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RESEARCH WITH SKIN CANCER CELLS AND R.H.E. TISSUES

(IN-VITRO STUDY OF THE EFFECTS OF UNIVERSAL AND HUMAN ENERGY ON BIOLOGICAL MODEL)

In-Vitro studies, done in France by a group of researchers (doctors, biologists, biochemists) specializing in skin biology, histology and cell culture, among others, began in January 1996, investigating the effects of Universal Energy (U.E.) on simple models: healthy normal cells and skin cancer cells. They also dealt with the reaction of these cells to chemical intoxication.

The research later focused on the effects of U.E. on a complex model: reconstituted human epidermis (RHE) (of the type used for therapeutic grafting on heavily burned patients). A whole series of skin studies were performed to observe the effects of U.E. on aging, intoxication and wound healing.

In order to ensure that scientific rules were strictly observed, all experiments were conducted blindly and all the persons involved were separated:

- Laboratory technicians never knew which samples were/were not treated by U.E.

- Researchers read the results.

- Universal Energy practitioners performed the U.E. transfer.

U.E. was transferred remotely (several kilometers to several hundred kilometers away) using the energy center Chakra 6 (C6) abilities, not only because the precision and power of U.E. are greater when sent by C6 than the “hands-on” method, but also because this preserves the bacteriological integrity of the models and the overall objectivity of the experiments.

It should be emphasized that this study was carried out for scientific research purposes only. Though they are quite sophisticated and are used in the field of biological and medical research, the in-vitro biological models used in these experiments differ greatly from the complex body of a human being in that:

- They are more fragile than the in-vivo models due to the absence of many defense and protection mechanisms.

- They lack a nervous system, a blood system, etc.

For the above-mentioned reasons, this study should not be taken as a medical study, since it goes beyond the objective of this project. However, the findings outlined here should serve as a useful basis for more detailed research in numerous fields.

MATERIALS AND METHODS USED

TREATMENT: U.E. treatments were performed in two ways; by a remote sender (C6) who did not see the cells nor the RHEs but was simply given the number of the labteks or RHEs to treat, and by hand contact on the plastic labtek or dish.

TECHNICAL WORK: Technical work was carried out by technicians who were not aware of the treatment involved. Coordination of the experiment (protocol and timing) was done by another person. The latter never interfered with the experiments at any time.

CELL CULTURES: Known standard culture methods and three cell types were used:

- normal human skin keratinocytes (HK) from primary culture

- normal human fibroblasts from primary culture

- human epidermal cancer cell line A-431 cells

Cells were cultured in flasks and in slide-size culture chambers called “labteks”.

CELL COUNT: Standard cell counting methods were used to determine the number of cells and proliferation activities using a “Malasser” chamber.

TIME STUDY COURSE: In these tests, cells grown in labteks and designated wells were treated by U.E. for various lengths of time and at different frequencies to determine the most effective duration and frequency for treatment of cells in culture. Cells were monitored each day, and after 3 to 4 days in culture (maximum time frame, due to the small size of wells), cells were stained and evaluated.

CELL STAINING: To examine cell morphology and confluency, cells were examined every day directly in the culture dishes or labteks using light microscopy, and pictures were taken. To improve viewing, H+E (hematoxylin + Eosin) standard coloration by staining methods was performed in most tests. This staining method gives a blue/pink color to the cells and allows for the estimation of their confluency.

The reading of cell confluency was done on the entire surface of each well, for every labtek. Pictures were taken only at one angle.

To further investigate U.E. influence on cells and to try to understand more about U.E.’s mechanism of action, immunohistochemical staining methods with specific markers were used. Immunoflourescence (IF) studies were performed using specific antibodies (Ab) directed against specific molecules in the cells. These reactions were observed by means of another fluorescein-conjugated Ab, which enabled viewing under a specialized microscope.

The following antibodies (from Immunotech, Marseilles/France) were used:

- CD95, a marker for cell apoptosis or “programmed cell death”

- Ki67, a nuclear marker, to study DNA activity and cell proliferation potential

- Pan keratin & filaggrin, to study cell differentiation

- EGFr (epidermal growth factor receptor), to study epidermal cell proliferation

- GB3 antigen (laminin-5/epiligrin), and collagen 4, to study basement membrane

zone formation, which is responsible for epidermal adhesion to the dermis in human skin

- ICAM (intercellular adhesion molecule), to study cell-to-cell adhesion

- Fibronectin, to study extra cellular matrix

RECONSTITUTED HUMAN EPIDERMIS (RHE): A full-thickness living model of RHE was used in these experiments. Only very few specialized labs in the world employ this technique. The RHE is made up of normal human skin cells, which have been cultured under very special conditions by which they multiply and differentiate in layers, thus reproducing the human epidermis in all its layers and features. This RHE living tissue is used in research laboratories for specialized in-vitro studies, and to replace animal testing for example. It is also the best grafting solution for heavily burned victims (particularly third degree burns), and patients with heavy skin loss or ulcers.

All RHEs were received at day 10 (d10) in our laboratories.

TOXICITY TEST PRODUCTS: Colchicine, an antimitotic (anticancer) substance, was used to give toxic doses for these in-vitro tests. The doses were: 1X (1 microgram/ ml), 5X, 10X, and 50X. Colchicine was used in 2 forms: normal colchicine solution (native), and U.E.-treated colchicine solution (30 seconds by C6 before application).

Toxic doses of detergent were also used for some of these in-vitro experiments.

(Detailed information of tests and experiments has been omitted here and may be made available to interested scientists and researchers. Please contact us at our E- mail address: Contact us. The following section is a summary of test results and conclusions.)

- NORMAL HUMAN KERATINOCYTES: did not reveal any significant changes on cell number and morphology in a 3-day experiment.

- NORMAL HUMAN FIBROBLAST: direct effect of U.E. on normal fibroblasts, helping their growth, improving their physiological conditions.

- EPIDERMAL CANCER CELLS: diminished the number of cancer cells by about 20% to 40% in 3 days if with 10 seconds of C6 U.E. treatment. Diminished cancer cells up to 100% if with 30 seconds of C6 U.E. treatment, once a day or every 18 hours. Hand contact treatment (on plastic container) showed best results if with 1 minute U.E. treatment, once a day. The cell surfaces after U.E. treatment appeared “granular,” altered and degenerated. The nuclei are less clear, with some dyskeratotic figures, which is a sign of cell stress and dysfunction in the dying cancer cells. Untreated cancer cells preserve their morphology and their dangerous proliferative potential without change.

- PROLIFERATION OF NORMAL AND CANCER CELLS: U.E. treated cells showed less positive staining with the Ki67 antibody, which indicates less mitotic activity of DNA in these cells compared to the untreated cancer cells. Stained with CD95 Ab, both normal keratinocytes and A-431 skin cancer cells, cell death condition showed up on day 2 and became clearer on day 4.

- CELL FRAGILITY: when A-431 cells were heavily trypsinized for a longer exposure time than needed with trypsine EDTA, which weakens cell membranes and puts their viability in danger, ALL U.E. treated cancer cells died after three days while only half of untreated cells were dead.

- INTOXICATED CANCER CELLS: when treated with U.E., colchicine intoxicated cancer cells became more sensitive to the antimitotic compound by increasing its effectiveness, and led to a 95-100% elimination of cells instead of the 30-40% of dead cells without U.E. treatment. Also, corroborated by immunofluorescence (IF) studies using the CD95 Ab, a specific marker of cell apoptosis (programmed cell death), show a significant increase in appotosis marker expression of dying cells after the antimitotic compound was applied to the U.E. treated cancer cells. While U.E. treated intoxicated NORMAL cells showed resistance and survival of healthy cells by approximately 50%, U.E. treated intoxicate CANCER cells resulted in a 95-100% increase in their death. This may be demonstrating the advantage of COMBINING both treatments: ANTICANCER DRUGS and U.E. transfers, because U.E. increased cancer cell elimination and also increased normal cell protection from intoxication. (Note: With in-vitro models, no other “repair mechanism” outside the cells exist. Thus, the toxic substances stay permanently in contact with the cells. As a result, U.E. only acts on the internal mechanisms of cells in these in-vitro tests. Hence, owing to diminished defenses, these adopted models are much more fragile than a human organism. Despite this, the results are significant. U.E. seems to act “intelligently” by inhibiting the toxicity when it is necessary (in the case of pollution on healthy cells), and also by activating the toxicity when it is required (improvement of the elimination of cancer cells).

- GROWTH AND AGING OF NORMAL RHE: marked differences resulted in RHE models with and without U.E. treatment. Daily examining of the models showed clear signs of “usual” aging in untreated models, while U.E. treated RHE did not show any sign of aging; the keratin layer was thick but not huge, the granular layer was normal, basal and other cells were normal with rare vacuoles, and no chromatin condensation.

- IMMUNOFLUORESCENCE STUDIES ON RHE: after 18 hours, with only one U.E. transfer, a dramatic protein increase was found, particularly in EGFr, keratin, filaggrin, Ki67, ICAM and fibronectin; indicating stimulation of tissue regeneration. U.E. stimulated basal cell proliferation, differentiation into keratin, and it increased cell adhesion and extra-cellular matrix to support the tissue. U.E. effectiveness at the molecular level is immediate, whereas at the macroscopic level, it may be detected only later. After 3-8 days, a gradual increase was seen in collagen 4, EGFr, ICAM, fibronectin, and Ki67, in all RHEs. On day 20, EGFr, ICAM, GB3 Ab, and Ki67 had a stable expression in the UE treated RHEs, which means a continuous balance in RHE regeneration and that the U.E. treated RHE did not feature the usual signs of aging.

- WOUND HEALING: These studies show that after only 6 days, the U.E. treated RHE healed 80-100% while untreated RHEs showed no healing at all except for one RHE at 20% maximum. U.E. treated RHEs showed an increase expression of Ki67 in basal cells and especially at wound edges, as well as in collagen 4 and laminin-5/epiligrin for epidermis-dermis adhesion, indicating the “real” healing process. Interestingly, it was also found that there was no defective scar formation in the U.E. treated RHEs.

- FUNGAL INFECTION AND WOUND HEALING: treated with U.E. 3 days prior to introduction with fungal contamination and everyday thereafter, RHEs continued to heal without signs of infection. It seems that U.E. enhanced the defense mechanisms in the treated tissues, making them resistant to infection, perhaps by producing high levels of protecting cytokines (to be further investigated).

The above summary seems to underscore the effects of Universal Energy (U.E.) in numerous biological fields as these results can be scientifically observed, analyzed, and often quantified. We hope to see that U.E. belongs in the domain of research rather than the field of fantasy. We however regret that the scientists who worked on the above experimentations no longer make their works available.  So at this point, if you are interested in the detailed information of the above studies, please contact the US MEL headquarters or the Australia Headquarters. 

 

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