International Journal of Research Studies in Medical and Health Sciences
Volume 6, Issue 5, 2021, PP 08-14
ISSN : 2456-6373
DOI: https://doi.org/10.22259/ijrsmhs.0605002

Phototherapy Induced Metabolism Change Produced by the
LifeWave X39 Non-transdermal Patch

Melinda H. Connor, D.D., Ph.D., AMP, FAM* [1], Caitlin A. Connor, MAcOM, DAOM [2], Naran Gombusuren, Ph.D. [3], Jens Eickhoff, Ph.D. [4], Nathan Anderson, MACOM, LAc.[5], Marsha Perry, RN, MA [6], Leonard Peugh, MAcOM [7]

1. Founder, Earth Songs Holistic Consulting
2. Post Doctoral Diploma Candidate, Health Sciences Research, Rewley House, Oxford University, UK
3. Global Operations Manager, Serametrix/Caprion
4. Senior Scientist, Biostatistics & Medical Informatics, University of Wisconsin Madison
5. Dean of Clinical Education, Arizona School of Acupuncture and Oriental Medicine
6. Study Manager, Earth Songs Holistic Consulting
7. Senior Research Assistant, Earth Songs Holistic Consulting

*Corresponding Author: Melinda H. Connor, D.D., Ph.D., AMP, FAM, Earth Songs Holistic
Consulting, 31907 South Davis Ranch Rd. Marana, AZ 85658, USA

ABSTRACT

Purpose: To determine X-39 patch impact in stimulation of Copper peptide biosynthesis and bio-available amino acid levels, neurotransmitters production, memory, sleep quality, vitality, muscle relaxation and blood pressure.

Materials: Biography Infinity physiology suite: Heart rate variability (HRV), GSR, EMG, EKG, blood volume pulse (BVP), temperature and respiration. Questionnaires: Marlow-Crowne, Global Mood Scale, Pittsburg Sleep Quality Index, Arizona Integrative Outcomes scale. WAIS III memory test. Amino acid and neurotransmitters testing of urine.

Method: Subjects were recruited (age 40 – 81), consented, randomized and scheduled. Data taken day 1, day 2, and day 7 except Marlow-Crowne taken day 1 and day 7.

Results: Improvements in short term memory p<0.001, sleep quality p<0.04, vitality p<0.03 day 2 and p<0.08 at day 7. Blood pressure change in VLF on day 7 at p<0.02, respiration on day 7 at p<0.04. Increase in amino acids: Creatinine, Normetanephrine, methionine, homocystine, isoleucine, glutamine, cysteine, 5-hydroxytrytophane, β-aminobutyric-acid.

Conclusion: The results of the double blind randomized controlled trial of 50 subjects with mean age 63 years, using the LifeWave non-transdermal X-39 phototherapy patch worn 8-12 hours per day for seven days produced an increase in 8 amino acids at significant levels. The study showed, there was an improvement in short term memory as measured by the WAIS III memory test at significant levels over 7 days, in Quality of Sleep at significant levels within 24 hours and a self reported increase in vitality at near significant levels in 7 days as measured by the Arizona Integrative Outcomes scale. Our findings suggest this patch is not only stimulating the biosynthesis of copper-peptide production, but also increases neurotransmitters production and improves metabolism. Additional studies could address underlying mechanisms of action to the phototherapy process and longer periods of study might be explored for additional potential physical changes and longevity of the demonstrated changes.

Keywords: Photobiology, Photobiomodulation, Amino Acid, Metabolism

Report

INTRODUCTION

The Lifewave X39 non-transdermal patch focuses on stimulating the copper tripeptide GHK- Cu [1,2,3]. “Copper tripeptide-1(GHKCu) is a small protein composed of the three amino acids glycine, histidine, and lysine combined in a specific geometric configuration with the physiologically beneficial mineral (copper)” [4]. The goal of this research was to determine X-39 patch impact in stimulation of Copper peptide biosynthesis and bio-available amino acid levels, neurotransmitters production, memory, sleep quality, vitality, muscle relaxation and blood pressure. This tripeptide was first isolated from human plasma albumin in 1973 by Dr. Loren Pickart. Pickart, noticed differences in the levels of fibrinogen based on age. He additionally noticed that these differences stopped when the older liver cells were incubated in blood from younger individuals. “In 1977, David Schlesinger of the Harvard University Chemistry Department confirmed that the growth modulating peptide isolated by Pickart was a glycyl-L-histidyl-Llysine peptide”[5].

It is interesting to note that this peptide has also been found in saliva, urine, and collagen. Additional research has established the strong affinity the GHK peptide has for copper, and exists in two forms, as this was not covered in the initial experiment. These two forms are GHK and GHK-Cu. It is also important to mention that none of the research around GHK has ever found it to cause an issue [4]. Research has identified that the peptide is used to signal the beginning of the natural repair process. This benefit has specifically been documented through research for post-laser or surgical wounds, ischemic, burns, skin or hair transplants, and diabetic ulcers. “Diabetic wounds healed three times faster in the presence of Copper tripeptide-1. Time to reepithelialization is shortened” [4].

The tripeptide has also been demonstrated to improve tissue remodeling. “It increases keratinocyte proliferation and normal collagen synthesis, improves skin thickness, skin elasticity and firmness, improves wrinkles, photodamage and uneven pigmentation, improves skin clarity, and tightens protective barrier proteins” [4]. This has an impact on both scars and other effects of damage to the skin, and natural aging processes. The effects of tissue remodeling also appear to have an impact on cancerous cells. “The fact that GHK was able to suppress 70% of genes involved in the development of an aggressive metastatic form of colon cancer indicates that GHK is capable of the regulation of various
biochemical pathways on a gene level and it seems to be resetting the gene activity back to
health, which leads to the improvement of tissue repair” [6].

GHK-Cu also has a demonstrated impact on other organs in the body after they have been damaged. “A collaborative study conducted by scientists from Boston University, University of Groningen, University of British Columbia, and University of Pennsylvania established that the GHK peptide reverses the gene expression signature of COPD, which is manifested by emphysema, inflammation, lung tissue destruction, and significant reduction of lung capacity” [6]. It is also important to note that “…the level of GHK is about 200 ng/mL(10−7M) at age 20 and it declines to 80 ng/mL by age 60” [6].

This likely explains the increasing effects of aging. It would also suggest that increased levels over time of GHKCu would have a positive effect on both life expectancy and aging. While phototherapy has been defined as “the use of ultraviolet (UV) light for its healing effects” [7], the LifeWave patches have been specifically developed to reflect light in the infrared and visible light bands back onto the skin where the patch has been placed. Supported by normal electro-dermal skin conductance [8,9], the human body gives off a number of materials biochemically including particulate release, gas emission, ultraviolet, infrared, near infrared, and visible spectrum light. This then stimulates the area of skin, producing improved physiological effects. Variations on phototherapy have been used for at least 100 years. In that time there has been little evidence of negative side effects. This suggests that this is a relatively untapped option for healing with relatively few risks.

MATERIALS AND METHODS
This study was done as a whole systems research randomized controlled double blind trial of 50 participants. Human studies ethics approval for this research was given NFFEH 03-31-19-02. Participants were recruited using flyers, emails and radio announcements. A sample of convenience drawn from those who responded and met inclusion/exclusion criterion of men and women age 40-81 with no major mental health issues who were then randomized into control and active groups via computer and to patch placements (GV14, CV6). Patches used in the trial were mailed prepackaged as groups A and B and the blind was not broken until after the statistical analysis was done. Participants were scheduled for the same time each day for each of the data points (day1/baseline, day2/24 hours, day 7). Participants were consented, and data was taken in the following order: questionnaires, urine sample, physiology measures. On day one the participant was given 9 days supply of the patches for use in the study. A one month supply of the active patch was given at the end of the week’s participation and $20 toward the participants time and gasoline. The trial was conducted at the Arizona School of Acupuncture and Oriental Medicine clinic in Tucson, AZ.

Non-transdermal Patch and Patch Placement

All X39 patches are sealed so that none of the substances in the patch actually penetrate the skin. This allows for consistent patch promotion of the light flow during the period of patch application. There is sufficient evidence of electrical-dermal response in orthopedic research to use previously measured acupuncture points as “strategic conductors of electromagnetic signals” [10]. Two points were selected for this research so the process of patch placement could be standardized across the participant population. Earlier studies of this patch selected acupuncture/acupressure points designated GV14 and CV6 [2, 3] and based on pilot results, the same patch placement was selected for consistency. Participants were randomized by computer into groups and the point usage was selected based on the randomization.

Metabolic Suite

Sabre Sciences Inc. conducted the urine analysis using their HPA1 metabolic panel using LCMS/MS tandem mass spectrometry. The metabolic panel measures excreditory level of amino acids and their metabolites, including catecholaminergic, serotonergic, glutamatergic pathways and metabolites, and trans-sulfuration and histidine pathways. Three urine data points were taken on each subject before 10am each of baseline/day1, at 24 hours/day 2 and on day 7. Samples were frozen and then shipped to the Sabre Science laboratory in Carlsbad, CA each day for analysis.

Physiology Suite
The Thought Technology Biography Infinity physiology suite was used for six minute measures of heart rate variability (HRV), galvanic skin response (GSR), EMG, EKG, blood volume pulse (BVP), temperature and respiration at each data point.

Questionnaires
Questionnaires and memory test were administered at each data point with the exception of the Marlow-Crowne which was administered day one and day seven.

Marlowe Crowne
The Marlowe Crowne is a 13 Item true/false short likert scale that measures political correctness. This instrument was selected to confirm accuracy of the data. The instrument takes about 2 minutes to do for most participants. (Norming: [11].)

Pittsburg Sleep Quality Index
Normed by Cole et al (2006) this scale is a mix of quantitative questions and five likert scale
questions 0-3 addressing the participants quality of sleep.

Arizona Integrative Outcome Scale, Visual Analogue Scale (AIOS-VAS) for Vitality

The AIOS- VAS (Normed: [12]) rates subject’s “overall sense of well-being and vitality.” It uses a 100mm one-line visual analogue scale on which the participant notes their sense of well being with an x on the line.

WAS III memory sub-test

A subsection of the WAS III was administered which looks at the level of short, mid and longer term memory. The test was administered concurrent to the filling out of other questionnaires. Ten numbers which were taken from a randomized list were repeated without inflection three times to each participant. Participants were asked to immediately repeat the numbers back at the end of the third repetition, then again asked to repeat the numbers back at 10 minutes and at twenty minutes. Count was taken of the number of correct digits in the order originally given. Count was stopped when an error in the order was made.

Global Mood Scale

Denollet normed the Global Mood Scale in 1993 It includes a twenty item, five factor likert
scale that assesses participants current mood and a one item, ten factor likert scale assessing well being at the current moment. It was selected both as a redundant measure comparison to the AIOS-VAS and an overall assessment of mood.

Statistical Analysis

All data results were entered into spreadsheets and they were then analyzed for significant
results. Questionnaires were analyzed for mean and standard deviation and stratified by
assessment time point. Paired t-test or nonparametric Wilcoxon Signed Rank tests were then done. Physiology measures were summarized stratified across the 6 study epochs looking at means and standard deviation. Pre-post changes were evaluated using a paired ttest. Distribution assumptions were then verified using normal probability plots. The values for p are two-sided and statistical significance was defined as p<0.05. The metabolic analysis included stratification by assessment point and summary means and standard deviation.

Analysis was done including changes from baseline to day 2, day 2 to day 7 and day 1 to day 7. These were evaluated using Paired t-test or nonparametric Wilcoxon Signed Rank tests. RESULTS AND DISCUSSION This was a double blind randomized controlled trial with a sample of convenience and which had a seven day intervention period. While this was a sample of convenience participants were recruited through five different methods including: radio announcements, email, posting in cafe’s and at university center’s, prior study participant announcement lists and through area community groups.

So while this could be considered a study weakness, the range of recruiting methods insured a diversity of population who decided to participate. There were 26 individuals in the Active group and 24 in the Control group, with an overall mean age of 63 and a range of 40-81 years. The Active groups mean age matched the overall mean, while the Control group was slightly lower at 62 years.

Despite the limitations of smaller participant sample size (N=50) in this study, there were several interesting significant changes, as well as some changes of near significance within the study test results. There were two adverse events, where two individuals in the study had headaches while using the patches. These headaches took place during monsoon season, and both were individuals who had a previous history of barometric pressure triggered migraine headaches.

The IRB reviewed the cases and determined that they were both consistent with the individuals past history and had nothing to do with the product. Two individuals were also dropped from the study when they did not complete the study visits. The specific impact of research location should also be recognized.

Aside from the barometric pressure effects of monsoon season, this study largely took place over the summer in Arizona. This meant that participant hydration was a constant issue, especially with the necessity of a urine sample. As such, it was determined that water had to be available and was offered to each participant at the beginning of each study appointment. Included are the significant and some near significance findings for the LifeWave non-transdermal X39 phototherapy patch.

The urine analysis data showed an increase in amino acids: Creatinine, Normetanephrine, methionine, homocystine, isoleucine, glutamine, cysteine, 5-hydroxytrytophane, β-aminobutyricacid.

These are important findings as branched chain amino acids like isoleucine, which is constantly oxidized in muscle, methionine and homocystine, are major ingredients of transsulfuration pathway. This pathway is responsible for methyl donor S-adenosylmethionine (SAMe) production.

SAMe is readily donates methyl groups to other substances, enabling cardiovascular, neurological, reproductive and detoxifying systems. 5-hydroxytrytophane is major building block of important neurological monoamine neurotransmitter, serotonin, which later methylates into melatonin, and is a main hormone which regulates sleep-wake cycle. Normetanephrine is a norephinephrine break down product in dopamine pathway and is major hormone of neuromodulatory system.

Table 1: Comparisons of amino acid concentrations between Control group vs. Active group at day baseline (1), day 2 (2) and day 7 (7) All amino acids concentration results were summarized in terms of means and standard deviations (SD), stratified by group (Control vs. Active) and assessment time point (1, 2, 7). Absolute changes from baseline (1) to 2 and baseline (1) to 7 were evaluated using a paired t-test or nonparametric Wilcoxon signed rank test. All reported p-values are two-sided.

Table 2: Comparisons of absolute changes in amino acid concentrations from baseline (1) to day 2 and
baseline (1) to day 7 between Control group vs. Active group

1: p-pvalue for evaluating changes from baseline to 2 and baseline to 7 within each arm
2: p-value for comparing changes from baseline to 2 and 7 between arms

All questionnaire scores were summarized in terms of means and standard deviations (SD), stratified by group (Control vs. Active) and assessment time point (1, 2, 7). Absolute changes from baseline (1) to day 2 and baseline (1) to day 7 were evaluated using a paired t-test or nonparametric Wilcoxon signed rank test. All reported p-values are two-sided. Questionnaire outcomes:

• WAIS III (three subscales #Short, # Mid, Long)
• Global Mood Scale (two subscale: Positive Affect (PA), Negative Affect (NA))
• Pittsburgh Sleep Quality Index (total score – higher score indicates worse sleep quality)
• Marlow-Crowne (total score)
• AIOS-VAS (total score)
  

Within the questionnaire results the shifts in sleep were particularly interesting, as they were significant in both groups, in opposite directions. The Active group had a significant improvement in sleep, and the control group had a significant decline in sleep. Vitality also showed a particularly interesting shift, with significant improvement within 24 hours which dropped slightly at day 7. The Global mood scale showed a change in negative affect in both groups. Additionally the Marlow-Crowne showed no significance so that responses on the questionnaires were not as a result of the desire to provide socially correct or desired answers.

Physiological measures showed significant normalization in blood pressure in VLF p<0.02 and near significance in LF p<0.09 and power p<0.06. Average respiration became deeper by day 7 at p<0.04 and neck and shoulder muscles showed improved relaxation effects by day 7 at p<0.08.

A comparison of this studies results with the pilot study showed some interesting differences. The first was the change in the level of significance in memory improvement. While the pilot study showed a near significant improvement in short-term memory, the double blind showed very clear significance, p<0.001, as well as near significance in mid-term memory. It would be very interesting to see if this improvement continued to strengthen with a longer intervention period or a population with a mean age of 70+. Of note, was the difference in which amino acids were significant between the pilot and the current study. The amino acids which has significance were further down the catecholaminergic, serotonergic, and glutamatergic pathways that the significant findings of the pilot study. This may be due to the difference in median age, with the double blind having an older median population or may be due to utilization by the body when higher concentrations of the amino acids were available for use. It should also be noted that while we saw more prevalent amino acid changes tied to antioxidant events in the pilot study, the data from the double blind shows an overall rebalancing of the gut and overall improvement in gut performance. It would be interesting to determine how long and at what continuing rate changes to gut performance, physiological and overall wellness support would be produced by wearing the LifeWave X-39 patch. Longer periods of study such as six to twelve weeks including comprehensive blood and metabolism testing might be considered in the future. CONCLUSION The results of the double blind randomized controlled trial of 50 subjects on the LifeWave non-transdermal X-39 phototherapy patch worn for 8-12 hours per day for seven days produced an increase in 8 amino acids at 10 significant levels over the 3 time periods. There was additional increase in 3 amino acids at near significance. These specific changes served to rebalance the gut toward a positive homeostatic balance. There was an increase in short term memory as measured by the WAIS III memory test at significant levels over 7 days. There was an improvement in Quality of Sleep at significant levels within 24 hours and a self reported increase in vitality at near significant levels in 7 days as measured by the Arizona Integrative Outcomes scale. Given the mean age of the population at 63 years this is a substantial improvement in overall quality of life. Additional studies could address underlying mechanisms of action to the phototherapy process and longer periods of study might be explored for additional potential physical changes.

ACKNOWLEDGMENTS

Funding for the study was provided by: LifeWave Corp. All author salaries for the research were provided by LifeWave. No author has a commercial interest in the company of any kind.

REFERENCES
[1] Connor, C., Connor, M., Yue, D., Eickhoff, J., Wagner, S., et al. (2021) Double-Blind Testing of the Lifewave X39 Patch to Determine GHKCu Production Levels. Internal Med Res Open J Volume 6(1):1-3.
[2] Connor, C., Connor, M ., Yue, D., Chang, CL., Eickhoff, J., Wagner, S. “Changes in Tripeptides Produced by the LifeWave X-39 patch”, International Journal of Healing and Caring Online, May 3, 2020
[3] Connor, M., Connor, C., Gombosuren, N., Eickhoff, J., “LifeWave X39 Pilot Demonstrates Light Triggered Changes”, International Journal of Healing and Caring Online, May 2020
[4] DeHaven, C., (2014) Copper Tripeptide-1. Science of Skincare.
[5] Schlesinger, DH; Pickart, L; Thaler, MM (1977). “Growth-modulating serum tripeptide is glycyl-histidyl-lysine”. Cellular and Molecular Life Sci 33 (3): 324–325. doi:10.1007/BF02002806.
[6] Pickart, L., Vasquez-Soltero, J., Margolina, A. (2015) GHK Peptide as a Natural Modulator of
Multiple Cellular Pathways in Skin Regeneration. Hindawi Publishing Corporation BioMed Research International Volume 2015, Article ID 648108, 7 pages http://dx.doi.org/10.1155/2015/648108.
[7] Kakimoto, C., (2017) What is phototherapy, and how does it work? https://www.dermatologistoncall.com/blog/what-isphototherapy-and-how-does-it-work/
[8] Flick, A. B., “Silver and Wound Healing”, CAM Research Symposium, Hershey Medical School, Hershey PA, July 2004.
[9] Becker, R., Selden, G., “The Body Electric”, William Morrow Pub., New York, NY, 1985.
[10] Feinstein, D., “Rapid Treatment of PTSD: Why Psychological Exposure with Acupoint Tapping May Be Effective”, Psychotherapy: Theory, Research, Practice, Training. 47(3), 385-402, 2010, American Psychological Association.
[11] Reynolds, W. (1982) Development of reliable and valid short forms of the Marlowe-Crowne Social Desirability Scale. Journal of Clinical Psychology. January 1982.
[12] Bell, I., Cunningham, V., Caspi, O., Meek, P., Ferro, L., (2004) “Development and validation of a new global well-being outcomes rating scale for integrative medicine research.” BMC Complementary and Alternative Medicine. 2004; 4: 1.

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As of December 2018, LifeWave has completed more than 80 clinical studies on its products since 2002. X39® is no exception, with clinical research comprising traditional blood, urine and saliva analysis along with various bioelectrical data such as Heart Rate Variability. We have also completed clinical work utilizing self assessment to determine how X39® improves such things as pain management and sleep.

The following is a summary of a study performed by Dr. Melinda Connor on the X39® patch. This study explores the metabolic implications and physiologic results of wearing the Lifewave X39® patch over the period of one week. Measures were taken at baseline, 24 hours and at 7 days of wearing the patch.

A sample of convenience of 15 subjects made up of both men and women aged 40-65 were selected to participate in this study. For the purposes of this study the Sabre Sciences HPA2 metabolic suite has been selected. The HPA 2 Profile includes both urine and saliva collection. The urine evaluates 5-HTP, Serotonin, Dopamine, Epinephrine, Norepinephrine, GABA, Glutamate, Histamine, L-DOPA, Normetanephrine, Metanephrine 3-Methoxytyramine. The saliva evaluates, Cortisol and DHEA.

Several additional measures have been added to the HPA2 suite, Glutathione, and a cysteine to cystine ratio will also be tested using the urine sample. All study participants had testing done at base line, 24 hours and 7 days. Six minute recordings of EKG, pulse, respiration, heart rate variability (HRV), temp,
blood volume pulse, galvanic skin response, 2 EMG (muscle) leads.

A series of questionnaires were administered to support metabolic findings. Marlowe Crowne (accuracy of findings), McGill Pain Scale, Pittsburg Sleep Quality Index, AIOS-VAS (for vitality and well-being), Profile of Mood States (POMS) (anxiety/depression), WAIS III (short and long term memory). After only 1 Day of wearing X39® people reported feeling better. After 1 week of using X39® there was statistical significance showing clear overall improvement in the feelings of vitality and wellness.

WAIS III (short and long term memory)
WAIS III is a well-established intelligence test which includes a standard memory test. There was a clear improvement in short term memory by day 7. It is also interesting that there was improvement in both mid and long term memory as well, though to a much lower degree

Pittsburg Sleep Quality Index
This questionnaire showed an immediate strong shift the first night, though it was not quite significant. The shift by day 7 was significant. This is particularly important as sleep strongly effects everything else related to health and wellbeing.

Metabolic outcomes urine testing
Amino acids and neurotransmitters play a critical role in health and wellbeing f an individual’s amino acid and neurotransmitter production is broken, the individual cannot maintain body health for long. The number of statistically significant changes demonstrated in this study shows the powerful impact of the X39®. Among the 20 of the biochemicals improved include: Epinephrine, GABA, Histamine, Normetanephrine, PEA (Phenylethylamine), Alpha-aminobutyric acid.

• Epinephrine – Marker of Response to Stress
• GABA – Quality of Sleep, Anxiety, Mood
• Histamine – Measures Inflammatory Response
• Normetanephrine – Measure for Adrenal Glands
• PEA (Phenylethylamine) – Indicator for Mood and Cognition
• Alpha-aminobutyric acid – Indicator for GABA Synthesis

A number of amino acids also improved:

• Cystine – Involved in Glutathione Formation
• Glutamine – Involved in Intestinal Repair
• Lysine – Immune Function
• Leucine – Muscle Repair
• Tryptophan – Involved in Sleep

Heart Rate Variability
HRV measures ratios of the low frequency and high frequency muscle contractions in the heart. Statistical significance was achieved in demonstrating that X39® helps to support the health of the heart.

BVP-HR, EMG, Skin-Condition, Temperature and Respiratory Rate
Reduction in blood pressure and improved muscle relaxation are consistent changes which are present in the physiology data. Greater flexibility in HRV in the over age 60 population is an important finding

Summary
The data from this study shows improvement in:

• Blood pressure and overall health of the heart
• 17 statistically significant amino acid changes over the 7 days
• Key neurotransmitters involved in overall health
• Improvement in sleep levels
• Improvement in short term memory
• Improvement in reported feelings of overall health and vitality

GHK-Cu may Prevent Oxidative Stress in Skin by Regulating Copper and Modifying Expression of Numerous Antioxidant Genes Loren Pickart, Jessica Michelle Vasquez-Soltero, and Anna Margolina Skin Biology, Research & Development Department, 4122 Factoria Boulevard SE, Suite No. 200, Bellevue, WA 98006, USA Received: 17 June 2015 / Accepted: 21 July 2015 / Published: 28 July 2015

The copper binding tripeptide GHK (glycyl-l-histidyl-l-lysine) is a naturally occurring plasma peptide that significantly declines during human aging. It has been established that GHK:Copper(2+) improves wound healing and tissue regeneration and stimulates collagen and decorin production. GHK-Cu also improves the condition of aging skin and hair, and possesses antioxidant and anti-inflammatory effects. In addition, it increases cellular stemness and secretion of trophic factors by mesenchymal stem cells. GHK’s antioxidant actions have been demonstrated in vitro and in animal studies. They include blocking the formation of reactive oxygen and carbonyl species, detoxifying toxic products of lipid peroxidation such as acrolein, protecting keratinocytes from lethal Ultraviolet B (UVB) radiation, and blocking hepatic damage by dichloromethane radicals. This paper reviews biological data demonstrating positive effects of GHK in skin and proposes interaction with antioxidant related genes as a possible explanation of its antioxidant activity.

One of the most common experiences people report with the X39® patch is an improvement in energy levels. When the energy patches were first developed in 2002, and then first sold in 2004, it was established with multiple clinical studies that the Energy patches were upregulating which is known as beta oxidation (fat burning) by around 23%. As the body derives more energy from fat than carbohydrates, it was easy to see why people were reporting the feelings of more energy.

The Aeon patch was released in 2011, and this patch was designed to reduce inflammation and increase antioxidant production with the benefit of providing stress relief. People that use Aeon often report feeling calmer during the day as well as having more energy from this stress reduction.

It is hypothesized at this point based on the data we have that the X39® is increasing the feeling of energy that people have through the reduction of inflammation, oxidative stress, and elevation of antioxidants. As we know that GHK-Cu provides these benefits, and we know that inflammation and
oxidative stress reduce a persons energy capacity, this is a reasonable conclusion. In the future, there may be studies that look at this specific benefit of the X39® patch. As of December 2018, a clinical study completed by Dr. Thornton Streeter did in fact demonstrate that continued use of the X39® patch resulted in an overall improvement of a person’s energy levels. As a review, following is what LifeWave has substantiated by its own clinical research as the benefits that come from wearing the X39® patch on a regular basis:

WOUND HEALING – Some of our most dramatic testimonial on the X39® have been in the area of wound healing. Wound healing is of course a natural process and requires the presence of stem cells. As we age, our stem cells become less effective though. Now, with X39®, you can activate you stem cells and provide support for your body’s natural healing process.

RAPID PAIN RELIEF – X39® users can experience dramatic reductions in pain. In fact, most people experience a reduction in the sensation of pain and inflammation within minutes of application.

GREATER ENERGY – Clinical studies performed by LifeWave show that the X39® patch improved a person’s energy levels. The results when compared to the baseline, see the improvement in overall energy of the body, organ balance, and L/R symmetry distribution of energy.

SLEEP BENEFITS – Clinical studies performed by LifeWave show that X39® improves the quality of sleep by altering levels of GABA, an important neurotransmitter in the brain. The benefit is a natural improvement in your quality and duration of sleep.

IMPROVED SKIN APPEARANCE – Would you like to not only feel younger but look younger as well? Now you can with X39®. An important part of activating stem cells is increasing the production of collagen. The result is that only a few weeks after applying X39® your skin will start to experience a significant reduction in the appearance of lines and wrinkles.

DRAMATIC ANTI-AGING – Initial clinical work performed by Dr. Loren Pickart, it was shown that when GHK-Cu is elevated in the body, we can see that cells start to behave like younger, healthier cells. Essentially helping to restore our cells to a younger state.

FASTER RECOVERY FROM EXERCISE – Users of X39® report significant improvements in their sports
performance and recovery. As stem cells are designed to repair damaged cells, this is not surprising.

REDUCED INFLAMMATION – Many users of X39® report that after several weeks of using the product, they notice a reduction in the appearance of scars. This is a well-known attribute of elevating copper peptide known as remodeling.

Abstract

Purpose: To determine if the LifeWave X39 non-transdermal photobiomodulation active patch would show improved production of GHK-Cu over controls in a double blind randomized controlled trial.

Materials: BD Vacutainer Safety Loc Blood Collection sets with Pre-attached holder sized 21GX0.75 or 23GX0.75 and lavender top tubes. Kendro Sorvall Biofuge Centrifuge 75005184+ and AB Sciex API4000 Qtrap. Analysis software included: Qtrap Analyst software 1.6.2 and R software version 3.5.1. Statistical analyses were conducted using R software (version 3.5.1; http://www.r-project.org/).

Method: Sixty people age 40-80 were computer randomized into two groups. One lavender top tube was drawn and then spun in Kendro Sorvall centrifuge for 10 minutes at 1300 rcf. The plasma was placed in cryo tubes and flash frozen to -22C then shipped in dry ice to laboratory for analysis. The filtrate was concentrated by speed-vac and reconstituted with de-ionized water to 50 ul and analyzed with AB Sciex API4000 Qtrap. Statistical assessments were evaluated using a nonparametric Wilcoxon signed rank test, p values are two-sided and p<0.05 was used to define statistical significance.

Results: A significant increase in GHK-Cu concentration in the blood of the active group was seen comparing changes from Day 2 to Day 7 between Group A vs. Group B in GHK-Cu Concentration (ng/ml) at p<0.035 and in Total GHK-Cu (ng) at p<0.03.

Conclusion: This study showed a significant increase in the GHK-Cu concentration present in the blood as a result of wearing the LifeWave X39 patch for 1 week in individuals age 40 to 80. This is seen from Day 2 to Day 7 between Active vs. Control in GHK-Cu Concentration (ng/ml) at p<0.035 and in Total GHK-Cu (ng) at p<0.03.

Keywords

GHK-Cu, Meridian, Non-transdermal, Photobiology, Phototherapy

Introduction

This study explores the impact of wearing the LifeWave X39 non-transdermal photobiomodulation patch over the period of one week on levels of glycyl-L-histidyl-L-lysine-copper(2+) (GHK-Cu) levels in the blood in a double-blind randomized controlled trial. This particular tripeptide was first isolated by Dr. Loren Pickart in 1973. GHK-Cu is important as the “copper tripeptide-1 belongs to a group of emergency response molecules which are released during injury and come to the body’s aid…” [1] It is naturally sent by the body to any type of injury to tissue. For example: the “copper tripeptide-1 has been suggested to have a potential therapeutic role in age-related neurodegeneration and cognitive decline. It improves axon survival and maintenance of nerves” [1]. It has been implicated in the resetting of 4000 genes [2]. Blood samples to determine GHK-Cu levels were taken at baseline, 24 hours and at 7 days of wearing the patch. A sample of convenience of 60 subjects made up of both men and women aged 40-81 were selected to participate in this study. Participants were randomized into Group A or Group B by computer.

Background

The LifeWave X39 patch uses phototherapy to stimulate a rebalancing of the body. Based on data from other studies, it was felt that a possible change in the copper tripeptide GHK-Cu might be a factor in the effects produced by the patch. As a follow on to prior studies it was determined that a double blind study was an appropriate method of testing this theory. The tripeptide has been demonstrated to improve tissue remodeling in previous research. “It increases keratinocyte proliferation and normal collagen synthesis, improves skin thickness, skin elasticity and firmness, improves wrinkles, photodamage and uneven pigmentation, improves skin clarity, and tightens protective barrier proteins” [3]. Research has identified that the peptide is used to signal the beginning of the natural repair process.

The Tripeptide

“Copper tripeptide-1(GHK-Cu) is a small protein composed of the three amino acids (protein building blocks) glycine, histidine, and lysine combined in a specific geometric configuration with the physiologically beneficial mineral (copper)” [4]. Later research established the strong affinity between the GHK peptide and copper, and the two forms (GHK and GHK-Cu) it exists in, as this was not covered in the initial experiment. It should also be mentioned that GHK has never been found to cause an issue in all of the research that has been done [1].

Non-transdermal Patch

All X39 patches are sealed to prevent the contact of any of the substances inside to the skin. The sealing of the patches allows for consistent light flow through the patch the entire time that the patch is worn. Patches are designed to reflect wavelengths of light in the infrared, near infrared, and visible light bands. Using the same adhesives as band-aids, this limits the level of irritation which might be developed through consistent daily use of the patch.

Phototherapy

Phototherapy has been used for over 100 years in various forms. There has been little evidence of negative side effects throughout that time period. This suggests that phototherapy is a relatively untapped option for healing, and one that has relatively few risks [5].

Purpose

To determine if the LifeWave X39 non-transdermal photobiomodulation active patch would show improved production of GHK-Cu over controls in a double blind randomized controlled trial.

Procedure

Once human research studies ethics board approval was received (NFFEH 01-16-20-01) recruitment was begun. Flyers advertising for interested research participants were posted at various local sites. Participants would call into the main study phone number and were assessed for inclusion and exclusion criterion. If appropriate they were scheduled for consenting. At the time of arrival at the study site, each participant was consented and then randomized into group A or B. Individual participants were then taken into the exam room and a blood sample was taken at baseline. Additional samples were taken at 24 hours and 7 days of patch placement.

For convenience, participants were asked to use what is a recognized meridian point, GV14 or CV6 [6], for the patch placement. BD Vacutainer Safety Loc Blood Collection sets were used with Pre-attached holder sized 21GX0.75 or 23GX0.75 and placed in lavender top tubes. Each blood sample was then placed in the Kendro Sorvall Biofuge centrifuge 75005184+ HERAEUS 7591 with a 4000 RPM rotor, spun for 10 minutes at 1300 rcf to separate the plasma, which was then placed in the cryo tubes, and then flash frozen using a medical freezer at -22C. Samples were then placed in 2″ thick polystyrene containers, wrapped in thermal box liners and placed in double walled boxes with dry ice for overnight shipping. Samples were sent to HT-Labs, a division of AxisPharm in San Diego, CA.

Analysis of Blood Samples

The blood samples were processed according to the original thesis of Dr. Pickard. The filtrate was concentrated by speed-vac and reconstituted with de-ionized water to 50 ul and analyzed with AB Sciex API4000 Qtrap. The data was analyzed with Analyst software 1.6.2. Values were placed in a spread sheet and then sent for statistical analysis. Both the blood analysis and statistical analysis was done at groups independent from the principle research laboratory.

Statistical Analysis

Absolute changes in GHK and GHK-Cu levels from baseline to the 24 hours and day 7 assessments were summarized in terms of means, standard deviations, medians and ranges. Changes from baseline to the 24 hours and day 7 assessments were evaluated using a nonparametric Wilcoxon signed rank test. All reported p values are two-sided and p<0.05 was used to define statistical significance. Statistical analyses were conducted using R software (version 3.5.1; http://www.r-project.org/). Once the statistical analysis was complete, the blind was broken.

Results

A sample of convenience of individuals consisted of 60 individuals randomized into two groups (A and B) with an age range of 41-80. Significant results of the LifeWave X39 patch testing are as follows:

Discussion

This was a randomized double blind trial which used a sample of convenience recruited from the general population of the greater Tucson, AZ area. Individuals were age 40-81. It should be noted that this trial was interrupted by the COVID SARS-2 pandemic in March of 2020 and resumed in Aug of 2020. At that time special procedures were put in place to be sure of the safety and health of all participants. This included separation of times for scheduled blood draws and special cleaning procedures between each participant: UV-C wanding of all hard surfaces, Clorox wipe of draw chair, changes in gloves and gowns for all study team members and the wearing of masks for both participants and study team members. Study team members were tested weekly to confirm no contagion. No study participant developed COVID SARS-2 through participation in this study process. This study confirmed that there was a significant change in the levels of GHK-Cu in 7 days in both concentration and total amount. This confirms data from earlier studies [7,8]. The repeated trials data supports promotion of positive benefits to the body through increased production of GHK-Cu by the wearing of the LifeWave X39 non-transdermal photobiomodulation patch.

Conclusion

This study explored the changes in amounts of GHK-Cu present in the blood as a result of wearing the LifeWave X39 patch 8-12 hours per day for 1 week. A significant increase in GHK-Cu concentration in the blood of the active group was seen in the p-value for comparing changes from Day 2 to Day 7 between Group A vs. Group B in GHK-Cu Concentration (ng/ml) at p<0.035 and in Total GHK-Cu (ng) at p<0.03 (Tables 1 and 2).

Table 1: A significant increase in GHK-CU concentration in the blood of the active group was seen comparing changes from Day 2 to Day 7 between Active vs. Control in GHK-Cu Concentration (ng/ml) at p< 0.035 and in Total GHK-Cu (ng) at p<0.03.

Table 2: Study X39 – Evaluation of Changes in Outcomes from Day 1 to Day 2, Day 1 to Day 7, and Day 2 to Day 7 within Groups and Comparisons of Changes between Groups.

¹p-value for evaluating changes from Day 1 to Day 2, and Day 2 to Day 7 within Control Group.

²p-value for evaluating changes from Day 1 to Day 2, and Day 2 to Day 7 within Active Group.

³p-value for comparing changes from Day 1 to Day 2, and Day 2 to Day 7 between Control
Group vs. Active Group.

References

1. DeHaven C (2014) Copper Tripeptide-1. Science of Skincare.
2. Pickart L, Vasquez-Soltero J, Margolina A (2014) GHK and DNA: resetting the human genome to health. BioMed Research International 2014: 151479. [crossref]
3. Pickart L, Vasquez-Soltero J, Margolina A (2015) GHK Peptide as a Natural Modulator of Multiple Cellular Pathways in Skin Regeneration. Hindawi Publishing Corporation BioMed Research International 2015: 648108.
4. Geo Peptides Staff (2015) What are Copper Peptides? https://www.geopeptides.com/copperpep.html
5. Kakimoto C (2017) What is phototherapy, and how does it work? https://www.dermatologistoncall.com/blog/what-is-phototherapy-and-how-does-it-work/
6. Deadman P, Al-Khafaji M, Baker K (2001) A Manual of Acupuncture. Eastland Press.
7. Connor M, Connor C, Gombosuran N, Eickhoff J, et al. (2020) LifeWave X39 Pilot Demonstrates Light Triggered Changes. International Journal of Healing and Careing ijhc.org
8. Connor C, Connor M, Yue D, Chang C, Eickhoff J, et al. (2020) Changes in Tripeptides Produced By the LifeWave X39 Patch. International Journal of Healing and Careing www.ijhc.org