Synopsis: Exposure of Normal Human Astrocytes Cells to Mobile Phone Radiation with and without MRET-Nylon Protection

Introduction: Astrocytes are a sub-type of the glial cells in the brain and spinal cord. They are also known as astrocytic glial cells. Star-shaped, their many processes envelope synapses made by neurons. Astrocytes are classically identified histologically as many of these cells express the intermediate filament glial fibrillary acidic protein (GFAP).

Fig. 1 Astrocytes can be visualized in culture because, like other glia, they express glial fibrillary acidic protein.[Wikipedia]

• Structural: involved in the physical structuring of the brain.
  
• Metabolic support: they provide neurons with nutrients such as lactate.
  
• Blood-brain barrier: the astrocyte end-feet encircling endothelial cells were thought to aid in the maintenance of the blood-brain barrier, but recent research indicates that they do not play a substantial role; instead it is the tight junctions and basal lamina of the cerebral endothelial cells that play the most substantial role in maintaining the barrier. However, it has recently been shown that astrocyte activity is linked to blood flow in the brain, and that this is what is actually being measured in fMRI.
  
• Transmitter reuptake and release: astrocytes express plasma membrane transporters such as glutamate transporters for several neurotransmitters, including glutamate, ATP and GABA. More recently, astrocytes were shown to release glutamate or ATP in a vesicular, Ca2+-dependent manner. But this glutamate release has not been proven yet.
  
• Regulation of ion concentration in the extracellular space: astrocytes express potassium channels at a high density. When neurons are active, they release potassium, increasing the local extracellular concentration. Because astrocytes are highly permeable to potassium, they rapidly clear the excess accumulation in the extracellular space. If this function is interfered with, the extracellular concentration of potassium will rise, leading to neuronal depolarization by the Goldman equation. Abnormal accumulation of extracellular potassium is well known to result in epileptic neuronal activity.
• Modulation of synaptic transmission: in the supraoptic nucleus of the hypothalamus, rapid changes in astrocyte morphology have been shown to affect heterosynaptic transmission between neurons. In the hippocampus, astrocytes suppress synaptic transmission by releasing ATP, which is hydrolyzed by ectonucliotidases to yield adenosine. Adenosine acts on neuronal adenosine receptors to inhibit synaptic transmission, thereby increasing the dynamic range available for LTP.
  
• Vasomodulation: astrocytes may serve as intermediaries in neuronal regulation of blood flow.
  
• Nervous system repair: upon injury to nerve cells within the central nervous system, astrocytes become phagocytic to ingest the injured nerve cells. The astrocytes then fill up the space to form a glial scar, repairing the area and replacing the CNS cells that cannot regenerate.

Recent studies have shown that astrocytes play an important function in the regulation of neural stem cells. Research from the Schepens Eye Research Institute at Harvard shows the human brain to abound in neural stem cells, which are kept in a dormant state by chemical signals (ephrin-A2 and ephrin-A3) from the astrocytes. The astrocytes are able to activate the stem cells to transform into working neurons by dampening the release of ephrin-A2 and ephrin-A3. Furthermore, studies are underway to determine whether astroglia play an instrumental role in depression, based on the link between diabetes and depression. Altered CNS glucose metabolism is seen in both these conditions, and the astroglial cells are the only cells with insulin receptors in the brain. [Wikipedia]

Experimental Protocol: The experiment was conducted at AltheaDx Technology, San Diego under supervision of Project Director: Qiang Xu, Ph.D., Project Scientist: Pat Pezzoli, B.S., Project Technician: Neil Tedeschi, M.S.

• Normal Human Astrocytes (NHA) (Lonza #CC-2565, Lot 80982) were grown in a humidified incubator at 37°C and 5% CO2 and were expanded until there were a sufficient number of cells for the experiment. The cells were harvested with trypsin and counted on a hemocytometer using trypan blue. The viability was 88.9% and 281,667 cells per well were plated in to six wells, two wells each on three six well plates. The cells were incubated overnight.
• An LG Verizon cell phone, Model # VX8350, FCC ID BEJVX8350, SW version # VX835V03, HW Rev. 1.1, MEID A000000C4F8FC5, using a AC power source was placed directly beneath and centered under one plate of duplicate NHA cell cultures at a distance of 0.5 inches below the growth surface (see Figure 3 and Figure 4).
• The MRET®-Nylon chip belongs to the new generation of electromagnetic radiation shielding materials based on Molecular Resonance Effect Technology developed by Igor Smirnov, Ph.D. The MRET®-Nylon polymer compound has a special fractal geometric structure. Due to the fractal nano-rings structure and enhanced piezoelectric properties of this compound, it generates random, subtle, low frequency oscillations when exposed to the external electromagnetic radiation (EMR). This polymer can significantly decrease the biological effects of electromagnetic radiation, both thermal and non-thermal, by imposing the random low frequency oscillations (noise field) on RF waves. The theoretical concept of the electromagnetic noise field is related to the ability of the noise field to offset the thermal effects.

The cell phone was called by a phone and the calling phone's hand set was placed next to the speaker of an operating radio so that the cell phone would be continuously active for duration of the exposure. The cells were exposed to phone radiation for one hour at room temperature. Following the one hour cell phone exposure, the cells were placed back in the incubator for 24 hours. A second identical NHA culture was then exposed similarly to the same cell phone and in the same geometry with the addition of the MRET-Nylon protection which was placed over the cell phone ear speaker as shown in Figure 5.

The cells were exposed to the cell phone radiation for one hour and then the cells were placed into the incubator for 24 hours. During the cell phone with the MRET-Nylon protection exposure, a third plate containing identical cells was placed in another room for one hour and was labeled Control Plate. Following one hour of incubation at room temperature without any cell phone exposure, it was placed back into the incubator for 24 hours.
After the 24 hour incubation period, the cells were harvested from each well using trypsin and counted on a hemocytometer with trypan blue dye to obtain cell counts and viability data. The cell count data consists of replicate wells for each treatment condition. Each well was harvested using the same volumes and each was subjected to the same pipetting action.
For each sample, RNA was extracted from duplicate one - the top well shown in the experimental setup. The RNA was processed according to the Affymetrix GeneChip Whole Transcript (WT) Sense Target Labeling Assay. The resultant labeled cDNA was hybridized to Affymetrix Human Gene 1.0 ST arrays and scanned. The data was normalized using RMA normalization with the Affymetrix Expression Console software.
This normalized data was used for the correlation analysis.

The in vitro experiment reveals that Normal Human Astrocyte cell counts after exposure to mobile phone radiation with MRET-Nylon protection decreased by 20% less compared to the cell samples exposed to the same mobile phone radiation without MRET-Nylon protection, and by 12% less compared to control samples not exposed to mobile phone radiation (Figure 6). The experiment also revealed that the viability of Normal Human Astrocytes cells in case of exposure to mobile phone radiation with MRET-Nylon protection was by 3% higher compared to the viability of cells exposed to the same mobile phone radiation without MRET-Nylon protection.

Fig. 6 Human Astrocytes Cell Counts: Before the experiment (before 1 hour exposure to mobile phone radiation at room temperature and 24 hours of post exposure incubation); Control (after 1 hour at room temperature without exposure to mobile phone radiation and 24 hours of post exposure incubation); Phone without MRET-Nylon (after 1 hour exposure to mobile phone radiation at room temperature without MRET protection and 24 hours of post exposure incubation); Phone with MRET-Nylon (after 1 hour exposure to mobile phone radiation at room temperature with MRET protection and 24 hours of post exposure incubation).

Data Comparison: We compared each sample using the Pearson correlation coefficient. A coefficient of 1 indicates perfect correlation while 0 indicates no correlation. All samples are correlated at a level of 0.99 or higher.

For further comparison, a ‘heat map’ was generated. The expression patterns are similar across the samples:

Conclusion: The in vitro experiment revealed that Normal Human Astrocyte cell counts after one hour exposure to mobile phone radiation with MRET-Nylon protection decreased by 20% less compared to the cell samples exposed to the same mobile phone radiation without MRET-Nylon protection, and by 12% less compared to control samples not exposed to mobile phone radiation. The experiment also revealed that the viability of Normal Human Astrocytes cells in case of exposure to mobile phone radiation with MRET-Nylon protection was 3% higher compared to the viability of cells exposed to the same mobile phone radiation without MRET-Nylon protection.
The visual inspection of cell samples with microscopy did not show a significant difference between the control and exposed samples. The microarray analysis showed no difference in mRNA expression patterns between the three sample types. Thus this study provides some evidence that one hour exposure of Normal Human Astrocytes cells to mobile phone radiation with 24 hours post exposure incubation did not affect cell genetics. On the other hand there was found measured effect of mobile phone radiation on cell counts and viability.
The study confirmed that the application of MRET-Nylon chip on mobile phone reduced the negative biological effect of microwave radiation by enhancing cell viability and resistivity to EMR thermal and non-thermal biological effects.

The Effect of MRET-Shield Material on SAR Values of Mobile Phones

DISCUSSION

Specific Absorption Rate (SAR) characterizes the level of absorption of EMR by the tissue of the body. The absorption of EMR by biological systems can lead to the distortion of cellular transduction mechanism, to the development of thermal effect in cells, their damage and, consequently, to the distortion of cellular function. Due to these reasons FCC (Federal Communication Committee) established the standards for allowed SAR values in the range of 0.2 – 2.0 W/kg. The reduction of SAR values obviously is beneficial for human health. The test evaluation revealed the two key results:

• The application of MRET-Shield to the RF phones does not lead to any significant distortion of transmitted RF signals.
• In each experiment SAR values were measured in 242 points of “phantom head.” The application of MRET-Shield to the RF phones showed that “Hot Spots” remained in the same location as without MRET polymer and their amplitudes decreased in 90% of data points. The application of MRET-Shield to the RF phones leaded to the reduction of the majority of meaningful SAR values in these experiments in the range of 0.3% - 29.0%.

Below are presented the “Hot Spot” Area Scan Diagrams that provide evidence that the application of MRET-Shield to the RF phone did not change the location of “Hot Spot” and significantly decreased SAR values in the scanned area.

“Hot Spot” Area Scan Diagrams

The Beneficial Effect of MRET-Shield on Blood Morphology in vitro Following the Exposure to Electromagnetic Radiation

The beneficial effect of EMR shielding material (MRET-Shield) on human blood in vitro was observed at the laboratory of Medical Center, Los Angeles, USA. The effect of the computer monitor radiation on human blood samples (22 samples in each group) was studied with and without installation of MRET-Shield and compared to control group not exposed to EMR. The blood samples were exposed to the computer monitor radiation at the distance of 15” (0.38 meter) for one hour. Test results are statistically valid with p<0.01.

The White Blood Cells (WBC) consist of Granulocytes (GRAN), Lymphocytes (LYM), and “Minimum Inhibitory Dilution,” a measure of rare cells and a number of precursor white cells (MID). The exposure of the blood samples to external EMR of the computer monitor leaded to the changes in the ratio of WBC components, particularly to the changes in Granulocytes, Lymphocytes and MID counts (measured in %). The installation of MRET-Shield EMR shielding device) on the computer monitor significantly reduced the level of changes of Granulocytes and Lymphocytes counts and almost did not affect the level of changes in MID count. These results confirm the reduction of stress following the exposure of blood samples to EMR in case of the installation of MRET-Shield on the computer monitor.

Granulocytes are a critical component of WBC and play an important role in the immune system reactions. Their decrease is not beneficial for the body homeostasis. The experiment revealed the decrease of Granulocytes in blood samples exposed to electromagnetic radiation. It also showed that the installation of MRET-Shield on the computer monitor reduced the level of changes in Granulocytes count by 29%.

An increase in the Lymphocytes count is associated with a physiological response to an antigenic or inflammatory stimulus and infection or other kind of external stress. The general consensus is that the increase of Lymphocytes above normal level in case of the absence of any infections increases the risk of leukemia, lymphomas, and other disorders. The experiment revealed the increase of Lymphocytes in blood samples exposed to the computer monitor radiation. It also showed that the installation of MRET-Shield on the computer monitor reduced the level of changes in Lymphocytes count by 38%.

The experiment revealed the increase of MID count for blood samples exposed to the computer monitor radiation. The reduction of the level of changes in MID count after the installation of MRET-Shield was insignificant.

CONCLUSION: The experiment revealed the decrease of Granulocytes count and the increase of Lymphocytes and MID (precursor white cells) counts in both cases. There were no significant changes in the total quantity of WBC. It also showed that the installation of MRET-Shield on the computer monitor significantly reduced the level of changes in the ratios of Granulocytes and Lymphocytes counts.

The changes in the ratio of GRAN were reduced by
(32 – 26.4)/(46 – 26.4) = 5.6/19.6 = 29%

The changes in the ratio of LYM were reduced by
(49.4 – 44.3)/(49.4 – 36) = 5.1/13.4 = 38%

The changes in the ratio of MID were insignificantly reduced by
(24.2 – 23.7)/(24.2 – 18) = 0.5/6.2 = 8%

Thus, the combined changes in the ratios of GRAN, LYM and MID following the exposure to EMR of the computer monitor were:
without MRET-Shield   19.6 (GRAN) + 13.4 (LYM)+ 6.2 (MID) = 39.2%
with MRET-Shield         14 (GRAN) +  8.3 (LYM) + 5.7 (MID) = 28%

This calculation shows that the installation of MRET-Shield on computer monitor reduced the level of combined changes in the content of White Blood Cells count (GRAN, LYM and MID) by (39.2 – 28)/39.2 = 29% in this experiment.

Thermography Test for MRET-Shield

Testing was conducted at the SAMEER Centre for Electromagnetic, a division of the Research and Design Institution of the Ministry of Communications and Information Technology, Department of Information for the Government of India, in Chennai on February 25, 2008. Thermographic imaging was conducted using standard thermography procedures, as per the guidelines from the Standards and Protocols in Clinical Thermographic Imaging released by the International Academy of Clinical Thermography. The subject was a forty year old male. The study was conducted using a Compix Infrared Scanner Model number 6000A. The first imaging established a baseline thermographic image and temperature of the subject (Figure 1). The subject then talked on a Nokia model 1600 type RH 64 cell phone model for thirty-three minutes and the second image and temperature were recorded (Figure 2). Then a MRET-Shield polymer device was placed on the cell phone near the internal antenna. The subject talked again for thirty-three minutes on the cell phone keeping the phone at the same standard position (Figure 3) and the third image and temperature were recorded.

The results are listed in Table I. The baseline (control) temperature was 36.7 degrees Celsius. The temperature increase caused by the radiation from the unprotected cell phone was 1.4 degrees Celsius. Following the installation of a MRET-Shield device the temperature change decreased by 0.9 degrees Celsius. Thus, the MRET-Shield device reduced the temperature increase by 64%. It can be seen from the thermographic images that there was a large scale heating of the tissues of the head and face compared to the baseline and a significant reduction of this bio-thermal effect following the installation of the MRET-Shield polymer device on the cell phone.

TABLE I TEMPERATURE OBSERVATIONS

Case Temperature Control 36.7° C Unprotected phone 38.1° C Protected phone 37.2° C

FIGURE 1 BASELINE TEMPERATURE

FIGURE 2 UNPROTECTED CELL PHONE TEMPERATURE

FIGURE 3 PROTECTED CELL PHONE TEMPERATURE

3D-MRA Test (Magnetic Resonance Angiography - Sectional Analysis of Human Brain Function)

The beneficial effect of MRET-Shield was confirmed by 3D-MRA test which was conducted by Tex Chu Technology Corp., Taipei, Taiwan. Test was conducted on human subject who was exposed to cell phone radiation (Sony Ericsson T630) with and without installation of MRET-Shield. Magnetic resonance angiography (MRA) is a technique for imaging blood vessels that contain flowing blood. Magnetic Resonance Angiography (MRA) is used to generate pictures of the arteries in order to evaluate them for stenosis (abnormal narrowing) or aneurysms (vessel wall dilatations, at risk of rupture). MRA is often used to evaluate the arteries of the neck and brain. A  technique known as "flow-related enhancement" (e.g.  3D time-of-flight sequences) was used to get images, where most of the signal on an image is due to blood which has recently moved into that plane. Test provides analysis for the following sections: Cerebrum, top view; Cross section of brain at the brain stem level; Arteries of brain, bottom view of brain; Left – Right hemisphere sequence function;

• First angiogram: Control, subject was not exposed to cell phone radiation;
• Second angiogram: After exposure to cell phone radiation without MRET-Shield protection;
• Third angiogram: After exposure to cell phone radiation with MRET-Shield protection.

The MRA angiograms clearly show the substantial negative effect of RF radiation generated by cellular phone on the state of blood vessels in all parts of the brain area. They also reveal that the installation of MRET-Shield on cellular phone compensates for this negative effect and helps to maintain the blood vessels of the brain in good state.

Live Blood Cell Analysis Conducted at Quantum Biotech Ltd. EEG Tests The EEG tests were conducted at the laboratory of SA Biomedical Instrumentation Co., California. These tests were conducted on the human subject introduced to the influence of cellular phone (SAMSUNG Model No: SCH-2000) without MRET-Shield protection and with MRET-Shield protection respectively. Each test was run for 3 minutes. The cellular phone was located in standard operating position at the right side of a head. The head of the subject was covered with a standard Electro-cap E1-L connected with a 4 channel SAI Bioelectric Amplifier that includes Analog Processor and Anti-aliasing Filters. The data were provided from the following electrodes: C4, Fz, F4, and T4. This precision Amplifier can record and analyze very lo-potential physiological data from surface electrodes. With this SAI BIOAMPS successful physiological data acquisition can be accomplished in most any environment without the use of a shielded “screen room. The comparison of the EEG charts that present the brain function wave patterns: The control chart of a subject not introduced to EMR of a cellular phone shows normal slow mode of brain function. The chart of a subject introduced to EMR of a cellular phone without any protection shows the excitation of the brain function wave patterns. The chart of a subject introduced to EMR with MRET-Shield protection is very close to the control chart. It means that the installation of MRET-Shield on the cellular phone synchronizes and slows down the brain function frequencies. It makes the brain function frequency spectrum more coherent and optimized from the physiological point of view. This fact proves that MRET-Shield neutralizes the electromagnetic stress and excitement caused by the effect of a cellular phone on the brain. The chart of a subject introduced to EMR of a cellular phone with competitive product (Pulse Clean 21) protection shows that this “protection device” makes the excitation of the brain function wave patterns even worse. It happens because the devices that claim to reduce EMR first of all suppress and disturb subtle electromagnetic processes in living cells. The installation of MRET-Shield on the cellular phone synchronizes and calms down the brain function frequencies. This fact proves that MRET-Shield neutralizes the electromagnetic stress and excitement caused by the effect of cellular phone on the brain functions. Cranial Nerve Cell Test Cranial Nerve Cell Test was conducted by Tex Chu, Inc., Taiwan on 30 human subjects to verify the immune system reactions to the exposure of human subjects to different sources of electromagnetic radiation with MRET-Shield protection (blue color bar) and without MRET-Shield protection (green color bar) respectively. The results are valid with p<0.01. This test prooves that the instalation of MRET-Shield on different sources of EMR actually supports and enhances immune system.

Comparative Study of the Effect of Microwave Radiation Neutralizers on Physiological State of Human Subjects

This research was conducted at Elsys Corp. in St. Petersburg, Russia with the help of Vibraimage method. It was designed to investigate whether microwave (MW) radiation of mobile phone can induce changes in the human's physiological state. This research included the testing of Electromagnetic Radiation Optimum Neutralizer MRET-Shield (United States Patent No. 6,369,399 B1) and three other competitive neutralizers: made in Japan Clean Pulse 21 (JP), made in Russia Aires (RU), and made in Taiwan Black But (TW), in order to determine their effects as protective devices against MW radiation negative physiological effects. MW effects were studied on mobile phone Nokia, model 3310 operating in the frequency range of 895-917MHz, with power output in the range of 0.002-2W, and SAR value less than 5.4mW/kg. Tested human subjects were controlled by Genius Video Cam Web (V1) connected with NB Samsung P30 (1.6GHz). The light illumination of each tested subject was set at the level of 500Lx.

EXAMPLES OF VIBRAIMAGE SYSTEM IMAGES AND HISTOGRAMS ACQUIRED DURING THE TESTING PROCEDURE:

Figure 1. The image of a subject in "FM" mode before and after 5 minutes exposure of the human subject to the mobile phone radiation in talk mode without neutralizer.
Figure 2. The image of a subject in "FM" mode before and after 5 minutes exposure of the human subject to the mobile radiation in talk mode with MRET-Shield.

The images on Figure 1 show substantial alterations in the form of Vibraimage color spectrum after the exposure of the human subject to the mobile phone radiation without any neutralizer. They show the increase of yellow and green areas in this spectrum, which identifies the increase of frequency values. This fact is confirmed by the substantial level of changes in the form of histograms on Fig.3. These histograms show the substantial increase of the mean values of frequencies and the increase of the percentage rate of the higher frequency values in the Frequency Distribution after the exposure of the human subject to the mobile phone.
The images on Figure 2 show insignificant alterations in the form of Vibraimage color spectrum after the exposure to the mobile phone with MRET-Shield. They show the slight increase of yellow and green color areas in this spectrum, which identifies the slight increase of frequency values. This fact is confirmed by the very slight level of changes in the form of histograms on Fig.4, comparing to the level of changes on Fig.3 for the mobile phone without MRET-Shield installation. The histograms on Fig.4 show the non substantial increase of the mean values of frequencies and the insignificant increase of percentage rate of the higher frequency values in the Distribution after the exposure of the human subject to the mobile phone with MRET-Shield.

Figure 3. "FM" mode: the histograms of human body Frequency Distribution before (red diagram) and after (blue diagram) 5 minutes exposure to the mobile phone in talk mode without any neutralizer.

Figure 4. "FM" mode: the histograms of human body Frequency Distribution before (red diagram) and after (blue diagram) 5 minutes exposure to the mobile phone in talk mode with MRET-Shield.

Figure 5 ."FM" mode: the histograms of human body Frequency Distribution before (red diagram) and after (blue diagram) 5 minutes exposure to the mobile phone in talk mode with Japanese (JP) neutralizer.

Figure 6."FM" mode: the histograms of human body Frequency Distribution before (red diagram) and after (blue diagram) 5 minutes exposure to the mobile phone in talk mode with Taiwanese (TW) neutralizer.

Figure 7. "FM" mode: the histograms of human body Frequency Distribution before (red diagram) and after (blue diagram) 5 minutes exposure to the mobile phone in talk mode with Russian (RU) neutralizer.

Normalized changes of the Mean Value of Frequencies of the human body micro vibrations following the 5 minutes exposure to the mobile phone radiation with and without MW radiation protective devices (neutralizers). The mean values are statistically significant with p<0.001.

Normalized changes of the Width of Frequency Range of the human body micro vibrations following the 5 minutes exposure of human body to the mobile phone radiation with and without MW radiation protective devices (neutralizers). The mean values are statistically significant with p<0.001.

CONCLUSIONS:

1. This research shows that there are statistically significant changes in the physiological state of the human subjects after the 5 minutes exposure to the mobile phone radiation in talk mode.
2. The installation of MRET-Shield on the mobile phone significantly reduces the level of changes in the physiological state of the human subjects and makes them insignificant.
3. The installation of all other tested neutralizers on the mobile phone, on the contrary, increases the level of changes in the physiological state of the human subjects.