PROTOCOL RESEARCH OBJECTIVE:

The purpose of this study was to quantify the effect of ingesting Activated Oxygen (ASO®) oxygen in a colorless aqueous medium on anaerobic threshold at moderate altitudes.

The first objective was to measure any changes in blood oxygen saturation with daily consumption of water treated with Activated Oxygen (ASO®).

The second objective was to determine if the ingestion of Activated Oxygen (ASO®) affects ventilatory threshold when performed at moderate altitudes.

The third objective was to determine if the ingestion of Activated Oxygen (ASO®) increases blood oxygen saturation levels (PaO2).

The fourth objective was to determine if the ingestion of Activated Oxygen (ASO®) affects blood pressure (systolic and/or diastolic).

The fifth objective was to determine if the ingestion of Activated Oxygen (ASO®) affects the nitric oxide (NO2) levels in the blood stream.

The hypothesis tested:

Does Activated Oxygen (ASO®) increase the partial pressure of oxygen in the blood? Two groups of 25 active military personnel, training and/or performing under identical conditions participated in the study.

One group received normal canteen water. The experimental group received the same water but was treated with Activated Oxygen (ASO®).

Fingertip pulse oximetry was utilized to monitor Pa02 as well as invasive blood sampling from arterial blood at precise intervals after consumption of Activated Oxygen (ASO®).

The Ventilatory threshold and blood lactate levels were also measured during treadmill exercise to determine if the anticipated increase in blood oxygen content has a beneficial effect on metabolic capacity.

An environmental symptoms questionnaire and profile of mood states was also administered to evaluate effects of Activated Oxygen (ASO®) on symptomatology and mood.

Analysis:

The following report is a review and assessment of the data obtained in the course of the Marine altitude field study conducted July 7–16, 1999 under the auspices of the Department of Human Performance, Naval Health Research Center, San Diego, CA, the Division of Foods and Nutrition, University of Utah, Salt Lake City, UT, and the Marine Corps Mountain Warfare Training Center, Bridgeport, CA:

“Effect of stabilized oxygen consumed with water on blood and urine markers of oxidative stress and blood oxygen saturation during extended military mountaineering training at moderate altitude.” Eldon W. Askew, Ph.D., Donald E. Roberts, Ph.D., James E. Reading, M.A., Jeffrey M. Pfeiffer, M.S., Lt. Lance Orr, MC, USNR.

This study was conducted at the United States Marine Corps Mountain Warfare Training Center, July 7-16, 1999. The purpose of the study was to investigate the effect of ingestion of stabilized oxygen (ASO®) in water, over the course of 8 days of military mountaineering training at roughly 8,000 feet above sea level.

Parameters studied included: 1) anaerobic threshold and performance as measured by cycling to exhaustion on a stationary bicycle, 2) blood oxygen saturation (SAO2), and, 3) indicators of oxidative stress: urine 8-hydroxydeoxyguanosine (8-OHdG), urine total alkenals, and blood lipid hydroperoxides.

The test subjects were instructed to put 4 TBS (60 ml) of the stabilized oxygen solution (ASO®) into each liter of their drinking water.

Positive Study Trends:

Some positive directional trends were observed in the treatment group, in terms of the percentage changes of group means between 7/09/99 and 7/16/99. Parameters measured where positive trends were observed included VO2 max (mL/min/kg body weight), VO2 max (L/min), and Ventilatory threshold (L/min):

(1) The treatment group showed a mean percentage improvement in VO2 max (mL/min/kg body weight) of 6.5% between 7/09/99 and 7/16/99, compared with a +2.1% mean change in the control group over the same time period.

(2) The treatment group showed a mean percentage improvement in VO2 max (L/min) of 4.7% between 7/09/99 and 7/16/99, compared with a +0.2% mean change in the control group over the same time period. Page 20 This means that the treatment group had roughly between a 5% to 7% improvement in the rate that their bodies used oxygen under aerobic exercise conditions that would “max” them out; in other words, training to exhaustion. With a greater amount of oxygen consumed in a given period of time, the body is able to produce a greater amount of energy for fueling muscles, other organs and tissues, giving greater stamina and endurance.

(3) The treatment group showed a mean % improvement in Ventilatory threshold (L/min) of 5.7% between 7/09/99 and 7/16/99, compared with a +0.8% mean change in the control group over the same time period.

The ventilatory threshold is associated with the lactate threshold, the point during muscular exertion at which there is not enough oxygen in the bloodstream to produce ATP energy in the normal fashion.

Aerobic Metabolism:

 ATP is normally produced within the mitochondria (energy powerhouses) in each cell of the body from glucose derived from sugars, other carbohydrates and foods that we consume. The glucose molecule is split into two molecules of pyruvic acid; normally the pyruvic acid enters the mitochondria of the cells, reacts with oxygen, producing ATP.

Anaerobic Metabolism:

When there is not enough oxygen available to meet the body’s (particularly muscles’) energy demands at any given moment, the pyruvic acid is converted into lactic acid, producing ATP without any oxygen at all. As lactic acid builds up in the muscles, though, fatigue sets in and performance becomes rapidly impaired.

Ventilatory Threshold:

By raising the ventilatory threshold by nearly 6%, the subjects in the treatment group would have been able to work out longer without muscle fatigue than those not receiving the oxygen supplement.

Negative Parameters:

Some parameters evidenced somewhat of negative percentage changes of group means between 7/09/99 and 7/16/99, in both the control as well as the treatment groups. These parameters included Time to exhaustion (seconds), percentage maximum heart rate achieved during test, blood oxygen saturation, blood lactic acid, Hematocrit, and body weight (kg; lbs).

The results indicate a slight general increase in oxidative stress indicators in both groups. The treatment group exhibited a significant increase with time for urine total alkenals and plasma lipid peroxides, but overall there were no significant differences in oxidative stress indicators between the two groups.

Urine Creatinine:

One interesting test result was in the area of Urine creatinine (mg/dL). The control group exhibited a significant increase in urine creatintine over time, while the treatment group’s mean percent change was a possible slight decrease. Although the reasons for these particular changes over time may not be known, the urine creatinine values are often used as indicators of relative average body mass differences between experimental groups. A higher urine creatinine value normally indicates a greater comparative lean body mass, as creatinine is a metabolic waste product from muscle creatine. Based on the Urine creatinine values of both groups at the start of the experiment, it is clear that the average lean body mass of the treatment group was significantly greater than that of the control group.

Lean Body Mass:

The difference in average lean body mass between groups may be one of many possible explanations for the lack of significant positive changes in the treatment group where they might have been anticipated. In this regard, it is helpful to look at the natural acclimatization of natives living at high altitudes:

“In all aspects of acclimatization, the natives are superior to even the bestacclimatized lowlanders, even though the lowlanders might also have lived at high altitudes for 10 or more years. Acclimatization of the natives begins in infancy. The chest size, especially, is greatly increased, whereas the body size is somewhat decreased, giving a high ratio of ventilatory capacity to body mass. In addition, their hearts, particularly the right side of the heart, which provides a high pulmonary arterial pressure to pump blood through a greatly expanded pulmonary capillary system, are considerably larger than the hearts of lowlanders.”

Given that the natural adaptation of the human body to high altitude (i.e. low oxygen) is to have a higher ventilatory capacity to body mass ratio, it is not unreasonable to assume that the treatment group in the experiment, due to their greater body mass, were at more of a disadvantage compared to the control group in terms of oxygen consumption; as they would tend to require and therefore consume more oxygen overall as opposed to the control group, the parameters measured would not indicate as great a benefit gained through the ingestion of the stabilized oxygen (ASO®) supplement as would have been anticipated.

Even though each group served as its own control from baseline to the end of the study, the dose of oxygen supplement (ASO®) given the treatment group may have been inadequate, based on body size as well as other possible variables, to show the greatest possible objective benefit.

Further research will be necessary, utilizing different test subject groups and experimental conditions, in order to fully evaluate parameters that are seen as inconclusive based on this field study. It is likely that the multitude of factors encountered in field studies (e.g. differences in training, fitness and activity among test subjects, supplement ingestion compliance) contributed to the variability in the data. A laboratory study where these variables can be more carefully controlled, and each test subject serves as his own control, is planned.

References:

1. Guyton, Arthur C., M.D., Hall, John E, Ph.D., Textbook of Medical Physiology, Ninth Edition, W.B. Saunders Co., 1996, p. 551

U.S. Marines Speak Out

PERCEIVED PHYSIOLOGICAL BENEFITS:

Although subjective responses from test subjects on questionnaires are not considered scientifically valid, they can indicate possible positive benefits derived from the supplement being studied. The subject responses can also highlight some of the possible weaknesses of the field study in terms of controlling the conditions of the experiment.

One case in point is found in the test subject responses to the question:

“Was the taste of the product when dissolved in water : — Noticeable? — Objectionable?”

In the group receiving the placebo supplement (consisting of sodium chloride and water), 69% found the taste noticeable, but 0% found the taste objectionable. In the group receiving the actual supplement (ASO®), 100% found the taste noticeable, and 29% found the taste objectionable. Additionally, there were no written comments regarding taste from the control group, whereas 35% of the subjects receiving the actual supplement (ASO®) volunteered written comments, all of them somewhat negative, in reference to the taste of the supplement dissolved in water. A couple of subjects in the treatment group went so far as to suggest that the objectionable taste made it likely that they weren’t drinking as much water as they would otherwise.

Obviously, if the test subjects in the treatment group are drinking varying amounts of the treated water, then they’re also receiving inconsistent levels of the oxygen supplement (ASO®); this probable variability in dosage of ingested supplement could be one of the most significant factors explaining the “less than earth-shaking” positive benefits noted in the treatment group.

All this being said, there were still significantly more positive benefits reported by the test subjects in the treatment group than by those in the control group. The following responses were in reference to question #3 of the Final Questionnaire given the test subjects:

“Did you notice any specific effects that you attribute to the supplement you dissolved in your water, such as:

TESTIMONIALS:

The following are written comments from a few of the test subjects in the treatment group (those that received the actual oxygen supplement (ASO®):

“more endurance towards strenuous workouts”

“I didn’t think I was ‘As’ out of breath as other Marines”

“I noticed some effects of the product. Like I was able to breathe more going up the hills and perform better.”

“In the beginning of the test it helped out a lot, I believe, but here at the end with all the hard work & training we’ve been through it’s gotten hard to tell…”

“I wish I could remain on the water. I did feel it helped me. I only wish it tasted a bit better!”

CONCLUSION:

In short, in both measurable parameters and subjective observations, the test subjects in the group treated with the oxygen supplement (ASO®) experienced the following to a greater degree than the control group:

• Greater stamina and endurance
• Reduced muscle fatigue
• More energy
• Less “out of breath”
• Greater feeling of strength
• Felt that the product helped them perform better.

ASO® and ASOC® are trademarks of BIO2 International, Inc. All international rights reserved. ©2006 BIO2 International, Inc.

These statements have not been evaluated by the U.S. Food and Drug Administration. This product is not intended to diagnose, treat, cure or prevent any disease or medical condition. Always consult with a professional medical practitioner before taking any dietary supplement, especially if pregnant, nursing, taking prescription medications or under a doctor’s medical care.

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