Detection of Electrophysical Relationships in the Use of Biologically Effective Electro-Stress Reduction Devices by Cell Culture Observations

König FM and Dartsch PC

Published on: 2020-12-13

Abstract

Mobile phone data transmission technologies like 4G, UMTS and GSM are fully established now. Furthermore, mobile phones have been considered to emit only a low intensity and minimal health damaging radiation when they are actively working more or less placed near to the human head. However, the safety aspects are not clearly immersive evaluated. To address this topic, we have already investigated again in human cellular effects related to a used smartphone field source and additionally used electro-stress relief systems (ERS). This ERS were placed around cell shells or were connected directly to the mobile phone headphone plug-in.

Connective tissue fibroblasts (cell line L-929) were exposed to the radiation of an actively transmitting commercially available mobile phone with 1.25 W/m2 at the level of the cells with and without the ERS. This was done and observed (due to cell vitality values) in four different experimental setups. Unexposed cells in a similar incubator served as corresponding controls. The resulting cell vitality was checked by measurement of the enzymatic activity of mitochondrial dehydrogenases by the color change of the sodium salt 2,3-bis[2-methoxy-4-nitro-5-sulfo-pheny]-2H-tetrazolium-5-carboxyanilide (XTT).

The investigation results clearly demonstrate that mobile phone radiations caused a significantly (bio-cellular or bio system or probably health conditions relevant) increase effect on the cell vitality by an additional use of ERS. This was accompanied by marked morphological changes in the cells such as intracellular vacuolization, rounding and detachment. Moreover, the increased of cell vitality was much more given if the ERS coil windings were near place to the cell culture holes (hint: effect hotspot) and if the wires of the ERS were directly connected to the mobile phone (see maximum increase of the cell vitality values). In an equally present research context, it was proven that the role of the ERS direction due to the special ERS coil arrangements directed or trimmed around 90 degrees in two axis ways related to the cell shells extracted a different effectiveness of the ERS (coil), which could be proven as an experimental starting hypothesis. Besides, this ERS coil direction and emitted probable bi-lateral coil field effect on cell culture seem to have a correlation to electro-physical laws like the so called “induction law” or “LENZ rule” regarding magnetic fields itself.

Keywords

Electromagnetic field; Mobile phone modulation; Cell vitality; Cell culture

Introduction

The use of electrotechnical devices and the resulting telecommunication devices have caused man-made, synthetic alternating field propagation for more than 100 years now. This has been observed for decades with regards to its biological effectiveness, to what extent such technical useful signals of, for example, 50/60 Hz voltage-supplied electrical engineering devices in the household or office up to modulated wireless data transmission technologies with their useful signal emissions whatever kind of impact on earth organisms [1-15]. In this context, a cell test method mainly used from the pharmaceutical industry has proven to be recommendable in the recent past [16,17,18], which can determine an electromagnetic compatibility of low / high-frequency man-made harmful or harmless to health radiations [19,20]. Thus, it is possible to identify the quality of radiation sources or to compare them to other sources as well as to correlate them in tests procedures around the electromagnetic compatibility or collect them epidemiological studies seeing the subject of "electro-smog". Or at least to compare and evaluate so-called portable "electro-stress reduction (relief) systems" (ERS; [21]) including so-called different types of harmonizers in their effectiveness bio-chemically [16,17,18]. In addition, another parallelly given correlating phenomena was discovered that exists electrophysical parallel to the electromagnetic radiation emission: Marginal μ-gram mass / weight changes which also allow evidences of the connection between an alternating field existence and possible human tactile alternating field influences and as well their field effect neutralizations on living beings. Thereby were used radiation sources like wireless telephones based on the DECT standard or cell phones / smartphones (see 4G, GSM). In summary, it was therefore firstly stated that alternating field emitters or their emitted health influencing biochemical effects on biological systems can also be “manipulated”. Second, depending on the ERS technology, it is not clear in which electrophysical or other manner, for example ERS or the above mentioned existing so-called “beneficial” harmonizers work; whether there “should exist” demonstrable correlations to electrophysical engineering textbook laws. For this purpose, LENZ rule [22], induction law [23], applied to directed field propagations according to a HELMHOLTZ coil [24] arrangement (and supposed bilateral incoming or outgoing alternating field mode interactions).

Exactly this experimental approach is pursued in the present investigations, since the applied ERS has its design origin in a high-frequency signal transmission technology use; see ERS based on copper waveguide elements and a so-called "wave swamp" [16,17,18].

Based on empirical observations it is postulated as an experimental starting hypothesis that the artificially coils generate alternating field action directions or interact with radiation sources. Thus, those ERS coils (as HELHOLTZ coils) standing vertically or tilted 90 degrees, i.e. arranged in a planar / vertical axis-like manner, can interact with action detectors, which in experimental form preferably takes place with exposed cell cultures (several cell samples in a group of holes in a cell dish).In the thought hypothesis, this should lead then to different cell vitality values ??as a reaction between four respective planned experiments. Anyway, as final experimental step, the ERS should even be galvanically connected directly to a mobile phone interface, which provokes a direct interaction between the ERS and the smartphone; not only an observation of bi-directionally effecting fields of sources and influencing elements. This provokes a differing observation view in alternating field influences and signal influences on exposed cell lines. It should also be emphasized, and it has already been proven in previous attempts, that in such experiments with the ERS no electromagnetically detectable changes in the alternating field form over the intensity, time or frequency (spectrum) is or was given. Nevertheless, there are often repeated changes in the effect that are now specifically to be checked on the basics of the field line direction (see ERS coil alignment towards the field or towards the cell shells) as a test manipulated variable factor, whereby variable or different effect would be assumed.

Materials and Methods

Cell Culture and Experimental Design

The background of recently presented studies [16,17,18] was based on cultured connective tissue fibroblasts (cell line L-929) as a standard cell line for toxicological studies. Cells were used over several passages and a total experimental period of approximately 3 months. The used cells were routinely cultivated in the moist atmosphere of an incubator at 37 °C and gassed with 5 % CO2 and 95 % air to yield a constant pH value of 7.4. The culture medium used was RPMI 1640 with 10 % growth mixture and standard amounts of gentamycin. All culture reagents were from Capricorn Scientific, 35085 Ebsdorfergrund, Germany.

For the earlier designated tests, cells were seeded from 80 to 90 % confluent mass cultures at a density of 20,000 cells/wells into at least 14 wells in the middle part of a 96 well plate. After 24 hours to ensure cell attachment and metabolization, culture medium was exchanged to Leibowitz L-15 medium containing 10 % growth mixture and standard amounts of gentamycin. The latter culture medium guaranteed a pH value at 7.4 at normal atmospheric conditions. Each plate used was transferred to an external mini incubator and cultivated further at 37 ± 1 °C without CO2 gassing. Directly into the mini incubator a commercially radiation source like a mobile phone having a continuous operation mode was placed. Radiation intensities were measured at the level of the cells at the same conditions as used later on for the assays with a measuring device (Spectran HF-4060; Aaronia, Strickscheid, Germany) with a calibrated area antenna of 1 cm2. An intensity of 1.8 W/m2 was measured for the actively transmitting mobile phone at the level of the cells. When a corrugated cardboard as already used in previous experiments [16,17,18] was placed between the mobile phone and the cells to avoid thermal influence by microwave radiation, the intensity was 1.25 W/m2. Hereby, when the corrugated cardboard was used, a value of 37.5 to 38 °C at the cover lid of the multiwell plates with the cells was measured. In conclusion it is to point out, that the experimental design omitted local thermal effects. The incubator temperature was kept constantly at 37 ± 1°C. Every cell test was conducted with unexposed control cells at the same cultivation conditions, but approximately 15 meters distant from the exposed cells.

Cells in multiwell plates were exposed to mobile phone radiation for 2 hours. Then, medium was exchanged to the primary culture medium and cells were cultivated for another 22 hours to allow cells to react after the radiation.

The main investigative factor cell vitality was checked by morphological observation of the cell cultures and by enzymatic activity. Regarding the second method, cell culture medium was finally replaced by fresh culture medium containing 10 % of 2,3-bis[2-methoxy-4-nitro-5-sulfo-pheny]-2H-tetrazolium-5-carboxyanilide (XTT; Xeno­metrix AG, Allschwil, Switzerland) and additionally incubated for 120 minutes in the incubator at 37 °C. By the mitochondrial activity oin viable cells, the yellowish XTT is cleaved to an orange formazan. Accordingly, the amount of formazan dye formed directly correlates to the number of metabolically active cells in the culture [25,26].

The optical density was measured as a differential measurement ΔOD = 450 – 690 nm after 4 seconds shaking interval using an ELISA reader (BioTek Slx808 with software Gen5 version 3.0; Bad Friedrichshall; Germany). Statistical analysis of all test assays was done using the two-tailed Wilcoxon-Mann-Whitney test.

Electro-Stress Relief Systems (ERS)

Fundamentally, another type of passive ERS consisted of filled up room compartments / waveguid with a length of around 35 cm and a tube diameter of 5 cm without any additional electronic parts. Thus, the previously described ERS [21] was provided with layers of material of iron, zinc, copper, carbon or carbon related granulate materials and varying quartz (SiO2) granulates. The utilization of these so-called hollow conductor elements (or ERS) was adopted from the usual high frequency electromagnetic signal transmission technologies [1-15,21]. Furthermore, this ERS construction was motivated by a so-called broadband signal absorbing “wave swump” [27,28] as it is common for high frequency circuit corner (antenna) element purposes.

Now for the present experiments was used an extended system in which a copper partition was inserted in the middle of the layered provided waveguide ERS (similar to the ERS filled up identically in two chambers inside the copper tube). Besides, two wires are inserted or embedded into one end of the filled copper tubes and separated into two chambers up to the opposite tube end; see figure Figure 1 [21]). Thus, two pieces of wire protrude from one end of the copper tube, which can either be used to form a continuous, coil-like element, such as the earlier named HELMHOLTZ coil in idle mode, or are directly connected to a radiation source (see a plug-in connection or interfaces on electronical devices like mobile phones).

Figure 1: An ERS is illustrated in a top view of a technical drawing. The element number 1 shows a copper tube (length: 5 to 10 centimeter’s), 2 a dividing copper partition element (realizes two chambers), 3 the filling different granulate layers in chamber I (see “wave swamp” materials like carbon or carbon related and varying quartz {SiO2} granulates). The element 4 shows different granulate layers in a chamber II having the same granulate materials. The element 5 and 6 illustrate the wires or wire ends inside the chambers I and II. Finally, the element 7 shows the case or device housing.

Four Different Cell Test Setups

For the present investigations, four different test arrangements were set up. Three of them contained an ERS with a HELMHOLTZ coil winding and one a direct cable connection, i.e. a galvanic connection to a smartphone, and finally irradiated dishes equipped with cells as follows:

Experiment 1: A coil was wrapped vertically or vertically in the center around the cell shell. The assumed main ERS (field influencing) action or interaction of the ERS coil (cf. red arrow) is perpendicular to the coil alignment and thus horizontal / parallel to the cell shell plane or to the shell perforations (see Figure. 2).

Figure 2: There are shown the setups of experiment 1 and 2 having vertical (A1) and horizontal coil winding (C1) around the cell shells; the green or blue colored coil was immersive built-in to the ERS for these experiments 1 and 2. On the other hand, it is illustrated the imaginable coils field effects direction on the cell cultures by a red colored arrow vector (A2) for the first experiment.

Experiment 2: A coil was wrapped horizontally around the cell shell. The assumed main ERS (field influencing) action or interaction of the ERS coil is perpendicular to the coil alignment and thus vertically crossing the cell shell plane or the shell perforations (see Figure. 2).

Experiment 3: A coil was wrapped vertically or vertically around the smartphone. The assumed main ERS (field influencing) action or interaction of the ERS coil (see red arrow) is horizontal to the coil orientation and thus horizontal and parallel to the cell shell plane or to the shell holes (see Figure 3).

Figure 3: It is shown a vertical coil winding of the experiment 3 and an illustrated imaginable coils field effect direction on cell cultures (see red colored arrow vector). The blue colored coil was connected to the ERS (brown color device; it is placed nearby the cell shell inside the mini incubator).

Experiment 4: From the two ERS chambers I and II (as a dipole cable element 3 and 4 in Fig. 1) two wires according to elements 5 and 6 are led out and directly connected to a smartphone interface in an electrically conductive manner. Thus, the dipole cable at first immersed in the ERS and secondly it was plugged in directly into the headphone socket of the smartphone (see Figure. 4).

Figure 4: It is shown a wired connection between the ERS directly to the headphone plug-in of a smartphone.

In the present four partial experiments and the associated following statistical measurement results evaluation, the cell vitality values obtained were compared with the initial state with additive irradiation via the mobile phone (and without ERS) using a difference value calculation. A measurement without additive irradiation via the mobile phone, let alone without using the ERS, was also implemented as control result basics. It should also be emphasized that extraordinary result parts were marked with regard to a hotspot in which the ERS coil or wire windings were in the immediate vicinity of the cell culture holes; relevance, see later by interpretations of the investigation results based for instance a proven existence of the LENZ rule.

Results and Discussion

Accordingly, a statistical evaluation of the cell vitality values / samples obtained was carried out using the four different test arrangements mentioned above. In the summarizing table were named the experiments in an ascending order of the mean values, which demonstrate the test tendencies in such a way that the ERS effectiveness correlates by an increase of the cell vitality with; hint: Given parallel comparison test "without / with ERS" related to the test design or the application of the ERS results as follows:

  1. Experiment 2 - A coil was wrapped horizontally around the cell shell; Mean = 3.71875%; see Figure 6.
  2. Experiment 1 - A coil was wrapped vertically or vertically in the center around the cell shell mean = 9.91875%; see Figure 5.
  3. Experiment 3 - A coil was wrapped vertically or vertically around the smartphone; Mean = 11.66875%; see Figure 7.
  4. Experiment 4 - The dipole cable immersed in the ERS was plugged directly into the headphone socket of the smartphone; Mean = 21.1875%; see Figures 8.

On the other hand, there are existing slight changes in the upper named sequence 1. To 4. when the focus is on the hotspot values of the evaluation considerations, i.e. where the ERS coil / wire is very close to the cell sample holes; this shows the following sequential listing of experiments in order:

  1. Experiment 3 - A coil was wrapped vertically or vertically around the smartphone; Mean = 6.225%; see Figure 7.
  2. Experiment 2 - A coil was wrapped horizontally around the cell shell; Mean = 11.9125%; see Figure 6.
  3. Experiment 4 - The dipole cable immersed in the ERS was plugged directly into the headphone socket of the smartphone; Mean = 21.1875% (see overall mean); see Figure 8.
  4. Experiment 1 - A coil was wrapped vertically or vertically in the center around the cell shell; Mean = 23.3625%; see Figure 5.

Experiment 4 (compare to the illustrated results of Figure 8) cannot or should not actually be counted because the direct wire connection to the smartphone prevents such a statement. However, the result should still be included in the "hotspot" sequence above; see relevance and consequences in the discussion of the results below.

The changes in the cell vitality values obtained recorded primary the connection between radiation effect of high-frequency mobile phone signals (4G smartphone as source) and their effects on human cell lines. In addition, in the four different test setup results were used ERS additionally / parallelly to an electro-magnetic field emitting smartphone. The presented results offer similar tendencies to earlier investigations [16,17,18,24] seeing beneficial effects on exposed human cell lines. “Similar” as well, because the ERS device constructions in the actual experimental use are not identic; hint for instance: The ERS now are smaller and they have differing made two chambers as illustrated in Figure 1. However, this given ERS for the actual given discussed experiments obviously demonstrates again: With a measurably unchanged electromagnetic radiation disposition by the used smartphone in the near field to cell shells it was generated a cell vitality-increasing effects at each four experiments. Essentially, depending on the transmission antenna cable harness and the different types of wire wrapping implemented, a different effectiveness or impact quality of the ERS emerged. Consequently, something like the HELMHOLTZ coil winding type arrangements formed “cell vitality beneficial” effects around the exposed cells. Furthermore a directly connected ERS on a smartphone have specifically different more intensive “cell vitality beneficial” effects, which in comparison to each other experimental setup can be clearly interpreted via the laws of the field line direction (see LENZ rule, induction law, etc.) or in its action depth; see comparative percentages of the ERS benefits by four different test setups in each of the following Figures 5 to 8.

Figure 5: It is shown the results of experiment 1; vertical coil. The abscissa illustrates each cell tests or probe number (1 to 16 / left to right); in addition, it is shown the “hot spot mean value” (going right after 16 bars at the graph) and finally right side at the bar graph the “total mean value” in percent again. Seeing this experiment results each cell test comparison value is determined at the ordinate in “%”; means it’s cell vitality difference calculation values in percent regarding “mobile phone radiated cells” compared to “mobile phone radiated cells plus ERS coil constructions added”.

Figure 6: It is shown the results of experiment 2; horizontal coil. The abscissa illustrates each cell tests or probe number (1 to 16 / left to right); in addition, it is shown the “hot spot mean value” (going right after 16 bars at the graph) and finally right side at the bar graph the “total mean value” in percent again. Seeing this experiment results each cell test comparison value is determined at the ordinate in “%”; means it’s cell vitality difference calculation values in percent regarding “mobile phone radiated cells” compared to “mobile phone radiated cells plus ERS coil constructions added”.

Figure 7: It is shown the results of experiment 3; vertical coil around the mobile phone. The abscissa illustrates each cell tests or probe number (1 to 16 / left to right); in addition, it is shown the “hot spot mean value” (going right after 16 bars at the graph) and finally right side at the bar graph the “total mean value” in percent again. Seeing this experiment results each cell test comparison value is determined at the ordinate in “%”; means it’s cell vitality difference calculation values in percent regarding “mobile phone radiated cells” compared to “mobile phone radiated cells plus ERS coil constructions added”.

Figure 8: It is shown the results of experiment 4; the ERS was connected directly to the smartphone / mobile phone. The abscissa illustrates each cell tests or probe number (1 to 16 / left to right at the bar graph). And finally, right side at the bar graph it is shown the “total mean value” in percent again. Seeing this experiment results each cell test comparison value is determined at the ordinate in “%”; means it’s cell vitality difference calculation values in percent regarding “mobile phone radiated cells” compared to “mobile phone radiated cells plus ERS directly connected to it”.

As explained at the beginning, the test results should be used to examine the extent to which the respective coil alignment, rotated by 90 degrees, correlates with the induction law and the electrotechnical LENZ rule; so also, the suspension hypothesis, which had to be confirmed. The postulated effect of the ERS coil winding could also be almost the same over experiments 1 to 3, which did not occur and especially the direct connection of the ERS to the actively radiating mobile phone reveal the maximum "benificial" effect on the exposed cell lines. In total, it was confirmed with Experiments 1 to 4 that

  • The explicitly applied coil alignment and the field direction created with it on the cells are most effective when there is complete penetration or extraordinary proximity to the wire of the coil (see hotspot position), i.e. the field line perpendicular to the cell lines. This effect of the coil wire winding in the vicinity of the cell perforations thus seems to be interpreted according to the LENZ rule or so-called "right hand rule", i.e. a wire / conductor through which a current flow and the field lines spreading around the coil wire / conductor. Otherwise the mean hotspot value the?? experiment 1 wouldn’t be the highest or higher than the existing at the experiments 2 and 3.
  • And at least this hotspot mean value (in experiment 1) is higher than the cell vitality over all mean value of experiment 4 having a direct wire connection to the smartphone headphone plug-in.

Besides, if the ERS is galvanically coupled directly to the radiation source (see headphone socket), the cellular electromagnetic field impact reduction quality of the ERS is the highest on average according to an increase in cell vitality. Thus, the highest effectiveness due to the smartphone field exposed cell lines seems to be found precisely here in this experimental set-up 4; see the highest "over all mean" here. In other words: Experiment 4 was the most cell-biological “beneficial” effective, which means that there was no ERS coil winding, but a galvanically conductive cable connection directly to headphone amplifier electronics of the probable health damaging mobile phone. As I said, the data transmission radiation intensity or spectrum did not decrease measurable in the present experiments as in earlier experiments too [16,17,18,24].

Conclusion

In summary, the hypothesis set out at the beginning was confirmed to be correct experimentally: Electrophysical laws were existing having different arrangements of ERS coils (and its field directions) in relation to smartphone / mobile phone irradiated cell shells. Therefore, exist in the experimental or practical application of used ERS including its coils around cell cultures plus said high-frequency mobile phone alternating field expositions. This procures an emphasis of a cell vitality value reduction between an increase of more than only 3% (on average) up to almost “cell vitality reduction neutralized” based on alternating field if a mobile phone and ERS were used optimized by a direct wired connection to the field source shown by Figure 9. In other words, regarding “neutralized”, the mobile phone field overall average of -23 % cell vitality reductions was excited and neutralized having an ERS effect of nearly +23%. This means as well, that the smartphone field impact was neutralized at a bio-cellular testing level; the electromagnetic field exposition was untouched (identic as before without ERS). Why this contradiction is allowed in electro-physics has already been examined in more detail with [24]; see marginal changes in mass as a possible equivalent for an effect transfer or electromagnetic impact neutralization.

Figure 9: It is shown the results of experiment 4; the ERS was connected directly to the smartphone / mobile phone. The abscissa illustrates each cell tests or probe number (1 to 16 / left to right at the bar graph). And finally, right side at the bar graph it is shown the “total mean value” in percent again. Seeing this experiment results each cell test comparison value is determined at the ordinate in “%”; means it’s cell vitality difference calculation values in percent regarding the “control case” (which is “no used mobile phone and no ERS”) compared to “an active mobile phone, which radiated cells plus an mobile phone directly wire connected ERS”.

The evaluation of the test results of the cell vitality values ??in the immediate vicinity of the ERS wire or coil windings is mostly interesting, because of a resulted higher increase in cell vitality at all experiments; see Figure 5 to 7. What kind of electromagnetic field corresponding relationships by ERS wires / coils and its correlation effects on cell culture can’t be described in teaching book physical legislations, but the evidence of an existing field directional effect (see vector of Figures 2 and 3) ERS coil arrangements around the cell and mobile phones was clearly confirmed as the initial declared experimental thesis. From a purely functional point of view, the ERS coil arrangement or the direct insertion of the ERS wires into a smartphone seem to work “like a kind of cell biological IMPACT vacuum cleaner”, which in the future must be followed up in terms of electrophysical system theory. Thus, the directional effect according to field line observations of the ERS coils (compare to [22,23,24]), on the other hand, has been demonstrated and is consequently highly relevant or significant as an bio-chemical or at cellular levels influencing efficiency or manipulated variable of the action effectiveness of ERS supposed on human health condition optimizations [21] at the modern wire-less data transmission technology using urban living areas. This is important as well because of the rising number of published international studies around the (wire-less / telecommunication) electro-magnetic field impact [1-15].

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