Published on 07/29/2008

THE INFLUENCE OF MODIFIED EMF ON THE VIABILITY OF HEAT SHOCKED DROSOPHILA MELANOGASTER

Introduction

THE INFLUENCE OF MODIFIED EMF ON THE VIABILITY OF HEAT SHOCKED DROSOPHILA MELANOGASTER

Lednyiczky G.1, Sakharov D.2, Vaiserman A.3, Koshel’ N. 3

1 Hippocampus Research Facilities, H-1092 Budapest, Ráday u. 8., Hungary;
Tel:+361.299.0200,
email: hippocam@hippocampus-brt.com
2 R.E. Kavetsky Institute of Pathology, Oncology and Radiobiology, The National Academy of Science of the Ukraine, Kiev, Ukraine
3 Institute of gerontology, The National Academy of medicine of the Ukraine, Kiev, Ukraine

The Information that formed background of the work

• Over the last two decades, numerous data have appeared that support the biological importance of extremely-low-frequency electromagnetic fields (ELF-EMF). Endogenous and Exogenous components of ambient electromagnetic fields seem to play a regulator role with regard to biological processes that help an organism to maintain homeostasis [Wever R.A., 1985; Choy R. et al., 1987, Recent Advances, 1992; Ho M.-W., 1994].
• Several investigations have shown that weak ELF magnetic fields can adversely affect the early embryonic development of a bio-object [Ho M.-W., 1992; Ubeda A., 1994], which increases the proportion of early deaths and developmental abnormalities.
• Finally, in some reports [Adair R.K., 1995, ORAU review 1992], authors argue that there is no convincing evidence to support the contention that exposure to weak ELF EMFs is a demonstrable health hazard nor that it can express beneficial influence.
• We consider that it is possible to help clear the current controversy over whether weak EMF are beneficial, inactive, or detrimental to biological systems if one supposes that artificial, biologically active EMFs only imitate the resonant parameters of Endogenous and/or Exogenous components of regulator EMF and thus produce the observed effects [Choy R. et al., 1987; Anninos P.A., 1992; Rein G., 1995].
• Also, device-generated EMFs can interfere with regulator EMFs, thus producing pathological effects [Ho M.-W., 1992; Ubeda A., 1994] and mitigating previously observed effects [Litovitz T., 1994].
• In addition it is noteworthy that “a healthy (normal) biological system residing in its homeostatic (i.e., stable-steady) state is not affected by currents of physiologically acceptable magnitudes. If the system resides in stressful (or unstable-steady) state, small perturbations may produce large changes in the state of the system” [Vodovnik L., 1995]. This is supported by theoretical [Vodovnik L., 1995] and clinical studies on the effects of weak EM stimuli [Andreev E., 1984].
• In order to confirm these suppositions and to shed light onto mechanisms of the interaction of weak EMF and biological systems, we study the influence of endogenous ELF EMF on the aging and vitality of fruitflies. Heat shock treated chrysalises were chosen as the object of influence because it is the unstable-steady stage of development that is established due to developmental processes of metamorphosis and heat-shock as well.

In this investigation, the vitality of adult flies (imago) is studied. During the early chrysalises developmental stage, flies underwent the influence of a heat-shock and the following influence of endogenous ELF EMF:

Methods

METHODS

An outbreed Oregon-R wild type laboratory population of Drosophila melanogaster was used in the experiments. The embryos were placed into one-liter glass vessels containing a standard medium and were kept under standard conditions. During the chrysalis stage, flies underwent the influence of two different heat shock regimes:
1) 120 min. at 370C - the “activating” heat shock regime (“A” groups); this influence increases the vitality of flies.
2) 120 min. at 400C - the “depressing” heat shock regime (“D” groups); this influence decreases the vitality of flies. We have included into the experiment additional “N” groups which consist of temperature intact (normal) chrysalises.
The groups were then divided into experimental subgroups, depending on the quality of electromagnetic information they experienced. On average, each subgroup consisted of >1000 chrysalises that continued to live in one-liter glass vessels.
The ‘BICOM’ device (Regumed GmbH, Germany, formerly Brugemann GmbH) [Brugemann H., 1993] was used to acquire the electromagnetic field of the chrysalises situated on the input electrode, modify these fields, and transduce them into the experimental group of chrysalises placed at the output electrode.
The programs of treatment are denoted as follows: The treatment mode is denoted by position 1, then in parenthesis (), the amplification or attenuation; position 2 indicates the time (in seconds) for a complete scan of frequency band from 10Hz to 150kHz; position 3 denotes either “discrete interval” (the BICOM has a feature to turn itself on and off) or “continuos” operation; Position 4 indicates the duration of treatment.

1. H (1), Frequency scan 15'', Continuos, 20 sec.
2. H (1), Frequency scan 15'', Continuos, 60 sec.
3. H (1), Frequency scan 15'', Continuos, 30 min.

Based on the results of a former experiment dealing with the effects of endogenous EMFs on the imago of Drosophila (Tab.1), the two first programs were considered to be the most effective ones. The last one is chosen in order to analyze the dose-dependence or the occurrence of a “dose window” [Lawrence A., 1982; Choy R., 1987; Webb S., 1984] of the influence.
The 1st-12th (duration of influence: 20 and 60 sec.) experimental groups experienced an influence of endogenous ELF-ELI EMFs for two days, four times daily, with a one hour interruption between treatment sessions. For the 13th-15th chrysalis groups, duration of influence was 30 min. for two days, once daily.

Fig. 1 The scheme of experimental setup

To generalize the results of the both applied influences - heat shock and endogenous EMFs - in terms of the vitality of chrysalises and flies, we analyzed following representative indices:

• The mortality of chrysalises in (%).
• The Fecundity of flies is considered normal if the eggs undergo complete metamorphosis, this is denoted in the tables below by a “+”. A successful embryonic development of approximately half of all produced eggs can be considered a reduced fecundity and is denoted by “±”. When produced eggs turn out to be non-viable, that is, almost no flies appeared from them, this can be considered a completely depressed fecundity and denoted by a “-”.
• The mean life-span (MLS) index (only for males from each experimental group). The mortality of the flies is estimated by observations performed daily on the vials by counting the number of dead Drosophila. On the basis of this data, the mean life span was calculated for each experimental group.
• The time of movement (sec) of flies towards a source of directional light stimulus, i.e., a positive phototaxis, was controlled since this index directly correlates with the integral viability of an organism [Mayer P. J., 1985].
• The resistance of adult flies to a heat shock (min.) is evaluated in terms of the time it takes for the death of 50 % of population by heating to 400C.
• The resistance of flies to starvation (hour) is evaluated in terms of the time it takes for the death of 50 % of population that is kept without food.

Mathematical analysis of the data was performed by using “Statgraphics” software [18].

Results and Conclusion

RESULTS
The transmission of EM signals from the heat-shock activated chrysalises of the “A” groups and the heat-shock depressed chrysalises of the “D” groups onto the “normal” chrysalises causes a significant decrease in the mobility of imago in three of five “N” groups. The flies’ mobility begin to be even lower than the reduced one of the flies from the control “A” group. It is worth to emphasize that: if in the control “A” group, the flies’ mobility is low because of a Heat-Shock treatment, then in the experimental “N” groups, the same is induced by the influence of weak endogenous ELF EMF. Interestingly, this effect of heat-shock gene activation under the influence of both temperature and very weak EMF was also achieved in a recently published study by Blank M. [1994] -- energetically, the influences differ 14 orders from each other.

Fig. 1: The time of positive phototaxis of flies from temperature intact experimental groups - the flies mobility
for all figures : * - p < 0.05; ** - p < 0.01; *** - p < 0.001
In the group D>N 20" the influence of EMF not altering flies’ mobility increases the starvation resistance of flies.

Table 1: The results of the transmission of EM signals from the “D” to the “N” groups
GROUPS Time of positive phototaxis (sec) Starvation resistance (hours)
N 13,89 ±0,42 30,83 ±0,52
DN (20") 13,67 ±0,47 33,98 ±0,92 *
In three groups -- A>N 20", A>N 60" and D>N 60", we observed a significant (p<0.05) increase in the longevity of flies in comparison with that of the “N” control (temperature and EMF intact).
Fig. 2: The Mean Life Span of flies from temperature intact groups after the influence of endogenous EMF

The treatment of chrysalises from the “A” groups with endogenous EMF in 4 groups from the 5 tested (Fig. 3), eliminates the negative influence of the “activating” Heat-Shock on the locomotion activity of flies - the mobility becomes the same as the mobility of flies from the temperature intact control. At the same time, the positive effect of the “activating” Heat-Shock - the increased starvation resistance - remains unchanged.

Fig. 3: The time of positive phototaxis of flies from groups of ‘Activating’ Heat-Shock

The magnitude of the changes in the flies’ vitality indices after the exposure to EMF is the highest amongst groups of the “depressing” Heat-Shock regime. Therefore it is presumed that chrysalises of these groups were the most sensitive and able to react. The exposure to EMF allows the improvement of the locomotion activity and starvation resistance of flies in all groups in the “depressing” Heat-Shock regime (Fig.4,5).

Fig. 4: The time of positive phototaxis of flies from the groups of the “Depressing” Heat-Shock Regime

Fig. 5: The starvation resistance of flies from the groups of the “Depressing” Heat-Shock Regime

Besides, after the applied EM influence, the high rate (25%) of the flies’ mortality on the stage of chrysalis is sufficiently decreased (in groups N>D 20", N>D 60", A>D 20", A>D 60" and A>D 30'), also female fertility is increased up to the level of the intact control (N control) (in groups N>D 60", A>D 60", A>D 20").

Table 2: The results of the transmission of EM signals from the “N” and “A” groups to the “D” groups

groups mortality of chrysalises % female fertility (alive eggs)
D control 25.5 -
ND (20") 5.4 -
ND (60") 0.5 +/-
AD (20") 0.2 +
AD (60") 0.1 +/-
AD (30') 2.0 -
Also, in groups N>D (20"), N>D (60") and A>D (20") we observed a considerable increase in longevity of flies.

Fig. 6: The Mean Life Span of flies from the groups of the “Depressing” Heat-Shock Regime

Conclusions

• Using of the low-frequency filter of the ‘BICOM’ device and the special design (closed metal cups) of antenna electrodes makes it possible to significantly decrease the probability of the undesirable influence of background electromagnetic noise on the experimental bio-objects. This fact and the complex dynamics of changes in various indices of flies’ vitality suggest that the main source of the influencing EMF might be solely the chrysalises’ organisms that were placed into the input electrode. The possibility to detect weak endogenous ELF EMF is discussed by Smith C.W. in [Smith C.W., 1994]
• Under the specified conditions, we obtained significant results that may indicate a strong biological importance of endogenous ELF EMF for the regulation of living systems. The experimental data show that the exposure to endogenous EMF may results in the normalization of the flies’ vitality that were earlier distorted by undergoing a stress influence (of temperature). Moreover, even a normal (healthy) organism is receptive to endogenous EMF that can optimize its vitality and thus considerably increases longevity of an organism.
• No doubt, the efficacy of endogenous EMF (that is its importance for a bio-object) increases especially under the circumstances when the homeostatic status of an organism is considerably distorted by the influence of a stress factor (a stressor).
• The effect of the influence on the flies’ genome has to be taken into consideration. During metamorphosis, a large quantity of the cells of a chrysalis’ body are eliminated and replaced by new ones that form the imago body. Despite such crucial changes in the functional activity of genes and the structures of Drosophila’s body, the effects of the EMF exposure are preserved and can be detected during any subsequent stage of the flies’ life -- even the integral index of the flies’ vitality and mean life span are affected. This suggests the participation of genes -- the structures of preservation and inheritance -- in the assimilation of endogenous EM stimuli. To some extent, it is proven in our experiments with tumor cells: we observed alterations in the rate of DNA synthesis of lymphoma U-937 cells after an influence from their endogenous ELF EMF [Sakharov D., 1993]. This DNA participation is also shown and discussed with regard to exogenous EMF in [Blank M., 1992; 1995]

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