Institute Research

Biological Research

The term biological research refers to the investigation of the basic biochemical and biophysical hypotheses concerning how radiowave therapy works.

The Institute recognizes the need for comprehensive biological research to investigate the biochemical action of radiowave therapy at a cellular level and to understand the biophysical effect of RF exposure on the human body in relation to antennae design and dosimetry. The requirements for this research includes studies of the:

  • biochemical effects on tumour cells in vitro, treated using the GMI/UHF and UHF/XRT modalities
  • delivery of radiofrequency with respect to different antenna designs, an extension to the first stage agreed with the University undertaking the research
  • absorbed dose
  • heating side effects.

This biological research will be conducted by an International University under the sponsorship of the Institute.

Scope of Biological Research

The University to undertake the Biological Research is one of Europe’s leading teaching and research institutions. The Centre to undertake the research within the University focuses on radiation biology and environmental toxicology, along with the effects of ionising and non-ionising radiation on mammalian and aquatic species. The laboratory has expertise in the investigation of in-vitro cultures with extensive facilities. The Centre also has access to a gamma ray source, for investigation of UHF/XRT.

The focus of this particular project is to investigate the biological basis for the GMI/UHF and UHF/XRT modalities. Radiowave therapy is based on hypotheses concerning the biochemistry of cancer cells, namely the ability to introduce drugs to inhibit glucose metabolism in these cells and possible non-thermal effects from RF energy on the physiology of cancer cells.

The term biological research refers to the investigation of the basic biochemical and biophysical hypotheses concerning how radiowave therapy works.

Bodies of work

The Institute has identified three key bodies of work that make up its biological research program:

  • GMI/RF research to study the action of various medications that have the potential to inhibit glycolysis metabolism in tumour cells and to study the possible synergistic effect RF radiation has in combination with pre-treatment by these drugs in killing cancer cells.
  • RF/x/y-ray research into the potential role of RF radiation as a radiosensitising agent to improve the effectiveness of radiotherapy (x-rays or y-rays) to kill cancer cells and the potential to do so at doses below the current therapeutic levels.
  • Antenna research investigating how the design of an RF transmitter system affects the dose of RF energy delivered to different tumour sites. In particular, studies of the effect of varying the type and arrangement of the antennae and the position and motion of a patient relative to the antennae.
Research proposals

The initial three years research projects have been approved the Board, and funding for this stage/period allocated.

Variations to treatment regimes

Notwithstanding the initial research foci based on Dr Holt’s protocols, the Institute is considering a number of additional stage 1 projects which will expand on the GMI protocols research and commence in vitro studies of radiowave therapy as an adjuvant treatment.

Biological hypotheses

The GMI/UHF and UHF/XRT radiowave therapies are based on seven biochemical and biophysical hypotheses:

  • H-ANAE: Anaerobic glycolysis. Cancer cells are reliant on the process of anaerobic glycolysis to provide the energy they require to grow and divide, even in the presence of oxygen.
  • H-GMI: Glycolysis inhibition. The infusion of certain agents (GMI) into cancer cells inhibits or blocks the production of energy by anaerobic glycolysis. The cancer cells are unable to obtain energy by means of aerobic respiration and they are effectively starved of energy.
  • H-DTH: Cancer cell death. Cancer cells in which the supply of energy from anaerobic glycolysis has been inhibited by GMI are less likely to divide (mitosis) or if they do attempt to divide are more likely to die or become damaged in the process.
  • H-DIVI: Cancer cell division. Exposure of cancer cells to RF waves increases the rate at which the cancer cells divide.
  • H-BIOL: Other biochemical effects. Exposure of cancer cells to RF waves has some non-hyperthermic biochemical and biophysical effect on the cells when compared to non-tumour cells other than the hypothesised increase in mitotic rate (see H-DIVI above). Possible effects include changes in cell membrane potential, mitosis of mitochondria, changes in DNA sensitivity and cell polarity.
  • H-SENS: Radiosensitisation. RF waves “sensitize” cancer cells so that they are more likely to be killed by subsequent radiotherapy. The cause of this sensitization is unknown other than the suggestion made by Dr Holt that it involves electrical resonance and fluorescence effects.
  • H-STEM: Increased rate of normal cell mitosis. RF waves increase the rate at which normal stem cells of the immune system divide.

With the exception of H-BIOL all of these are hypotheses are based on Dr John Holt’s experiences and assertions. The H-BIOL hypothesis has been added in light of the literature reviews conducted to date by the Institute.

In-vivo research

The biological hypotheses listed above summarise the science of how radiowave therapy is believed to work and are the necessary first things to prove by means of in vitro research. Once some of the hypotheses have been proved further research will be required in vivo to investigate how the results scale up from cells and tissue samples to whole living organisms. The results from both the in vitro and in vivo research will provide the biological evidence for the effectiveness and safety of the therapy. The in vivo research will entail investigating:

  • RF penetration. How far radiowaves can penetrate through the patient’s body until they are completely absorbed or reduced in intensity to such a degree that they lack sufficient power to initiate the hypothesised biological processes (H-DIVI, H-BIOL, H-SENS and H-STEM);
  • Tumour response. The extent to which tumour cells in the patient are killed by the treatment, providing a clinical response measured by a reduction in tumour size, rate of tumour growth or improvement in tumour markers;
  • Side effects of radiowaves. Identify any side effects for the patient from the dose of RF waves required to produce the cellular and tumour responses
  • Toxicity. Confirm there are no toxicity effects of the ingredients of the GMI in the doses required to inhibit glucose metabolism.
  • Resistance. If repeated exposure of cancer cells to GMI/RF cause the tumour cells to build up a resistance to these agents. Acquired (adaptive) resistance is an issue in chemotherapy and radiotherapy and may limit the effectiveness of these cancer therapies. One advantage of radiowave therapy advanced by Dr Holt is that it does not suffer from these limitations. This is an important consideration if it is confirmed that large tumours require multiple courses of GMI/UHF radiowave therapy for there to be any positive tumour response.
Research subjects

The GMI/RF and RF/x/y-ray research works in Stage One shall be conducted primarily (but not exclusively) at a cellular level, in vitro. The antenna research will require a range of subject material including computer modeling, physical tissue-specific simulation and experiments using bulk tissue samples.

Output from research

The output from the research must be suitable for publication in a peer-reviewed journal, although the timing of the publication is a matter for agreement between the Institute and the University sponsored to conduct the works.

Cell endpoints

Cell endpoints refer to the outcomes from the in vitro studies. These are measurements of the proportion of cells that survive or are killed by the treatment as well as the proportion that stop growing or slow down dividing. Hence, the studies shall measure these endpoints:

  • cell death – the rate at which cells die either due to apoptosis, a type of programmed cell death (PCD) or by necrosis
  • cell survival
  • cell senescence – the number of cells that are no longer capable of dividing
  • cell viability – the number of healthy cells in a sample
  • cell proliferation – the number of cells that are dividing in a culture.
Measurement of changes in cell physiology

In addition to the above endpoints the following changes in the physiology of normal and cancer cells shall be measured over time before, during and following the infusion of GMI agents and RF, x-ray or y-ray exposure:

  • cell metabolic activity
  • cell membrane integrity
  • cell hypoxia. This refers to inadequate oxygen supply to a cell. Tumour hypoxia is associated with tumour propogation, malignant progression and resistance to therapy
  • damage to nuclear DNA (nDNA) and to mitochondrial DNA (mDNA)
  • O2 consumption
  • mitochondrial oxygen consumption
  • stress proteins
  • mitochondrial membrane potential
  • redox cycling of glutathione (for GMI studies only).
Assays

Below are some common biochemical assays and analyses required in the research:

  • clonogenic cell survival assay to measure cell survival;
  • COMET assay to detect DNA strand breaks (single and double) in nuclear DNA;
  • deletion analysis (polymerase chain reaction or PCR) and mutation analysis to measure mitochondrial DNA damage;
  • fluroscence using Annexin A5 assay to measure apoptosis;
  • oxygen scavenger such as sodium hydrosulfite, Na2S2O4 to measure cell hypoxia;
  • Propidium Iodide to measure necrosis;
  • polarography (voltammetry) to measure mitochondrial oxygen consumption This uses the Clark oxygen electrode;
  • reactive oxygen species (ROS) including oxygen ions, free radicals and peroxides. These species have been proposed as a regulator of programmed cell death. Measured using ROS scavengers (PDTC) and ROS donors (NDPO2);
  • upregulation of heat shock proteins (HSP) to measure thermal effect, including cell hyperthermia.
Abstracts of the Biological Research

Confidentiality arrangements exist in relation to the organization contracted to conduct the biological research and the specifics of the topics for research that have been outlined herein. However it is important to note that there exists timelines for outcomes that are to be published.

Mathematical Simulation

Mathematical modelling (computer simulation) using finite-element modelling (FEM) shall be used to investigate different antenna designs and patient/antenna geometries and the effect on the RF dose received by the patient and the absorption of RF by the human body.

The predictions from a mathematical model will need to be tested against experimental data.

The parameters of the human body tissue on which the model is based shall be identified and the source of the values for these parameters. These parameters which affect the energy absorbed by the patient’s body include:

  • relative dielectric constant (permittivity) (εr)
  • dielectric loss factor (δ)
  • impedance (Z0)
  • mass density of tissue (ρ)
  • electrical conductivity (σ)
  • thermal conductivity (k)
  • specific heat (C).

The parameters are frequency-dependent and tissue-dependent, so there will be different values for fat, fibro-connective tissue, muscle and bone, and possibly differences between normal and tumour tissue.

Software for 3D electromagnetic field simulation has been identified.