Update September 2008

Mal's Musings

Malcolm A Traill

IN  THE  PUBLIC  INTEREST
Updated 23/9/2008

 

    SUBMISSION to the NH&MRC

 

         REVIEW OF THE USE OF MICROWAVE THERAPY

       FOR THE TREATMENT OF CANCER

 

 

Click to E-mail me

 

Brief Historical: During August to October 2004, television Channel 9 (Melbourne) presented a number of segments dealing with the treatments for cancer given by Dr John Holt of Western Australia. In the summary at the channel's internet site, there was reference to 'radio waves - - at a specific frequency'. That frequency was not mentioned in the web-site but was, in fact, ~434 MHz. To the public, the treatment is regarded as microwave radiation, no doubt because of the now ubiquitous microwave ovens (which operate at about 2.4 GHz) to which the public can relate. As I understand the Minister's request, the major purpose of the current examination seems to be an assessment of Dr Holt's treatment using ~434 MHz and associated agents. As presented, other forms of 'microwave' treatment (applied loosely) may be considered. 

 

Clarification of Terms: Unless otherwise stated, I shall refer to electromagnetic waves (radio waves) of a non-ionising type. These can be classified here as :

 

               Term                         Abbreviation             Frequency              Wavelength                                        

   a) Very High Frequency         VHF                     30 - 300 MHz 

   b) Ultra High Frequency        UHF                     300 - 3000 MHz

                                                                                       = 3 GHz

   c) Microwave                                                        600* MHz -          0.5 m - 1 mm*

                                                                                    1 GHz -                                                           *Macquarie

                                                                                                                                                                Dictionary

 

From this one can see that 434 MHz is UHF and not, strictly speaking, 'Microwave'  but, from the context and the loose use of the word 'microwave', I assume that you propose to examine the effects of UHF and, in particular, the frequency 434 MHz upon cancers in patients. It is this aspect that I shall address primarily.

The Tronado machine was originally brought to Australia with the belief that it would supply heat. It used the frequency of ~434 MHz applied through multiple dipoles.  I am not aware of any Tronado machine still in existence in Australia now - machines at that frequency now appear to be custom built and use loop antennae. The use of the term Tronado is passé, and I will not use it again. I shall refer to 434 MHz as the (primary) treatment form. 

Radiations absorbed by tissues will produce heat. This heating is referred to as thermotherapy or, more recently, fever-range hyperthermia (FRHT) if the heating of the tissue is between 40-42oC, hyperthermia if the temperature is above 42oC. Where the heating is confined to microscopic points so small and for such limited times that the surrounding cells/tissues do not heat significantly, it is referred-to as athermal heat (ie molecular agitation limited to a very small site) as discussed by Lawrence et al., 20001#.

 

#Thesuperscript numberrefers to the consecutive numbering of the Abstracts/Summaries attached. If not numbered, they will be found in Alphabetical order following.

Holt has written a lot over the years and, during this time, he has referred to the 434 MHz as Hyperthermia, VHF, UHF and Microwave. All these terms should, in this context, be regarded as synonymous with 434 MHz (+/- 1-2 MHz).

 

My Position: I am a Specialist General Clinical Pathologist (FRCPA) with extensive experience in laboratory and clinical pathology, having become Vocationally Registered. I generally self-determined my own pathology testing. I have published or read over 50 papers and letters on diverse topics, with particular emphasis upon serum lipoproteins. More recently, I have examined the specialist-level, non-psychiatric treatments with lithium for such diverse and chronic, disabling conditions as Multiple Sclerosis, Hepatitis C, Rheumatoid Arthritis, Sarcoidosis and Lupus Erythematosus. I present 3 cases :

 

Multiple Sclerosis (AB): Female born 15/5/1965, symptoms May 1991, diagnosed by Neurologist August 1991, with MRI in September 1991. Had been employee in laboratory with knowledge and understanding of issues. Tried slow release Lithium from August with limited positive effect. Later used standard tablets and found that best effect was abortive treatment, provided it was applied, early before vomiting started. Tried beta-interferon when it became available on the PBS, but discarded it after about three months, claiming unacceptable side effects. Compliance with Lithium only fair, so had a number of bad relapses requiring hospitalisation and Prednisolone, the last bad relapse in 1998, for which she needed bladder catheterization and rehabilitation. MRI 18/5/1998 noted 'Findings consistent with multiple sclerosis. There is evidence of active plaque particularly in the right centrum semiovale'. This scared her and her compliance was more punctilious. Over the years there has been minimal clinical progression, with good bladder and bowel control and no need for appliances. She holds a job, working in real estate/financing 'seven days a week'. Photo taken 20/7/2003 having the patient balancing with one foot on a curb (left). Another June 2006 (right).

                                         July 2003                                                 

Hepatitis C (BL): Male born 14/4/1958. Had past history of alcohol and iv drug use. Was rejected from Interferon trial by Fairfield Hospital because of mild cirrhosis on liver biopsy. Options fully discussed and he signed a declaration that he knew Lithium treatment in this context was experimental.

The graph of his first course of treatment in 1993 is attached. There was a rapid decline of his ALT enzyme. About 10 days into the course he developed an antibody negative T3 thyrotoxicosis, self-limited to several days. As the T3 became normal he developed paranoid ideation, being unable to sing in a church choir because he felt that everyone was staring at him. This lasted several days. Compliance and interest waned and he was lost. He reappeared three years later. Lithium was prescribed again at a lower dose (because of the thyroid/ideation effects). His ALT enzyme again fell and once again there seemed a short episode of paranoid ideation about 32 day into the course. (Both episodes seemed to be related to a rise in the ALT enzyme.) Compliance and interest waned after a while. He has been lost to follow-up since. The thyroid and ideation effects would seem consistent with cytokine actions. These side effects have not been noted in any other patient, nor have they been found in the literature associated with Lithium.

 

                                   

 

Rheumatoid Arthritis (JH): Female born 13/3/1923. She had active disease in September 1996. Lithium treatment was fully discussed. It seemed to induce remissions to acute flares, and when administered in prophylaxis mode, seemed to prevent flares (see graphs: ESR, Haemoglobin and Platelet counts are represented). When she tired of prophylaxis mode, a relapse followed in July 1999. Over the years her course has been similar, but with the addition of Polymyalgia rheumatica late 2003, which did not seem to respond to Lithium, so she was treated with Prednisolone. Late 2004 she had another flare of Rheumatoid arthritis, which seemed to respond to Lithium. More recently she has noticed 'shakes' which seem due to Myasthenia gravis (anti-acetylcholine receptor antibody positive). Her antinuclear antibody test has been non-reactive throughout.  Her right hand shows only minimal ulnar deviation (photograph).

 

                                                     

 

                 

 

Sheet 1 continues into Sheet 2. Haemoglobin = red lines/circles; ESR = black lines/squares; total platelet count = dotted lines, crosses. Treatments with Li+ are indicated by Rx and vertical lines.

 

                       

         Relapses (with peaks in ESR) abated with Rx, and were prevented by regular Rx

 

Accordingly, based upon my clinical experience and related literature reading, I present myself as a scientific-based clinical Specialist and expert in certain aspects of lipoprotein biochemistry and the non-psychiatric uses of Lithium. It is with this background that I became interested in hyperthermia and UHF, also having had an amateur wireless licence (VK3BI).

 

In early 2000, equipment capable of delivering 434 MHz and UHF at other frequencies for local or regional hyperthermia was brought to Fairfield, Victoria, and I was able to apply these to patients. Patients were almost always referred to me as a Specialist Consulting Pathologist (HIC Categorization Code 029); patients being able to claim rebates at Specialist rates (Items 104-5).

 

From the outset, I predicted that there would be professional problems, given the controversial nature of some of the modalities. One complaint by an Associate Professor was lodged with the Medical Practitioners Board of Victoria in mid 2002. There has been notice of an Open Hearing, but nothing more to date.  Accordingly, I set out to establish evidence that the 434 MHz had effects upon patients &/or their tumours (to be detailed later). (The Medical Practitioners Board advised me early 2003 that the Medical Board of Western Australia had not had cause to investigate Dr Holt and his treatment - he should be so lucky ! This position may be because there were enough witnesses to his early work to believe that he may be on to something, even if they could not understand it.)

 

I have also given considerable time and thought to consider the nature of the treatment, and how its presumed effectiveness could be explained (to be detailed later).

 

In preparing this report, not only have I had the background outlined above, I have read most of Holt's writings (published and unpublished, which includes two monographs of personal communication type, which he may show to the enquiry if he sees fit.), and many of his references. I have spoken and written to him for his thoughts on particular issues, and spoken to, and obtained a brief report from David Spall, who claims to have been attending Holt's clinic in the 1970s when many of the observations were made and witnessed the work. His testimonial is attached. He related that those in the work environment at the time showed intense interest in the phenomena observed, and further stated that he witnessed the saga of the German technician, there to install equipment, volunteering to be a normal control. To everybody's amazement, he radiated emissions when under the 434 MHz. Shortly afterwards, a Chest X-Ray displayed a mediastinal mass ! (I understand he is included in Holt's published tables).

 

An Overview of Holt's Work as I see it.

1. Holt's writings (general comments).

 

a)      His writings are based upon a massive foundation of clinical experience, with him claiming some 10,000 patients treated with 434 MHz over some 30 years.

b)      He has tended to publish in lesser known journals

c)    His format in publishing has been to present observations and hypotheses as he saw them, generally without marshaling material orderly together to 'sell' the treatment to the medical profession (or any particular group)

d)     There seems no index to his writings. This means that looking up a particular point can be difficult. For this reason I have prepared the attached Table 1.

e)       He has failed to identify a cellular organelle which can be equated to his unit of power, ERex

f)     Without an identifiable organelle, there is difficulty relating his hypotheses with current biochemical and ultra-structural knowledge

g)     There are certain points that I cannot follow (eg 1 electron into and 2 electrons out of the ERex indefinitely. I have read the reference by Kosower and Kosower, 1976. I shall look at this later

h)   He has had limited histopathology experience. I understand that he was reliant upon reports and photographs from histopathologists. Apparently he never had the slides, being unable to supply them when I requested them. His criteria for mitotic activity seem to differ from mine. Photomicrographs are frequently at unrecorded magnification and comparison pairs may seemingly be at differing magnifications. The absence of electron micrographs is noted with regret

i)       He has conducted some remarkable trials with his own patients in an attempt to understand better the phenomena he had observed. He has produced awealth of documentation deserving closer scrutiny (eg his observation of insulin resistance in cancer patients. This is timely, considering the current interest in 'Syndrome-X' and insulin resistance in some Hepatitis C patients [Hui et al 20032]).

j)      Holt often includes in his publications formulae with logarithmic functions. These are almost guaranteed to 'turn off' most medical practitioners attempting to read his writings, unless they are acquainted with radiotherapy theory and its relevance.

k)     I do not agree with his use of the term 'fluorescence'. I stick to the term 'resonance' as the phenomenon. 'Phosphorescence' would seem more appropriate for the emission frequencies.

l)       A notable proportion of his references are pre-1970. These undoubtedly stimulated his research direction in the 1970s but, with an inability to relate his work to the increasing knowledge since 1980, has resulted in a 'burn-out'. Some references are hard to obtain.

m)      His reverence to epsilon has been intrusive.

n)    Undoubtedly, he has incredible achievements, which I admire and cannot underrate. How he, for the most part a solo practitioner while achieving such patient treatment numbers and performing the studies that he has documented, has shown remarkable staying powers.

o)       I have difficulty understanding his explanation of energy transfer to his ERex.

 

                                                                 Table 1

                                                  HOLT'S WRITINGS

                                                     Biochemical Emphasis

                    Med. Hypoth.          Med. Hypoth.          Progress in           Med. Hypoth.                

                    5: 109-144               6: 145-192                Radio-Oncol.       12; 359-367                                  

                    1979                       1980                        1982                     1983________

Themes       Glucose                   Resonance/UHF         UHF & X-ray        D-Glucose                

                    O2/ANO2                      Glucose Metab.           & Combined          L-Lactic acid

                    Histology                                                   Prev. reports          Pasteur effect

                    Healing                                                      Radiosensitivity

                    Growth

                    Differentiation

                    Heat/Temp.

RESONANCE & HEAT

Enhance                                      D-Glucose                                               D-Glucose

                                                     D-Fructose                                              GSH

 

Antagonist                                   Ethanol                                                    L-Glucose

                                                     L-Glucose                                               D-Gluc. analogues

                                                     Insulin                                                      Oxidant for GSH                                                     

                                                     5-thio-D-Glucose                                     n-Tert-hydroperoxide

                                                     Hyperbaric                                               Glyoxalase Inhib.

                                                     (? Steroids)                                              Ophthalmic acid

                                                     (cyclophosphamide)

 

No effect                                      2-Deoxy-D-Glucose

                                                      D-& L-Mannose

                                                      D- & L-Fucose

                                                      Azaserine

                                                      DON

                                                      Streptokinase

                                                      Glutamine

                                                      Biguanide

                                                      Sulphonyl-urea

 TO L-LACTIC ACID

Enhance                                                                                                        D-Glucose

                                                                                                                      (D-Fructose)

 

No effect                                                                                                       L-Glucose

                                                                                                                      2-Deoxy-D-Glucose

                                                                                                                       5-Thio-D Glucose

                                                                                                                       D-&L-Glyceraldehyde

                                                                                                                       D-&L-Ribose

                                                                                                                       D-Xylose

                                                                                                                       D-&L-Mannose

                                                                                                                       D-Galactose

                                                                                                                       D-Allose

                                                                                                                       D-&L-Fucose

 

2. Holt's writings on UHF at 434 MHz and Cancer

            a)     In establishing 434 MHz as a treatment of cancer, Holt has put almost total emphasis upon
                  three criteria :

i.                                             i.                       The 434 MHz power absorbed over cancers, evidenced by changes in plate output
                                    current on the transmitters and/or core temperature rises in patients.

ii.            A fascinating resonance phenomenon. This has been witnessed (by David Spall at least - but not by me), photographed and published. This seeming emission phenomenon would be consistent with i. above. There are certain features of importance to note: firstly, there are short-term bursts of frequencies emitted both below and above the incident 434 MHz. This can only be explained by energy being supplied by the living cancer tissue; secondly, there may be a tendency for these emissions to occur at about 250-260 kHz intervals, to be discussed later. The features do not seem consistent with typical fluorescence, and there is difficulty explaining the phenomenon as being due to external interference. From Holt's account, the emitted signals are of significant power - not weak.

For credibility, those who wish to disparage Holt and his work must first explain this resonance phenomenon :

 

 

                              SOME OF HOLT'S SPECTRAL PHOTOGRAPHS

 

Photographs 2 and 3 occurred on page 100 of a monograph entitled "Your Cancer is Unique, the Cure is Common" (undated and possibly not published) but acquired by me about late 2000. Note that the photographs are "stills" of constantly changing patterns, having new patterns with each horizontal sweep of the cathode ray oscilloscope. All of Holt's numerous photographs show individual differences in the patterns. He generally seems to have used exposure times of 1/8th of a second, with the horizontal sweep 50 sec-1. The  left side horizontal limit to the tracings  would seem to be included within Photograph 1. Calculations derived from these parameters have been presented later. (Other photographs have been published more recently in  The Journal of Molecular Liquids,2004; 114; 193-206. [See p. 195])

 

                                                                Photograph 1

                                                                    434 MHz

This tracing is from page 65 of Holt's monograph (see Photograph 2) and is said to have been obtained from a patient without cancer (ie a normal control). There are small resonance patters on both the high and low frequency sides of the main 434 MHz peak.

 

                                                               Photograph 2

                                                                    434 MHz

Holt's caption states that the spectrum is typical for the UHF reflected from a moderately sized deposit of cancer (eg a pancreatic mass 5 cm in diameter. The central peak is at 434 MHz, with higher frequencies shown on the left and lower on the right, with 0.5 MHz major scale divisions. This spectrum extends from 436.5 to 432 MHz.

 

                                                                 Photograph 3

                                                                    434 MHz

"Immediately after the intravenous injection of 10 g of L-Glyceraldehyle, the spectrum (in Photograph 2) changed abruptly to this pattern (Photograph 3) which is closer to normal (ie no tumour, as in Photograph 1)." After about 45 minutes, the pattern as seen in Photograph 2 returned. (Slightly edited by MAT.)

            Lessons to be learned from the Photographs. Let us now carefully inspect the photographs to see what can be deduced by reason :

 

a)     Confining attention to those photographs associated with cancer (Photographs 2 &3), we cannot discern any gross or microscopic symmetry. This would make extracorporeal electrical interference or deception very unlikely, if not impossible. That is because modulation of a carrier frequency produces symmetrical sidebands about the carrier frequency at +/- the modulating frequency (eg 434 MHz +/- 10 kHz for a 10 kHz modulating frequency).

b)     There are emitted frequencies above and below the incident frequency. These do not show the typical features of fluorescence, where there is a set fluorescence frequency at a new energy level (because of energy losses) below and clearly separated from the incident frequency. Those above the 434 MHz must have energy added by an endogenous energy source, because the energy of an electromagnetic radiation is directly proportional to the frequency. Those below the 434 MHz incident frequency must also have added energy in order to overcome the energy losses and bring them back towards 434 MHz.

c)     The emissions are sporadic, consistent with coincidental activations (the two types that seem to be present shall be discussed later).

d)     The effect of L-Glyceraldehyde in quenching the emission would be consistent with the blocking of the endogenous energy source. Loss of the emission above 434 MHz has been most affected, consistent with the loss of the additional energy needed for the higher energy levels.

e)   The appearance of small emissions both above and below from the normal subject in Photograph 1 would indicate that the resonance phenomenon is not specific for cancer - rather, it is produced much more effectively from cancers than normal tissues.   

          

  iii.              Clinical observations of the tumours in his patients. To have substance, these observations have to be compared with reference or control groups. The types of patients he will have treated form such an heterogeneous group that establishing any control group for any one subgroup of his huge throughput is very difficult, if not nearly impossible. That was the problem I faced in the short term, and it will be the major problem for any critical examination. Irrespective of the side any assessor ultimately favours, criticisms will arise that the control groups used were not reasonably comparable with the treated groups.

 

b)     Having found the resonance phenomenon, Holt set out examining the energy supply and what would influence the excitation. He placed reliance upon biochemistry studies in the 1920-1930, ultimately quoting the works of Joseph Needham et al. (1937) for the first time in 1983. This extensive work was on a form of anaerobic glucolysis found in brie from 3-4 day old chick embryos. It did not require phosphate and seemed only able to metabolise glucose or mannitol. I understand the work was well respected in its time but, as the authors acknowledged, did not sit easily with the Embden-Meyerhof scheme; glucose was metabolised in the cellular sediment (grana) in the presence of glutathione toL-Lactate, without any coupling of energy to any biological system. There was no identifiable organelle for the reaction. The work was committed to oblivion until rescued by Holt. Having resurrected it, Holt has not found a convincing role for it in feeding electrons to his ERex units, nor the structural relationship it has with his ERex.

 

c)        The 'Glucose Blocker' intravenous infusion consists usually of oxidized glutathione (disulphide) +/- S-methylL-Cysteine Sulphoxide. Both occur naturally. The use of oxidized glutathione is predicated by his published trials in 1983. Why a chemical which does not gain ready access into cells should reduce the resonance effect is not clear. Why this should inhibit cancers is also unclear. Holt believes that it blocks the energy production by ERex for mitosis The Sulphoxide occurs in Brassica vegetables (eg cabbage) and is a non-essential amino acid. The adult Japanese on a traditional diet were claimed to consume some 300 mg per day. Said to lower blood cholesterol, interest in this aspect has waned. Holt claimed (2001, personal communication) that its role as a cancer inhibitor in the protocol was as a 'spanner in the works' and depended upon its Nitrogen being available; that N-acetyl Cysteine stimulated the growth of cancers. Holt suggests that its effect accumulates, to resemble the effects ofL-Glyceraldehyde (which inhibited the resonance).Again, why that should be beneficial in cancer treatment is obscure.  Documentation on the Sulphoxide is scanty, and its use seems based upon clinical effects. The term 'glucose blocker' seems unsatisfactory, but no-one has thought of anything better. The most likely explanation for the effects of these chemicals is that they reduce the available intracellular reduced glutathione. N-acetyl Cysteine however, is a precursor of glutathione.

There must be a distinct separation of Holt's trials and treatment results, and Holt's hypotheses.

 

3. Publications by others on the treatment. These are limited. Supportive are:

                         i.                    Caldwell 19793, who came to observe the treatment, then returned to the USA to die
                                    in an hunting accident. He never unpacked the equipment and never used the treatment.

ii.            Hornback's group. This group published a preliminary study (19774) of Deep X-ray Therapy (DXRT) and hyperthermia using 434 MHz. The group then published a follow-up report in 19795 based upon a presentation to a medico-scientific meeting in 1977 using apparently different patients classified on what would seem to be similar criteria.  In the first report, the 434 MHz was given 'immediately prior' to the radiotherapy. In the second report 'one [heating] unit was used for 20 minutes prior' to irradiation. (Later the exposure time was longer.) My examination of these details was prompted by an analysis of the results, because the second published group, supposedly selected and treated similarly, with 434 MHz, seemed to do worse than the preliminary group, showing a trend through Subjective improvement and Objective improvement, with the difference for Survival significant (Chi sq. = 3.97; 0.05>p>0.025). In view of these anomalies, I wrote to Holt for background information. He stated (2004, personal communication) that Hornback had fallen foul of the FDA for using 434 MHz on humans and had taken to sending patients to a veterinary clinic nearby which had the equipment. Here the 434 MHz was applied the day before the DXRT ! So it seems that the 'for 20 minutes prior' was actually 'for 20 minutes ~24 h prior' to. I suspect that Holt became confused by all this, because he came to believe that the 434 MHz could be given 24 h before the DXRT and still augment the latter. When the two publications are compared with respect to DXRT given before the 'heat', there appeared no significant difference for each assessment type, consistent with the selection criteria otherwise being similar.

 

I believe that the preliminary report (1977), in which the timing and sequence is clear, is the only paper by Hornback's group to be considered reliable with regard to 434 MHz as an augmenter given prior to DXRT, and it reports a favourable, improved response. 

 

Neutral cell culture studies were by Sapareto et al. 19826. Here, cells grown on discs were subjected to 434 MHz in a special container. The cells were in exponential growth and in culture media. The authors concluded that there was no evidence for any selective effect of the frequency used. However, whether dispersed, single cells, very small with respect to the wavelength, and in a medium which may conduct electricity better than the cells, can be compared with a compact block of cells, say 5 cm across, in tissue not conducting electricity as well as the tumour, has difficulties.

 

4. Other Experimental Work

 

Dr Peter French, better known for publications on the effects of 835 MHz on cell cultures (a frequency close to twice 434 MHz), funded in part (at least) by Holt, apparently did some studies for him, but I have not seen a specific publication using 434 MHz. Some information formed a personal communication (2000). There is presented a photomicrograph of A172 human glioma cells stained to show actin. 'After 10 min 434 MHz there was break-up of the actin, with a calculated 60 % loss of actin mass per cell. After 30 min culture, there was actin regeneration ~40 %'. Whilst this would be consistent with the cell shape changes (rounding) seen after 434 MHz in biopsy histology, and documentation of some effect, the changes are otherwise too non-specific and short-lived to be particularly helpful.

We are then left with two options - either the resonance phenomenon and all the work related to it has a real basis or else it is all an extended aberration. In favouring the former, recognizing the near impossibility of establishing control groups, and with the likelihood of flak, I set out to seek surrogate evidence that 434 MHz with the Holt protocol had effects we may regard as desirable, and to be obtained as quickly as possible.

5. My own clinical experience.This is limited. Brain tumours form a more identifiable group:

 α       A male, DoB 13/2/1953, with Glioblastoma multiforme. Had surgery and DXRT, the latter considered of no benefit. Came for 434MHz+* some 2 months after DXRT in Oct 2000. A CT showed midline shift (Right third ventricular wall on left side as % of diameter =53%.) After treatment he reported feeling fine, more mobile, being able to get around and shower on his own. Repeat CT showed the right ventricular wall was 50% of diameter. (He was persuaded by others to have more surgery which proved disastrous, making further follow-up of little value.)

  (*434 MHz+ indicates both 434 MHz and 'glucose blocker')

 

 β       A female, DoB 1/9/1954 had a fit in Aug 2000. Craniotomy and biopsy Astrocytoma grade 2 (1-4). Declined DXRT and opted for 434 MHz+ as only major treatment. Treated Sept 2000, one course only. MRI 3/6/2002 concluded 'Compared with the previous MRI study of 6/11/2001 shows no significant change with no evidence of tumour recurrence. . '.  Repeat MRI 25/11/2003 showed no change. That done 13/11/2004 concluded 'The left temporal astrocytoma is stable in size and appearance when compared to last year's scan'. She is well currently (but declined to make a submission because she 'does not want the stress'). (Note Dec. 2005: MRI 9/10/2005 - 'No interval change in size compared to the films of five years ago'. Note Nov. 2006: MRI 15/11/2006 - "Comparison with previous imaging from October 2005 shows no significant interval change in the size of the intra-axial neoplasm in the left posterior temporal lobe." Note added December 2011; she was reported to have some enlargement of the tumour on MRI. Biopsy showed no markers of proliferation abnormally increased.)

 

 γ      Male, born 28/8/56 biopsy Jan 2001, GBM. Had 434MHz + and DXRT following within 25 minutes, starting March 2001. Little further feedback. Died 31/10/2002, some 22 month survival.

 

 δ       Male, born 20/9/49, biopsy 22/11/2001, GBM. Had 434 MHz+ and DXRT following within 25 minutes, starting 14/1/2002. Had regular MRI follow-ups and Chemo (?Temozolamide); mild/no change to 23/8/2002. MRI 6/11/2002 showed marked increase in abnormality. Was still alive (at over 1 year) when last contacted. The last two cases illustrate collaborative and adjunctive use of the therapy.

 

 ε        Male, born 20/11/1950, with Glioblastoma Multiforme diagnosed June 2001. Had DXRT and Temozolamide, with subsequent recurrence. Had 434 MHz+ starting 6/9/2002. On 22/9/2002 he felt better and his foot was not dragging as much. Telephone report from wife 5/12/2002 'a lot better - very happy'.

 

b)     Other observations. With a mixed collection of patients having had innumerable treatments already and often at a late stage of their disease, survival figures would be almost meaningless. I have examined the effects of 434 MHz+ on blood tumour markers. The limited series will be presented, but first a graph, Patient VD (see attached) :

 

       The graph of serum alpha-foeto protein from a patient with an Hepatoma - see below. (The graph is on semi-logarithmic graph paper showing AFP in the vertical axis, weeks on the horizontal axis. The patient, an elderly female, born in Italy, with longstanding Hepatitis C, leading to cirrhosis and later a multifocal hepatoma.)

 

         

 

                                          Features of note are:

 

α      A rising slope (probably due to the appearance of satellite primaries) lines A-B, B-C

β       A fall after commencement of 434 MHz+ C-D. The first part of this has a half life (α-FP1/2) of about 29 days. (? A [partial]
    switch-off)

γ      A slow rise and fall D-G (? Breakdown after ? apoptosis [cf necrosis])

δ      Re-activation of growth G-H. The slope to the last rise was not maintained in a subsequent report from interstate (it was less
    steep).

ε      The slope pre-treatment is steeper than at reactivation - doubling time pre-treatment is approx. 43 days, that at H-I, about
    73 days, the growth is some 60% slower.

ζ      The interval from pre-treatment peak to same level post treatment is approximately 3.5 months. I could claim that treatment may
    have added this interval to her life - for a condition generally considered to be untreatable by any accepted form of therapy.

η      The change in slopes would, if maintained, be expected to add further to her survival, increasing with time.

θ      The change in slopes would seem to indicate that there has been a change in the growth behaviour of the tumour.

 

ii       The short - medium effects of 434 MHz on blood tumour markers. These are presented in Table 2.

 

 

 

                         TABLE 2

 

Initials   Condition                     Marker          Graph Features

 

  RL         Ca prostate to bones    PSA                 Plateaux. New slopes less steep

  NS         Ca breast to brain        CEA                 Fall, CEA t1/2~26 day

  IG          Ca pancreas - local      CA19.9            Fall, CA19.9 t1/2~7 day

  CD        Ca bowel to liver          CEA                Plateau. New slope less steep

                                                                                  CEA doubling pre~23 d, post~62 d

  RG         Ca prostate to bones    PSA                 Fall, PSA t1/2~10 day

  JM         Ca bowel to lungs        CEA                 Plateau. New slope less steep

  TB         Ca bowel to liver         CEA                 Fall. CEA t1/2~23 day

  EW        Ca parotid to neck       CEA                  Fall. CEA t1/2~28 day

  VD        Ca liver                          AFP                   α-FP t1/2~29 day.

                                                                                   α-FP doubling pre~43 d, post~73 d

  FK         Ca bowel to liver         CEA                  Fall, CEA t1/2~8 day

  MR        Ca breast to nodes       CA15.3             Plateau - flat

Ca=carcinoma; CEA=carcinoembryonic antigen; PSA=Prostate specific antigen; α-FP= Alpha foetoprotein; t1/2=estimated, extrapolated half-life; pre/post 434 MHz+.                                       

 

ιιι        The patterns noted to date fall into two types :

              α     Rapid fall of the marker concerned. The half-lives are presented (median 23 days). [I understand that most
                                              glycopeptides are broken down by the kidneys with a half-life of about 7 days.]
A fall occurred in 7 of 11 cases.

β     A plateau, lasting about 1-2 weeks, with the continuing growth which, when assessable, was at a slower rate (see above), meaning that increased survival is largely influenced by the slower growth rate.

 

        The graphs indicate that 434 MHz+ has an acute effect upon tumour marker levels that would be regarded as desirable. There appears to be a late effect, where the tumour re-grows at a slower rate, which would also seem to be desirable.

 

c)     Other blood tests: Patients with large tumour loads can be expected to show rises in serum uric acid, reaching hyperuricaemia. I have also noted rising corrected serum calcium, with 2 cases of non-Hodgkin's lymphoma carrying large tumour loads developing severe clinical hypercalcaemia in the second week of treatment, requiring Pamidronate iv. David Spall noted a similar case in Queensland. Since raised serum calcium can be found in about 10% of hospital admissions for non-Hodgkin's lymphoma (excluding Burkitt's) in North America (Burt & Brennan 19807), I have difficulty believing that these severe clinical changes during therapy can be by chance. [I suspected that the tumours may have been releasing Lymphotoxin (tumour necrosis factor beta) to stimulate osteoclastic action, but that would require clarification. They may have been releasing their own intracellular calcium stores.]

 

d)     Because lymphoma patients developed hypercalcaemia I watched blood levels of other patients more closely. Cancers seemed to show mild rises within the reference ranges, not normally noticeable. In the dying days of the clinic, I examined the Calcium-to-Creatinine levels and Urate-to-Creatinine levels in spot urine specimens. These seemed to show inverse relationships, so Calcium-to-Urate levels were plotted for 4 patients* (AM, a child with desmoplastic small cell tumour, received 434 MHz alone). Three graphs all show a rise in the urinary Calcium to Urate ratio, peaking about 5-12 days after the commencement of 434 MHz exposure. One, without the initial baseline reading showed a fall some time after reaching 9 days. These results (four out of four) would seem to be in accord with the observations on the lymphoma patients: there seems to be a rise in mobilized Calcium late in the first week of treatment (and possibly a small reduction in urate produced). Based upon these very preliminary results, which seem consistent with the 434 MHz +/- 'glucose blocker' having some measurable effect, there appears some potential in following such changes with a larger series and in a more detailed way.

* AM's baseline specimen was missed. The extrapolation back to Day 1 was to make the graphing software work. The Calcium determination for TB's baseline reading was missed.

 

e)     A female HK, born 1/3/1947 presented 22/3/2002 with a biopsy diagnosis of leiomyosarcoma of the uterus. She had 434 MHz+ followed by a single fever range hyperthermia (FRHT) with chemotherapy. She had a recurrence biopsied on 2/5/2002. The biopsies are presented below shown at the same magnification (approximately x 100). One could be excused for wondering if they were from different lesions. Noticeable is the transition from spindle cells (in 2 slides from 2   blocks) to rounded cells with numerous mitoses (metaphase). There are also some multinucleated giant cells - some may be tumour, others may be foreign body type (conspicuous). There were numerous areas of necrosis and regressive changes in the post treatment blocks (poor differentiation in 6 slides from 6 blocks). Such changes in isolation have limited meaning, given the possible variations within tumours and the problems of fixation and processing. However, the general pattern is consistent with the changes reported by Holt.

 

                  

                       Leiomyosarcoma pre-treatments. Note spindle cells and interlacing fascicles. Reasonably good
                         differentiation

                  

                         Leiomyosarcoma post 434 MHz+ & chemo. + FLHT. Now seems poorly differentiated to anaplastic.

                         Cells have rounded and become more compact. Cells have shrunk due to processing.

 

  6.Unifying Hypothesis

Since Holt's observations and clinical trials, there have been spectacular advances in the understanding of cellular structures and biochemistry. I shall try to unify his work within the current knowledge.  

The resonance emission. This has been photographed by Holt. From the photographs and documented background, there can be little doubt that :

 

a)      Energy is produced within the tumour to augment the incident emission

b)     This energy requires living tissue

c)     The central frequency is relatively stable (tumour to tumour) but with brief, intermittent, close variations of the resonance
   frequency to the incident frequency during each exposure throughout the tumour populations.

d)     The pattern is sensitive to biochemical agents administered intravenously.

 

The resonance is not typical of fluorescence. With the latter there is usually a resonance created by electrons changing orbits (visual) or by molecules resonating (eg as dipoles) involving Ultra-violet, Visual or Infra-red incident radiation, producing Infra-red radiation. 434 MHz is far too low a frequency for such resonating electron or molecular mechanisms. The resonance must be created by physical structures with very fast and simple reaction mechanisms.   

 

The Organelle. Of the major organelles to consider, I shall dismiss the nucleus and the endoplasmic reticulum (ER), because neither has a firm and stable structure and no significant internal energy source. The mitochondria have an ill-defined reticular shape in interphase, becoming globular through mitosis (Rossignol et al. 20048). They are a significant energy source, but require sequential enzyme reactions and shuttles to negotiate the mitochondrial wall, which would slow activity. Their ability to function as UHF resonators at a relatively fixed frequency seems improbable and there appears no clear coupling mechanism to detect and emit UHF.

 

This leaves the centriole as the only significant, identifiable organelle within cells that has the rigidity, uniformity of size and regularity of structure to be capable of being a resonator at 434 MHz (Paintrand et al. 19929, Urbani & Stearns 199910, Marshall 200111). On this basis, let us look at the feasibility of this conclusion :

 

a)     Emission detection and radiation. Associated with the mother centriole (in particular) are the minus ends of the microtubules that are nucleated out of the pericentriolar material (PCM). There is a microtubule concentration associated with the sub-distal appendages of the mother centriole. Microtubules radiate from the centrioles to the periphery of the cells, to be associated with the cell (plasma) membrane. The microtubules are considered to provide electrical conduction pathways for both electrons and protons. The 'aromatic' amino acids, with tryptophane the best, provide electron mobility (Hameroff et al. 200212).  This means that a tumour can be viewed as providing an 'aerial' by its parallel and sequential microtubules-centriole-microtubules-(cell plasma membrane junctions)-microtubules-centriole-microtubules- in series right across the tumour, forming a macroscopical detection unit with a size relative to the 434 MHz wavelength to be effective in signal transfer. Normal tissues may only show minimal effects, possibly due to the reduced parenchymal cell-to-cell compactness of such tissues, reducing the lengths of good conducting paths.

 

                 

              

b)     Power. The centriole is not associated with any known power creator for the cell. However, we have the anaerobic glucolysis described by Needham et al. 1937. This lacks a recognized organelle. If we accept that Holt is correct in showing that some features of this pathway apply, we can assume further that this form of glucolysis must occur in the centriole - its biochemistry now has a home and is no longer a forgotten, unwanted vagabond :

 

                                                         i.                       The process of degrading glucose to L-Lactate was suggested by Needham et al. ' - - glucose give first two molecules of d-glyceraldehyde by a movement of one hydrogen atom from carbon atom 3 to 4, and that these two molecules then produce two molecules of lactic acid by rearrangements of the hydrogen and hydroxyl groups at carbon atoms 1, 3, 4 and 6. The considerable intramolecular oxidation-reductions involved in this scheme might explain the high demand for glutathione'. They provide further elaboration later (p 1924).

 

                                                       ii.                      Such a reaction is associated with Free Energy Changes of Glucose (-686 kcal/mol) to 2 x Lactate (-326 kcal/mol) = (652 kcal/mol), leaving a difference  (~34 kcal/mol of glucose) for the Glucolysis pathway. This energy does not seem to be coupled to any significant cellular chemical energy transfer system (ATP, NAD or NADP etc.). Accordingly, it must be dissipated by atomic, electron or molecular agitation (heat or radiation). Thus, when a cell uses such a form of glucolysis, it loses the opportunity to create 2 of the theoretical maximum of 38 ATP molecules per molecule of glucose. This seeming waste would need to be offset by particular advantages - the speed of response (seconds/minutes - as opposed to minutes/hours) and (possibly desirable) heat production.

 

                                                      iii.                     The catalytic unit (enzyme site) would be created by the apposition of the walls of the glutamylated tubulin forming the rods of the centriole. I would see the organelle in the natural state as constantly writhing due to the contractions of its centrin(s), the contractile, EF hand, acidic proteins of approximately 20 kDa that bind and are stimulated by Calcium. These are found within the distal lumen of the centriole (and between the microtubules of the closely related cilia/flagella), would maintain its tone, and probably show fast responses to both intracellular calcium and calcium waves respectively (Metcalfe et al. 198613). By constantly writhing, similar to the action of related mutant sperm axonemes without radial spoke head protein (Gringras et al. 199814) the catalytic site would be ever changing, minimizing the local heat problem and possibly leaving a 'singe' track (see the helical features in Paintrand et al. 19929). Under the influence of 434 MHz, this situation would change drastically, with electrons &/or ions being pulled around the x 9 triplet/doublet microtubule 'blades' forming the circumference of the centriole(s), providing a 'ratchet' effect to explain the 250-260 kHz minor emission peaks, and the energy supply being triggered to maintain an approximate frequency match with the triggering impulses (see Appendix). This would be expected to release heat as well as radiant energy matching the incident radiation, together with breakdown products, probablyL-Lactate and other 3 carbon compounds at least.L-Lactate produced would be in close proximity to the Lactate-dehydrogenase (LDH) attached to the PCM (Gosti et al. 198715) making pyruvate available (provided the LDH is the right type).

 

Problem 1: Is the intracellular level of glucose adequate, given that tumours have an up-regulated Hexokinase II with a low Km ? (Wilson 200316, Mazurek & Eigenbrodt 200317.) However, intracellular glucose in the green monkey kidney epithelial cell line COS-7 has been found to be about half the level outside the cell (Fehr et al. 200318) with no clear intracellular pooling and no plateaux referable to the Hexokinase II Km. Transformed cells typically have a block in the glycolytic pathway, with a build-up of phosphorylated intermediates. Glucose-6-phosphate and inorganic phosphate may build up enough to suppress the Hexokinase II catalytic action, which is partially compartmentalized by its attachment to, and dependence upon mitochondria for ATP.

 

Problem 2: There is a possibility that glucose may access the centriole via Glucose-6-phosphate entering the ER by the phosphatase transporters, be de-phosphorylated by the Glucose-6-phosphatase and reach the vicinity of the centrioles via the trans-Golgi apparatus, possibly bringing glutathione with it. Needham et al. (1937) provide some support for this in the chick embryo, when Glucose-6-phosphate appeared to be metabolised in intact embryos, but not when brei, obtained by grinding embryos with sand and distilled water in a pestle and mortar, was used - presumably the ER/Golgi was disrupted. A similar fate would occur during mitosis, when the Golgi disintegrates (Sutterlin et al. 200219). In transformed cells the Glucose-6-phosphatase is down-regulated (an important feature in PET scanning). All told, this possible pathway does not seem likely to be significant in transformed cells.

 

Problem 3: Why has this pathway not been noticed in studies since 1937 ? Lactate formed through the Embden-Meyerhoff pathway and Lactate formed through this glucolysis cannot be distinguished. If the latter is only activated intermittently (eg calcium waves, transduction, in embryology, cell stress) it would be difficult to detect, considering that only recently have full 'energy budgets' been assessed, with some surprising results (Guppy et al. 199720, 200221). (See Appendix)

 

Problem 4: What is the point of creating L-Lactate rapidly ? In recent years, there has been the realization that many cells do not particularly like glucose as the major nutrient - some find it toxic (eg the effect of chronic insulin resistance on the beta cells of the Islets of Langerhans). Neurons receive Lactate stoichiometrically from the astroglia 'nursemaids' in response to stimulation by neuronal glutamate transmission* (Pellerin & Magistretti 200322; Magistretti & Pellerin  199923; Meeks & Mennerick 200324), with glioma cells showing a compartmentation of Lactate (Bouzier et al. 199825). Human ova and embryos prefer Lactate &/or Pyruvate (Gardner et al. 199626, 200127), presented by cumulus cells and oviduct cells respectively. Antigen presenting cells also present Lactate to T-cells (Dröge et al. 198728), and one wonders about Sertoli cells# and any other nursemaid relationships.(#Jutte NH, Jansen R et al. J Reprod Fertil. 1983  68:219-26)

*β-amyloid, a player in Alzheimer's Dementia, affects intracellular calcium transport which, in turn, may impact adversely upon the rapid lactate response and production, and contribute to neuronal glutathione starvation (Kasparova et al. 200129; Canevari et al. 200430.)     

 

The mechanism outlined above would represent a mechano-chemical transducer that may convert short ionic pulses into metabolic pathway changes.

 

 

7. The Centriole and 434 MHz

 

a)     The most likely effect of 434 MHz on the centriole is the production offocal heat (athermal heat = confined focal molecular agitation) :

 

                                 i.           Centrioles show a poorly studied 'melting' phenomenon when heated above 40o C (Paoletti et al. 199631), it having been noted earlier with regard to axonemes of gill cilia (Stephens et al. 198932). In some respects, this may be like the effects of hydrostatic pressure upon centrioles (Rousselet et al. 200133). Centrin, tubulin, presumably hSfi1 (Kilmartin, 200334) and calcium are expected to be released into the cytosol inside the Golgi by such heat. Centrin is relatively heat stable (unlike tubulin) and the wrong isoforms (of some 12) in a compartment may perturb cellular and nuclear function (Paoletti et al. 199631; Araki et al. 200135; Popescu et al. 200336) which may be important in the cDNA repair post γ-irradiation. Denatured &/or fragmented tubulin could overload the centriolar proteosome (Fabunmi et al. 200037) blocking substrates p53, Cyclins A, B1, E, Presenelin, IκB and CFTR &/or overload the near saturated chaparonins, TCP (Kubota 200238; Brown et al. 199639; Brown et al. 199640; Won et al. 199841; Sternlicht et al. 199342; Clark & Meyer 199243; Gao et al. 199344) possibly blocking the folding of α and β tubulins, actin and centractin, Cyclins B & E, enolase and transducin. Perturbation of tubulin may stimulate phosphorylation of Blc-2 (Haldar et al. 199745) (see later). The release of Calcium from tubulin and centrin may stimulate the calpains (see later). The overproduction of Lactate would cause a pH drop and may accentuate Calcium toxicity (Kiang & Koenig 199646). There are other structural centriolar components, such as Sp77, Sp83, Rib43 and Tektin and, no doubt, more. The effects of 'melting' or other insults upon these are little studied.

 

ii Pericentriolar material is sensitive to heat. This is particularly so in Drosophila (Debec & Marcaillou 199747) and has been studied in humans (Barrau et al. 197848; Malawista & de Boisfleury-Chevance 198349, Vidair et al. 199350) with clumping of the PCM being the major effect noted. Translation of this change to specific biochemical defects appears limited, but vulnerable could be AKAP450 with its link by Ran to the nucleus (Keryer et al 200351), proteosomes, TCP-1, PCM-1, pericentrins A & B, Hsp70 and LDH at least. Hyperthermia affects microtubule organization (Knox et al. 199152). A general overview is presented by Dewey (198953). Of interest, heat is believed to cause centrosomal dysfunction, resulting in non-apoptotic mitotic catastrophe (Nakahara et al. 200254). Perturbation of the nucleation of microtubules in the pericentriolar material will be expected to impair mitosis and the morphology of differentiated cells.

 

b)  There may be other effects produced by ionic &/or redox perturbations. Little information on these seems available at present. Glutathione may be oxidized, reducing its protective effect upon γ-irradiation, with increased cellular sensitivity. (Refer to Dröge 200255 for a Review on redox reactions.)

 

8. The Centriole and ERex. By now, the centriole should seem to share some features with Holt's ERex unit: re-reading his writings with this thought opens new perspectives and concepts :

 

a)     The number of centrioles/ERexs per cell. A typical mammalian cell contains 2 centrioles within the centrosome (diplosome) during interphase (prior to G1-S): the number being linked to ploidy status (Bessis 1977). Transformed cells (probably) contain many more: Sharp & Osborn (198256) reported that rat pup dorsal root ganglia contained 2 centrioles/cell, whereas neuroblastoma cell strains contained an average of between 3.4 to 5 centrioles per cell, with even numbers preferred. There are those who believe that centrosomal and centriolar abnormalities figure prominently in malignant transformation (Lingle & Salisbury 199957, Lingle et al. 200258, Pihan et al. 200159). In the 2002 report, there were 1.5 centrosomes/cell registered for normal tissues (some may have been dividing), 2.8 for diploid tumours, 2.7 with monosomy tumours, 3.6 in stable, aneuploid tumours and 6.8 in unstable, aneuploid tumours. If we assume each centrosome in a malignant cell contains ~2 centrioles, the above numbers can be doubled, giving an average of about 13.5 centrioles/unstable aneuploid cell (the scatter is unclear).

 

 b) Holt estimates, using radiotherapy theory, that there are 2-3 units (representing x, n or m [depending upon author] 'hits' required, derived from graphical extrapolation, Andrews 1978) per lymphoma cell (sensitive), 2-4 (occasionally up to 7) for squamous cell cancer of head and neck, 3-20+ for cervical cancer, 15 for limb 'cancers' (probably sarcomata).

 

Such figures are similar to those presented by Elkind (198860) using data from other authors, such as Deacon et al. (198461). Here, n (= previous x, above) for sensitive tumours has a mode of ~1.5, rising to a mode of ~8 for resistant tumours, with a scatter from about 1.5-100. So, the numbers appear consistent with the idea that ERex may equate with the centriole.

 

c)     Holt presents the view that the lethal 'hits' are to his ERex units. But many of the 'inactive' ERex units are resistant to γ-radiation. Those 'inactive' units are 'activated' by the 434 MHz and become less resistant, so that the effective number drops to ~1 per cell.  (eg x drops from say ~ 5 to ~1). The x simply represents the (metaphorical) lethal 'hits' per cell that are required for clonogenic failure, without consideration of the size or sensitivity of the tumour cell 'target'. If the target becomes effectively larger, or more sensitive, the chance of a lethal 'hit' from the same radiation dose increases, so x becomes smaller per given tumour, and may be able to approach one, the radiation dose that would normally produce one lethal 'hit'.

 

In that the treatments are conducted over time in courses, there is time for cellular processes to be moulded by therapies. For instance, the mother centriole is more clearly attached ('wired in') to the microtubule mesh. The daughter(s) go walkabout on actomyosin leashes, whilst mother's duties are more domestic, tied by the microtubule apron strings (Piel et al. 200162; 200063). Daughters may thus be regarded as 'inactive' as far as 434 MHz is concerned. If the mothers are disabled by 434 MHz +/- γ-irradiation, and no more daughters can be replicated by the mothers due to a block in the cell cycle or maternal damage, the existing daughters may mature in time to be more 'active' later in the treatment course. This may explain in part the 'activation' effect of 434 MHz on Holt's ERex. Another explanation may be that the 'melted' centrioles leave a vulnerable template/skeleton for a time which, if 'hit', aborts functional reconstitution of the organelle or formation of the pro-centriole. Total ablation of a centriole would not seem desirable, given that transformed cells stripped of centrioles may generate replacements by de novo replication (La Terra et al. 200564). Whilst maimed centrioles remain in cells, the de novo replication could be blocked by the unknown factor(s).

           d)   γ-radiation has been presented as causing centrosomal/centriole defects (Sato et al. 198365,
                 200066). These seem to affect nuclei secondarily

All that has been noted so far is consistent with Holt's ERex effectively being the centriole, but with different interpretations and implications.

 

9.The centriole 'removed' by catabolism can regenerate over about 12 hours (Bobinnec et al. 199867), provided its 'template' skeleton is not damaged. Heat/electrical assaults listed above are little more than nuisance value to the centrosome unless there is damage to the 'template' or other structures that prevent centriole reconstitution, duplication, or precipitates cell death by apoptosis or necrosis.  The centriolar and pro-centriolar 'templates' would seem to be particularly vulnerable, given that untransformed mammalian cells cannot easily replicate centrioles de novo and are not free to replicate (Wong & Stearns 200368). Untransformed cells without centrioles cannot progress through G1 to S (Hinchcliffe et al. 200169), and HeLa cells (transformed cells) deprived of Centrin-2 for their centrioles continue to divide, but with decreasing centriole numbers, and ultimately develop giant cells and die (Salisbury et al 200270) - (the death process was not defined).

 

10.  Cell Death. There are numerous pathways for cell apoptosis, with many interacting. Traditional is a separation into 'extrinsic' and 'intrinsic' pathways, the former starting with plasma membrane receptors and involves c-Jun, the latter is triggered by intracellular perturbations and the release of one or more BH3-only pro-apoptotic factors, which inactivate the Bcl-2 pathway. Both ultimately involve Bax, cytochrome-c and Apaf-1, (apoptosome), activation of Caspase-3, producing endonucleases which cleave the DNA into oligo-nucleotides.

 

There seems scope for both these pathways to be activated by 434 MHz +/- γ-radiation. A summary of pro-apoptotic factors in relation to the 'intrinsic' pathway is presented in the Table 3 :

 

                                                                TABLE 3

 

                         PRO-APOPTOTIC FACTORS - 'INTRINSIC' PATHWAY

 

   Cyto-                          Light               BH3-   Chromo-    Stimulus            Absence

   Skeleton    Motor    Chain 8 kDa  only      some           apoptosis           affects

 

   Actin#        Myosin   DLC2              Bmf*   15q14          Cyto-                  Metastases

                                                                                                    chalasin D/         lose Bmf

                                                                                                    actin

                                                                                                    Cl difficile 

                                                                                                    UV

                                                                                                    Anoikis

 

   Micro-      Dynein     DLC1              Bim*    2q12/13      Cell cytokine     Myeloidup-reg.

     Tubules#                  LC8/PIN                                           loss (eg IL-3)     Lymphoid up-reg.   

                                                                                                    Gamma rays      Tumour suppressor

                                                                                                    Staurosporine         loss

                                                                                                    Anoikis

                                                                                                    UV

                                                                                                    Taxol

                                                                                                    Glucocorticoid

                                                                                                    Calcium flux

                                                                                                    Ionomycin

 

#Linked by Spectrin (De Matteis & Morrow 200071) and by engagement (Cao et al. 200372)

*Phosphorylation by JNK releases Bmf & Bim from motor complexes (Lei K & Davis 200373)
 

 The topic has been reviewed by Puthalakath & Strasser (200274), with more recent work by Day et al. (200475). There is the impression that 434 MHz is more inclined to produce 'gratifying' results in those tumours that have not yet metastasised or are not inclined to do so until late (eg mesotheliomata, gliomata and hepatomata). If this impression is correct, the involvement of Bmf may be suspected. The report by Peter French (see earlier) describing actin changes may be in accord. Bim, transcribed from a different chromosome, may be more generally available (Bouillet et al. 200176).

 

Histology changes. Of interest, with respect to 434 MHz, is that histology seems to indicate a rounding and dedifferentiation of cells (see earlier). Such cytoskeletal changes would be expected to trigger anoikis and those factors more closely related with integrins, actin and microtubules, Bmf & Bim.

 

Other pathways include :

 

i.  Calpain (Jayanthi et al. 200477), which shows some attraction, since m-Calpain is triggered by (substantial) Calcium waves. However, to function, reducing conditions are needed (McCollum et al 200478), so the oxidized glutathione in the iv infused 'glucose blocker' may block this pathway for the time the more oxidizing conditions prevail. The activated m-Calpain cleaves Bcl-xL and Pro-Caspase 12 (Nakagawa & Yuan, 200079).

 

ii   Nuclear Clusterin is relatively recently described, being unlike the main component that resides in the cytoplasm, and which is anti-apoptotic (Debure et al. 200380). It is induced by ionising radiations, translocates into the nucleus and brings about apoptosis (Yang et al. 200081; Leskov et al. 200382). Clusterin(s) seem particularly involved in the culling of spermatozoa in the testis (Barone et al. 200483) by the Sertoli cell nursemaids.

 

      There can be some interest in this pathway, because Holt claimed that he and Dr Nelson noted that males undergoing 434 MHz became sterile (personal communication 2003). Females seemed unaffected. Both Holt and Nelson subjected themselves to the 434 MHz and Holt reported that the result was 'complete aspermia'. Apparently there was recovery (in part at least) one year later. The nuclear Clusterin pathway may have been involved.

 

iii.     Ceramide. This relatively poorly studied pathway seems to be triggered by reactive oxygen species (ROS) related to mitochondria (Quillet-Mary et al.199784) with ceramide translocating and activating Protein Kinase Cd in the Golgi (Kajimoto et al. 200485; Jackson & Foster 200486) Diversion of glucose metabolism through the centriole may deplete glucose, encourage the production of ROS (Aulwurm & Brand 200087) and induce apoptosis.

 

iv.    There appear to be others less studied.

 

11. The 'glucose blocker'. Needham et al (1937) describe the need for glutathione (GSH) to enable the glucolysis to proceed. In general, GSH is not easily taken up by cells, being broken down and reassembled. Its disulphide (oxidized glutathione, GSSG) is hardly taken up at all. GSH seems important in the early phases of an immune response (Lacombe et al. 198788, Dröge 199489) and the binding of NFkB to cDNA (Mihm et al 199590). Having entered the ER, possibly against a gradient, a significant proportion of GSH forms mixed disulphides with protein (Bass et al. 200491) and is oxidized to GSSH, apparently as a buffer for the protein disulphide isomerase/Ero1 reactions which fold nascent   proteins in the ER (Moltini et al. 200492, Ostergaard et al. 200493). There is some evidence that in some specialized circumstances, GSH may pass from one cell to another within minutes (Schuttle & Werner 199894). With glutamic acid a terminal amino acid, and glutamylated tubulin forming the centriolar microtubules, glutamate groups may be able to move around the microtubular structure. How GSSG may help treatment with 434 MHz is unclear, unless by favouring the oxidized equilibrium state, it inhibits cellular uptake or, if intracellular, causes a depletion of intracellular free GSH89. By depleting mobile intracellular electron/proton donors, the electrons/protons may be pulled from structural proteins, with resulting damage or cross linkages. The S-methyl-L-Cysteine Sulphoxide role seems largely based on empirical assessment. It may be a competitive blocker, perhaps by blocking the formation of glutathione, but why that should assist cancer treatment responses is not clear, unless it diverts the electron/proton movements described above to involve structural proteins in a more lasting way.

  

12. Safety of 434 MHz+

 

a)     434 MHz, when delivered at power levels (plate current) of 1600-2000 Watt produces 'comfortably warm' heat. Patients are usually heated for 8 - 10 min then given a short break, before another 8-10 min. They usually develop a sweat.  They are fully awake so that, in the worse case scenario, they could slide out of the heat field if there were any failure of staff. I have never seen this, have never seen any burn or distress, and believe it to be quite safe. Holt reported (personal communication) that he had a problem with a child in about 1975. The child had a large intracranial cystic fluid collection. The presumed problem (and there was some doubt) was expansion of the fluid by the heating, causing brainstem/aqueduct obstruction. Care in patient selection with this regard is important. Exclusion of electronic pacemakers, implanted TENS machines etc. is mandatory. Artificial (metal) hips etc. do not seem to be a problem - the policy being to use less power for longer intervals. No patient has complained, noted local heat or deep pain, nor indicated that there was any problem. Holt has referred to a patient from the Middle East who, I understand, seemed to haemolyse as a result of the 434 MHz+. I believe that this was attributed to thalassaemia. I find this explanation difficult to accept. Assuming the patient had a form of thalassaemia, I believe that the cause was most likely due to Paroxysmal Nocturnal Haemoglobinuria as well. The latter can be triggered by minor stresses, glutathione, mild acidity and peroxide - all involved in the 434 MHz+ (Wintrobe 1981*)

 

*Wintrobe MM et al. in 'Clinical Hematology', 8th Edition., 1981. Editor M Wintrobe, page 3 978-990. Lea & Febiger, Philadelphia.  (The 10th  Edition [1999] seems to deal less with physical and chemical causes of haemolysis.)

 

b)     The 'glucose blocker' of oxidized glutathione and S-methyl-L-Cysteine Sulphoxide is hypertonic and needs large veins. Even then it can cause pain in the upper arm and shoulder if it is infused too quickly. I have never seen venous thrombosis as a result. It gives a taste of 'boiled cabbage'. If there is extravasation there is a swift complaint by the patient. By the next day there is only mild erythema and oedema. I have never seen ulceration or skin necrosis. It seems as safe as any hypertonic saline solution. I have never seen any systemic reaction - shock, faint, bronchospasm, laryngeal spasm, deep tissue oedema, generalized rash etc.  It seems to pose no risk to the vast majority patients.

  

13. 434 MHz+ and my patients. Referring to the graph of VD's α-FP values, the likely explanation is that the 434 MHz+ induced a slow-down through mitosis, so that there was a build up of cells with the Golgi apparatus dissipated. This would block secretion of molecules such as α-FP. Later, there may have been a period of cell breakdown, with α-FP being released, and the subsequent rise less steep because there had been created centriole defects similar to the lack of Centrin-2, producing impaired or limited mitosis.

 

The urine excretion of Calcium may reflect release of Calcium from bone and cell stores (chiefly the ER, but also the mitochondria and possibly centrosome and microtubules. The seemingly mild fall in urate excretion may also reflect Golgi dissipation during mitosis. (Whilst the biochemistry of purine breakdown seems well worked out, the organelles involved are not so clear, with two pathways described, one oxidizing, the other reducing, with probably most breakdown in or associated with hepatic parenchymal peroxisomes and cytoplasm  Frederiks & Vreeling-Sindelarova 200295.)

 

14. Future Research.

 

a)  The first aim should be to reproduce Holt's resonance phenomenon in humans and animals. The small size of animals and their  tumours in relation to the incident wavelength may preclude adequate absorption of the energy. Aerials will need to be dipoles and the frequency analyses needs to be analogue (according to Holt)

b)   Good morphological studies at both the light microscope and the electron microscope levels should be sought.

c)  Why the cancers show this resonance phenomenon apparently much better than normal cellular tissues (brain, liver, spleen) should be sought - is it because more centrioles can be 'wired-in' or some biochemical explanation specific to the cancer cells ?

d)  The roles of glutathione and calcium (in particular) would seem worth pursuing.

e)   Whether the responses of tumour markers &/or calcium may have some prognostic significance.

f)    434 MHz and γ-radiation. Confirm the augmentation, seek explanations and improvements.

 

I would suggest that any unit set up to research the phenomena should be close to, but independent of major cancer treatment facilities.

 

Malcolm A Traill

Copyright © MA Traill 2004, 2005, 2008