Study On Immune Cells

Beneficial Cell Effects Of QiOne® 2 Pro

Increased Cell Metabolism

Protection from E-Smog

1. Background And Question Of The Present Study

According to the manufacturer, Qi Blanco UG (haftungsbeschränkt) from Germany, QiOne® 2 Pro contains a GitterChip™ which forms a static field that stimulates water molecules to undergo a transition into the coherent state. Since our body consists of about 70 to 90 % of water (depending on age), the coherent state of the water molecules influences the cells of our whole body. In this present in vitro study, current cell biological methods were used to investigate whether the use of QiOne® 2 Pro results in any beneficial effects which can be proved by scientifically accepted bioassays.

2. Description Of Cells Used For This Study

Neutrophils are the most abundant type of granulocytes which make up about 60 % of all white blood cells in humans and are normally found in the bloodstream. They form an essential part of the innate immune system and, therefore, play a key role in the front-line defense against invading microbial pathogens. They are also one of the first responders of inflammation. They are able to generate highly reactive superoxide anion radicals in the course of a so-called oxidative or respiratory burst.

3. Basic Experimental Design

The investigations presented here were conducted with human promyelocytes (HL-60 cell line; ACC-3; ECACC 98070106; Leibniz Institut, DSMZ, Braunschweig, Germany). The cells were routinely grown in RPMI 1640 medium with 10 % growth mixture and 0.5 % gentamycin and cultivated in an incubator at 37 °C in an atmosphere of 5 % CO2 and 95 % air at 98 % humidity.

The non-adherent cells were routinely cultivated as suspension mass cultures and were regularly subcultured twice a week with fresh culture medium. By adding 1.5 % dimethyl sulfoxide to the culture medium, the cells were differentiated over a period of 6 days into functional neutrophils which are capable of generating superoxide anion radicals after the addition of a phorbol ester. The radicals result in a cleavage of the tetrazolium dye WST-1 (Roche Diagnostics, Mannheim), which was also added to the reaction mixture. The amount of oxygen radicals present is directly proportional to the cleavage of the dye, i.e. the higher the amount of reactive radicals, the stronger is the cleavage of the dye and the change of the optical density in the reaction mixture.

The optical density (= color change) of the reaction was measured by a differential measurement ∆OD = 450 – 690 nm at definite time points by an Elisareader (BioTek SLx 808 with software Gen 5/3.00) and calculated with Microsoft Excel 2016. Moreover, the basal metabolism of the functional neutrophils was checked in the same manner, but without phorbol ester stimulation.

4. Long-term Effect Of QiOne® 2 Pro

HL-60 cells which were differentiated to become functional neutrophils as described above were continuously exposed to QiOne® 2 Pro during the differentiation process for a period of 6 days. The distance between the cell suspension in plastic culture flasks and QiOne® 2 Pro was about 1 mm. Thereafter, basal cell metabolism and the superoxide anion radical generation was calculated. Four independent experiments with duplicate wells were con- ducted.

As depicted in Fig. 1, the test results showed that the basal cell metabolism was significantly increased in comparison to the untreated control by 12.5 ± 2.9 % (mean value ± standard deviation; p ≤ 0.05, two-tailed Wilcoxon-Mann-Whitney test). The generation of radicals was even more increased vs. untreated control by 27.7 ± 7.7 % (mean value ± standard deviation; p ≤ 0.01, two-tailed Wilcoxon-Mann-Whitney test).

Both results clearly demonstrate that the long-term use of QiOne® 2 Pro might increase the efficiency of the innate immune system in vivo by stimulating the basal metabolism and the radical generation of neutrophils which are floating in the blood, i.e. in an aqueous environment which is primarily influenced by QiOne® 2 Pro to undergo a transition into the coherent state.

Fig. 1: Presentation of the long-term results with QiOne® 2 Pro present during the differentiation process of promyelocytes to functional neutrophils for 6 days. Note the significant increase in basal cell metabolism and superoxide anion radical generation for the frontline defense against invading microbial pathogens. Controls are set as “0”. Data represent mean values ± standard deviations of 4 independent experiments with duplicate wells.

*p ≤ 0.05 vs. untreated control and **p ≤ 0.01 vs. untreated control; two-tailed Wilcoxon-Mann-Whitney test.

5. Inactivation Of Mobile Phone Radiation By QiOne® 2 Pro

Promyelocytes which have been differentiated to functional neutrophils for 6 days were ex- posed to the radiation of a current and commercially available mobile phone from a leading brand manufacturer with a SAR value of 0.76 W/kg. No distinction was made between thermal and non-thermal radiation, because both are also present in reality when making a call and have an effect on the human body.

The experimental setup was conducted in such a way that two culture flasks with the cells were placed on the display of the mobile phone and two culture bottles were placed beneath its back cover near the energy cells and the antenna. The mobile phone transmitted speech and environmental sounds discontinuously; the WLAN function was switched on and the display was switched-off. For clarification of the setup and the positions of the flasks and their nomenclature, see Fig. 2. This corresponded to the real situation of a radiation direction towards the user or away from the user. By this experimental design also varying radiation intensities at different local areas of the mobile phone could be examined. Three independent experiments with quadruplicate culture flasks were conducted.

The total exposure time to the actively transmitting mobile phone was 4 hours at 37 °C for each independent experiment (with and without the presence of QiOne® 2 Pro) in a temperature-controlled external mini incubator. Thereafter, the cells were transferred back to the standard incubator and incubated for another 20 hours before the intensity of superoxide anion radical generation in the course of an induced oxidative burst was examined.

Fig. 2: Experimental setup during exposure with quadruplicate samples placed on the mobile phone display (= direction of radiation towards the user) and placed beneath its back cover (= direction of radiation away from the user). QiOne® 2 Pro was placed between the culture flasks at each radiation level.

The positions of the flasks are marked as UL ( = upper left), UR (= upper right), LL (= lower left), LR (= lower right).

Fig. 3: Presentation of the results on the superoxide anion radical generation by functional neutrophils after 4 hours of exposure to the radiation of an actively transmitting mobile phone without protection and after protection by QiOne® 2 Pro. The untreated controls are set as „100 %“. The positions of the flasks are marked as UL ( = upper left), UR (= upper right), LL (= lower left), LR (= lower right). Data represent mean values ± standard deviations of 3 independent experiments. **p ≤ 0.01 vs. untreated control; two-tailed Wilcoxon-Mann-Whitney test.

6. Summary And Conclusions

The present study with cultured cells of the innate immune system (so-called functional neutrophils) has shown that the long-term use of QiOne® 2 Pro is able to promote basal cell metabolism and superoxide anion radical generation by these cells. This might result in a better front-line defense against invading microbial pathogens in the bloodstream. Moreover, QiOne® 2 Pro was also very effective in the inactivation of mobile phone radiation when the generation of reactive superoxide anion radicals was chosen as the measurement criterion. From our results the use of QiOne® 2 Pro can be highly recommended.

Study On Intestinal Cells

Protective Effect Of QiOne® 2 Pro On Cultured Intestinal Epithelial Cells After Mobile Phone Radiation

1. Introduction

The intestinal epithelium with single cell layer thickness has two essential tasks. The first one is to create a physical barrier between the contents of the intestinal lumen and the rest of our body. The second one is to ensure an efficient absorption of essential nutrients from the gut lumen and to produce mucus, anti-microbial peptides and cytokines with both protective and immune-regulatory properties. Thus, a reduced barrier function may have far reaching consequences, not only for intestine, but also for systemic health [1]. This barrier function can be destroyed by oxidative stress [2] as generated by mobile phone radiation. Despite the ubiquity of electromagnetic field emitters, the influence on living organisms and cells has become the subject of numerous research studies with contradictory results [3-6].

However, it has been demonstrated that oxidative stress plays a key role in the events causing cell death [7-11]. Although the gastrointestinal tract does not play a key role in the mobile phone radiation studies, there are some articles dealing with the effect of electromagnetic radiation on the intestine. For example, in an experimental animal study Sieroń et al. [12] demonstrated that electromagnetic radiations modified the redox balance in the gastrointestinal tract of rats. This might enhance the sensitivity against intestinal oxidative stress by injury or disease with all its unwanted health consequences. According to the manufacturer, Qi Blanco UG (haftungsbeschränkt) from Germany, the QiOne® 2 Pro device contains a GitterChip™ which forms a static field that stimulates water molecules to undergo a transition into the coherent state. Since our body consists of about 70-85% of water (depending on age), the coherent state of the water molecules positively influence the cells of our whole body. Moreover, QiOne® 2 Pro is stated to neutralize the unwanted effects of mobile phone radiation. In this present in vitro study, current bioassays were used to investigate whether the use of QiOne® 2 Pro might result in a protective effect on cultured intestinal epithelial cells against mobile phone radiation and the arising oxidative stress.

2. Material And Methods

QiOne® 2 Pro device

QiOne® 2 Pro was provided by Qi Blanco UG (haftungsbeschränkt), D-97711 Maßbach, Germany, for the duration of the experiments. The device is a second generation GitterChip™ of a specifically developed gold alloy with eightfold strength which creates a static field that stimulates water molecules to enter a coherent state in your body.

Mobile Phone

A current and commercially available mobile phone from a leading brand manufacturer with a SAR value of 0.76 W/kg was used. No distinction was made between thermal and nonthermal radiation, because both are also present in reality when making a call and have an effect on the human body 37°C in an atmosphere of 5% CO2 and 95% air at nearly 100% humidity. The cells were cultivated as mass cultures and were regularly subcultured twice a week with fresh culture medium. For the experiments, cells were taken from 80-90% confluent mass cultures.

Cell Regeneration After Mobile Phone Radiation

Cells were seeded at a density of 100,000 cells/ml into the four individual compartments of a silicone 4 well-culture insert made (ibidi, Gräfelfing, Germany). The single compartments of the inserts are separated by a 500 μm thick silicone bar with an outer silicone frame of 700 μm. Due to the special adhesion area, an insert adheres firmly to the bottom of a culture dish and forms a distinct cell-free area (artificial wound) in which the cells can colonize by migration and proliferation. Upon reaching confluency within 48 hours after cell seeding, the cells were exposed to the radiation of an actively transmitting mobile phone & WLAN with and without QiOne® 2 Pro. The cell culture dish was placed on the mobile phone display (direction of radiation towards the user) in a mini-incubator for 4 hours at 37°C without gassing (Figure 1). To avoid any pH changes during exposure at normal air conditions, the routine culture medium was replaced by Leibowitz L-15 medium with 1% growth mixture and 0.5% gentamycin. Cells which were cultured in the same way, but without mobile phone radiation, served as corresponding controls. After 4 hours of radiation, cells were incubated for another 8 hours in the same culture medium. Finally, cells were fixed with methanol, stained with Giemsa’s azur eosin methylene blue solution (Merck, Darmstadt, Germany), air-dried and examined by micrographs and a specialized software with artificial intelligence from KML Vision, Graz, Austria (IKOSA AI software).

Fig. 1: Arrangement of cell culture dish which was placed on the mobile phone display (direction of radiation towards the user) together with a QiOne® 2 Pro device on the right side.

Transepithelial Electrical Resistance (TEER) After Mobile Phone Radiation

IPEC-J2 cells were grown for 5 days on the surface of a 0.4 µm porous membrane (transwell plate, Corning, SigmaAldrich, Deisenhofen, Germany) which yields two seperated compartments within the cell culture dish. As a matter of fact, the cells covering the surface of the membrane represent a physical barrier against the lower compartment. TEER was measured by placing one electrode in the culture medium in the upper compartment and by placing one electrode in the culture medium in the lower compartment. Electrical resistance was directly measured by a portable voltmeter (Millicell ERS-2 Voltmeter, Millipore/Merck, Darmstadt, Germany) as described in detail elsewhere [1,13,14].

Epithelial cell layers were taken for the experiments at an electrical resistance of at least 2,000 Ω/cm2 representing an intact physical barrier with very good integrity and were exposed to the activel transmitting mobile phone mobile phone & WLAN with and without QiOne® 2 for 4 hours at 37°C as already described. After another 24 hours of cultivation, TEER was measured again and data were compared to each other. Three independent experiments were conducted. As a reference, TEER of the porous membranes without any cell barrier was measured to be 150 to 180 Ω/cm2.

Fig. 2: Original micrographs (left part) as well as the evaluation by IKOSA AI software (right part) with marked calculated frontlines of the IPEC-J2 cells closing the cell-free area. (A) Untreated control at the same culture conditions as below, but without any exposure to mobile phone radiation. (B) Unprotected cells after 4 hours of mobile phone radiation with subsequent cultivation for 8 hours. (C) QiOne® 2 Pro protected cells after 4 hours of mobile phone radiation with subsequent cultivation for 8 hours. Note the largely decreased colonization of the cell-free area in (B). Untreated control and QiOne® 2 Pro protected cells show a similar colonization pattern.

3. Results

Cell Regeneration

As seen in Figures 2 and 3, mobile phone radiation caused a reduction in cell regenerative activity by leaving a cell-free area which was 43.7 ± 9.0% (mean value ± standard deviation; n=4) in comparison to the total colonized cell area. In contrast, the residual cell-free area was only 13.8 ± 2.6% for the untreated control and 14.5 ± 1.6% for the QiOne® 2 Pro protected cells (both mean values ± standard deviations; n=4). The difference between the unprotected cells and QiOne® 2 Pro protected cells was statistically significant (p ≤ 0,01; two-tailed WilcoxonMann-Whitney test) demonstrating the effectiveness of the device against mobile phone radiation. Moreover, there was no statistically relevant difference between the QiOne® 2 Pro protected cells and the cells which were not exposed to mobile phone radiation at all.

Fig 3: Presentation of the results on cell regeneration after mobile phone exposure with and without QiOne® 2 Pro for 4 hours and subsequent cultivation for 8 hours. Note that the residual cell-free areas of untreated control and QiOne® 2 Pro protected cells do not differ from each other, whereas unprotected cells have a significant higher cell-free area due to a markedly reduced cell activity and even cell death. Data represent mean value ± standard deviation of four independent experiments.

Transepithelial Electrical Resistance (TEER)

As seen in Figure 4, mobile phone radiation caused a rupture on the epithelial barrier by causing cell death due to oxidative stress. In contrast, untreated controls and QiOne® 2 Pro protected cells did not show such massive morphological changes of the cell layers. This situation is reflected by the TEER values measured. For the unprotected cells a TEER of 152 ± 16 Ω/cm2 was measured demonstrating the complete loss of barrier integrity. The TEER value for the QiOne® 2 Pro protected cells was 1,837 ± 349 Ω/cm2 and for the untreated controls 2,542 ± 389 Ω/cm2 (all mean value ± standard deviation). The difference between protected and untreated cells was statistically not significant, whereas the difference to the unprotected cells was highly significant (p ≤ 0,01; twotailed Wilcoxon-Mann-Whitney test; n=3). When calculating the relative values by setting the TEER of the untreated controls as 100 ± 13.8%, the QiOne® 2 Pro protected cells had an value of 72.6 ± 14.5% and the unprotected cells of 6.0 ± 3.2% (all mean values ± standard deviations; Figure 5). Again, the data demonstrate the effectiveness of the device against mobile phone radiation.

Fig. 4: Original micrographs of IPEC-J2 cells establishing a physical barrier on the porous membranes of transwells after 4 hours of mobile phone radiation with subsequent cultivation for 24 hours. (A) Untreated control. (B) Unprotected cells. (C) QiOne® 2 Pro protected cells. The small dark points are the 0.4 µm pores in the membrane and the purple stained structures are the cell nuclei. The cytoplasm of the cells is only weekly stained by the dye. Note the rupture of the epithelial barrier in (B), whereas the barrier integrity in QiOne® 2 Pro protected cells in (C) is nearly similar to the untreated controls in (A).

Fig. 5: Presentation of the results on transepithelial electrical resistance (TEER) after mobile phone exposure with and without QiOne® 2 Pro for 4 hours and subsequent cultivation for 8 hours. Note that the TEER of untreated control and QiOne® 2 Pro protected cells do not differ from each other, whereas unprotected cells have a significant decreased TEER due to a rupture of the epithelial barrier. Data represent mean value ± standard deviation of three independent experiments.

4. Discussion

Bhattacharyya et al. [15] have summarized in their review that reactive oxygen species are generated as by-products of normal cellular metabolic activities which can be inactivated by endogenous enzymes such as superoxide dismutase, glutathione peroxidase and catalase. However, reactive oxygen species are generated as a response by a number of traumatic influences acting on our body. Among these are ultraviolet radiation, cigarette smoking, alcohol, nonsteroidal anti-inflammatory drugs, ischemia-reperfusion injury, chronic infections and inflammatory disorders.

Moreover, mobile phone radiation is also known to produce an excess of reactive oxygen radicals [10-13,16-20] which cannot be inactivated by the enzymes mentioned above. Since the epithelial cells of the intestinal barrier have a high turnover rate, they are more sensitive against oxidative stress which can induce oxidative injury and inflammatory responses involving a deficiency of the epithelium and immune/inflammation mediating cells [15]. Prompted by this background cultured intestinal cells were used to examine the effect of oxidative stress on the regenerative potential and the integrity of the epithelial barrier. According to Vergauwen [21] “IPEC-J2 cells are intestinal porcine enterocytes isolated from the jejunum of a neonatal piglet. The IPEC-J2 cell line is unique as it is derived from the small intestine and is neither transformed nor tumorigenic in nature. IPEC-J2 cells mimic the human physiology more closely than any other cell line of non-human origin”. The cells were originally isolated in 1989 by Helen Berschneider at the University of North Carolina [22]. The advantage of the IPEC-J2 cell line as an in vitro model originates from its morphological and functional similarities with intestinal epithelial cells in vivo. IPEC-J2 cells have microvilli on their apical side and tight junctions to act as a barrier and reflecting epithelial functionality [23]. The determination of TEER is a technique that provides information about the uniformity of the IPEC-J2 cell layer on the microporous filter membrane and the integrity of the tight junctions formed between the polarized cells. Therefore, TEER measurements are often used to study epithelial barrier function [1].

The present results with QiOne® 2 Pro complement confirm our previous findings in which we demonstrated that this device was able to protect functional neutrophils against mobile phone radiation [24]. In these experiments, cell viability was checked by the generation of superoxide anion radicals in the course of an oxidative or respiratory burst. Mobile phone radiation in unprotected functional neutrophils caused a reduction in superoxide anion radical generation by approximately 40% in comparison to untreated cells. In contrast, QiOne® 2 Pro protected cells showed a reduction in superoxide anion radical generation by only 16% in comparison to untreated cells. In the present experiments with IPEC-J2 cells the regeneration values for unprotected and QiOne® 2 Pro protected cells were in the same range as in the previous experiments with functional neutrophils. However, the examination of the integrity of a three-dimensional intestinal barrier demonstrated a much higher sensitivity against oxidative stress from mobile phone radiation. This is in accordance to the findings reviewed by Bhattacharyya et al. [15]. The use of QiOne® 2 Pro protected the cells in a significant manner as shown here.

Although the principles of quantum electrodynamics (QED) are not really accepted in conventional medicine as a method to influence the state of water, the present investigation has shown that coherent water as generated by use of QiOne® 2 Pro obviously has a definite positive impact on cells by increasing their resistance against exogenous traumatic influences such as mobile phone radiation. It has been stated that electromagnetic fields can be coupled to coherent systems resulting in a “self-trapping” which causes a common in phase dynamical oscillation [25-28]. As a matter of fact, the coherent system might be protected against exogenous electromagnetic fields like the inner part of a Faraday cage.

HRV Measurements

1. Rank Diagram

Measurement 1 – Start:
Stressor – inactive
Qi Blanco Systems – inactive

Biol. HRV Age (short-term HRV): 56 years

Measurement 2 after 13min:
Stressor – active
Qi Blanco Systems – inactive

Biol. HRV Age (short-term HRV): 61 years

Measurement 7 after 60 min:
Stressor – active
Qi Blanco systems – active

Biol. HRV Age (short-term HRV): 47 years

Despite WLAN signals (stressor) from a distance < 1m, the biological HRV age decreased by 14 years when the Qi Blanco systems were activated. This corresponds to a reduction of 23% (measurement 2 / measurement 7).

2. Autonomous Nervous System [ANS] Status

Measurement 1 – Start:
Stressor – inactive
Qi Blanco Systems – inactive

Measurement 2:
Stressor – active
Qi Blanco Systems – inactive

Measurement 7 after 60 min:
Stressor – active
Qi Blanco systems – active

3. Test Setup

4. Experimental Procedure

A total of 7 measurements were conducted

Test subject: male, 32 years old

Test duration: 1:11h

During the first measurement, neither the stressor (see point 3.3) nor the Qi Blanco systems (see point 3.1) were in use. Here the HRV values of the test person were recorded without external influences (except for the background noise, see point 3.5).

The stressor was switched on from the 2nd measurement and used continuously until the 7th measurement was completed.

The Qi Blanco systems were used during the 3rd, 4th, 5th and 7th measurement.

Measurement, Date, Time, Duration, Rest Period, Stress Index, Median HF, Stressor active, QiBlanco System active, HRV-Age (short time age), Heartbeat per 5 min

5. Validation

Test subject: male, 43 years old

Test duration: 1:40h

The rest period between measurements was deliberately increased to emphasize the effect of the stressor and the Qi Blanco systems.

Features of the validation:

 1) The low RF band function increases continuously over the test duration until it reaches a normal level of approx. 250 ms².
2) The deactivation of the Qi Blanco systems in test series 6 shows a jump in body activity in both cases.
3) The activation of the stressor is clearly visible in the records. The body is subject to high stress.
4) With active Qi Blanco systems, the body functions improve with increasing duration of exposure.

The validation is therefore successful.

6. Further Measurements

Test subject: male, 32 years old

Test duration: 2:29h

7. Note On The Tests

For any further questions, please feel free to contact us at info@qiblanco.com.

These are individual certificates.  Every person is different and the respective effects are therefore individual. The tests presented here serve to qualitatively classify the effects of the Qi Blanco systems on the human body.

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Treatment of neurological and mental disorders. Patent No. 5, 470, 846

Jaffe L. F. 1981
The role of ionic currents in establishing developmental pattern. Philosophical Transactions of the Royal Society of London B 295:553-566]

Bassett C. A. L. 1995
Bioeletcromagnetics in the service of medicine. American Chemical Society, Washington DC

Oschman J. L. 2000
Energy Medicine. The Scientific Basis. – Churchill Livingstone