SCIENTISTS DISCOVER THAT HUMANS HAVE A ‘MAGNETIC SENSE’ TO DETECT things WE CAN’T SEE
It’s called magnetoreception, and it refers to the ability to perceive magnetic fields. Several animals use it to find their way over long distances by aligning themselves with the Earth’s magnetic field. Sea turtles. honeybees, spiny lobsters, dolphins, migratory birds, and more all have a magnetic compass which allows them to use the information that’s coded into magnetic fields. We know little beyond that, however. How they use them, how they sense them, and what information they are getting from them remains up for speculation. For all we know, these magnetic fields could be used for much more than navigation for certain species.
According to Joe Kirschvink, the geophysicist at the California Institute of Technology who is currently testing humans for a magnetic sense, “it’s part of our evolutionary history. Magnetoreception may be the primal sense.” (source)
A recent study published by Kirschvink in the journal Nature Communications suggests that a protein in the human retina, when placed into fruit flies, has the ability to detect magnetic fields. The research claims that it can serve as a magneto sensor, but whether or not humans actually use it in this way is unknown.
“It poses the question, ‘maybe we should rethink about this sixth sense,'” University of Massachusetts Medical School researcher Steven Reppert told LiveScience. “It is thought to be very important for how animals migrate. Perhaps this protein is also fulfilling an important function for sensing magnetic fields in humans.”
In one of Kirschvink’s recent experiment, a rotating magnetic field was passed through study participants while their brainwaves were measured. He discovered that when the magnetic field was rotated counterclockwise, certain neutrons responded to this change which, in turn, generated a spike in electrical activity. This suggests a possible magnetic sense in humans.
Yet multiple questions still remain. For example, was this neural activity evidence of a magnetic sense or something else? Even if the human brain responds to these fields in some way, that doesn’t mean that information is being processed by the brain. There is still the question of what mechanisms are in place within the brain or body that receive these signals. If the body does indeed have magneto receptors, where are they? The next step for researchers is to identify them.
Kirschvink’s study is one of many publications delving into the mysteries of magnetic fields and what impact they have on human beings. The leaders in this area of research will most likely be found at the HeartMath institute. An internationally recognized nonprofit research and education organization dedicated to helping people reduce stress, self-regulate emotions, and build energy and resilience for healthy, happy lives, HeartMath tools, technology, and training teach people to rely on the intelligence of their hearts in concert with that of their minds at home, school, work, and play.
Researchers at HeartMath have begun what’s called the The Global Coherence Initiative (GCI), an international cooperative effort to help activate the heart of humanity and facilitate a shift in global consciousness. It aims primarily to invite people to participate by actively adding more heart-coherent love, care, and compassion into the planetary field. The second focus is scientific research into how we are all energetically connected with each other and the planet, and how we can utilize this interconnectivity to raise our personal vibration and thereby help create a better world.
The hypotheses of the researchers and scientists behind this project are as follows:
Scientific literature is also clear on the fact that several physiological rhythms and global collective behaviours are not only synchronized with solar and geomagnetic activity, but also that disruptions in these fields can create adverse effects on human health and behaviour
When the Earth’s magnetic field environment is distributed it can cause sleep problems, mental confusion, usual lack of energy or a feeling of being on edge or overwhelmed for no apparent reason. At other times, when the Earth’s fields are stable and certain measures of solar activity are increased, people report increased positive feelings and more creativity and inspiration. This is likely due to a coupling between the human brain, cardiovascular and nervous system with resonating geomagnetic frequencies. (source)(source)(source)
The Earth and ionosphere generate frequencies that range from 0.01 hertz to 300 hertz, some of which are in the exact same frequency range as the one happening in our brain, cardiovascular system, and autonomic nervous system. This offers one way to explain how fluctuations in the Earth’s and Sun’s magnetic fields can influence us. Changes in these fields have also been shown to affect our brainwaves, heart rhythms, memory, athletics performance, and overall health.
Changes in the Earth’s fields from extreme solar activity have been linked to some of humanity’s greatest creations of art, as well as some of its most tragic events. (source)
We know how these fields affect us, but what about how we affect these fields? That’s the real question here. GCI scientists believe that because brain wave and heart rhythm frequencies overlap the Earth’s field resonance, we are not just receivers of biologically relevant information, but also senders of it. We feed information into the global field, thus creating a feedback loop with the Earth’s magnetic fields.
Human emotions and consciousness interact with and encode information into the geomagnetic field and this information is distributed globally. . . . We are suggesting in essence that this encoded information is communicated nonlocally between people at a subconscious level, in effect linking all living systems. Magnetic fields act as carrier waves for this information, which can influence all living systems – positively or negatively – within the field environment as well as our collective consciousness.
If we look at the heart, for example, it emits electromagnetic fields which change according to our emotions, and these can actually be measured up to several feet away from the human body.
These fields have been shown to affect not only ourselves, but those around us. You can read more about that here.
This research on this topic, which is still in its infancy, has immense ramifications for our world. It will further prove and highlight the great extent to which our attitudes, emotions, and intentions matter, and that these factors within the realm of non-material science can affect all life on Earth. Coherent, cooperative intention could impact global events and improve the quality of life on Earth. Practicing love, gratitude, and appreciation, as well as bettering ourselves as individuals, are some of the many crucial action steps towards changing our planet for the better.
Sources used not highlighted in the article:
(1) Heart Intelligence: Connecting With The Intuitive Guidance Of The Heart
NY PTA Calls for Wi-Fi To Be Turned Off:
MAJOR WIFI CONTROVERY AS SCHOOL DISTRICT IS SERVED WITH LEGAL LIABILITY NOTICE
In this video an Onteora School District Trustee reads the Phoenicia PTA's Letter
where the PTA calls for the Wi-Fi to be turned OFF.
Facts About the New York State Onteora School District Wi-Fi Debate
1. Parents initiated a dialogue on the health risks of Wireless radiation. The Board grappled with the decision to turn the Wi-Fi off while gathering data. The local Woodstock Times paper covered some of the debate.
2. The District created a Wi-Fi Task Force
According to the Board minutes, the Task force was charged to "Review studies and determine if precautionary principle should be done for our youngest students, which is to remove Wi-Fi from the elementary building until the industry proves it is safe. Look at both sides of the research and most up-to-date research. They were to consider the health implications and pros and cons in terms of technology in 21stcentury learning and get Input from administrators and school personnel." They were to make a recommendation to the Board. Read about it in the Onteroa meeting minutes of August 11, 2015 (page 27, point 11) here.
Watch a video where the Board discusses the WiFi Task Force here.
3. In March a Notice of Wi-Fi Radiation Legal Liability Was Served to New York School District Board of Education Members
The Wi-Fi Task Force was formed to gather information. Then it was disbanded by the Board of Education without any transparency in the process.
Parents served the New York Onteroa School District Superintendent Victoria Mclaren and Board of Education members a notice of their liability concerning the health risks from the wi-fi installation in this New York school district on the March 15, 2016 Board of Education Meeting.
"You have been urged to hardwire all your computers in the schools."
" Ignoring the above documented information and documented statements and proceeding with the use of WiFi routers in the schools, you are committing willful blindness and can be liable for child negligence, or possibly even "negligent endangerment of child" which carries with it criminal law repercussions if any student gets injured from the schools WiFi system and a parent decides to press charges."
Watch the Board Meeting in this video.
Please watch below the video where parents served the Board members with this notice If you are wondering why we cannot see the parents, we were wondering the same thing. You can watch the full Board meeting at this link to the School District website.
THIS NOTICE STATES:
Concerned parents in the United States can also send notarized " No-Consent to WiFi " letters to their BOE. This an important step towards accountability.
Examples of A Parents Do NOT CONSENT Letter For Schools:
Is EMF or RF Safe? Most people assume that all this electromagnetic radiation is safe for us. Younger generations have literally grown up in it and can’t remember a time when they weren’t surrounded by it. It “feels” safe. We can’t see the radiation and it’s everywhere—so the assumption is that it must be okay for us.
And besides, aren’t there research studies that have concluded that cell phones and WiFi are safe—or at least claim there is no evidence that they are unsafe?
The answer is yes, there are those studies. However, if you locate the funding source for most of them, you’ll find that the corporations that funded them were the very companies who produce cell phones and wireless devices.
And, very importantly, there now exists an overwhelming abundance of independent, peer-reviewed studies from all over the world that have conclusively linked electromagnetic radiation with a wide variety of adverse health effects and serious diseases.
Here is a great video on 70% independent studies show danger:
Video: Erica Mallery-Blythe, MD at the Commonwealth Club of CA, June 22, 2015 (hd)
Erica Mallery-Blythe, MD is Founder of Physicians’ Health Initiative for Radiation and Environment (PHIRE) in the U.K. where she is also Trustee of the Radiation Research Trust, Medical Advisor to Electrosensitivity UK and Member of the British Society of Ecological Medicine. She has worked in emergency medicine, led trauma teams and taught trauma medicine throughout the UK and abroad. She has a broad base of medical experience, including surgery, anesthesiology and intensive care (both neonatal and adult). For many years, Dr. Mallery-Blythe has worked intensively with adults and children suffering from the little-understood biological and health effects of wireless technologies, working closely with their doctors. She is author of a resource for parents, teachers, schools and policymakers called “Electromagnetic Health for Children”. Here, Dr. Mallery-Blythe presents on risks to children at a program organized by ElectromagneticHealth.org and Environmental Health Trust held at the Commonwealth Club on June 22, 2015. To see all videos, please go to: tinyurl.com/pazyhl5
Experiments with Cress [and Wi-Fi]
Original post from www.MastSanity.org
http://www.dr.dk/Nyheder/Indland/2013/05/16/131324.htm (in Danish)
Foreign researchers are extremely excited for a biology project from five 9th grade girls.
Researchers from England, Holland and Sweden have shown great interest in the five girls' biology experiments.
Take 400 Cress seeds and place them into 12 trays. Then place six trays in two rooms at the same temperature. Give them the same amount of water and sun over 12 days, and remember to expose half of them to mobile [Wi-Fi] radiation.
It is a recipe for a biology test so brilliant that it has attracted international attention among acknowledged biologists and radiation experts. Behind the experiment are five girls from 9b in Hjallerup School in North Jutland, and it all started because they found it difficult to concentrate during the school day:
- We all think we have experienced difficulty concentrating in school, if we had slept with the phone next to our head, and sometimes also experienced having difficulty sleeping, explains Lea Nielsen, who is one of the five aspiring researchers.
The school was not equipped to test the effect of mobile phone radiation on them. Therefore, the girls had to find an alternative. And the answer was Cress.
Six trays of seeds were put into a room without radiation, and six trays were put into another room next to two [Wi-Fi] routers. Such routers broadcast the same type of radiation as an ordinary mobile.
The "healthy" cress without the influence of the router. Photo: The girls from 9b
Then it was just necessary to wait 12 days, observe, measure, weigh and take pictures along the way. And the result spoke was clear: cress seeds next to the router did not grow, and some of them were even mutated or dead.
- It is truly frightening that there is so much affect, so we were very shocked by the result, says Lea Nielsen.
The "sick" cress exposed to the [Wi-Fi] router.
The experiment secured the girls the finals in the competition "Young Scientists", but it was only the beginning. Renowned scientists from England, Holland and Sweden have since shown great interest in the girls' project so far.
From left: Lea Nielsen, Mathilde Nielsen, Signe Nielsen, Sisse Coltau and Rikke Holm. Photo: Kim Horsevad
The renowned professor at the Karolinska Institute in Stockholm, Olle Johansson, is one of the impressed researchers. He will now repeat the experiment with a Belgian research colleague, Professor Marie-Claire Cammaert at the Université libre de Bruxelles, for the trial, according to him, is absolutely brilliant:
- The girls stayed within the scope of their knowledge, skilfully implemented and developed a very elegant experiment. The wealth of detail and accuracy is exemplary, choosing cress was very intelligent, and I could go on, he says.
He is not slow to send them an invitation to go on the road:
- I sincerely hope that they spend their future professional life in researching, because I definitely think they have a natural aptitude for it. Personally, I would love to see these people in my team!
No mobile by the bed
The five girls from northern Jutland have not yet decided their future careers. They are still very surprised by all the sudden attention.
- It has been such a rollercoaster ride. I still cannot believe it, says Lea Nielsen.
And Mathilde Nielsen added:
- It's totally overwhelming and exciting. It's just not something you experience every day.
But there have also been other consequences of the cress trial, which is quite low-tech in nature.
- None of us sleep with the mobile next to the bed anymore. Either the phone is put far away, or it is put in another room. And the computer is always off, says Lea Nielsen.
See Live wi-fi on this new App
Architecture of Radio is a data visualization, based on global open datasets of cell tower, Wi-Fi and satellite locations. Based on your GPS location the app shows a 360 degree visualization of signals around you. The dataset includes almost 7 million cell towers, 19 million Wi-Fi routers and hundreds of satellites. A site specific version of the app includes wired communication infrastructure embedded in the exhibition space. It's aim is to provide a comprehensive window into the infosphere.
*The infosphere refers to an interdependent environment, like a biosphere, that is populated by informational entities. While an example of the sphere of information is cyberspace, infospheres are not limited to purely online environments.
Wi-Fi is convenient but many have raised doubts concerning the safety of unseen forces that permeate everything around us. Since the introduction of Wi-Fi in 1997, researchers have performed dozens of studies to explore the subject. The results are clear and shocking — Wifi can negatively affect overall health and brain health, especially in children.
Perhaps most shocking is that this information is not new or even that controversial. In fact, in 2008 the well-renowned publication Scientific American ran a piece called “Mind Control by Cell Phone” which explained the danger Wi-Fi has on the human brain.  Let’s further explore the potential dangers of Wi-Fi with these 10 facts.
1. Contributes to the Development of Insomnia
Have you ever felt more awake after using Wi-Fi or even struggled to sleep through the night? Reports of these phenomena have been frequent and even prompted a study in 2007 that evaluated low-frequency modulation from cell phones and its impact on sleep. Participants were exposed to the electromagnetic signals from real phones, or no signal from fake phones. Those exposed to the electromagnetic radiation had a significantly more difficult time falling asleep and changes in brainwave patterns were observed. 
It’s been suggested that sleeping near a phone, in a home with Wi-Fi, or in an apartment building with many Wi-Fi signals can create chronic sleep problems as the constant bombardment of Wi-Fi pollution interferes with falling asleep and sleep patterns. For many, sleep deprivation is just the start for larger problems. The development of depression and hypertension have also been linked to inadequate sleep. 
2. Damaging to Childhood Development
Exposure to non-thermal radio frequency radiation from Wi-Fi and cellular phones can disrupt normal cellular development, especially fetal development. A 2004 animal study linked exposure to delayed kidney development.  These findings were supported by a 2009 Austrian study. In fact, the disruption of protein synthesis is so severe that authors specifically noted, “this cell property is especially pronounced in growing tissues, that is, in children and youth. Consequently, these population groups would be more susceptible than average to the described effects.”  In short, bathing the developmentally young in Wi-Fi increases their risk of developmental issues.
3. Affects Cell Growth
When a group of Danish ninth graders experienced difficulty concentrating after sleeping with their cell phones by their head, they performed an experiment to test the effect of wireless Wi-Fi routers on garden cress. One set of plants was grown in a room free of wireless radiation; the other group grown next to two routers that released the same amount of radiation as a cell phone. The results? The plants nearest the radiation didn’t grow. 
4. Derails Brain Function
Just as the Danish high schoolers noticed problems with concentration, scientists have begun to look at the impact of 4G radiation on brain function. Using MRI technology, research performed just last year found that persons exposed to 4G radiation had several areas of reduced brain activity. 
5. Reduces Brain Activity
in FemalesA group of 30 healthy volunteers, 15 men and 15 women, were given a simple memory test. First, the entire group was tested without any exposure to Wi-Fi radiation — no problem. Then, they were exposed to 2.4 GHz Wi-Fi from a wireless access point for about 45 minutes. During that portion of the testing, brain activity was measured and the women had a noticeable change in brain activity and energy levels.  Sorry ladies! But guys, don’t get too comfortable…
6. Neutralizes Sperm…
Because we’ve known for a long time that the heat generated by laptops kills sperm. Well, now it turns out that heat isn’t the only threat to a man’s virility. Research has found exposure to Wi-Fi frequencies reduce sperm movement and cause DNA fragmentation. Both human and animal testing has confirmed that exposure negatively affects sperm. 
7. May Impact Fertility
And, it’s not just sperm. The results of an animal study suggest that some wireless frequencies may prevent egg implantation. During the study, mice exposed 2 hours a day for 45 days had significantly increased oxidative stress levels. The cellular damage and impact on DNA structure from exposure suggests a strong possibility of abnormal pregnancy or failure of the egg to implant. 
The Karolinska Institute in Sweden released a warning in 2011, stating:
8. Provokes Cardiac Stress
If you think your heart races when surrounded by wireless networks or 3G or LTE cell phones, it may not be in your head. A study involving 69 subjects reported that many of them experienced a real physical response to electromagnetic frequencies. Exactly what was the physical response? Increased heart rate — similar to the heart rate of an individual under stress. 
9. Linked to Cancer?
This is extremely controversial but we can’t ignore that plenty of animal models indicate that exposure to electromagnetic radiation increases the risk of tumor development. While human studies are rare, reports and case studies abound. One such case involves a young 21-year-old woman who developed breast cancer. What makes this case unique was that her family did not have a predisposition to breast cancer… and she developed the tumor right on the spot she carried her cell phone in her bra. 
10. You Can Protect Yourself
Although mainstream outlets may ignore the proven dangers, especially in the US and Canada, researchers have identified several methods that can offer a level of defense. First off, reduced melatonin seems to correspond with exposure. Thus, increasing melatonin through supplementation may help offset some of the effects.    In animal tests, L-Carnitine provides antioxidant support for nutrients negatively affected by 2.4 GHz radiation.  
Limiting Exposure and Staying Healthy
Although melatonin and L-Carnitine offer nutritional defense, they don’t block exposure. And that’s very hard to accomplish anyway. Look at coverage maps from cell phone companies, or notice how many Wi-Fi networks your smart phone prompts for you to join. We’re surrounded and bombarded by electromagnetic radiation. Blocking exposure is difficult but there are a few small steps you can take. For one, do not keep cell phones, laptops, and tablets close to your body. And if it’s not being used, shut them off (your wireless router too). There are also a number of devices available to counteract electromagnetic frequencies. Check out these ways to protect yourself from laptop radiation and cell phone radiation, too.
Have you experienced negative side effects of Wi-Fi? What measures have you taken to protect yourself and your family? Please leave a comment below and share your experience with us!
†Results may vary. Information and statements made are for education purposes and are not intended to replace the advice of your doctor. Global Healing Center does not dispense medical advice, prescribe, or diagnose illness. The views and nutritional advice expressed by Global Healing Center are not intended to be a substitute for conventional medical service. If you have a severe medical condition or health concern, see your physician.
Earthing: Health Implications
OF RECONNECTING THE HUMAN BODY TO THE EARTH'S SURFACE ELECTRONS
AbstractEnvironmental medicine generally addresses environmental factors with a negative impact on human health. However, emerging scientific research has revealed a surprisingly positive and overlooked environmental factor on health: direct physical contact with the vast supply of electrons on the surface of the Earth. Modern lifestyle separates humans from such contact. The research suggests that this disconnect may be a major contributor to physiological dysfunction and unwellness. Reconnection with the Earth's electrons has been found to promote intriguing physiological changes and subjective reports of well-being. Earthing (or grounding) refers to the discovery of benefits—including better sleep and reduced pain—from walking barefoot outside or sitting, working, or sleeping indoors connected to conductive systems that transfer the Earth's electrons from the ground into the body. This paper reviews the earthing research and the potential of earthing as a simple and easily accessed global modality of significant clinical importance.
Go to:1. IntroductionEnvironmental medicine focuses on interactions between human health and the environment, including factors such as compromised air and water and toxic chemicals, and how they cause or mediate disease. Omnipresent throughout the environment is a surprisingly beneficial, yet overlooked global resource for health maintenance, disease prevention, and clinical therapy: the surface of the Earth itself. It is an established, though not widely appreciated fact, that the Earth's surface possesses a limitless and continuously renewed supply of free or mobile electrons. The surface of the planet is electrically conductive (except in limited ultradry areas such as deserts), and its negative potential is maintained (i.e., its electron supply replenished) by the global atmospheric electrical circuit [1, 2].
Mounting evidence suggests that the Earth's negative potential can create a stable internal bioelectrical environment for the normal functioning of all body systems. Moreover, oscillations of the intensity of the Earth's potential may be important for setting the biological clocks regulating diurnal body rhythms, such as cortisol secretion .
It is also well established that electrons from antioxidant molecules neutralize reactive oxygen species (ROS, or in popular terms, free radicals) involved in the body's immune and inflammatory responses. The National Library of Medicine's online resource PubMed lists 7021 studies and 522 review articles from a search of “antioxidant + electron + free radical” . It is assumed that the influx of free electrons absorbed into the body through direct contact with the Earth likely neutralize ROS and thereby reduce acute and chronic inflammation . Throughout history, humans mostly walked barefoot or with footwear made of animal skins. They slept on the ground or on skins. Through direct contact or through perspiration-moistened animal skins used as footwear or sleeping mats, the ground's abundant free electrons were able to enter the body, which is electrically conductive . Through this mechanism, every part of the body could equilibrate with the electrical potential of the Earth, thereby stabilizing the electrical environment of all organs, tissues, and cells.
Modern lifestyle has increasingly separated humans from the primordial flow of Earth's electrons. For example, since the 1960s, we have increasingly worn insulating rubber or plastic soled shoes, instead of the traditional leather fashioned from hides. Rossi has lamented that the use of insulating materials in post-World War II shoes has separated us from the Earth's energy field . Obviously, we no longer sleep on the ground as we did in times past.
During recent decades, chronic illness, immune disorders, and inflammatory diseases have increased dramatically, and some researchers have cited environmental factors as the cause . However, the possibility of modern disconnection with the Earth's surface as a cause has not been considered. Much of the research reviewed in this paper points in that direction.
In the late 19th century, a back-to-nature movement in Germany claimed many health benefits from being barefoot outdoors, even in cold weather . In the 1920s, White, a medical doctor, investigated the practice of sleeping grounded after being informed by some individuals that they could not sleep properly “unless they were on the ground or connected to the ground in some way,” such as with copper wires attached to grounded-to-Earth water, gas, or radiator pipes. He reported improved sleeping using these techniques . However, these ideas never caught on in mainstream society.
At the end of the last century, experiments initiated independently by Ober in the USA  and K. Sokal and P. Sokal  in Poland revealed distinct physiological and health benefits with the use of conductive bed pads, mats, EKG- and TENS-type electrode patches, and plates connected indoors to the Earth outside. Ober, a retired cable television executive, found a similarity between the human body (a bioelectrical, signal-transmitting organism) and the cable used to transmit cable television signals. When cables are “grounded” to the Earth, interference is virtually eliminated from the signal. Furthermore, all electrical systems are stabilized by grounding them to the Earth. K. Sokal and P. Sokal, meanwhile, discovered that grounding the human body represents a “universal regulating factor in Nature” that strongly influences bioelectrical, bioenergetic, and biochemical processes and appears to offer a significant modulating effect on chronic illnesses encountered daily in their clinical practices.
Earthing (also known as grounding) refers to contact with the Earth's surface electrons by walking barefoot outside or sitting, working, or sleeping indoors connected to conductive systems, some of them patented, that transfer the energy from the ground into the body. Emerging scientific research supports the concept that the Earth's electrons induce multiple physiological changes of clinical significance, including reduced pain, better sleep, a shift from sympathetic to parasympathetic tone in the autonomic nervous system (ANS), and a blood-thinning effect. The research, along with many anecdotal reports, is presented in a new book entitled Earthing .
Go to:2. Review of Earthing PapersThe studies summarized below involve indoor-testing methods under controlled conditions that simulate being barefoot outdoors.
2.1. Sleep and Chronic PainIn a blinded pilot study, Ober recruited 60 subjects (22 males and 28 females) who suffered from self-described sleep disturbances and chronic muscle and joint pain for at least six months . Subjects were randomly divided for the month-long study in which both groups slept on conductive carbon fiber mattress pads provided by Ober. Half the pads were connected to a dedicated Earth ground outside each subject's bedroom window, while the other half were “sham” grounded—not connected to the Earth. Results are presented in Table 1.
Subjective sleep, pain, and well-being feedback.Most grounded subjects described symptomatic improvement while most in the control group did not. Some subjects reported significant relief from asthmatic and respiratory conditions, rheumatoid arthritis, PMS, sleep apnea, and hypertension while sleeping grounded. These results indicated that the effects of earthing go beyond reduction of pain and improvements in sleep.
2.2. Sleep, Stress, Pain, and CortisolA pilot study evaluated diurnal rhythms in cortisol correlated with changes in sleep, pain, and stress (anxiety, depression, and irritability), as monitored by subjective reporting . Twelve subjects with complaints of sleep dysfunction, pain, and stress were grounded to Earth during sleep in their own beds using a conductive mattress pad for 8 weeks.
In order to obtain a baseline measurement of cortisol, subjects chewed Dacron salvettes for 2 minutes and then placed them in time-labeled sampling tubes that were stored in a refrigerator. Self-administered sample collections began at 8 AM and were repeated every 4 hours. After 6 weeks of being grounded, subjects repeated this 24-hour saliva test. The samples were processed using a standard radioimmunoassay. A composite of the results is shown in Figure 1.
Cortisol levels before and after grounding. In unstressed individuals, the normal 24-hour cortisol secretion profile follows a predictable pattern: lowest around midnight and highest around 8 a.m. Graph (a) illustrates the wide variation of patterns among ...Subjective symptoms of sleep dysfunction, pain, and stress were reported daily throughout the 8-week test period. The majority of subjects with high- to out-of-range nighttime secretion levels experienced improvements by sleeping grounded. This is demonstrated by the restoration of normal day-night cortisol secretion profiles.
Eleven of 12 participants reported falling asleep more quickly, and all 12 reported waking up fewer times at night. Grounding the body at night during sleep also appears to positively affect morning fatigue levels, daytime energy, and nighttime pain levels.
About 30 percent of the general American adult population complain of sleep disruption, while approximately 10 percent have associated symptoms of daytime functional impairment consistent with the diagnosis of insomnia. Insomnia often correlates with major depression, generalized anxiety, substance abuse, dementia, and a variety of pain and physical problems. The direct and indirect costs of chronic insomnia have been estimated at tens of billions of dollars annually in the USA alone . In view of the burdens of personal discomfort and health care costs, grounding the body during sleep seems to have much to offer.
2.3. Earthing Reduces Electric Fields Induced on the BodyVoltage induced on a human body from the electrical environment was measured using a high-impedance measurement head. Applewhite, an electrical engineer and expert in the design of electrostatic discharge systems in the electronic industry, was both subject and author of the study . Measurements were taken while ungrounded and then grounded using a conductive patch and conductive bed pad. The author measured the induced fields at three positions: left breast, abdomen, and left thigh.
Each method (patch and sheet) immediately reduced the common alternating current (AC) 60 Hz ambient voltage induced on the body by a highly significant factor of about 70 on average. Figure 2 shows this effect.
Effect of bed pad grounding on 60 Hz mode.The study showed that when the body is grounded, its electrical potential becomes equalized with the Earth's electrical potential through a transfer of electrons from the Earth to the body. This, in turn, prevents the 60 Hz mode from producing an AC electric potential at the surface of the body and from producing perturbations of the electric charges of the molecules inside the body. The study confirms the “umbrella” effect of earthing the body explained by Nobel Prize winner Richard Feynman in his lectures on electromagnetism . Feynman said that when the body potential is the same as the Earth's electric potential (and thus grounded), it becomes an extension of the Earth's gigantic electric system. The Earth's potential thus becomes the “working agent that cancels, reduces, or pushes away electric fields from the body.”
Applewhite was able to document changes in the ambient voltage induced on the body by monitoring the voltage drop across a resistor. This effect clearly showed the “umbrella effect” described above. The body of the grounded person is not subject to the perturbation of electrons and electrical systems.
Jamieson asks whether the failure to appropriately ground humans is a factor contributing to the potential consequences of electropollution in office settings . Considerable debate exists on whether electromagnetic fields in our environment cause a risk to health , but there is no question that the body reacts to the presence of environmental electric fields. This study demonstrates that grounding essentially eliminates the ambient voltage induced on the body from common electricity power sources.
2.4. Physiological and Electrophysiological Effects
2.4.1. Reductions in Overall Stress Levels and Tension and Shift in ANS Balance Fifty-eight healthy adult subjects (including 30 controls) participated in a randomized double-blind pilot study investigating earthing effects on human physiology . Earthing was accomplished with a conductive adhesive patch placed on the sole of each foot. A biofeedback system recorded electrophysiological and physiological parameters. Experimental subjects were exposed to 28 minutes in the unearthed condition followed by 28 minutes with the earthing wire connected. Controls were unearthed for 56 minutes.
Upon earthing, about half the subjects showed an abrupt, almost instantaneous change in root mean square (rms) values of electroencephalograms (EEGs) from the left hemisphere (but not the right hemisphere) at all frequencies analyzed by the biofeedback system (beta, alpha, theta, and delta).
All grounded subjects presented an abrupt change in rms values of surface electromyograms (SEMGs) from right and left upper trapezius muscles. Earthing decreased blood volume pulse (BVP) in 19 of 22 experimental subjects (statistically significant) and in 8 of 30 controls (not significant). Earthing the human body showed significant effects on electrophysiological properties of the brain and musculature, on the BVP, and on the noise and stability of electrophysiological recordings. Taken together, the changes in EEG, EMG, and BVP suggest reductions in overall stress levels and tensions and a shift in ANS balance upon earthing. The results extend the conclusions of previous studies.
2.4.2. Confirming Shift from Sympathetic to Parasympathetic Activation A multiparameter double-blind study was designed to reproduce and expand on previous electrophysiological and physiological parameters measured immediately after grounding with an improved methodology and state-of-the-art equipment . Fourteen men and 14 women, in good health, ages 18–80, were tested while seated in a comfortable recliner during 2-hour grounding sessions, leaving time for signals to stabilize before, during, and after grounding (40 minutes for each period). Sham 2-hour grounding sessions were also recorded with the same subjects as controls. For each session, statistical analyses were performed on four 10-minute segments: before and after grounding (sham grounding for control sessions) and before and after ungrounding (sham ungrounding for control sessions). The following results were documented:
2.4.3. Immune Cell and Pain Responses with Delayed-Onset Muscle Soreness Induction Pain reduction from sleeping grounded has been documented in previous studies [10, 13]. This pilot study looked for blood markers that might differentiate between grounded and ungrounded subjects who completed a single session of intense, eccentric exercise resulting in delayed-onset muscle soreness (DOMS) of the gastrocnemius . If markers were able to differentiate these groups, future studies could be done in greater detail with a larger subject base. DOMS is a common complaint in the fitness and athletic world following excessive physical activity and involves acute inflammation in overtaxed muscles. It develops in 14 to 48 hours and persists for more than 96 hours . No known treatment reduces the recovery period, but apparently massage and hydrotherapy [23–25] and acupuncture  can reduce pain.
Eight healthy men ages 20–23 were put through a similar routine of toe raises while carrying on their shoulders a barbell equal to one-third of their body weight. Each participant was exercised individually on a Monday morning and then monitored for the rest of the week while following a similar eating, sleeping, and living schedule in a hotel. The group was randomly divided in half and either grounded or sham grounded with the use of a conductive patch placed at the sole of each foot during active hours and a conductive sheet at night. Complete blood counts, blood chemistry, enzyme chemistry, serum and saliva cortisol, magnetic resonance imaging and spectroscopy, and pain levels (a total of 48 parameters) were taken at the same time of day before the eccentric exercise and at 24, 48, and 72 hours afterwards. Parameters consistently differing by 10 percent or more, normalized to baseline, were considered worthy of further study.
Parameters that differed by these criteria included white blood cell counts, bilirubin, creatine kinase, phosphocreatine/inorganic phosphate ratios, glycerolphosphorylcholine, phosphorylcholine, the visual analogue pain scale, and pressure measurements on the right gastrocnemius.
The results showed that grounding the body to the Earth alters measures of immune system activity and pain. Among the ungrounded men, for instance, there was an expected, sharp increase in white blood cells at the stage when DOMS is known to reach its peak and greater perception of pain (see Figure 3). This effect demonstrates a typical inflammatory response. In comparison, the grounded men had only a slight decrease in white blood cells, indicating scant inflammation, and, for the first time ever observed, a shorter recovery time. Brown later commented that there were “significant differences” in the pain these men reported .
Delayed onset muscle soreness and grounding. Consistent with all measurements, ungrounded subjects expressed the perception of greater pain. Related to the pain finding was evidence of a muted white blood cell response indicating that a grounded body ...
2.4.4. Heart Rate Variability The rapid change in skin conductance reported in an earlier study led to the hypothesis that grounding may also improve heart rate variability (HRV), a measurement of the heart's response to ANS regulation. A double-blind study was designed with 27 participants . Subjects sat in a comfortable reclining chair. Four transcutaneous electrical nerve stimulation (TENS) type adhesive electrode patches were placed on the sole of each foot and on each palm.
Participants served as their own controls. Each participant's data from a 2-hour session (40 minutes of which was grounded) were compared with another 2-hour sham-grounded session. The sequence of grounding versus sham-grounding sessions was assigned randomly.
During the grounded sessions, participants had statistically significant improvements in HRV that went way beyond basic relaxation results (which were shown by the nongrounded sessions). Since improved HRV is a significant positive indicator on cardiovascular status, it is suggested that simple grounding techniques be utilized as a basic integrative strategy in supporting the cardiovascular system, especially under situations of heightened autonomic tone when the sympathetic nervous system is more activated than the parasympathetic nervous system.
2.4.5. Reduction of Primary Indicators of Osteoporosis, Improvement of Glucose Regulation, and Immune Response K. Sokal and P. Sokal, cardiologist and neurosurgeon father and son on the medical staff of a military clinic in Poland, conducted a series of experiments to determine whether contact with the Earth via a copper conductor can affect physiological processes . Their investigations were prompted by the question as to whether the natural electric charge on the surface of the Earth influences the regulation of human physiological processes.
Double-blind experiments were conducted on groups ranging from 12 to 84 subjects who followed similar physical activity, diet, and fluid intake during the trial periods. Grounding was achieved with a copper plate (30 mm × 80 mm) placed on the lower part of the leg, attached with a strip so that it would not come off during the night. The plate was connected by a conductive wire to a larger plate (60 mm × 250 mm) placed in contact with the Earth outside.
In one experiment with nonmedicated subjects, grounding during a single night of sleep resulted in statistically significant changes in concentrations of minerals and electrolytes in the blood serum: iron, ionized calcium, inorganic phosphorus, sodium, potassium, and magnesium. Renal excretion of both calcium and phosphorus was reduced significantly. The observed reductions in blood and urinary calcium and phosphorus directly relate to osteoporosis. The results suggest that Earthing for a single night reduces primary indicators of osteoporosis.
Earthing continually during rest and physical activity over a 72-hour period decreased fasting glucose among patients with non-insulin-dependent diabetes mellitus. Patients had been well controlled with glibenclamide, an antidiabetic drug, for about 6 months, but at the time of study had unsatisfactory glycemic control despite dietary and exercise advice and glibenclamide doses of 10 mg/day.
K. Sokal and P. Sokal drew blood samples from 6 male and 6 female adults with no history of thyroid disease. A single night of grounding produced a significant decrease of free tri-iodothyronine and an increase of free thyroxin and thyroid-stimulating hormone. The meaning of these results is unclear but suggests an earthing influence on hepatic, hypothalamus, and pituitary relationships with thyroid function. Ober et al.  have observed that many individuals on thyroid medication reported symptoms of hyperthyroid, such as heart palpitations, after starting grounding. Such symptoms typically vanish after medication is adjusted downward under medical supervision. Through a series of feedback regulations, thyroid hormones affect almost every physiological process in the body, including growth and development, metabolism, body temperature, and heart rate. Clearly, further study of earthing effects on thyroid function is needed.
In another experiment, the effect of grounding on the classic immune response following vaccination was examined. Earthing accelerated the immune response, as demonstrated by increases in gamma globulin concentration. This result confirms an association between earthing and the immune response, as was suggested in the DOMS study .
K. Sokal and P. Sokal conclude that earthing the human body influences human physiological processes, including increasing the activity of catabolic processes and may be “the primary factor regulating endocrine and nervous systems.”
2.4.6. Altered Blood Electrodynamics Since grounding produces changes in many electrical properties of the body [1, 15, 19, 28], a next logical step was to evaluate the electrical property of the blood. A suitable measure is the zeta potential of red blood cells (RBCs) and RBC aggregation. Zeta potential is a parameter closely related to the number of negative charges on the surface of an RBC. The higher the number, the greater the ability of the RBC to repel other RBCs. Thus, the greater the zeta potential the less coagulable is the blood.
Ten relatively healthy subjects participated in the study . They were seated comfortably in a reclining chair and were grounded for two hours with electrode patches placed on their feet and hands, as in previous studies. Blood samples were taken before and after.
Grounding the body to the earth substantially increases the zeta potential and decreases RBC aggregation, thereby reducing blood viscosity. Subjects in pain reported reduction to the point that it was almost unnoticeable. The results strongly suggest that earthing is a natural solution for patients with excessive blood viscosity, an option of great interest not just for cardiologists, but also for any physician concerned about the relationship of blood viscosity, clotting, and inflammation. In 2008, Adak and colleagues reported the presence of both hypercoagulable blood and poor RBC zeta potential among diabetics. Zeta potential was particularly poor among diabetics with cardiovascular disease .
Go to:3. DiscussionUntil now, the physiological significance and possible health effects of stabilizing the internal bioelectrical environment of an organism have not been a significant topic of research. Some aspects of this, however, are relatively obvious. In the absence of Earth contact, internal charge distribution will not be uniform, but instead will be subject to a variety of electrical perturbations in the environment. It is well known that many important regulations and physiological processes involve events taking place on cell and tissue surfaces. In the absence of a common reference point, or “ground,” electrical gradients, due to uneven charge distribution, can build up along tissue surfaces and cell membranes.
We can predict that such charge differentials will influence biochemical and physiological processes. First, the structure and functioning of many enzymes are sensitive to local environmental conditions. Each enzyme has an optimal pH that favors maximal activity. A change in the electrical environment can alter the pH of biological fluids and the charge distribution on molecules and thereby affect reaction rates. The pH effect results because of critical charged amino acids at the active site of the enzyme that participate in substrate binding and catalysis. In addition, the ability of a substrate or enzyme to donate or accept hydrogen ions is influenced by pH.
Another example is provided by voltage-gated ion channels, which play critical biophysical roles in excitable cells such as neurons. Local alterations in the charge profiles around these channels can lead to electrical instability of the cell membrane and to the inappropriate spontaneous activity observed during certain pathological states .
Earthing research offers insights into the clinical potential of barefoot contact with the Earth, or simulated barefoot contact indoors via simple conductive systems, on the stability of internal bioelectrical function and human physiology. Initial experiments resulted in subjective reports of improved sleep and reduced pain . Subsequent research showed that improved sleep was correlated with a normalization of the cortisol day-night profile . The results are significant in light of the extensive research showing that lack of sleep stresses the body and contributes to many detrimental health consequences. Lack of sleep is often the result of pain. Hence, reduction of pain might be one reason for the benefits just described.
Pain reduction from sleeping grounded has been confirmed in a controlled study on DOMS. Earthing is the first intervention known to speed recovery from DOMS . Painful conditions are often the result of various kinds of acute or chronic inflammation conditions caused in part by ROS generated by normal metabolism and also by the immune system as part of the response to injury or trauma. Inflammation can cause pain and loss of range of motion in joints. Inflammatory swelling can put pressure on pain receptors (nocireceptors) and can compromise the microcirculation, leading to ischemic pain. Inflammation can cause the release of toxic molecules that also activate pain receptors. Modern biomedical research has also documented a close relationship between chronic inflammation and virtually all chronic diseases, including the diseases of aging, and the aging process itself. The steep rise in inflammatory diseases, in fact, has been recently called “inflamm-aging” to describe a progressive inflammatory status and a loss of stress-coping ability as major components of the aging process .
Reduction in inflammation as a result of earthing has been documented with infrared medical imaging  and with measurements of blood chemistry and white blood cell counts . The logical explanation for the anti-inflammatory effects is that grounding the body allows negatively charged antioxidant electrons from the Earth to enter the body and neutralize positively charged free radicals at sites of inflammation . Flow of electrons from the Earth to the body has been documented .
A pilot study on the electrodynamics of red blood cells (zeta potential) has revealed that earthing significantly reduces blood viscosity, an important but neglected parameter in cardiovascular diseases and diabetes , and circulation in general. Thus, thinning the blood may allow for more oxygen delivery to tissues and further support the reduction of inflammation.
Stress reduction has been confirmed with various measures showing rapid shifts in the ANS from sympathetic to parasympathetic dominance, improvement in heart rate variability, and normalization of muscle tension [19, 20, 27].
Not reported here are many observations over more than two decades by Ober et al.  and K. Sokal and P. Sokal  indicating that regular earthing may improve blood pressure, cardiovascular arrhythmias, and autoimmune conditions such as lupus, multiple sclerosis, and rheumatoid arthritis. Some effects of earthing on medication are described by Ober et al.  and at the website: http://www.earthinginstitute.net/. As an example, the combination of earthing and coumadin has the potential to exert a compounded blood thinning effect and must be supervised by a physician. Multiple anecdotes of elevated INR have been reported. INR (international normalized ratio) is a widely used measurement of coagulation. The influence of earthing on thyroid function and medication has been described earlier.
From a practical standpoint, clinicians could recommend outdoor “barefoot sessions” to patients, weather, and conditions permitting. Ober et al.  have observed that going barefoot as little as 30 or 40 minutes daily can significantly reduce pain and stress, and the studies summarized here explain why this is the case. Obviously, there is no cost for barefoot grounding. However, the use of conductive systems while sleeping, working, or relaxing indoors offer a more convenient and routine-friendly approach.
Go to:4. ConclusionDe Flora et al. wrote the following: “Since the late 20th century, chronic degenerative diseases have overcome infectious disease as the major causes of death in the 21st century, so an increase in human longevity will depend on finding an intervention that inhibits the development of these diseases and slows their progress” .
Could such an intervention be located right beneath our feet? Earthing research, observations, and related theories raise an intriguing possibility about the Earth's surface electrons as an untapped health resource—the Earth as a “global treatment table.” Emerging evidence shows that contact with the Earth—whether being outside barefoot or indoors connected to grounded conductive systems—may be a simple, natural, and yet profoundly effective environmental strategy against chronic stress, ANS dysfunction, inflammation, pain, poor sleep, disturbed HRV, hypercoagulable blood, and many common health disorders, including cardiovascular disease. The research done to date supports the concept that grounding or earthing the human body may be an essential element in the health equation along with sunshine, clean air and water, nutritious food, and physical activity.
Go to:DisclosureG. Chevalier, S. T. Sinatra, and J. L. Oschman are independent contractors for Earthx L. Inc., the company sponsoring earthing research, and own a small percentage of shares in the company.
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Articles from Journal of Environmental and Public Health are provided here courtesy of Hindawi Publishing Corporation
Gaétan Chevalier, Stephen T. Sinatra, James L. Oschman, Karol Sokal,
and Pawel Sokal, “Earthing: Health Implications of Reconnecting the
Human Body to the Earth's Surface Electrons,” Journal of Environmental
and Public Health, vol. 2012, Article ID 291541, 8 pages, 2012.
Benefits of Earthing (Grounding)
ELECTRONS and Living Things:
Every chemical reaction in a living body involves some kind of change in the distribution of electrons. Electrons can move in response to electrical current, to light, to sound, to magnetic forces – it all depends on the nature of the substances the electrons are associated with.
In our modern life of technology, living in high rises and wearing shoes, we tend to accumulate a positive charge. Various substances in the body are at times competing for electrons, causing chemical reactions that are not necessarily helpful to our well-being. Several studies indicate that excess positive charge is associated with inflammation. Inflammation is the underlying cause of most pain. Thus, if we can find ways to neutralize the excess positive charge, certain chemical reactions are less likely to occur producing less inflammation and less pain.
Being isolated from the Earth by non-conductive materials such as rubber and plastic (our shoes), wood, plastic, laminate, and asphalt (flooring surfaces) can take it's toll on our health. Most footwear is electrically insulating, as are building materials. Furthermore, many of these materials rob us of electrons. Consequently, we spend most of our time having a shortage of electrons. This affects the chemistry of our body in ways that are only beginning to be understood. Studies done in the last decade have shown than excess positive charge can cause inflammation that can be reduced by making regular contact with a source of free electrons.
One way of tapping into the earth’s abundance of electrons is to go barefoot on soil, sand, or even concrete. Another way is to wear footwear that allows electrons to pass through it. (Such as Pluggz: see their link on many of our pages)
But it doesn't have to be your feet! Gardening, swimming in an ocean/lake or hugging a tree are all ways to easily ground, too! As long as you touch Mother Earth directly, your grounding! The wetter the environment the better, so If you live by a body of water or it's raining outside, that's the easiest!
The Nature of Electrons
Everything is made of atoms and all atoms have electrons and protons. Electrons have a negative charge and protons have a positive charge. Electrons can be in perfect balance with protons, in which case there is no “static charge.” However, various natural and man-made phenomena can cause electrons to become in excess, or in short supply. This happens to our bodies as well as objects. The imbalance of protons and electrons results in a “static charge." Static charge can be reduced simply by making contact with the earth. The earth has an abundance of electrons and if an object holds a net positive charge these electrons are attracted to that object or person through any pathway that is electrically conductive (like water).
How long before noticing positive effects?
The period of grounding in current studies is 28 minutes or longer to feel the positive effects, but many changes may be noticed within a few seconds. The most important change is that the whole body voltage can change greatly within one minute. Many people have reported diminished feelings of stress during and after grounding. You will benefit from a reduced static charge, regardless of whether the effects can be measured.
Just as the sun gives us warmth and vitamin D, the Earth underfoot gives us a natural, and gentle energy. Receiving Earth's energy by touching her may be the difference between feeling good and not so good, of having little or a lot of energy, or sleeping well or not so well.
~Its free, so take those shoes and socks off and get out there!
Published on Sep 27, 2012Watch "Take Back Your Power" now: http://takebackyourpower.net
An 2012 interview with Dr. Dietrich Klinghardt about evidence of damage to human health from "smart" meters. This interview is from the making of the revelatory documentary Take Back Your Power (2013).
Utility companies are racing to replace electricity, gas and water meters worldwide with new generation "smart" meters at an unprecedented rate. With compelling insight from insiders, researchers, government representatives, lawyers, doctors and environmentalists, Take Back Your Power investigates claimed benefits and apparent risks of this ubiquitous "smart" grid program. Transparency advocate Josh del Sol takes us on a journey of revelation and discovery, as we question corporate practices of surveillance, extortion and causing harm in the name of "green".
What you discover will surprise you, unsettle you, and inspire you to challenge the status quo.
Produced & Directed by: Josh Del Sol
Featuring: B. Blake Levitt, Foster Gamble, Jeffrey Armstrong, Dr. Dietrich Klinghardt, James Turner esq, Dr. David O. Carptenter, Bill Vander Zalm, Duncan Campbell esq, Viginia Farver, Ian R. Crane, Edward Snowden, Michael Mitcham, Cal Washington, Anura Lawson.
This film clip contains material used pursuant to fair use doctrine under 17 U.S.C. 107 and has been vetted by an experienced clearance attorney.
Mediators of Inflammation Volume 2014 (2014), Article ID 924184, 14 pages http://dx.doi.org/10.1155/2014/924184
Clinical StudyMetabolic and Genetic Screening of Electromagnetic Hypersensitive Subjects as a Feasible Tool for Diagnostics and InterventionChiara De Luca,1,2 Jeffrey Chung Sheun Thai,3 Desanka Raskovic,4 Eleonora Cesareo,4 Daniela Caccamo,5 Arseny Trukhanov,2 and Liudmila Korkina1,21Centre of Innovative Biotechnological Investigations (Cibi-Nanolab), Novoslobodskaya Street 36/1, Moscow 127055, Russia
2Active Longevity Clinic “Institut Krasoty na Arbate”, 8 Maly Nikolopeskovsky lane, Moscow 119002, Russia
3Natural Health Farm, 39 Jln Pengacara U1/48, Seksyen U1, Temasya Industrial Park, 40150 Shah Alam, Selangor, Malaysia
42nd Dermatology Division, Dermatology Institute (IDI IRCCS), Via Monti di Creta 104, 00167 Rome, Italy
5Department of Biomedical Sciences and Morpho-Functional Imaging, Polyclinic University of Messina, 98125 Messina, Italy
Received 28 November 2013; Accepted 26 February 2014; Published 9 April 2014
Academic Editor: Beatriz De las Heras
Copyright © 2014 Chiara De Luca et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
AbstractGrowing numbers of “electromagnetic hypersensitive” (EHS) people worldwide self-report severely disabling, multiorgan, non-specific symptoms when exposed to low-dose electromagnetic radiations, often associated with symptoms of multiple chemical sensitivity (MCS) and/or other environmental “sensitivity-related illnesses” (SRI). This cluster of chronic inflammatory disorders still lacks validated pathogenetic mechanism, diagnostic biomarkers, and management guidelines. We hypothesized that SRI, not being merely psychogenic, may share organic determinants of impaired detoxification of common physic-chemical stressors. Based on our previous MCS studies, we tested a panel of 12 metabolic blood redox-related parameters and of selected drug-metabolizing-enzyme gene polymorphisms, on 153 EHS, 147 MCS, and 132 control Italians, confirming MCS altered –0.0001) glutathione-(GSH), GSH-peroxidase/S-transferase, and catalase erythrocyte activities. We first described comparable—though milder—metabolic pro-oxidant/proinflammatory alterations in EHS with distinctively increased plasma coenzyme-Q10 oxidation ratio. Severe depletion of erythrocyte membrane polyunsaturated fatty acids with increased ω6/ω3 ratio was confirmed in MCS, but not in EHS. We also identified significantly altered distribution-versus-control of the CYP2C19*1/*2 SNP variants in EHS, and a 9.7-fold increased risk (OR: 95% C.–74.5) of developing EHS for the haplotype (null)GSTT1 + (null)GSTM1 variants. Altogether, results on MCS and EHS strengthen our proposal to adopt this blood metabolic/genetic biomarkers’ panel as suitable diagnostic tool for SRI.
1. IntroductionThe term electromagnetic hypersensitivity or electrosensitivity (EHS) referred to a clinical condition characterized by a complex array of symptoms typically occurring following exposure to electromagnetic fields (EMFs) even below recommended reference levels and is followed by remission through the complete isolation [1, 2]. The most frequently claimed trigger factors include video display units, radio, televisions, electrical installations, extremely low-frequency ranges of electromagnetic fields or radio-frequencies—including the so-called dirty electricity due to poor isolation of electric wires and telephonic lines, wireless devices, and wi-fi—fluorescent lamps and low-energy lights, appliances with motors, photocopiers, microwave transmitters, and high tension power lines (reviewed in [3, 4]). EHS is characterized by a broad range of nonspecific multiple-organ symptoms implying both acute and chronic inflammatory processes, involving mainly skin and nervous, respiratory, cardiovascular, musculoskeletal, and gastrointestinal systems, in most cases self-reported in absence of organic pathological signs except skin manifestations (reviewed in [2, 5]).
Many efforts have been made to determine if a causal relationship between exposure to EMFs and claimed health symptoms does exist and to identify biologically plausible mechanisms underlying this syndrome (for review, see [2, 6, 7]). Despite the growing wealth of evidences gathered both in vitro and in vivo on animal models, data from human case-control and double-blind trials attempting to correlate EMFs exposure and claimed symptoms, resulted so far controversial [8–10]. Nowadays, wide gaps still exist in understanding EHS, which most often remains neglected by the medical community or confined within the frame of mere psychogenic etiology [11, 12]. In the persistent lack of a proven pathogenetic mechanism for electromagnetic hypersensitivity and of clinical consensus on the few proposed diagnostic and therapeutic approaches hypothesized, no guideline for safe and efficient validated treatments has been made available until now to the patients worldwide [13, 14].
Nevertheless, the number of subjects self-reporting EHS is progressively increasing, especially in European countries [15–17], with symptoms that are often strongly disabling both professionally and socially, motivating patients to leave home and job to find rescue in “electromagnetic pollution-free” environmental settings. Because of the huge socioeconomic impact anticipated for EHS syndrome worldwide, the World Health Organization has devoted considerable attention to EHS, acknowledging this condition and recommending that people self-reporting sensitivities receive a comprehensive health evaluation .
Clinical similarities and frequent comorbidity between EHS and the other medically unexplained multisystem conditions of environmental origin, like multiple chemical sensitivity (MCS), fibromyalgia (FM), chronic fatigue syndrome (CFS), sick building syndrome, Persian Gulf War veteran syndrome, and amalgam disease, to which EHS is often associated [19, 20], have induced many authors to hypothesize that these so-called idiopathic environmental intolerances (IEI), more extensively also defined as sensitivity-related illnesses (SRI) , may share common genetic and/or metabolic molecular determinants connected with an impaired capability to detoxify xenobiotics (for review, see [19, 22]). Our group has evidenced for the first time a set of altered metabolic blood parameters—comprising selected redox-active and detoxifying enzymes, low-molecular weight antioxidants and oxidation markers, membrane polyunsaturated fatty acid, and proinflammatory cytokine patterns—specifically and selectively compatible with the MCS condition . Recently, we contributed to the still open issue of possible genetic polymorphic patterns associated with MCS proneness, proposing a pattern of genotypic alterations of the cytochrome P450 isoenzymes CYP2C9, CYP2C19, and CYP2D6, as candidate risk factors for this specific condition, also being potentially able to discriminate different environmental-borne hypersensitivities (MCS, FM, and CFS), depending on specific combinations of their mutated alleles .
In this study, the working hypothesis was that EHS, as previously proposed for MCS and other environmental SRI [19, 22], may as well be based on aberrant responses to physic or chemical xenobiotic stressors through airborne or other routes of exposure, due to inherited or/and acquired dysfunction of the chemical defensive system, that is the interrelated network of phase I and II xenobiotic-metabolizing and antioxidant enzymes . Based on the results of our past clinical studies on MCS, FM, and CFS, we sought to assess if similar profiles of metabolic or genetic dysfunctions could be found in those subjects self-reporting EHS phenotype. To this purpose, we measured possible alterations of a previously identified panel of twelve blood redox and lipid parameters and frequencies of selected genetic mutated variants of a set of drug-metabolizing enzymes and transcription factors with first-line roles in the detoxification of physical and chemical xenobiotics, in a group of 153 patients self-reporting EHS symptoms, co-morbid in most cases with different degrees of MCS symptoms. Results were compared to those obtained on 147 MCS patients without EHS symptoms and on a healthy control group of 132 age- and sex-matched subjects, all groups enrolled within the Italian population.
2. Materials and Methods
2.1. PatientsA group of 153 Italian Caucasian consecutive subjects self-reporting hypersensitivity to electro-magnetic fields (EHS group) as described in Figure 1 were enrolled in the study at a specialized Diagnostic Unit for Redox Balance of Istituto Dermopatico dell’Immacolata, IDI IRCCS, Rome, Italy. Age ranged from 16 to 75 years of age (mean ± SD: ) and female sex represented 85.6% (131 subjects). This group was compared with a size-matched group of 147 patients (age range 19–72 y, mean ± SD: , 129F (87.8%)/18M), diagnosed with MCS, but not reporting any symptom of EHS (MCS group). MCS diagnosis was set in both groups according to Cullen’s criteria  and modified Quick environmental Exposure and Sensitivity Inventory (QEESI) questionnaire scoring [26, 27]. Cullen’s criteria refer to a disorder characterized by symptoms that involve more than one organ system and are regularly elicited by chemically unrelated compounds at doses far below those known to cause adverse effects in the general population. Symptoms typically improve considerable or heal completely after trigger withdrawal . QEESI is a validated self-administered questionnaire developed as a screening tool for patients with multiple chemical sensitivity. It is based on five different scales of assessment: symptoms severity, chemical triggers, other triggers, life impact, and finally a masking index to ongoing exposures [26, 27]. A modified QEESI score of 10 common environmental exposures and 10 major symptoms enabled the diagnosis of MCS: full diagnosis (20 ≤ Score ≤ 30) or strongly suspected diagnosis (sMCS, suspected MCS), that is subjects fulfilling diagnostic criteria only partially (10 ≤ Score ≤ 20), or subjects excluded from enrollment (0 ≤ Score ≤ 10) . As commonly seen by our group occurring in the Italian patient population, the large majority (94.7%) of the EHS group was also affected with multiple chemical sensitivity (fully diagnosed or suspected MCS).
Figure 1: Electromagnetic field sources reported as symptom triggers in the group of patients self-reporting electromagnetic hypersensitivity (EHS, ). Data are expressed as percent of patients affected on the total number of patients.A cohort of 132 healthy age- and sex-matched subjects was enrolled as the control group (CTR group), (age range 18–74 y, mean ± SD: , 109F (82.6%)/23M), according to the established criteria of (i) absence of any clinically diagnosed disease, in particular allergic or immunologic disturbances, (ii) no drug or nutraceutical supplement since at least six weeks, at the time of blood sampling, and (iii) whole blood total production of reactive oxygen and nitrogen species (ROS/RNS) below 650 cps/μL, as determined by luminol-dependent chemiluminescent response to phorbol 12-myristate 13-acetate (PMA)  (Study protocol approval by Istituto Dermopatico dell’Immacolata—IDI IRCCS, Rome, Italy—Ethical Committee, n.52/CE/2010).
All patients and controls entering the study had taken no drugs or nutraceutical supplements known to interfere with metabolizing/antioxidant enzymes activity since at least six weeks, at the time of blood sampling. Nonsmokers in the patient groups were, respectively, 89.3% in EHS and 81.8% in MCS, and 85.2% in the CTR group; undetermined smoking habits were registered in 2% of EHS and 7% of MCS patients, and in 5% of controls. Patients and controls were selected from different Italian regions in the attempt to minimize the historical genetic variability in this country . Demographic information (age, race, weight, and height) and a detailed medical history were recorded in a standardized questionnaire-assisted interview, by trained medical personnel. In particular, subjects were asked to report age at onset of symptoms, agents or events likely to initiate EHS and MCS condition, if recognized, and those capable of triggering symptoms once the condition was established. No alcohol or drug abusers were present in any of the three cohorts studied.
The study protocol was reviewed and approved by the Hospital Ethical Committee Board (IDI IRCCS n.121/CE/2008). All subjects gave informed consent to personal and anamnestic data collection, blood sampling for the specific sets of analyses, and blood fraction’s banking.
2.2. Reagents and Assay KitsMajority of chemical reagents, HPLC standards, mediums, fluorogenic probes, and reverse transcription polymerase chain reaction (RT PCR) primers for gene polymorphism analyses were from Sigma Chemical Co. (St. Louis, MO, USA); kits were from Cayman Chem. Co. (Ann Arbor, MI, USA)—enzyme activities are from Qiagen (Hilden, Germany)—DNA extraction is from Applied Biosystems Inc. (Foster City, CA, USA)—polymerase chain reaction is from PCR Kit for CYPs.
2.3. Redox StudiesComplete differential blood cell counts and metabolic/genetic analyses were performed on fresh EDTA-anticoagulated venous blood of 12-hour fasting subjects. Biochemical assays were performed on plasma or erythrocytes (RBC) either immediately (coenzyme Q10—CoQ10) or within 72 hr. on sample aliquots stored at −80°C under argon. Whole blood luminol-dependent chemiluminescence (CL) response to phorbol 12-myristate 13-acetate (PMA) was quantified by chemiluminescence according to , levels of (nitrites/nitrates) by Griess reagent . Plasmatic total antioxidant capacity (TAC) was determined as described previously . Reduced and oxidised glutathione (GSH and GSSG) levels in erythrocytes , reduced and oxidized CoQ10, and alpha-tocopherol levels in plasma  were quantified by HPLC equipped with array photodiode and electrochemical detection. Activities of CuZn superoxide dismutase (CuZn-SOD) , catalase , glutathione S-transferase (GST) , and glutathione peroxidase (GPX)  in erythrocytes were measured spectrophotometrically.
2.4. Erythrocyte Membrane Fatty Acid ProfilingThe fatty acid (FA) pattern of erythrocyte membrane phospholipids was analyzed by gas-chromatography coupled with mass spectrometry with the selected ion monitoring technique, set to identify C16:0, C16:1, C18:0, C18:1cis, C18:1trans, C18:2ω6, C18:3ω6, C20:4ω6, C20:5ω3, C22:4ω3, C22:5ω3, and C22:6ω3 peaks . Results were expressed as percent of the total fatty acid content of membrane phospholipids for saturated + monounsaturated FA (SFA), polyunsaturated FA (PUFA), and single representative FA of the ω3 and ω6 series.
2.5. Genotyping of Drug Metabolism-Related EnzymesTargeted genotype analysis was performed on subgroups of EHS () and MCS patients () and of controls (), with reduced due to financial limitations—but yet representative—group sizes for single genotype. Genomic DNA was purified from 400 μL of human whole blood using the QIAamp DNA Blood Mini Kit (Qiagen, Hilden, Germany) according to the manufacturer’s instructions. DNA was quantified spectrophotometrically at 260 nm, aliquoted, and stored at −20°C until being assayed. Genotyping and controls for eight single nucleotide polymorphisms in drug metabolism- and inflammation-related genes were carried out by real-time PCR allelic discrimination using predesigned TaqMan single nucleotide polymorphism (SNP) genotyping assays available from Applied Biosystems (Applera Italia, Monza, Italy). The polymorphisms analyzed were those of genes coding for the following: cytochrome P450 (CYP), family 2, subfamily C, polypeptides 9 and 19, namely, CYP2C9*2 (C>T, rs1799853; assay ID: C_25625805_10), CYP2C9*3 (A>C, rs1057910; assay ID: C_27104892_10), and CYP2C19*2 (G>A, rs4244285; assay ID: C_25986767_70); CYP2 subfamily D, polypeptide 6, namely, CYP2D6*4 (1846G>A, rs3892097; assay ID: C_27102431_D0) and CYP2D6*41 (C>T, rs28371725; assay ID: C_34816116_20); aryl hydrocarbon receptor (AHR) Arg554Lys variant (G>A, rs2066853; assay ID: C_11170747_20). Genotyping reactions were set up in a 96-well plate on a 7900HT fast real-time PCR System (Applied Biosystems, Foster City, CA) and were carried out in a final volume of 20 μL containing 1× TaqMan Genotyping Master Mix, 1× TaqMan-specific assay, and 10 ng genomic DNA, using thermal cycling conditions suggested by manufacturer’s protocols.
The GSTP1 polymorphisms resulting in an Ile (wild type) to Val (mutant) substitution at residue 104 in exon 5 and Ala (Wild Type) to Val (mutant) substitution at residue 113 in exon 6 were determined by real time PCR using two different fluorogenic probes for the wild type and the mutant. By combining the results of the analysis of exon 5 and exon 6, the allelic setup was determined (GSTP1*A = Ile104/Ala113; GSTP1*B = Val104/Ala113; GSTP1*C = Val 104/Val113). The deletion polymorphisms for the GSTM1 and the GSTT1 genes were determined simultaneously in a single assay using a multiplex PCR approach with the amplification of the GSTM1 and the GSTT1 genes from genomic DNA and using β-globin as internal control .
2.6. Statistical AnalysisStatistic significance of redox and fatty acid parameters was evaluated using STATISTICA 6.0 program (StatSoft Inc., Tulsa, OK, USA). Normality of data was checked using the Shapiro-Wilk test. Since the distribution of the data in the three groups was significantly different from normal, nonparametric statistics was used. Values were presented as mean, standard error of the mean, and 1.96× standard error. Mann-Whitney U-test for independent samples was employed for comparison between case groups and controls. All reported P values are from two-tailed tests, and P values of less than 0.05 were considered to indicate statistical significance. If necessary, Pvalues were adjusted for multiple comparisons using the Bonferroni adjustment.
The comparison of allele and genotype frequencies between patients and controls, or in-between patient cohorts, was performed using the GraphPad Prism 4 software (San Diego, CA, USA). Genotypes frequencies of patients’ and control groups were compared with Fisher’s exact test. AP value ≤0.05 or lower was regarded as statistically significant. Odds ratio (OR) and 95% confidence interval (CI) were used to analyze the frequency of genotypes since they provide a measure of the strength of association, compared to the control population.
3.1. Anamnestic and Lifestyle DataAmong EMFs emissions recognized as trigger factors in the group of 153 patients self-reporting electromagnetic hypersensitivity-EHS, video display units and television were the most frequently reported sources (75% of patients), followed by mobile and landline phones (53%) and by domestic appliances (48%), while 25% of the electrosensitive population studied could not indicate a specific triggering factor (Figure 1). Potential exposure patterns to indoor EMFs can be inferred from the analysis of the percent distribution of occupational features in the EHS group, described in Figure 2.
Figure 2: Occupational features in the group of patients self-reporting electromagnetic hypersensitivity (EHS, ). Data are expressed as percentage of the total number of patients.The percent distribution of concomitant organ diseases (comorbidities) in the EHS patient cohort, as obtained by clinical anamnestic evaluation, is presented in Figure 3(a). Body mass index (BMI) in the EHS subjects ranged between 15 and 37 (mean ± SD: ), while in the group of MCS without electro-hypersensitivity there were 20% overweight patients (BMI: 25.00–29.99), 11% obese (BMI: 30.00–34.99), 2% severely obese (BMI: 35.00–39.99), 11% underweight (BMI: 18.49–16.00), and only 56% normal-weight patients (BMI: 18.50–24.99). Figure 3(b) shows the percent distribution of the other sensitivity-related illness-SRI coexisting with electromagnetic hypersensitivity in the EHS study cohort, where the 52.7% of MCS cases and the 42% of suspected MCS cases sum up clearly predominant 94.7% of multiple chemical sensitivity symptomatic subjects, within the patients self-reporting EHS symptoms.
Figure 3: Distribution of specific organ comorbidities (a) and sensitivity-related illness-SRI comorbidities (b) registered in the case history of the group of patients self-reporting electromagnetic hypersensitivity (EHS, ). Data are expressed as percentage of the total patient group, for patients affected by each single category of organ pathologies (a), and by each SRI (b), specifically multiple chemical sensitivity (MCS) or suspected MCS (sMCS), chronic fatigue syndrome (CFS), fibromyalgia (FM), and posttraumatic stress disorders (PTSD).In Figure 4, the main classes of cutaneous symptoms or specific diseases recorded by the clinical operators through questionnaire-assisted anamnestic interview are represented, evidencing remarkable prevalence of acute dermatitis or chronic eczema conditions (both symptoms referable to different etiologies) among EHS subjects, whilst in the MCS group without electro-hypersensitivity urticaria and itching referable to (different etiologies) represented the most common findings.
Figure 4: Skin manifestations (common symptoms and specific diseases) registered in the case histories of the groups of patients self-reporting electromagnetic hypersensitivity (EHS, ) and of patients affected by multiple chemical sensitivity without EHS symptoms (MCS, ). Data are expressed as percentage of patients affected by each specific class of cutaneous manifestations.
3.2. Blood Metabolic ParametersCandidate metabolic biomarkers of electrhypersensitivity, as compared to multiple chemical sensitivity without EHS manifestations and to the corresponding values of the same blood parameters in the group of healthy controls, are shown in Figures 5–8.
Figure 5: Metabolic redox parameters: the antioxidant/detoxification enzymatic activities of erythrocyte GST (a), GPX (b), CuZnSOD, (c) and catalase (d), in the groups of patients self-reporting electromagnetic hypersensitivity (EHS, ), of patients affected by multiple chemical sensitivity without EHS symptoms (MCS, ), and of control healthy subjects (CTR, ). Values are represented as mean (□), standard error of the mean (upper and lower limits of the box), 1.96× standard error (upper and lower whiskers). Intergroup significant differences (P) are reported under each panel. RBC: red blood cells; SOD (CuZn superoxide dismutase); GST: glutathione S-transferase; GPX: glutathione peroxidase; prot.: proteins; Hb: haemoglobin.
Figure 6: Metabolic redox parameters: levels of the low-molecular weight antioxidants/cofactors, erythrocyte glutathione ((a) and (b)), and plasma coenzyme Q10 ((c) and (d)), in the groups of patients self-reporting electromagnetic hypersensitivity (EHS, ), of patients affected by multiple chemical sensitivity without EHS symptoms (MCS, ), and of control healthy subjects (CTR, ). Values are represented as mean (□), standard error of the mean (upper and lower limits of the box), and 1.96× standard error (upper and lower whiskers). Intergroup significant differences (P) are reported under each panel. RBC: red blood cells: GSH: glutathione reduced form; GSSG: glutathione oxidized form; GS TOT: total glutathione; CoQ10: coenzyme Q10.
Figure 7: Selected representative parameters describing fatty acid (FA) patterns of erythrocyte membrane phospholipids, in the groups of patients self-reporting electromagnetic hypersensitivity (EHS, ), of patients affected by multiple chemical sensitivity without EHS symptoms (MCS, ), and of control healthy subjects (CTR, ). (a) % saturated and monounsaturated acid (SFA) on total FA content of phospholipids, (b) % polyunsaturated fatty acids (PUFA) on total FA content of phospholipids, and (c) ratio omega-6/omega3 PUFA. Values are represented as mean (□), standard error of the mean (upper and lower limits of the box), and 1.96× standard error (upper and lower whiskers). Intergroup significant differences (P) are reported under each panel. RBC: red blood cells.
Figure 8: Selected representative omega-6 and omega-3 polyunsaturated fatty acids (PUFA) of erythrocytes membrane phospholipid fatty acids (FA), in the groups of patients self-reporting electromagnetic hypersensitivity (EHS, ), of patients affected by multiple chemical sensitivity without EHS symptoms (MCS, ), and of control healthy subjects (CTR, ). (a) % C18:26; (b) C20:46; (c) C18:36; (d) C22:63 FAs, on total FA content of phospholipids. Values are represented as mean (□), standard error of the mean (upper and lower limits of the box), and 1.96× standard error (upper and lower whiskers). Intergroup significant differences (P) are reported under each panel. RBC: red blood cells. 18:26 (linoleic acid), 18:36 (alpha linolenic acid), 20:46 (arachidonic acid), and 22:63 (docosahexaenoic acid).A set of 12 metabolic enzymatic and nonenzymatic redox parameters were measured in the blood of the 153 EHS patients, 147 patients with MCS reporting no EHS, and in the 132 healthy age- and sex-matched CTR subjects. Figure 5 shows the respective alterations of all four enzymatic activities studied in the EHS group, compared to MCS and to control values. More specifically, GST activity in erythrocytes was severely decreased in both EHS and MCS groups, compared to the CTR group (), with no significant difference between the patients’ subgroups (Figure 5(a)). A clearly uprisen erythrocyte GPX activity was registered in the EHS and more markedly in the MCS groups versus controls ( and resp.) (Figure 5(b)), and the same was true for RBC CuZnSOD activity of MCS group versus CTR (), while EHS patients showed only a trend towards increased activity ( versus MCS) (Figure 5(c)). Finally, Figure 5(d) shows how catalase activity rate in RBC was found decreased in both EHS and MCS patients as compared to healthy CTR, though reaching a clear-cut and elevated statistical significance only in the MCS group (), as previously already reported .
Figure 6 describes the alteration of the blood levels of four redox-active low-molecular weight parameters investigated as suitable biomarkers of EHS condition, in comparison to the uncomplicated MCS and the healthy control study cohorts. The levels of both reduced (GSH) and oxidized (GSSG) glutathione forms (data shown in the figure only for GSH (Figure 6(a))) were strongly decreased in the RBC of EHS and MCS environmentally sensitive groups as compared to CTR subjects (GSH: for both groups; GSSG: and , resp., for EHS and MCS), although decrease scores for both glutathione forms were inferior in the EHS than in the MCS subgroup (GSH: ; GSSG: in EHS versus MCS). Also the ratio of GSSG/GSH (Figure 6(b)), indicating the relative oxidation grade of the erythrocyte glutathione marker, displayed a trend to elevation in the two patient subgroups versus control, although data were too scattered to reach any statistical value.
The plasmatic levels of coenzyme Q10 and alpha-tocopherol displayed a similar trend-to-depletion in both patient subgroups versus controls. Figure 6(c) reports results of ubiquinol (CoQ10H2, the reduced form of coenzyme Q10) analysis which, together with levels of total CoQ10 (reduced + oxidized forms) and of alpha-tocopherol (both groups of data not shown)—showed similar trend of reduction for EHS as well as MCS subgroups, as compared to CTR group, though lacking statistical significance. Indeed, we found a higher percent coenzyme Q10oxidation (ratio oxidized-CoQ10/total-CoQ10), significant versus CTR at in EHS patients, not confirmed for MCS patients, as reported in Figure 6(d).
Although a trend-to-increase in the values of whole blood chemiluminescence (CL) and to decreased levels of plasmatic total antioxidant capacity (TAC) were recorded for both patient subgroups compared to controls, differences were unable to reach any statistical significance (data not shown). The increase of/plasma levels of MCS patients obtained in our previous study  was not confirmed in this new MCS subgroup, as well as in the EHS group of the present study, respectively, averaging or being inferior to control values (data not shown).
Since the majority of the above metabolic data were similar for EHS and MCS subgroups, the costly and time-consuming analyses of fatty acid profiles were carried out on a more limited subgroup of patients who fully corresponded to all diagnostic criteria. Representative fatty acid profiles in the phosholipid fraction of the erythrocyte membranes of EHS (), MCS () and CTR () patients are shown in Figures 7 and 8. The comparative analysis of the fatty acid (FA) profiles in the erythrocyte membranes of the 3 studied groups showed elevated levels of the saturated and monounsaturated fatty acid fraction (SFA) for both environmental-sensitive patients (Figure 7(a)) and correspondingly depleted levels of the polyunsaturated fatty acid fraction (PUFA) (Figure 7(b)), with both parameters statistically significant at for MCS patients versus controls, whilst the EHS group differed sensibly from MCS in displaying only a mild trend-to-alteration of fatty acid patterns versus control group. In detail, the percent levels of the omega-6 FA linoleic (18:2ω6), alpha linolenic (18:3ω6), arachidonic (C20:4ω6), and the omega-3 FA docosahexaenoic (C22:6ω3) (Figures 8(a)–8(d)) were lower than control values in both EHS and MCS cohorts, although the clear-cut statistical significance registered for the MCS group (–0.001 for all 4 parameters) was confirmed in EHS patients only for linoleic acid fraction () (Figure 8(a)). Finally, the range of the ω6/ω3 PUFA ratio in electrosensitive subjects practically equalled that of controls, whilst MCS patients showed significantly increased values versus both CTR () and EHS group (), as reported in Figure 7(c).
3.3. Genetic Parameters
The main results of genotype analysis for a selected panel of detoxifying enzymes, obtained on limited subgroups of EHS, MCS, and controls, are illustrated in Table 1. Having previously demonstrated in the MCS population a significantly higher-versus-CTR frequency of the homozygous mutated *1 allele and a CYP2C19*2 heterozygous genotype *1/*2, with a lower frequency of the *2 allele in the homozygous and heterozygous forms , we here confronted the panel of previously investigated CYP isozymes in the EHS versus the already studied MCS cohort previously studied. Genotype frequencies for cytochrome P450 CYP2C19 SNP variants in EHS and MCS patients’ groups showed that the CYP2C19*1/*1 and the CYP2C19*1/*2, *2/*2 genotypes differed with statistical significance at between EHS () and MCS () groups. The other gene polymorphisms of CYPs studied (CYP2C9 and CYP2D6), as well as the aryl hydrocarbon receptor (AHR) variant Arg554Lys, displayed similar frequency distributions for EHS and MCS patients (data not shown).
Table 1: Statistical analysis of genotype distribution of cytochrome P450 (CYP) isoenzymes in EHS-patients self-reporting electromagnetic hypersensitivity () versus MCS-multiple chemical sensitivity patients without EHS () and of glutathione S-transferase P1 (GSTP1), glutathione S-transferase M1 (GSTM1), and glutathione S-transferase T1 (GSTT1) isoenzymes in CTR-healthy control subjects () versus EHS-patients ().
Genotype frequencies of the glutathione S-transferase (GST) isoenzymes GSTP1, GSTM1, and GSTT1, previously found not significantly differing in MCS versus healthy control populations , were compared in 127 EHS patients versus 68 CTR subjects. No statistically significant differences were observed for GSTP1 in the frequency of the GSTP1*A, GSTP1*B, or GSTP1*C homozygous and heterozygous variants between the EHS patient and control groups (Table 1).
The statistical analysis of the distribution of GSTM1 and GSTT1 isoenzymes showed no statistical difference in homozygous + heterozygous and null genotype variants neither in GSTM1 nor in GSTT1, when analyzed independently. Conversely, the combined GSTM1 (*0/*0) + GSTT1 (*0/*0) null genotypes differed significantly (13% versus 1.5%, resp.), with , in EHS patients versus CTR subjects, conferring to this association of gene variants 9.7 times higher risk (OR: 95% C.I. = 1.3–74.5) of developing EHS compared to other GSTM1 and GSTT1 combinations of genotypes examined (Table 1).
4. DiscussionTill now, no causal relationship between electromagnetic fields exposure and onset of clinical symptoms has been clearly proven. Nevertheless, the term electric hypersensitivity is currently used both by patients who claim health effects of environmental electromagnetic pollution and doctors to define patient clusters of symptoms . Most of the evidences about altered organic parameters due to EMF exposure have been so far obtained on cell or animal models. Very few human studies investigated possible organic parameters distinctive of the hypersensitivity to electromagnetic stressors ([41, 42]; for review, see ).
Main difficulties for clinical studies’ implementation arise from the necessity to deal with patients in a protected environment, sheltered from EMF sources and also free of chemical barriers, since the majority of electrosensitive patients are also intolerant to a multiple array of chemical triggers . Indeed, in the group of 153 EHS subjects enrolled for this study, 145 were also affected at different degrees by MCS symptoms (Figure 3(b)). The experimental group of EHS patients was exposed by lifestyle to the most common electromagnetic sources deriving mainly from indoor or outdoor urban electromagnetic pollution and no heavy professional exposure in industrial settings was recorded in the group (Figure 2). In addition, EHS patients shared with MCS patients the sensitivity to the most frequent organic chemical triggers initiating and sustaining MCS.
Another relevant issue complicating human studies is connected with the difficulties encountered in provocation studies, aimed at connecting the electromagnetic trigger with electrohypersensitivity symptoms’ onset. These difficulties arise generally from the necessity to standardize types and dosages of EMF sources, from the broad qualitative and quantitative range of individual multiorgan responses to trigger, difficult to measure objectively, and also from heavy psychoemotional bias factors affecting experimental protocols and their repeatability [44, 45]. Notably, provocation studies commonly proposed as the main milestone for EHS assessment and validation are based on the questionable assumption that the individual capability to directly perceive EMFs at low or very low intensities below established toxicological thresholds, claimed by EHS subjects in analogy with MCS odor perception, may be conditio sine qua non for EHS symptom manifestation [40, 46]. Waiting for a consensus on a standardized methodology for an objective clinical assessment of electro-sensitivity, our present work referred to self-reported EHS as registered in the course of the anamnestic evaluation performed by trained medical personnel.
Data concerning the involvement of organic causes connected with chronic oxidative damage as a key factor in the induction and perpetuating of symptoms in functional SRI syndromes has been growing in the last decade (reviewed in: ). Our previous studies provided evidence of a specific and peculiar metabolic disease-marker profile in multiple chemical sensitivity, the prototype of all medically unexplained environmental illnesses so far described. In fact, moving from published data accounting for the altered redox balance in favor of a prooxidative and proinflammatory state in patients with fibromyalgia or chronic fatigue symptoms [7, 22], we identified a profile of 12 specifically altered blood parameters connected with systemic oxidative stress and impaired detoxification, in a representative sample of the Italian population fully or partially complying with MCS diagnosis . In the same line, the present study was conceived to verify if analogous alterations of this pattern of MCS reliable organic biomarkers may also apply to EHS condition, in order to seek evidences of the organic etiology of this group of environmental sensitivity disorders and provide the clinicians with suitable tools for laboratory diagnosis and treatment follow-up.
The profiles of metabolic parameters’ alteration observed in EHS subjects were comparable to those of the “pure MCS” group, though generally less pronounced (Figures 5–8). Similarly to those MCS patients self-reportedly nonelectrosensitive, the EHS cohort showed a highly significant-versus-control decrease in the erythrocyte GST activity and an increase in GPX activity levels (Figure 5), coupled with a marked decrease of GSH levels (Figure 6). Again in line with MCS, EHS group showed a trend to the increase in erythrocyte CuZnSOD activity and to the depletion of the main lipophilic antioxidants in plasma-reduced coenzyme Q10 and alpha-tocopherol (vitamin E) (Figures 5 and 6). The most striking difference between the two patient subgroups was recorded, instead, for erythrocyte catalase. Enzymatic activity was in fact only slightly and not significantly, reduced in EHS as compared to control values, while the highly significant () reduction recorded in the MCS group (Figure 5) confirmed our previous reports, validating the relevance and selectivity of this blood metabolic marker specifically for the MCS condition , being previously confirmed also in those patients only partially complying with MCS criteria (suspected MCS group).
We also calculated the ratios between oxidized and reduced forms of glutathione and coenzyme Q10 as suitable indicators of a systemic oxidative and proinflammatory status . Relative oxidation of the two redox molecules was increased, though not significantly, in both EHS and MCS groups versus CTR (Figure 6). Interestingly, only in electrosensitive subjects, the oxidized/total CoQ10 ratio reached statistical significance () versus normal values. Due to its marked lipophilicity, coenzyme Q10 is essential, along with alpha-tocopherol and squalene, for skin protection against oxidizing environmental physicochemical stressors, and it is able to efficiently reach the skin from the blood compartment [48, 49]. The elevated oxidation of plasma coenzyme Q10 observed in EHS appears to be consistent with the higher frequency of cutaneous involvement in EHS (40.7%) symptoms self-reported by our experimental group (Figure 3(a)), as compared to the minor relative clinical relevance assessed in the classical MCS condition, previously described . Accordingly, Figure 4 shows how the prevalent skin symptom, in the EHS but not in the MCS cohort, resulted in being acute or chronic dermatitis (eczema), a group of inflammatory skin diseases where systemic and local lipophilic antioxidant depletion is strongly implicated .
A second parameter proved to be significantly different () between EHS and MCS groups that is the ratio omega-6/omega-3 polyunsaturated fatty acids in the erythrocyte membrane phospholipid fraction (Figure 7(c)). The ratio showed a remarkable elevation versus CTR in favor of the more proinflammatory ω6 PUFA in the MCS group (), while EHS values were instead nearly overlapping CTR values, data that appears consistent with the overall less pronounced prooxidative and proinflammatory state evidenced in EHS versus MCS, from the whole pattern of redox parameters investigated in this study. Again, this molecular marker difference between the two environmental hypersensitivities can possibly be connected with the clinical setting, where, for example, a higher frequency of pathological obesity with metabolic syndrome is observed in MCS , whereas EHS condition features a milder chronic inflammatory status .
As a whole, MCS values of all metabolic parameters studied confirmed our previous results obtained in a larger cohort of 226 MCS + sMCS patients , highlighting the reliability of the selected redox-marker panel on this additional study cohort. With two exceptions, (a) erythrocyte CuZnSOD activity, now found significantly increased () in MCS versus CTR (Figure 5(c)) whilst nonsignificant in the first study, and (b) plasma nitrites/nitrates values, significantly elevated in the previous study MCS cohort , a finding not confirmed in the present study (data not shown). These differences may possibly be related to the extreme individual genetic and metabolic variability characterizing MCS populations, even within the same ethnic, geographic, lifestyle, and cultural setting, which represented one of the difficulties facing SRI human studies .
The question as to whether genetic background may determine a proneness to environmental hypersensitive syndromes remains still unanswered, from the time of the first pioneer studies on multiple chemical sensitivity [53, 54], followed by a wealth of extensive investigations on MCS, FM, and CFS western populations worldwide [19, 23, 55]. We attempted to contribute to this unresolved issue of utmost relevance for diagnostic purposes in these poorly defined clinical settings. In previous works, we had investigated gene and allele frequencies of selected polymorphisms of a wide array of phase I and II xeno- and endobiotic metabolizing enzymes, GST (M1, T1 and P1), UDP-glucuronosyl transferase (UGT), and cytochrome P450 (CYP) variants belonging to the CYP2C9, CYP2C19, CYP2D6, and CYP3A5*3 isoenzymes. After a first study not showing any significant prevalence of the studied CYP, UGT, and GST gene polymorphisms in a group of 110 MCS patients , we proceeded to a second investigation on a clinically better characterized MCS group of 156 patients and of 113 matched controls, where we identified significantly (–0.0001) higher frequencies versus CTR for the polymorphisms CYP2C9∗2, CYP2C9∗3, CYP2C19∗2, CYP2D6∗4, and CYP2D6∗41, confirming other studies indicating these genetic variants as a risk factor for SRI . Starting from these results, in the present study, genotyping for the CYP2C19 single nucleotide variants showed that the frequency of the homozygous mutated *1 allele was significantly higher in EHS, than in MCS cases, whilst the *2 allele in the homozygous and heterozygous forms was less frequent in EHS than in MCS () (Table 1). Moreover, our previous work had shown that the CYP2C19*2 heterozygous genotype *1/*2 was significantly more frequent () in MCS cases, not only versus controls but also versus FM + CFS cases . The same study showed for the first time that the Arg554Lys mutated variant of the aryl hydrocarbon receptor-AHR gene did not reach significant differences in distribution between SRIs and controls when analyzed alone but showed in specific haplotype combinations with CYP variants promising implications for in-between group discrimination within SRI comorbidities, namely, MCS versus sMCS and FC + FM versus controls . In the present work, we were able to confirm the absence of significant differences for AHR genotype between EHS and CTR groups (data not shown).
Having previously found no significant difference between MCS patients and controls, in the distribution of GST isoenzyme genotypes , in the GST study we now compared EHS and healthy controls. Differently from our previous results on MCS, we here identified a mutated (null) allele combination of GSTT1 and GSTM1 variants able to predict risk of developing EHS by a 9.7 fold versus CTR (Table 1).
Taken together, our genetic results obtained on a number of cases due to be enlarged in the studies to come, although being far to be conclusive on such a controversial matter, can at least contribute additional indications to the complex mosaic of genetic risk factors in environmental hypersensitivities, still waiting to be correlated with individual metabolic phenotypes.
The outcomes of this work confirmed, in the whole, our previous results on MCS and provided additional evidences for the validity of the selected panel of metabolic blood parameters also in the self-reported EHS condition. Further developments must necessarily include a more objective and standardized classification of individual electromagnetic sensitivity scores, to conclusively assess the proposed parameters as a distinctive and specific panel of disease biomarkers for EHS. Our findings will hopefully contribute, in combination with the so-far putative genetic-risk factors, a better molecular definition of environmental-borne sensitivity-related illnesses and a tool to discriminate single SRI comorbidities, based on sufficiently proven molecular evidences able to gain clinical consensus.
Conflict of InterestsThe authors declare that they have no conflict of interests.
AcknowledgmentsThe authors acknowledge the generous participations of Dr. M. Grazia Bruccheri, MD, from IRMA-Istituto Ricerca Medica e Ambientale, Acireale (CT), Italy, for patient diagnosis and enrolment in Sicily, Francesca Romana Orlando, from the Patient Association A.M.I.C.A.—Associazione per le Malattie da Intossicazione Cronica e/o Ambientale, Rome, Italy, for excellent documental support, and Andrea Stancato, from IDI IRCCS for skilled technical assistance.