Synopsis: Exposure of Normal Human Astrocytes Cells to Mobile Phone Radiation with and without MRET-Nylon Protection
Igor Smirnov, Ph.D.
Global Quantech Inc., California, USA
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
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: