Project Proposal for International Science and Technology Center (ISTC G-989)
http://www.istc.ru/ISTC/sc.nsf/html/projects.htm?open&id=G-989
Foreign Collaborators:
University of Washington / Department of Bioengineering
National Institute of Public Health
COMPLEX STUDY OF NEUROLOGICAL, CARDIOLOGICAL AND HEMATOLOGICAL CORRELATES OF EXTREMELY LOW FREQUENCE ELRCTROMAGNETIC FIELD EXPOSURE IN ANIMALES
Summary of the Project
The main purpose of the Project is to reveal new criteria for the estimation of acute and persistent exposure to extremely low frequency electromagnetic fields (ELF EMF) in laboratory animals and their offspring, as well as to develop new- and improve known noninvasive and inexpensive methods for examination of population of high electromagnetic risk. It is supposed to use power and EEG D, q, a, b and g rhythms frequency and to deliver various shape of signal including the feedback of flux density by own heart beat or respiration rate of the exposed animal. Implementing the original methods for evaluation the neurobiological, cardiological and hematological effects of ELF EMF exposure, we intend to identify dynamics of system deviations from the point of view of the risk degree of biological and social value in animals’ population and their offspring. Three basic principles, which underlie the Project ideology are: 1) Feasibility of extrapolating the results of experimental studies for estimation the acute and remote risk-effects of ELF EMF exposure and hazards to the humans; 2) Development of new- and improvement of known noninvasive and inexpensive methods for human testing and solution of other similar tasks, including ecological and epidemiological ones; 3) Possibility of a maximal automation of the methods implemented for facilitating and standardizing the studies, as well as creating databases and archives for further computer generalization of large massifs of information.
Analysis of available reference data and long-standing experience in this field enable to contemplate with high probability that in order to fulfill the Project task and experimental motivation of the effective recommendations, the most promising is application of new criteria for estimation of the acute and chronic effects of ELF EMF exposures in laboratory animals. Such are in particular: 1) Complex analysis of the results of studies of the neurobiological mechanisms in modification of inborn and induced aggressiveness, learning and memory, as well as adaptive behavior in extreme conditions and disturbances in the sleep-waking cycle, 2) Analysis of changes of mechanisms regulating the vago-sympathetic balance and neuro-autonomic status by studies of heart rate variability (HRV), and 3) Original method of complex analysis of changes in the red blood system (RBS), based on the study of volume relaxation in conditions of gradual reversible spherulation in peripheral blood erythrocytes (PBE).
The Project will be realized by specially appointed group of scientists, recruited from the Laboratories of Radiation Physiology, Cell Radiobiology, Physical-Chemical Radiobiology, Systems Engineering and Informatics of the Scientific Center of Radiobiology and Radiation Ecology, Georgian Academy of Sciences. Information about Laboratories, scientific activity and possibilities of our Center is available at: http://www.radiobiology.org.ge
Introduction and Overview
More than 20000 works have been published for the last two decades dedicated to the studies of different aspects in biological effect of EMF exposure of wide spectrum. These studies cover all levels of biological system organization - from molecular to population - and are published in editions of different information value - from newspaper article to monographic generalizations and specialized web-sites in various languages, for example EMF Link(http://infoventures.com/emf/) and other sites and specialized editions [11, 19, 31 and etc.]. These works have been implemented within the framework of various international, national and university programs of most developed countries. On some estimation, a general costs invested for these purposes have amounted to over US$ 1.5 billion for the last five years.
However, many key aspects of biological effect of EMF exposure have not been solved as yet or are disputable. Besides the carcinogenesis, the most controversial issues are whether induces or not ELF EMF exposure: 1) Changes in cellular functions, tissues, and organs; 2) Decrease in the melatonin production; 3) Alterations of immune system; 4) Acceleration of tumor growth; 5) Changes in biorhythms; 6) Changes in human brain activity and heart rate; 7) Alterations of reproductive processes, etc. Scientometric (meta) analysis of representative literature, conducted in our Center has shown that according to some data EMF exposure to any biological systems causes an effect but according to other data there is no effect. This pattern is usually interpreted as an absence of the effect. However, any response of biological system to any external stimulus of threshold- or even over-threshold intensity carries out in a nonlinearly low mode and produce practically similarly probable deterministically positive or negative answer. Except for intra-species diversity there is an individual diversity in sensitivity to external factors, depended on genotype or phenotype. Moreover, according to our data, the animals with initial prevailing of adrenergic mechanisms of regulation of homeostasis are significantly more sensitive to various external factors, in particular to ionizing radiation, gravitation load, pharmacological substances, etc., than the ones, in which adaptation optimum is reached by prevailing of cholinenergic mechanisms of regulation of physiological processes [14, 15, 29, 30, 32, 33].
Considering significant scientific and social value of clarification of the physiological mechanisms of EMF effects and under the above general assumptions, we have proposed the current Project for ISTC, concerned with complex study of neurobiological, cardiological and hematological correlates of ELF EMF exposure in animals. Methodical and information support, as well as long experience of studying this problem in our Center enable to guarantee a successful realization of the Project and revealing new, unknown earlier facts completing the conception about modification of physiological process, caused by ELF EMF exposure.
Proposed Project includes three ideologically associated sections:
1. Study of neurobiological effects of ELF EMF exposure is the subject of numerous studies worldwide. Positive or negative effects are revealed in a range from separate nerve cells to cognitive functions of different complexity. However, aggressive and other types of emotional-motivational behavior have not been sufficiently studied so far. At the same time, this problem is actual from the standpoint of possible influence on social stability and creation of adaptation compensatory background for epidemiological consequences of EMF exposure. Our studies will be directed at determining the changes of integrating functions of the brain and their neurochemical correlates according to individual characteristics of emotional-motivational behavior of each animal. We intend to study the neurobiological mechanisms of modification of inborn and induced aggressiveness, learning and memory, depending on the initial neuro-autonomic status, adaptive behavir in extreme conditions, modification of sleep-waking cycle in rats and cats, as well as imprinting mechanisms in chicks. The special attention will be given to study of physiological and neurochemical mechanisms of prey/predator aggressiveness. This is concerned with the fact that ELF EMF exposure alters melatonin production, which in turn influences neurotransmitter/neuromodulator systems, especially the serotonergic mechanisms - the key system in organization of aggressive behavior. We intend also to use an alternative advance - a study of above processes after exposure the animals and cultivated nerve- and glial cells to the hypomagnetic environment. For each of the above-mentioned directions we have a certain scientific reserve, including information maintenance on global trends in these problems. For example, among Project executive specialists there are scientists, who have been studying the ELF EMF effects on the ethological parameters of rats’ behaviour [11, 12, 22, 24], modification of inborn aggressive behavior and memory [13, 23, 24], effect of antenatal ELF EMF effects on emotional-motivational behaviour, learning and memory in the offspring of rats and rabbits [53], effect of hypomagnetic environment on animal behavir, proliferation and differentiation of nerve and glial cells in tissue culture of different brain structures in rats, and imprinting in chicks [27, 28, 44-46]. It was revealed in these works that the changes of integrative brain activity have distinct neurochemical correlates such as characteristic change in quantitative contents, distribution and correlation of some biologically active substances (transmitters, mediator amino acids, biogenic amines, receptors, etc.) in different structures of the brain and in the sera. According to our data, out of the neurobiological criteria the most sensitive to the ELF EMF exposure are the mechanisms of short-term memory [22, 23], related to the protein synthesis. This hypothesis was reviewed in the model experiments in various mutant strains of E.coli - it was found that an acute inhibition of protein synthesis did occur following the ELF EMF exposure [1]. It is also necessary to note that application of EMF was shown to be feasible as a conditioned stimulus in classical Pavlovian reflexes [42]. Besides the electrostatic movements of whiskers and hair in the rats and signal perceptions through piloerectory system, a directl sensation of single-domain magnetite crystals as the basic sensing element, or excitation of various interoceptive and nerve cells in deep structures of brain was supposed, that is similar to the sensation, which occurs in non-visual perception of light by septal, hypothalamic, and pineal cells of the brain [56].
Considering the above-mentioned, i.e. informational maintenance and scientific- methodical potential of executives of this section of the Project, there are all reasons to consider that it will be successfully realized.
2. Study of EMF effects on cardiovascular system began many years ago and was stimulated by the known fact of increased sensitivity of some persons to geomagnetic fluctuations. These reactions are not always linked with observable somatic or autonomic pathology. Our studies will be directed at revealing the change of central and peripheral mechanisms of heart rate regulation and neuro-autonomic status. There are data that the ELF EMF-exposed population (8 h/d for 1-5 y, flux density - 0,2-6,6 mT) had a significant increase in degree of certain neuro-autonomic disorders i.e., physical fatigue, psychical asthenia, decreased libido, melancholy, depressive tendency, irritability, etc. [2]. According to modern conception, heart rate variability (HRV) reflects neuro-autonomic status and fluctuations of metabolic supply of the processes of vital activity and adaptive resources of organism, as well as emotional and motivational intensification and adreno-/cholinergic balance [51]. Taking into account that ELF EMF exposure causes a reduction of cholinergic activity in different brain structures [20-22] and exposure of hypomagnetic environment during embriogenesis increases a noradrenaline and dopamine content in the hippocampus [46], its revealing by analysis of HRV should be expected. The relevance of these studies is also concerned with the fact that according to one group of authors ELF EMF exposure changes the heart rate, but according to the others, there are no changes whatsoever [4-10, 16-18, 43- 45, 49, 50].
The Project executives have special experience in study of ELF EMF effects on the animals’ cardiovascular system [3]. These studies have shown that a single total 50 Hz EMF exposure for one hour (cranio-caudal direction of field vector and induction of 3 mT) caused a hyperdynamic phase syndrome in cardiovascular system, which represented an alarm reaction of stress course. These changes held within a day and revealed itself in increased electrical stability of the heart, acceleration of repolarization and atrio-ventricular conductivity, and in intensification of system blood supply. An increase of systolic and minute volumes and reduction of common peripheral resistance, without observable changing of mean heart rate indicates the same. Regrettably, because of heavy social and economic conditions in Georgia, it was impossible to continue the studies in this field.
Within the framework of the Project it is expected to continue these studies, as well as those concerned with alternations of HRV after ELF EMF exposure of various duration and induction.
3. Studies of hematological effects of ELF EMF exposure are intensively conducted all over the world and directed mainly at studying the mutagenesis of bone marrow stem cells. Purpose of these studies is to reveal a possible induction of leukemia and other cancer diseases. This material is extremely contradictory. Experimental simulation of these processes in vitro has shown that ELF EMF exposure inhibits cells proliferation and accelerates terminal differentiation [51, 54, 56], changes the lymphoid phenotype in mice [26], etc. There are data that ELF EMF-exposed population experienced a significant fall in total lymphocytes and in CD4, CD3, and CD2 lymphocytes, as well as a rise in NK cells. Leukopenia and neutropenia were also observed [2]. Somatic mutations, as well as changes of proliferation and differentiation in erythroid pool of the bone marrow stem cells practically was not studied. At the same time, a posession of this information is of significant interest from the point of view of understanding the common mechanisms of biological effects of ELF EMF exposure. Our studies will be aimed at revealing the dynamics of possible specific and nonspecific changes in the red blood system (RBS).
It is known that changes in the hemopoietic system are universal criteria for estimating any reactive, pathological and pre-pathologic processes. The RBS from position of ELF ELF exposure has not sufficiently studied yet. Our investigations have shown that analysis of population spectra of peripheral blood erythrocytes (PBE) provides far greater diagnostically and prognostically important information, than it has been known before. Current Project section intends to create a principally new methodology on the basis of developed in our Center methods for estimation of RBS functional state by characteristic dynamics of changes in the PBE population spectra. The methodology relies on an original principle of study the relaxation processes in the erythrocyte volume variation under conditions of their gradual and reversible spherulation. The general theoretical background, mathematical tools and algorithmic base of the principle are described in a number of papers and in the monograph [34-41], and is also available at our Center’s Internet Web-site: http://www.radiobiology.org.ge (Section - Editions and Publications) or http://geocities.com/radiobiology_ge/ (Other links - Review).
The methodology rests on the theoretically sound and experimentally verified assumption that the families of PBE population spectra and the dynamics alteration of the pattern of erythrocyte volumes relaxation under conditions of gradual reversible spherulation reflect adequately not only the degree of tension of functional activity of adaptive, compensatory, and reparatory mechanisms of the red blood system, but also of the whole body. This methodology will make it possible:
1) to determine the speed and quality of erythrocyte maturing and to evaluate the state of erythropoiesis of the pool of the stem cells, not resorting to the exploration of bone marrow punctates and complex hematological studies,
2) to determine the rate of physiological aging of cells (biological age), the extent of functional and pathological modification (wearing) of the erythocytes' structural elements (physical or chronological age), to estimate the rate of aging and elimination of cells having exhausted the biological resource, as well as to judge about the metabolic demands and abilities of the body in order to renovate and maintain biological full-value of the circulating and deposited erythrocyte population,
3) to estimate the functional state of the red cell membranes and oxygen density of erythrocyte hemoglobin in various age groups and judge on the rheological properties and metabolic efficiency of circulating blood,
4) to evaluate behavior of interdependence depot-circulation and define the sum reserve possibilities of adaptation mechanisms during functional loads on metabolism,
5) to define the RBS response during pharmacological or any other activation or depression of erythropoiesis,
6) to define variations of average life span of erythrocyte dependent on exposure intensity and duration, observation period, etc.
The methodology can be widely applied not only in hematological, but also in general medical practice, including diagnostics and evaluation of efficiency of therapeutic measures in various diseases, doping control with the use of erythropoietins or hemotransfusion, for the estimation of physical condition of schoolchildren, training of sportsmen, professional preparation of service-men and, in the case of selection of adequate representative groups - it can be applied for objective estimation of the population life quality of different states and regions. The methodology under consideration shall be realized as an integral complex of engineering hardware-software systems. Moreover, on the basis of experimental evidences, an experimental model was made of RBE, which will enable a computer simulation of adaptive changes.
The biomedical procedure of the method developed is maximally simplified, standardized and may be combined with the routine hematological study, because it requires no more than 15 ml of blood. The methodology may have a multipurpose application. Experimentally founded biomedical premises, a correct actuarial mathematics, a distinct algorithm basis and the accessible hardware-software complex securing the method enables to use the given approach not only for tracing the course and evaluating the efficiency of treatment of radiation injury or other forms of pathological processes, but also to carry out mass investigation of population for the quality of life and the degree of ecological risk. In the world literature there are no analogous systems mentioned so far.
Effective use of the above-mentioned set of behavioral, neurochemical, neuro-autonomic and hematological criteria including a complex analysis of the results, as well as taking into account our experience and availability of preliminary figures in this field, professional degree of investigators, informational provisions and equipment, all this give grounds to believe that the Project, as a whole, will be successfully realized. The warranty to this assumption is given by the fact that the institution of the Project executors has a vast experience in carrying out similarly large projects concerned with studying the mechanisms of biological effect, as well as of other factors of risk such as ionizing radiation, noise and vibration, infra- and ultrasound, etc.
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Organizational Structure Diagram