English

Development, Growth, Behaviour and Differentiation of Living Materials Exposed in Front of a Cathodic Ray Tube Screen. A Review.
SURBECK, Jacques, LAVERDURE, Anne-Marie, NORTH, Marie-Odile and TRITTO, Giuseppe

- Surbeck J., CIST / ICOH Member, S.E.I.C. sa, 3 rue du Léman, CH-1201 Genève. E-mail: surbeck@pingnet.ch
- Laverdure A.M., CIST / ICOH Member, A_Nox Technology Laboratory, 41-43 rue de Reuilly, F-75012, Paris, France. Tel. [33] 1 43 46 92 34 Fax: [33] 1 43 42 52 72. E-mail: ALaverdure@aol.com
- North M.O., CNRS, UMR 147, Institut Curie, 24 rue d'Ulm, F-75005 Paris. E-mail: madile@aol.com
- Tritto G., Service d'Urologie, Hpital Saint-Louis, rue C. Vellefaux, F-75010 Paris. E-mail: gtritto@magic.fr

Presented at the 2nd Conference of the Victorian Chapter of the IEEE Engineering in Medicine and Biology Society
Melbourne (Australia), 19-20 February 2001

Published in the Proceedings of The 2nd Conference of the Victorian of the IEEE Engineering in Medicine and Biology Society - “Biomedical Research in 2001” - held at Monash University, Clayton, Victoria, Australia, pp. 195-202


ABSTRACT

Cathodic Ray Tubes Screen (CRT's) equipping personal computers and televisions emit a large spectrum of radiations responsible of some biological actions. Development disturbances in Drosophila (increase of the total dead ratio and of the development time) and chick embryos (malformations) have been observed. Behavioural and physiological disorders appeared in rats exposed during their neonative period and studied after their puberty (micturitions, locomotor activity). In vitro cultured biopsies showed important mitotic and meiosis perturbations (increase in apoptosis and cell death). Causes of those disturbances have to be determined. Note a particular sensibility of embryonic, prepubertary and cellular maturity periods, marked by numerous mitotic crisis, and efficiency of the tested protections. With the aim of protecting the users, several living biological materials and human testicular biopsies were breeded or cultivated in front of a CRT equipped or not with the EMF-Bioshield® device. Measured electric (13 V/M) and magnetic (50 nT) fields (EMF) were identical with and without EMF-Bioshield®. An inadaptation of the actual measuring devices can be deducted from absence of differences in the physical measures. But we can also hypothesize a modification in the emitted spectrum of radiations in presence of the EMF-Bioshield® device occurs.

Key words : Cathodic screens, EMF-Bioshield®, development, behaviour, meiosis.


INTRODUCTION

Before 1990, only a few number of papers related to physiological actions of the CRTs total spectrum of radiations in living beings were available. Among the reported results, an increase of the percent of malformations in chick embryos exposed to weak extremely low frequency pulsed electromagnetic fields, has been demonstrated [1].

The CRTs are known to emit electromagnetic waves, ionizing ones such as low energy X rays (1,2 kKeV, 0,1 mSv/Yr) and non ionizing ones such as ELF (extremely low frequency: 0 to 3 KHz, 65 V/M, 0,2 A/M), VLF (very low frequency: 3 kHz to 3 MHz, 150 V/M, 0,1 A/M), and sometimes HF, VHF (hyperfrequencies: 3 to 300 MHz, 0,5 V/M, 0,0002 A/M), photonic rays such ultraviolets (200 to 400 nM, 0,1 W/M2), infrareds (700 nM to 1 mM, 4 W/M2), light radiations (400 to 700 nM, 2,5 W/M2), electrostatic fields (0 Hz, 1,5 kV/M) and various sound waves, particularly ultra-sounds [2]. These measurements, performed on 200 to 3000 CRTs from different origins indicate the existence of very complex low power radiations. In addition, the contribution of harmonics has been estimated to a little less than 13% of the total radiation spectrum [3]. More recently, it was admitted that some of the electromagnetic radiations emitted by the CRTs are undetectable by our actual measuring devices, but are able to provoke some biological effects on living organisms: the obtained results have to be considered [4].

Considering the worldwide proliferation of the cathodic devices, including the video games screens particularly used by children, the choice to study a possible effect of the total spectrum of CRTs radiations has been taken. The biological actions of those radiations have been first tested on living organisms in which growth, development, reproduction and behavioural modifications were studied, then in human testicular biopsies cultured in vitro. An experimental apparatus, the EMF-Bioshield® device, has been developed, in the aim of interfering the CRT radiations effects and tested to evaluate its possible protective action.


MATERIALS AND METHODS

The computers were directly connected on the electrical system (a 50 Hz frequency alternative current under 220 V in Europe and a 60 Hz under 110 V in U.S.A.).

During the Drosophila and chick embryos tests, we used an IBM-PC color monitor, model 4863002, lighting at its maximum. For the experiments performed with rats, we used current CRT (Wang Model 2236 DE) from which the physical measurements (with a HI3604 - HI3603 Survey Meter Hololay Industries Inc.) indicated 30 nT in the VLF and 200 nT in the ELF and 300 nT in the VLF. The corresponding electric fields were 3,2 and 50 V/M. For the biopsies cultures, we used TCO-95 norm MPR2 CRT screens (low radiations) (Philips design type 7CP 5279/30T). Intensities of the electric and magnetic fields (EMF) of the CRTs, equipped or not by the tested EMF-Bioshield® device, were measured with a digital multimeter METEX 3800 at 30 cM from the central point of the screen. The obtained values corresponding to the total spectrum of radiations were 13 V/M and 50 nT respectively.

Three groups of experimented organisms have been considered: "Controls", "Exposed", "Exposed + Bioshield". "Control" breedings (drosophilas), animals (chicken embryos, rats), and samples (human testicular biopsies) were kept in normal controlled breeding or culture conditions (feeding, constant temperature, relative humidity and light), out of the CRT radiations. The tested organisms and samples: "Exposed" and "Exposed + Bioshield", were rigourously maintained in the same conditions as controls but continuously exposed at 50 cM in front of a current CRTs equipped (said "Exposed + Bioshield") or not (said "Exposed") with the EMF-Bioshield® system.

Drosophilas from the Charolles stock (Cha-C-) were maintained in standard conditions. Before experiments, a large number of 2-days adults were picked from the breeding and transferred on a large egg-laying space. Few hours later, it was replaced by a new one and left for 1 hour. The eggs were gathered and spread in groups of 50 in 3x10 sterile tubes containing feeding material. They were kept in 3 separated incubators (10 tubes per incubator) in which hygrometric conditions were under control (80% RH) and the temperature maintained at 25ÉC and continuously registered for eliminating an eventual warming due to the computer. The "Unexposed" 10 control tubes were placed into the first incubator, far away from any electromagnetic perturbations. The 10 "Exposed" tubes were placed in the second one and the screen was placed inside the incubator. The 10 "Exposed + Bioshield" tubes were placed in the third one and the screen equipped with the tested device was placed as described. The Drosophilas were placed at 50 cM from the axial point of the screen and exposed 24 hours a day to the radiations. Embryonic mortality % was determined by counting the white unfertilized eggs (14,2%) and the residual chorions after the first larvae hatching. Comparison between the number of adult flies compared with the number of hatched eggs permitted us to determine the total mortality %. Confidence limits for each percentage were determined and the percentages compared by calculating the chi square. The sex ratio value was reached by comparing the number of males to the total number of adults obtained.

Chicken embryos were classically breeded in a thermostated incubator and exposed to the radiations during the first 5 days of their embryonic life. The screen was placed on the transparent cap of the incubator. Then, the embryos were studied, the number of dead determined and the developmental and teratological disturbances described.

The rats (White Wisstar) were maintained under artificial 12/12 photoperiod at 22 ± 2ÉC. The animals were exposed in plexiglas cages 60 cM away the axial point of the screen. The exposure, from 7.00AM to 7.00 PM, took place during the days 1 to 15 of their neonative period. Then, the animals were breeded in normal conditions and studied after their puberty (70 days old). Behavioural and physiological open field-tests were realized: Animals were carefully picked up from its cage, gently dropped in the center of the cleaned experimental field and observed during 5 min. Locomotor activity was determined by calculating the number of crossed squares and a classical determination of the number of micturitions is a good test to have a first approach of eventual physiological modifications.Analysis of our quantitative results was done through the Student-Fisher tests, adjusted to the case of small samples (n Ê 30). The study of the comparison between the different averages was done by calculating the t of Student.

The possible action of the CRT radiations on humans was studied in testicular biopsies. We used testicular samples originated from five 25-35 years-old patients who presented a varicocele. Specific consent form was agreed by the patients and the protocol approved by the Bioethical Committee of the Centro Internazionale of Andrologia in Roma where experiments were realized. Biopsies were taken off in the two testis and divided in four fragments : one was immediately fixed in Bouin's solution as a control (A) and the three other parts ( B, C, D) were submitted to a touch imprint, then immersed in a standard culture medium (Ham F10), and maintained for 24 hrs in thermostated incubators [5, 6]. Temperature was continuously measured by an AMR Type 2290-3 and a Testoterm Type 451 registering aparatus. The sample (B) was cultivated without any particular treatment. Testicular biopsies controls were obtained at the start of the experiment (sample A = T0) and after 24 hrs culture (sample B = T0+24 h). In the same experimental conditions, each of the two C and D samples were exposed at 50 cm in front of a Philips computer "low radiations" (MPR II-TCO95), 14 inches, design type 7CM 5279/30T, serial numbers HD 0097 062 18 910 and HD 0097 062 18 900, but one of the two screens was equipped by EMF-Bioshield®. At the end of the experiment, the explants were cut in two pieces. The first pieces were fixed as before, dehydrated, embedded in paraffin and 5 µm-sectionned. The sections were mounted in a 0,2% gelatin (Sigma)/0,05% chrome-alun and dried. After deparaffinization in xylene and rehydration, the slides were coloured by hematoxylin-eosin or treated for apoptosis study according to the instruction of the supplier (Boehringer Mannheim). The slides with touch-imprint were fixed into 70É alcohol and coloured with eosin-hematoxylin. With the second pieces, air-dried meiotic preparations were done according to standard [7]. All the cells were identified and counted, each meiotic abnormality indexed and counted. Meiotic abnormalities entered in the class IIC of the Meiotic Abnormalities Classification [8, 9].

The tested experimental device, supposed to protect the screen users against the harmful effects of the screen radiations is constituted by 2 minispheres filled with rare earth oxides solutions and has been previously described [10].


RESULTS

An important peak of mortality has been observed in Drosophila, where the ratio of adult mortality significantly increased from 14,8 ± 1,4 to 32 ± 3,5. Two successive experiments were carried out with 500 eggs positionned in front of the screen. Observation of the imagos realized on the hatching day showed that this significant increase in total mortality was not accompanied by a significant alteration of the sex ratio. The living adults appear morphologically normal. In females, a microscopic observation of their ovaries did not show any evidence of modification.

In chicken embryos exposed to a CRT screen, a 60% increase of teratological damages and infertility were observed. Important anatomical perturbations, wing and foot lethal malformations have been observed.

Importance of the irradiation time was studied. In Drosophila, embryonic mortality (observed at the embryos hatching) was determined by an exposure to the CRT radiations during the embryonic life. After a 500 eggs exposure during the whole embryogenesis, embryonic mortality did not present any significative disturbance when compared with the control value (2,1 ± 0,19 vs 2,6 ± 0,23). But after an analogous short exposure, the ratio of adult mortality (measured 10 days later) directly reached the maximum value precedently described: the embryonic life appears as a sensitive period and the effect seems to act a posteriori, as an after-effect.

A few studies were related to behavioural and physiological disturbances : in male and female rats exposed during their neonative period, significative modifications of the number of micturitions and locomotor activity were observed over puberty (Fig. 3A, 3B). Obtained with groups of 12 animals, a significant increase in the number of micturitions appeared in males and females, indifferently. In the same time, their locomotor activity decreased, as demonstrated by the reduced number of the crossed squares.

In normal testis, compared with the uncultured control piece (Fig. 4, A), the cultured biopsy showed a good survival after 24 hrs of culture (Fig. 4, B) with presence of all kind of cells with normal distribution rates and normal meiotic intercourse. The rate of apoptosis remained extremely low in both (Ê5%). After 24 hrs of culture in front of a VDT screen, some degenerating signals appeared (Fig. 4, C). The meiotic abnormalities were typical impairment secondary to unfavourable physiological background : autosomal synaptic failure (asynapsis, breakdown, high level of autosome-sex vesicule association), degeneration (high level of background silver deposition, fragmentation of synaptonemal complex, XY excredence). The rate of meiotic abnormalities increased from 5 to 20% of all the meiotic cells (Table 1). Histopathological degenerating signals consisted in appearance of slight seminiferous tubules modification (thickening of wallsmonash1.html), low pycnotic cell rate, low spermatid number decreasing. The rate of apoptosis became significantly higher (~20%) correlated to histopathological degenerating aspects and to meiotic abnormalities.

In the varicoceled testis, the two controls (T0 and T0+24) showed the same effect of varicocele on spermatogenesis (Fig. 4, E, F) : high rate of meiotic abnormalities (~50%), low rate of spermatid cells with rare spermatozoa, seminiferous tubules disorganization, high rate of apoptosis (~50% of the cells). In the cultured sample in front of the screen (Fig. 4, G) , the degradation of spermatogenesis was amplified with 80% of meiotic abnormalities, high rate of seminiferous tubule disorganization, high number of pycnotic cells, rare spermatids and no spermatozo, and ~90% of apoptotic cells.

As a complement to our experiments, an experimental apparatus, EMF-Bioshield® has been tested in the same experimental conditions to evaluate the possible protective effect of this system set placed on the screen.

In Drosophila exposed during the whole development, no significant difference with unexposed controls carefully kept away from the radiations could be observed : this device ensures an effective protection against the noxious effect of the radiations. An analogous observation has been realized in chick embryos, accomplishing in a normal way their development in front of a screen equipped with the tested device. Also in rats, the presence of the EMF-Bioshield® system on the screen had not permitted to observe the parameter modifications precedently described. In the normal testicular biopsies cultivated in front of an equipped screen with the tested system, meiotic, histopathological and apoptosis results were not significantly different from those of the controls T0 and T0+24. For the varicoceled sample, we found again the same aspect of the controls T0 and T0+24.


DISCUSSION AND CONCLUSION

All these experiments clearly demonstrate the noxious action of the CRT radiations on living organisms and confirmed previous results. Our physical measures of the electric and magnetic fields intensities can appear worthless, as it is in our modern buildings. But there, the ambient fields are not so directional. On the contrary, the radiations emitted by the cathodic tubes equipping all the computer screens are focused by the electronic gun and these radiations are pulsed as they are interrupted at the scanning spot disappearing and at its reappearance. In these conditions, the biological effects appear more noxious than in an ambient field.

Intensity values of the electro- and magnetic fields are strictly idencical with and without the EMF-Bioshield® device. An inadaptation of the actual measuring devices can be deducted from the absence of differences in the physical measures. The spectrum of radiations emitted by a cathodic screen is complex and we can also hypothesize a modification in this spectrum of radiations in presence of the EMF-Bioshield® device, which would change a noxious crude radiation spectrum in a compatible field with the cellular electromagnetic field. Further investigations are required for resolving this problem.

In Drosophila (Oregon R breeding), a significant delay of the developmental processing from 10 to 12/13 days was reported [11] and also observed in an unicellular Alga culture in which a 24 hrs exposure was sufficient [12]. In chicken embryos, our results are in agreement with an irreversible development alteration in embryos exposed to weak extremely-low-frequency magnetic fields [13], and an increase of the abnormal development rate and of the embryonic mortality [14]. Peaks of mortality have been observed in Daphnias, 9 and 15 days after the experiment starting [15] and by us in Drosophila. An interesting observation concerns the importance of the irradiation time observed in Drosophila. An apparent innocuity was observed after the embryonic life irradiation since first larvae normally hatched. The noxious action appears 10 days later, with the adult hatchings, simulating a delayed effect : the embryonic life, with his numerous mitotic crisis, appears as a sensitive period. The development processing of the tested materials appears sensitive to the radiations.

Physiological and behavioural disturbances appeared in rats exposed during their neonative life. In mice placed in front of a color TV screen, it was described some important hematologic evolution alterations [16]. Furthermore, a decrease of immunoglobin G and corticosterone levels after a porcine thyroglobulin immunization was reported. They are also in agreement with some of those precedently described [17, 18, 19] in humans.

In in vitro culture of human testicular biopsies, histopathological and metabolic parameters can be measured and repeated in reproductible manner with standard conditions. After eliminating a possible effect of culture by the two controls (T0 and T0+24), our results permit us to show mitotic and meiotic disturbances, degenerating cells, seminiferous tubules disorganization. They also demonstrate that a testis submitted to an unfavourable micro-environment (a slight increase of temperature secondary to a varicocele in our study) was more sensitive to an exterior aggression.

In conclusion, all these results obtained in various living beings and human tissues showed a harmful effect of all the cathodic screens on development, behaviour and physiology. In the human tissue, normal and weakened by a varicocele, important physiological perturbations have been described. All these results demonstrate a harmful effect of the radiations emitted by a CRT on users. There is scientific evidence that physical sources can interfere with workers and children [20, 21]. On this basis, it is possible to test materials designed for protecting the living organisms. An experimental apparatus, EMF-Bioshield® by A_Nox Technology (developed by S.E.I.C., ISO 9001 qualified for Research and Development) has been tested in the same experimental conditions to evaluate its possible protective effect. In all our experimental cases, living organisms, the same experiments performed again gave observations and results analogous to those of the controls.


ACKNOWLEDGEMENTS

The authors thank Prof. LE RUZ (Rennes University, France), and Prof. B. SISKEN (Lexington University, U.S.A.) for their active participation to the experiments on rats and chicken embryos, respectively. This work was sponsored by S.E.I.C., 3 rue du Léman, CH-1201 Geneva, Switzerland (E-mail: seic@pingnet.ch.


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