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44MITOGENETIC EFFECT (STIMULATION OF MITOSES WITH ULTRA-WEAK UV-RADIATION) AND ITS APPLICATIONS TO CANCER RESEARCH AND DIAGNOSTICS
ILYA VOLODYAEV1 AND ELENA NAUMOVA2
1Biological faculty, M. V. Lomonosov Moscow State University, Russia 2D Rzhanov Institute of Semiconductor Physics, Russian Academy of Science, Russia
a_naumova@isp.nsc.ru
Abstract
The report briefly reviews early researches on the mitogenetic effect (1923-1948) including the cancer diagnostics based on this effect and the claimed tumor marker. The key methodical details of mitogenetic effect observation are pointed outon the ground of the relevantliteratureandpracticalexperienceofoneoftheauthors(IV). Prospects and necessity of the unambiguous verification of mitogenetic effect and related cancer diagnostics are shown. The review covers both published and unpublished materials from the unique archive by A.G. Gurwitsch and his scientific dynasty.
The phenomena of ultraweak endogenous radiation of biological systems and mitotic rate stimulation with ultra-weak UV-radiation were discovered by A.G. Gurwitsch in 1923 [1]. He directed the root of one onion (inducer) onto the meristem zone of the other root (detector) and observed that the inducer locally stimulated mitoses in the detector. This phenomenon was called a mitogenetic effect. The factor that mediated mitogenetic effect was shown to be ultraviolet radiation (chemical isolation of the inducer from the detector and insertion of quartz plate did not influence the effect, while a glass plate eliminated it). It was called mitogenetic radiation. Mitogenetic effect was observed in microbe and cell cultures, and tissues. When an inducer and a detector belonged to different families or even kingdoms the effect was also observed. There were more than 700 publications devoted to mitogenetic effect in 1923-1948. This effect was proved by more than hundred researchers including Nobel Prize Winner D. Gabor, academician G.M. Frank, famous microbiologist C. Wolff and others [2-6].
The spectra of mitogenetic radiation of various inducers were studied with spectrographs having rows of biological detectors instead of photographic plates [7, 8]. Spectra of most of the biological inducers were reported to be within the range of 190-250 nm, mitogenetic effect was induced also by a number of biochemical and even inorganic chemical reactions [9]. It was suggested that an energy released in chemical reactions involving free radicals resulted in the excitation of specific molecules or groups and could be emitted as quanta of mitogenetic radiation [10]. Intensity of mitogenetic radiation was estimated with modified Geiger-Müller counters as 10-1000 photons/cm2 s [9].
The study of cancer cells and early cancer diagnostics were the most developed practical applications of mitogenetic effect. Whole blood and blood serum of young, healthy animals and people were proved to be good inducers of mitogenetic effect, while blood of cancer patients and animals did not induce it [11, 12]. The lacking of mitogenetic effect from blood was actually not an utterly specific indicator of cancer. For instance, blood of exhausted people did not induce mitogenetic effect also [13]. It was an ability of blood being added to any other known inducer to inhibit mitogenetic effect, that was really specific for cancer patients [14-16]. The substance responsible for the luminescence quenching in inducers was highly specific for cancer diseases and was called a "cancer quencher". Nowadays, it is alpha-fetoprotein that is generally recognized as the first tumor marker. However, an existence of a tumor marker in blood, which was called cancer quencher, was claimed almost thirty years earlier. The physical and chemical properties of the cancer quencher were studied and it was shown to be a peptide. An important fact shown in [17] and other works, was that blood of animals with experimental tumors (induced with chemical carcinogens, injection of suspension of malignant cells, or tumor transplantation) quenched luminescence of other inducers before the malignant process could be diagnosed clinically. Compliant results were obtained in clinics, for instance, the cancer quencher was found in blood of women with myoma only in those cases when they were to get cancer later, and before tumor could be diagnosed by any other means [17]. According to the ample clinical data of the leading medical institutions of the USSR gathered in 1940s the cancer diagnostics based on the detection of this tumor marker had specificity and sensitivity >95%. The detection was based on the lacking of mitogenetic effect from the yeast inducers to which the studied blood was added.
Researches on mitogenetic effect were conducted in 1928-1935 years mainly in the USSR and in Germany, and also in the USA, France, the Netherlands, Italy, Japan and other countries. In the USSR the works were persecuted altogether with genetics after the decisions of the All-Union Academy of Agricultural Sciences in 1948, see details in [18]. In Western countries these researches declined even earlier and were completely abandoned in the beginning of the World War II. It happened mainly due to historical reasons and a few influential negative publications (e.g. [19, 20]). Detailed analysis of methodical aspects of these works reveal severe violation of the methods suggested in the "positive" works (See [18]). The main points there are: (1) the recipient culture state (it should be a certain part of the lag-period or post-diauxic phase); (2) the time and duration of exposure (it should be optimized individually for each inducer-recipient pair;
the effect of over-exposure was observed in a many works); (3) some physical limitations, like the recipient culture density less than a certain limit, very thin suspension layers, semidarkness, lack of any external UV.
Further experiments on the subject were rather sporadic and gave no final proof or disproof of the key results of 1923-1948. One of us (IV) participated in attempts to reproduce mitogenetic effect, his scientific team managed to get a good reproducible effect from the yeast-inducer culture on the yeast-detector culture which was in the lag phase during the exposure [21]. Yet, the effect appeared to be caused by CO2, which was secreted by the inducer and accepted by the detector (it was also shown to be signal-like action, but not purely metabolic). Later, analyzing their results in the context of publications of 1923-1948, the authors came to the conclusion that they paid not enough attention to methodical details of early works, and if the mitogenetic effect existed, it was sure not to manifest itself under used experimental conditions. First of all, it were the culture state and the medium composition that made mitogenetic effect observation impossible, and the effect, which was observed [21], had nothing to do with mitogenetic radiation that was the initial goal of the work (see [18]). After this failure the authors of paper [21] including the author of this report (IV) came to the deep reading of the methodical conditions of early works and besides their own mistakes, concluded that all the well-known "negative" works [19, 20, 22-25] also contained critical deviations from these methods, that would have guarantee their failure in the mitogenetic effect verification. Detailed methodical recommendations were given in a number of early works e.g. [26, 27], see also both positive and negative works analyzed from methodical viewpoint in [18] (Fig. 1).
A
c O <J
Bacteria
Inoculum culture:
E.coli. Suspension culture (-5 * 10B Cells/ml).
Age: 48 h. 24 h - no effect!
Recipient suspension layer <0.6 mm
Inductor quartz
Medium: broth 1:10
Induction
OR
3 Recipient suspension layer <0.6 mm
Inductor
quartz
Delude in medium (1:10 broth) to 5—5000 cells/ml, 105 cells/ml - no effect!
Incubation: 373C Sampling —
Growth curve
Ä
Yeast
Inoculum: Burgundy yeast 24h suspension culture Medium: raisin extract, 30°C
24h agar culture Medium: raisin agar
Wash off: 5 ml H20
Delude 1:100 with H20
Seed: cells are far apart
Induction (30°C)
^ Inductor -quartz -Recipient
I
Fix on agar (iodine) Budding curve
Figure 1: Modern analysis of the most effective methods for obtaining the mitogenetic effect on bacterial (left) and yeast (right)
cultures (after [18])
Many conclusions made in experiments on MGE were proved to be correct and much ahead of their time. These are: an existence of tumor peptide markers in blood, photo-reactivation, anti-Stokes luminescence, free-radical mechanism of ultra-weak photon emission etc. Yet, the phenomenon itself remains an unresolved problem in both fundamental and applied biology till now in spite of the progress in UV-sensing. Particularly, photomultiplier tubes with AlGaN photocathodes have a high spectral sensitivity in the whole mitogenetic range and extremely low dark counts that makes signal/noise ratio high enough for the unambiguous verification of experiments on mitogenetic radiation and related cancer diagnostic.
Verification of experimental results on mitogenetic effect and cancer quencher with the use of present-day techniques, knowledge and level of evidence is very promising both for basic science and practical applications and requires a thorough revision of early publications. More details about mitogenetic effect and cancer quencher can be found in monographs [8-10, 27-30] and reviews [18, 31, 32].
References
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[2] T. Reiter, and D. Gabor, Ultraviolette Strahlen und Zellteilung, Strahlentherapie, (28), 125-131, 1928.
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