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| PROTECTIVE EFFECT ON HUMAN PERIPHERAL BLOOD LYMPHOCYTES OF A DEVICE (TECNO AO) CLAIMING TO PROTECT FROM ADVERSE EFFECTS OF CELLPHONE RADIATION | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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(note: this study is not peer reviewed) Roger Coghill and Tamara Galonja-Coghill SUMMARY The objective of this study was to test the manufacturer’s claim that a device called Tecno AO has a beneficial biological effect on human health. The cell model chosen was the lymphocyte because of its easy availability and its well characterised importance to immune defence against infection and in tumour oncostasis. Moreover there are a large number of studies reporting sensitivity of lymphocytes to biophysical agents, including electromagnetic fields and radiations. The adverse effects on human peripheral blood lymphocytes of RF/MW exposure from cellphones was first verified. Viability of lymphocytes following exposure to the device was tested by trypan blue exclusion. It was found that the device had a significantly protective effect against overnight exposure to electromagnetic fields and radiations from a mobile phone on standby, compared both with cellphone-exposed cells not exposed to the device, and with controls. The mechanism remains unknown and further research is necessary to verify this effect. METHOD AND MATERIALS Human peripheral blood lymphocytes were isolated from 20ml fresh whole blood drawn from vena cubitale into anti-coagulated vacutainerrs (Becton Dickinson, EDTA, K3), transported into four 5ml sterile test tubes, and differentially centrifuged at 450g. The serum was removed for heat inactivation, and then the buffy coat was detached by micropipetter with as little disturbance as possible of the red blood cells and platlets which were discarded. The collected buffy coat (approx 2ml) was mixed with an equal amount of density gradient Ficoll-Triosil prepared accordng to standard procedures, and centrifuged for 5 minutes at 800g. The lymphocytes were removed as the layer between the density gradient and the remaining serum, which contained the platelets. The pellet was resuspended in balanced saline solution with added glucose, and washed twice by centrifugation at 100g for five minutes. To the final pellet was added a culture medium (RPMI-1640 plus antibiotics and antimycotics). The medium was divided into three samples and one of these were exposed to a device known as Tecno AO. Another sample was not exposed to the device, whilst the other served as control. One Tecno AO-exposed sample and one unexposed sample were placed in separate mu-metal boxes and then exposed overnight to a Philips 301 mobile phone on standby, by means of a separate 30cm gold wire leading into the box. For the same period one sample was placed in a separate mu-metal box, into which separate but adjacent gold wires were introduced, but kept in a separate room and not near the cellphone. The final (third) sample was enclosed in a mu-metal box and then placed inside a double skinned mu-metal container for the duration of the experiment and maintained at 20 ° C. On the day following exposure the cells were mixed sequentially prior to counting with trypan blue dye left for 15 minutes and then counted double-blind in a haemocytometer (Brightline, Hausser-Scientific ) in accordance with the method recommended by the suppliers (Sigma-Aldrich Co., Poole Dorset, UK). At least 400 cells were counted from each sample. After counting the codes were broken and the results analysed statistically. Results It was first necessary to establish whether cellphones do actually adversely affect the tissues of interest (human peripheral blood lymphocytes). This was done by exposing them in various conditions, including two different cellphone models (Nokia 2410 and Philips 301). These results are shown below Table 1: Lymphocytes exposed to Endogenous & RF/MW electric fields Cell numbers counted blind 6 hrs after exposure, 28 October 1998 A. RF/MW -exposed for 8 hrs. to cellphone in standby mode
B. Endogenous electric field-exposed for 8 hrs.
C. Sham-exposed for 8 hrs
D. Controls (unexposed for 8 hrs, and also enclosed in mu-metal container)
SUMMARY
Statistical analysis of the data One must first establish whether there is any statistically significant difference between viability of the RF/MW exposed cells and the unexposed controls. The nul hypothesis is that there should be no difference between cells exposed to RF/MW and those not so exposed. Clearly if there is no difference then there is no need for a protective device, and this is the argument sometimes advanced by those in the cell phone industry, who say that there is no convincing evidence of adverse health from use of mobile phones. This however ignores a number of cellular, live animal, human and epidemiological studies all suggesting the existence of ill health from excessive use of cellphones. These studies are discussed elsewhere. In this study we have predicated that adverse effects on lymphocytes, which are vital for immune competence and tumour immunity, constitutes an ill health effect. The proper statistical test is to examine the difference between proportions (z test) of RF/MW exposed and the sham & unexposed. The calculations are: RF/MW exposed Sham/unexposed Totals
Thus p = 1472/ 3517 = 0.4185 and so q = 1 - 0.4185 = 0.5815. Since nT = 858 and nC = 2659, and each of nTp, nTq, nCp and nCq are all well in excess of 5, the critical ratio (z) becomes: 445-0.5 - 1027 + 0.5 --------- ------------- zC = 858 2659 = 0.132/Ö 0.0004 = 6.6 --------------------------------------------------- Ö (0.4185) (0.5815) (1/858 + 1/2659) which is significant (p>0.001). Therefore the RF/MW exposed cells are significantly less viable than the sham/unexposed cells. CHART ONE  Thus these results, based on some 4000 cells counted blind, confirm that the donor’s endogenous field is protective, and that RF/MW radiations adversely affect the viability of lymphocytes as measured by trypan blue exclusion. The next step was to see whether there is any protective effect of the Tecno AO device. The results are shown graphically below in chart format (Chart Two): One sees in the first column how the normal endogenous fields of the donor are protective of the lymphocytes’ viability. In the second column the effects of exposure to RF/MW radiations from the cellphone on standby have lowered the viability, but not so much as when the Tecno AO device is not present: CHART TWO    Statistical Test of Tecno AO effectiveness The question now arises whether the difference in viability between the RF/MW exposed cells and those protected by the Tecno AO device is sufficient to be statistically significant. Again, the appropriate test is the difference between proportions (z-test). The further question is whether the correct data is that from day one after exposure or day two. We decided that the more chronic effects should be more important, since an early recovery from the exposure would plausibly be less biologically significant. The calculations are as follows: Tecno AO protected RF/MW exposed Total
p = 171/1001 = 0.171 and q = 1- 0.171 = 0.829 nT =, 435 and nC = 566. Since each of nTp, nTq, nCp, and nCq are certainly well over 5, the critical ratio becomes: 96-0.5 - 75 + 0.5 0.2195 - 0.1334 --------- ------------ == ----------------------- = 3.58 435 566 Ö 0.1418 x 0.0041 Zc = ------------------------------ Ö (0.171) (0.829) (1/435 + 1/566) which is significantly different (p= 0.0002). Therefore there is a significantly protective effect from the Tecno AO device against RF/MW radiation from the cellphone on standby. DISCUSSION The statistical analysis of these data indicates a significant protective effect when the Tecno AO -protected cells are compared for viability with the RF/MW exposed cells. Our previous studies of lymphocyte viability (and indeed this study also) showed that there is also a protective effect when similar cultures are exposed to the donor’s own endogenous fields, which viability exceeding 70 percent in such conditions. It would appear that the device is in some way able to mimic this beneficial effect, though not completely so. The mechanism of interaction is not known, and any hypothesis of the possible mechanisms must at this stage be considered speculative. It would be useful to replicate this experiment to confirm that the effect reported here is robust. A number of possibilities present themselves based on what is known currently of the biological effects of weak energies. For example it is noted that the cell numbers of the energised sample per unit volume were higher than unenergised or control samples, suggesting that cell surface markers leading to apoptosis were not being expressed by the cells cultured in the protected culture. The feature distinguishing endogenous field-exposed cells from RF/MW-exposed (whether or not they were Tecno AO protected) is that not only were the absolute cell numbers in the RF/MW exposed cells lower, but the non-cviable cell numbers were also higher. In both regards the Tecno AO-protected cells fared better, but not so well as the unexposed benefiting from the endogenous fields. We speculate that one possible way in which the device may operate is by transducing the radiations into frequencies which do not disturb lymphocyte viability. Further work is necessary to investigate this possibility. CONCLUSIONS The study confirmed that a donor’s endogenous electric fields have a protective effect on lymphocyte viability. Using the same model we found that weak RF/MW radiation from cellphones on standby have an evidently adverse effect. Finally although viable Tecno AO -exposed cell numbers were still lower than when the cells were exposed to the body’s natural fields, the effect of the Tecno AO device afforded some protection against these radiations even two days after cessation of the RF/MW exposure. |
