______________________________________________________________________ Project no. SSPE-CT-2004-502173 Project title EMF-NET: EFFECTS OF THE EXPOSURE TO ELECTROMAGNETIC FIELDS: FROM SCIENCE TO PUBLIC HEALTH AND SAFER WORKPLACE Instrument: Co-ordination action Thematic Priority: Priority 8, POLICY ORIENTED RESEARCH – AREA 2.3, Call Identifier FP6-2002-SSP-1 Deliverable report D43: Report estimating approximate upper and lower bounds on ‘cost/benefit ratios’ for the services provided Due date of deliverable: february 2007, Actual submission date: October 2008 Start date of project: March 2004 Duration 54 months Organisation name of lead contractor for this deliverable: UGOA – Università degli Studi di Genova Person in charge: Dr. Gugliemo D’Inzeo (Email: [email protected]) Project co-funded by the European Commission within the Sixth Framework Programme (2002-2006) PU PP RE CO Dissemination Level Public Restricted to other programme participants (including the Commission Services) Restricted to a group specified by the consortium (including the Commission Services) Confidential, only for members of the consortium (including the Commission Services) PU _______________________________________________________________________________ Page 1 (of 38) EMF-NET Deliverable Report D43 ______________________________________________________________________ CONTENTS Introduction Methodological approach Plausible interaction mechanisms Conclusion References _______________________________________________________________________________ Page 2 (of 38) EMF-NET Deliverable Report D43 ______________________________________________________________________ Introduction Literature on possible non thermal interaction mechanisms and resulting specific effects is wide and variegate, mainly due to: - the variability of exposure conditions in terms of frequency content and modulation scheme of the EM signals; - the complexity of biological systems including a great variety of structures possibly sensitive to the EM fields. However, even if a mechanism of interaction exists between the exogenous field and any biological structure, it will not necessarily lead to an adverse health effect. In fact, for electric or magnetic fields to initiate or promote adverse health effects in an organism, they must trigger a series of steps, through different levels of complexity of biological systems, from molecular level up to cell, organ and organism ones, that ultimately leads to some health outcome [1]. Methodological approach Bioelectromagnetic investigation has to take into account the complex organization typical of living systems. All biological systems may be considered, from a logical point of view, as a stratification of complexity levels, from the microscopic one of atoms and molecules, up to the macroscopic one of the whole organism, going through sub-cellular structures, cells, tissues, organs and systems. The functionality of a level is related to those of all lower levels but is not completely determined by them, i.e. each upper level shows the so called emergent proprieties. In order to deal with such a complex organization, the typical approach used in computational biology requires an initial stage of analysis of the system followed by the integration of mathematical models developed for each its component. This is the philosophy underlying the “Physiome Project”, an international effort to provide a framework for modelling the human body, using computational models that incorporate biochemical, biophysical and anatomical information on cells, tissues and organs [2]. Any attempt to link molecular and cellular events with physiological function must deal with length scales that range from 1 nm, typical of a protein, to the 1 m scale of an intact body. Similarly, the range of timescales must encompass the 1 ms, characteristic of Brownian motion, to the 109 s, characteristic of a human life. No single model can cover a factor of 109 in a spatial scale and a factor of 1015 in a timescale. The suitable approach is therefore to develop models for a more limited range of spatial and temporal scales and to develop _______________________________________________________________________________ Page 3 (of 38) EMF-NET Deliverable Report D43 ______________________________________________________________________ techniques to link the parameters of this hierarchy of models. This means that, at any one level, there is a “black box” that groups all of the detail of the level below [2]. The same kind of approach was proposed by Chiabrera and d’Inzeo ever since 1993 [3] for the study of mechanisms of interactions and effects of EM fields on biosystems. The aim was to reach, even in the case of non thermal mechanisms, the understanding of the overall chain of effects, from the cell membrane up to the organ level. Proposed methodology is schematized in Fig. 1. Fig. 1 Schematic representation of a multilevel methodology proposed by Chiabrera and d’Inzeo in 1993. The concept of chain of events, from the exposure of a subject to the onset of a possible health effect was then developed by Repacholi and Greenebaum in 1999 [4]. This chain of events commences with some crucial aspect of the field interacting with biological molecules or structures. The field may alter their size, shape, charge, chemical state or energy. This energy “transduction” step involves some transfer of energy for an effect to occur in the biological molecule or structure. The change can then be sensed and amplified within the biological system to produce subsequent responses that might have consequences for the organism. Fig. 2 shows the described causal chain of effects and identifies the multiple points in the chain for which changes might be within normal homeostasis and so produce no functional consequences. _______________________________________________________________________________ Page 4 (of 38) EMF-NET Deliverable Report D43 ______________________________________________________________________ Fig. 2 Links of casual steps connecting disease outcome with EM field exposure. The term “mechanism of interaction” is generally considered to include the full chain of events, although it is sometimes used only for the initial transduction step, referred to as “first interaction step”. For a mechanism to be plausible it should link electric and magnetic field exposure by means of a biophysical mechanism to the beginning of the aforementioned chain of events [4]. But, above all, theories for mechanisms of interaction should make sufficiently concrete predictions that they can be tested experimentally and be capable of being verified, if correct [4]. A unifying methodology was formulated by Apollonio et al. in 2000 [5], [6], taking into account requirements of integration, plausibility and testability. It allows one to approach the bioelectromagnetic problem in a comprehensive way, without losing in specificity of different possible interaction mechanisms involved. A schematic view of proposed integrated methodology is reported in Fig. 3, in the case of neuronal system. _______________________________________________________________________________ Page 5 (of 38) EMF-NET Deliverable Report D43 ______________________________________________________________________ Fig. 3 Schematic representation of the integrated methodological approach, evidencing both dosimetric (red on the left) and modelling (blue on the right) pathways at different levels of biological complexity. On the right spatial scale and temporal scale are reported. In order to establish a quantitative relationship between the exogenous EM field and a biological variable, the approach encompasses the dosimetric pathway (Fig. 3, red on the left) and the modellistic one (Fig. 3, blue on the right). Regarding dosimetry, it should be noticed that, while going down towards microscopic structures, the evaluation of the average exposure of tissues and organs is not sufficient, but microdosimetric techniques are needed for calculating the real distribution of the field on sub-cellular structures such as cell membranes. In fact, while for the thermal mechanism biological tissues are considered as a macroscopically homogenous material with an average permittivity, in the study of specific mechanisms the non homogeneous structure of tissues at microscopic level must be accounted. This implies a consequent non uniform distribution of the field on cell compartments. Microdosimetry research is considered as a future need in the World Health Organization (WHO) agenda since it may yield new insights concerning biological relevant targets [7]. Moreover, more research is needed in microdosimetry not only for a better understanding of the interaction mechanisms, but also for its implications on the quality of the “in vitro” experimental investigation [8]. Moving to the modellistic pathway, different levels of biological complexity require different theoretical tools and involve specific mechanisms characterized by their own timings. Therefore, a precise knowledge of each sub-system is important in order to include all realistic details that are necessary. _______________________________________________________________________________ Page 6 (of 38) EMF-NET Deliverable Report D43 ______________________________________________________________________ The EM field estimated at each biological level represents an input for the corresponding model. The output of each model is one input for the model of the following level in the biological scale of complexity. This approach implies that the whole system could be sensitive to EM fields in a large range of frequencies, up to tens of GHz. In order to reach an in depth understanding of the bioelectromagnetic interaction and to link mechanisms with biological effects, the whole framework of proposed methodology should be clarified. Plausible interaction mechanisms In the understanding of the interaction modeling of EM fields with biological tissues, the only well assessed mechanism is the energy transfer to the molecules (essentially water) and the transformation of this energy in heating after a short time [9], [10]. However this process considers the biological tissue as an averaged material with an average dielectric constant representing how the cells are globally reacting to an applied EM field. On the contrary, the non-uniform distribution of cells inside the tissue and of the cellular compartments inside a cell shows a highly inhomogeneous distribution of different molecules inside the cellular space. This consideration has implication even in the calculation of the EM field on the cells and on the cell compartments: the microdosimetry. In the context of the identification of a mechanism that can modify cellular behavior in a non thermal or specific mode, several models have been proposed in literature [11]-[39], but most of them are facing the problem of overcoming local water viscosity and the thermal agitation which tend to distribute and damp the energy among the molecules [40]. Starting from the lowest level of biological organization, where the “first interaction step” takes place, the dimensions of the involved structures and the importance of the electronic distribution in the biochemical processes controlling the cell functioning indicate the total Quanto-Mechanic (QM) approach to be the most suitable one [40]. Such an approach has the advantage of being totally exact from a theoretical point of view; the drawback is the huge complexity of the problem and the high computational cost of these studies. However the recent mixed QM/MD (molecular dynamics) techniques [41]-[44] enable us to carry out a totally quantitative approach to the problem of EM interaction at molecular level. Such a method combines classical and quantum descriptions. The site of reaction in the protein is described using the QM approach, whereas surrounding environment, including the exogenous field, is considered as a perturbation at the site of chemical reaction [42]. This allows the exact description of the interaction target and the evaluation of possible EM induced changes in the energy barriers of binding and unbinding reactions [41]-[44]. _______________________________________________________________________________ Page 7 (of 38) EMF-NET Deliverable Report D43 ______________________________________________________________________ The aforementioned method has been recently applied to study of the covalent binding process of a ligand (carbon monoxide) to a protein (myoglobin) under the action of a 1 GHz microwave signal [44]. Although, in this case, the biochemical process is not affected by fields well below those involved in atomic/molecular interactions [44], the followed methodology is robust to investigate protein behavior and biochemical reactions, and shows a possible way to study the basic mechanism of microwave bio-effects, investigating at the basis of the possible molecular transduction of the field [44]. Another important approach at molecular level is based on the action of magnetic field on Radical Pairs (RP) involved in some biochemical reactions with induction of free radicals, such as lipid peroxidation and enzymatic reactions. To date, it is the only well-established mechanism by which magnetic fields are known to influence the rates and yields of chemical reactions. In particular, the hyperfine interaction with an exogenous magnetic field can change the probability of RP recombination, thus altering the reaction equilibrium [45]-[48]. The reason why it can occur even at low intensity fields is that the spins of radicals are very weakly coupled to the thermal bath. Such a mechanism is considered the most plausible model for some ELF and static effects [45], [48] and has been identified in a recent review as the only possible coupling mechanism up to 150 MHz [49]. Moreover, some recent papers have developed theories and experimentally confirmed, through in vitro experiments, the action of low magnetic fields on RP even in the RF range [50], [51]. The radical pair model does not produce large changes at the initial field detection step, so we should expect cooperation mechanisms for biologically relevant detection of magnetic fields [52]. Different investigations have suggested that avian compass relies on the action of the geomagnetic field on radical pairs formed by photoinduced intramolecular electron transfer reactions in an array of aligned photoreceptors in the retina [53], [54]. In a recent paper, the feasibility of chemical detection of magnetic fields as low as the Earth’s has been experimentally proved [55]. All proposed interaction theories need experimental confirmation both at biochemical and biological level. The idea is to promote a set of experiments evaluating effects at low level of complexity in the biological scale, e.g. on enzymatic reactions and membrane channels conductivity. These experiments must be conducted in high interdisciplinary laboratories in order to verify and control all the parameters involved both with biological and physical-engineering competences. Once theories have been experimentally checked, the next step is to connect models at different levels of biological complexity to account for macroscopic effects, such as possible adverse health effects. In this context, RP mechanism and MD techniques could explain the transduction step of a _______________________________________________________________________________ Page 8 (of 38) EMF-NET Deliverable Report D43 ______________________________________________________________________ magnetic or EM field into variations in kinetics of chemical reactions of biological interest. The case of the avian compass mechanism is a good example on how the chain of effects can be reconstructed to explain the action of the geomagnetic field on a macroscopic behavior such as bird navigation. Conclusions To date, a debate is still open in the scientific community about possible health effects due to non-thermal or specific mechanisms of interaction with low level EM field. As discussed in D42, present knowledge does not allow us to quantify a threshold below guidelines, above which the EM field could induce adverse biological effects. As an example, a recent study [44] on the binding process of the carbon monoxide to the myoglobin under a 1 GHz microwave field indicates alterations in the biochemical process only for E field intensities comparable with the atomic/molecular ones, well above any possible value induced by the environmental fields. Therefore, to date, a cost/benefit analysis for the services provided seems to be unnecessary. However, although it has not been established that any of proposed specific mechanism could result in adverse health effects at levels below guidelines, one cannot conclude that long term effects will not be seen. Therefore, in depth investigation on specific mechanisms is advisable, in order to reconstruct the whole chain of effect, from the “first interaction step” at molecular level up the whole organism behaviour. References [1] M. H. Repacholi, "Low-Level Exposure to Radiofrequency Electromagnetic Fields: Health Effects and Research Needs," Bioelectromagnetics, vol. 19, pp. 1-19, 1998.IEC, "Technical Report, Type 2 Effects of Current Passing Through the Human Body", 1983. [2] P. J. Hunter and T. K. Borg, "Integration from proteins to organs: the Physiome Project", Nature Reviews, vol. 4, pp. 237-243, 2003. [3] A. Chiabrera, G. d'Inzeo, European COST Action 244, Modelling of Interaction, 1993. [4] M. H. Repacholi and B. Greenebaum, "Interaction of Static and Extremely Low Frequency Electric and Magnetic Fields With Living Systems; Health Effects and Research Needs," Bioelectromagnetics, vol. 20, pp. 133-160, 1999. [5] F.Apollonio, M. Liberti, G. D'Inzeo, and L. Tarricone, "Integrated Models for the Analysis of Biological Effects of EM Fields Used for Mobile Communications," IEEE Transactions on Microwave Theory and Techniques, pp. 2082-2094, November 2000. _______________________________________________________________________________ Page 9 (of 38) EMF-NET Deliverable Report D43 ______________________________________________________________________ [6] M. Liberti, F. Apollonio, A. Paffi, and G. D'Inzeo, "Overview of bioelectromagnetic interactions models: proposal for a unifying integrated methodology," presented at XXVII General Assembly URSI (invited paper), August 2002. [7] World Health Organization (WHO), http://www.who.int/en/. [8] M. 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