently published.(1) However, several issues about this manuscript deserves special attention: (i) the article was published the next day after submission; (ii) samples of RNS60 were provided by Revalesio; and (iii) authors acknowledge the important contribution of corporate leaders of Revalesio Corporation for their useful review to the manuscript. Some of this publications have been duplicated and the corporate leaders of Revalesio are listed as authors in all of the published manuscripts.

The articles published in Journal of Biological Chemistry and Plos One have also some issues to consider (Table 3).

Publication title Authors Year Journal Comments
Suppression of nuclear factor-κB activation and inflammation in microglia by physically modified saline. Khasnavis S, Jana A, Roy A, Mazumder M, Bhushan B, Wood T, Ghosh S, Watson R, Pahan K. 2012 J Biol Chem Mazumder M and Brusham B are not listed as authors in the same abstract published on  Neurology (2012)
Protection of Tregs, suppression of Th1 and Th17 cells, and amelioration of experimental allergic encephalomyelitis by a physically-modified saline. Mondal S, Martinson JA, Ghosh S, Watson R, Pahan K. 2012 Plos One Martinson JA are not listed as author in the same abstract published on  Neurology (2012)
Do not describe the analysis and characterization of RNS60

Table 3. Publications of RNS60 (PubMed)

Revalesio technique for the production of RNS60 have already been protected by patent  (US8410182 B2) and their use for several diseases have already been submitted to patenting agencies. There is no reason to hide the results of these studies from the scientific community and the public opinion, especially considering the possibility of recruiting people for a phase II clinical trial without a proper knowledge on the effects RNS60 on animals, tissues, and phase I studies in humans.

During the last two months there was a very high news media coverage reporting the beneficial effects of RNS60 humans with alzheimer, parkinson, stroke, and other neurological degenerative diseases (especially in Colombia – see links below). Misleading advertisement of RNS60 and their properties deceits patients and their families.

As a US citizen my concerns are the corporate practices promoting the use of food supplements with dubious beneficial effects and the intentions of these companies to test these products in ethically questionable human clinical trials.

As a Colombian citizen my main concern is the recruitment of hundreds of patients in a phase II clinical trial to test an intervention not appropriately characterized (i.e., the existence of nanobubbles on RNS60 have not been adequately proved) with insufficient evidence of their effectiveness and safety in human beings.

As a biomedical researcher, University professor and president of the human ethical research committee my concerns are the conflict of interests between scientists with the pharmaceutical industry and misleading advertising of ineffective therapies.

Available information suggests that a multicentric phase II study in Colombia designed to test the effects of RNS60 in stroke is unethical. I am requesting information regarding the approval by the FDA of phase I and phase II studies of RNS60. Furthermore, if the studies of RNS60 were in fact approved by the FDA, I will also like to know the scientific basis for this approval, how the existence of this nanobubbles was confirmed, and information regarding the review processes of phase I clinical trial results before the approval of phase II studies.

My initial FOIA request was denied because it could damage corporation interests. However, human health, public interests, and research ethics are above any corporate, governmental, and personal interests.

Notifications: ———– ERASED | CONFIDENTIAL ————–

Sincerely yours,

Jorge Hernán Ramírez Escobar. MD, MSc, PhD.
Professor of Pharmacology at Icesi University
US Citizen – Passport Number ———– ERASED | CONFIDENTIAL ————–
Colombian Citizen – ID ———– ERASED | CONFIDENTIAL ————–

Scientific references

Mondal S, Martinson JA, Ghosh S, Watson R, Pahan K.Protection of Tregs, suppression of Th1 and Th17 cells, and amelioration of experimental allergic encephalomyelitis by a physically-modified saline. PLoS One. 2012;7(12):e51869.

Khasnavis S, Jana A, Roy A, Mazumder M, Bhushan B, Wood T, Ghosh S, Watson R, Pahan K. Suppression of nuclear factor-κB activation and inflammation in microglia by physically modified saline. J Biol Chem. 2012 Aug 24;287(35):29529-42.

Bhushan BB, Pan Y, Daniels S. AFM characterization of nanobubble formation and slip condition in oxygenated and electrokinetically altered fluids. Journal of Colloid and Interface Science 2013; 392:105–116.

News media


















Scientific blogs


Revalesio corporation and Reliant Water








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