Objective of the project. Within the framework of the project it is planned:
· To investigate changes in size and shape of erythrocytes, as well as changes in the structural and functional properties and crystallinity of cellular membranes, induced by incident non-ionizing radiation of various wavelengths (in the superhigh frequencies (SHF), extremely high frequencies (EHF) and THz ranges);
· To reveal mechanisms and modes of action of electromagnetic radiation (EMR) of various ranges, in particular, non-ionizing radiation (NIR) on the structural and functional state of erythrocytes;
· To identify the resonant frequencies effectively influencing change of size and shape of erythrocytes, in order to clarify effects that occur in the mechanisms of EMR impact as well as to assess the prospects of their application;
· To investigate the impact of electromagnetic radiation on the innate resistance and acquired immunity factors of mammals, and to assess the effects of medicinal plants with targeted immunomodulatory and anti-proliferative properties toward ameliorating such impact;
· To research the direct, indirect and combined effects of electromagnetic radiation on the osmotic resistance of blood erythrocytes;
· To study the qualitative and quantitative composition of the main groups of biologically active substances in erythrocytes;
· To study the antioxidant properties of substances isolated from various plants;
· On the basis of the data obtained, to investigate the problem of human electromagnetic safety, specifically the potential physiological harm presented by common non-ionizing radiation sources (cellular communications, computers, Wi-Fi, microwave ovens, etc.), and to propose a innovative methods for diagnosis and detection of cellular damage and malignancies induced by non-ionizing radiation sources. By methods of X-ray structural analysis, scanning tunneling microscopy, optical polarizing microscopy and by numerical investigation electronic work function properties, a comprehensive study of the mechanisms that mediate the effects of non-ionizing radiation (low-intensity millimeter and terahertz electromagnetic radiation) on biological membranes will be undertaken.
Within the framework of the proposed project, basic mechanisms that may lead to various manifestations of harmful effects of electromagnetic waves will be explored, and the mechanisms of action of non-ionizing radiation on biological systems will be researched. The study will proceed to identify specific irreversible biological perturbations induced by radiation at various wavelengths and dosages, such as those that are common in clinical use for medical imaging and therapeutic purposes. A deliverable at the end of the project will be the evaluation of novel and preferably inexpensive rapid physical diagnostic methods for assessing the severity of lesions induced by non-ionizing radiation on living systems. Effective schemes will be developed for lesion assessment, as well as for prediction of adverse effects of electromagnetic waves and environmental changes in an affected environment.
Experimental methodologies for the proposed research will comprise methods of hematology, enzyme immunoassay and immuno-physiological analysis of blood and blood products, including leucocytic materials. Comparative studies will involve analyses prior to and after irradiation, and also in conditions of administration of different doses of various potentially biologically active compounds derived from natural products. Screening of the sets of biologically active substances for antioxidant protective activity will be a primary focus. The primary experimental system will be the wildtype Rattus norvegicus (brown rat) species.
Libraries of candidate biologically active compounds derived from natural products will be generated by harvesting and milling plant materials followed by ethanol precipitation and extraction. Qualitative and quantitative characteristics of the major groups of these compounds will be assessed using liquid chromatography and spectrophotometry analytical techniques.
For preliminary assessment of qualitative composition of the isolated natural compounds, the method of laboratory pharmacopoeia selection of essential oils using a Clevenger-type apparatus will be used, followed by identification of the essential oils components using chromatography-coupled mass spectrometry. For identification of the essential oils and lipids and extract components, contemporary spectroscopic research methods will be used, including proton nuclear magnetic resonance, infrared and ultraviolet spectroscopy, column chromatography, high performance liquid chromatography (HPLC), thin layer chromatography, elemental analysis, X-ray diffraction (XRD) analysis, and gas-liquid chromatography techniques.
In the development of the specialized food products and biologically active compounds on the basis of the isolated and characterized substances from plant materials, technological and microbiological methods of investigation will be used to evaluate experimental models. To assess the immune status the major subpopulations of peripheral blood lymphocytes will be identified by the reaction of indirect immunofluorescence using commercial research assay kits for CD3, CD4, CD8, CD25, CD16 and SD72 antibodies. Non-specific functional activity of T-helper lymphocyte subpopulation will be studied in the standard reaction of inhibition of leukocyte migration from capillaries. The non-specific humoral indicators of immunological reactivity of the organism the levels of major classes of serum immunoglobulins (M, G, A) will be determined in a standard reaction of radial immune diffusion in agar and circulating immune complexes (CIC) in the precipitation reaction with polyethylene glycol (PEG) 6000 («Pharmacia», Sweden).
From the in vivo experiments in the rat model, sensitive rats will be isolated in which low dose radiation has induced physiological stress. A comprehensive immune-physiological study of the organism after exposure to lower frequency non-ionizing EMR will be carried out with assessment of induced immunodeficiency status.
An overarching objective of these research projects is the development and evaluation of various methodologies for monitoring and identifying adverse effects in mammalian systems due to non-ionizing radiative sources of anthropogenic origin, such as telecommunications, background and diagnostic imaging sources. The proposed project will amass a large body of data for the assessment of non-ionizing radiation safety in humans, and will contribute toward the development of technologies for the facile and rapid assessment and identification of adverse effects that are attributable to such radiation, such as malignancies and other disorders. A primary aim is to establish and propose safe dose thresholds for non-ionizing radiation in various contexts, including medical imaging and therapy, toward avoiding pathophysiological effects.
