Method of biochemical -biophysical indication and organisms’ "life’s quality” forecast in conditions of radiation pollution

Khlebnyy E.S.

Institute of biological problems cryolitozone the Siberian Branch of the Russian Academy of Science, Yakutsk

Responses of living organisms to any natural stress factors are formed due to changes of genome activity regulation in replication processes, the reparations, directed on translation. And in these processes, as well as in reactions of specific irritants reception, the big role play conformational changes of DNAand the protein being an obligatory stage of their synthesis and functioning [1], andpermolecularthe basic medium-forming clusters: waters and membrane lipids [2]. In turn these processes underlie adaptive potential of a living organism and cause, together with activity of first echelon of protective systems - antioxidants, its change “life’s quality" at action of different stress factor of environment.

The purpose of work - development of the biochemical-biophysical integrated approach of an estimation of organisms’ “life’s quality” in polluted environments which includes cytological, biochemical and biophysical methods of an estimation of stress factors action on a homeostasis of growing cells of various organisms.

Materials and Methods

Simultaneous definition of inclusion 3Н-thymidine in DNA, 14С-leucine in synthesized fibers and mitotic index of growing cells for an estimation of differential processes activity of replicative and reparative DNA synthesis, translations of fibers withτ½> 24 hours, and also definition of low-molecular (LMAO) maintenance and activity of high-molecular antioxidants (SOD) is carried out. All characteristics are estimated in relative units in relation to the control. The biophysical component consists in registration of secondary radiation parameters (S) of cellular culture in an aquatic environment, describing a degree of structurization and conformational alteration in system water clusters of mediums - permolecular biopolymer structures of cells.

The method is approved on yeast cultureCuf liverat action on them of radiating stress factors, for example, at reception of a total absorbed dose by themγradiation100÷350μRentgen (μR). For creation of a background have been usedγ-quantum243Am in a range of total dozes from 100 up to 350 μR.

Results and Discussion

Results of consequences of influence γ-radiation in three dozes are introduced on tab. 1.

Table 1. Cytology-biochemical and biophysical characteristics of Cuf livere culture after influence γ-radiations. (normalized in relation to the control)
radiation dose, μRentgen / mitotic index / 3Н-thymidine inclusion / 14С-leucine inclusion / Superoxide dismutase activity / Content of low-molecular antioxidant / Area of secondary luminescence
0 / 1,0 / 1,00 / 1,0 / 1,0 / 1,0 / 1,00
100 / 0,9 / 1,3 / 1,3 / 0,8 / 1,1 / 1,17
250 / 0,9 / 0,8 / 0,8 / 1,0 / 1,0 / 1,40
350 / 0,8 / 1,2 / 1,2 / 0,8 / 1,3 / 1,22

Primary results allow to calculate in relative unitprocesses speed of replicative(krepl)and reparative (krep)synthesis of DNA, and also translation(ktrans), the total activity and efficiency of genome, and also a degree of conformational changes, and structures of DNA, proteins and water clusters (fig. 1)

kaop (antioxidant protection) = (SOD activity + [LMAO]) /2 / krep (reparation) = [3Н-thymidine inclusion]N - [mitotic index]N
ktga (general genome activity) = [14С-leucine inclusion]N + [3Н-thymidine inclusion]N. / krepl (replication) = [mitotic index]N
kgst (general genome stability) = kaop +krep/ kgga / ktrans (translation) = [14С-leucine inclusion]
kprod (productivity) = (ktrans +krepl)/ 2 / kcc (conformational changes) = [S secondary luminescence]N
Fig.1. Used factors calculation formulas.

Values of coefficient (table. 2.) testify, that dependences of genome functional activity and stability, as well as characteristics of conformation transposition and structure degrees of biopolymers and water clusters, from a doze γ-radiations, nonlinear.

Table 2.Calculated valueskоаз,kтрансл,kрепл,kреп,kоаг,kуст,kпрод, R and biophysical characteristics of cells of yeast штаммаCuf livereafter influenceγ-radiations (normalized to the control)
radiation dose, μRentgen / kaop / ktrans / krepl / krep / ktga / kgst / kprod / Area of secondary luminescence / R
0 / 1,00 / 1,00 / 1,00 / 1,00 / 1,00 / 1,00 / 1,00 / 1,00 / 1,00
100 / 0,92 / 1,25 / 0,85 / 1,42 / 1,17 / 1,07 / 1,05 / 1,17 / 1,12
250 / 1,00 / 0,78 / 0,88 / 0,94 / 0,873 / 1,04 / 0,83 / 1,40 / 0,86
350 / 1,01 / 1,23 / 0,84 / 1,39 / 1,15 / 1,11 / 1,04 / 1,22 / 1,15

Main of them, connected to protein synthesis, DNA replication, the total activity, genome efficiency and stability achieve a maximum at a total doze 100 μR and show the tendency to presence of the second maximum at a doze 350 μR. At the same time, the maximum of conformational intensitythe transitions resulting in increase of structure degree, first of all water clusters falls at a doze 250 μR (tab. 3).

Apparently, at this doze the resonance between parameters of the given form ionizing radiation and own frequency characteristics of fluctuations water dissipative structures is reached. As provides the maximal structurization of water, it is similar to increase in intensity of a geomagnetic field, characteristic for small indignations causes increase in a degree of structure of water in 2 times [3].

Table. 3. Reorganizations water clusters depending on γ-radiations
radiation dose, μRentgen / replication
contribution / translation
contribution / Reparation contribution / Water structures contribution
100 / 1,0 / 0,070,01 / 0,050,01 / 0,16
250 / 1,0 / 0,070,01 / 0,050,01 / 0,42
350 / 1,0 / 0,070,01 / 0,050,01 / 0,22

Due to that the degree "reception" ofγ-radiationsby cells (more precisely structures of their aquatic environment raises. The last, apparently is the reason of decrease, at a doze 250 μR, activity of DNA reparation, as consequence of genome efficiency and a parameter of "life's quality" (R =kprod*kgst)

Conclusion

Carried out research shows efficiency of suggested method at an estimation "life's quality" organisms in conditions of radiation pollution connected with change of stability and activity of cellular genomein processes of replicativeand reparative synthesis of DNA and directed on translation, and also with induced conformational reorganizations not only biopolymers, but also clusterstructures of water.

Acknowledgements

We appreciate very much the comments on the manuscript of Zhuravskaja A.N. and Kershengolts B.M.

References

  1. Alexeys V.G. Stability of plants in conditions of the North: ecology-biochemical aspects. Novosibirsk: VO the Science, 1994.152 p.
  2. Hlebnyy E.S., Kershengolts B.M. To a question on physical and chemical mechanisms of formation of reciprocal adaptive reactions of unicells on action stress –factors of environment // the Science and education. №2, 2005. P. 65-74.
  3. Kershengolts B.M., Chernobrovkina T.V., Nebrat V.V., Rabinovich E.V., Hlebnyy E.S., Sheyin А.А, KershengoltsE.B. Actionof hydroalcoholic systems on dissipative conditions of the person. Hypothetical model of biogenic and narcogenic spirit-basedproducts // Narcology. №8, 2004. P. 64-76.