N. A. Karnaukhova

Information theory of ageing: Studying the effect of bone marrow transplantation on the life span of mice

In this paper the method of life span extension of multicellular organisms (human) using reservation of stem cells followed by autotransplantation has been proposed. As the efficiency of this method results from the information theory of ageing, it is important to verify it experimentally testing the basic concepts of the theory. Taking it into consideration, the experiment on bone marrow transplantation to old mice from young closely related donors of the inbred line was carried out. It has been shown that transplanted animals exhibited a survival advantage, a mean life span increased by 34% as compared to the control. This result not only demonstrates the efficiency of the proposed method for life span extension of multicellular organisms, but also confirms the basis of the information theory of ageing.

Effect of ionizing radiation on the synthetic activity of blood system cells in ground squirrels in different physiological states

The functional (synthetic) activity of blood lymphocytes and bone marrow hematopoietic cells in ground squirrels was studied in different seasons and at different stages of the torpor-arousal cycle. The effect of γ-irradiation on animals in different physiological states was also studied. The synthetic activity of cells was estimated from the amount of active RNA per unit DNA in the cell (parameter α). The α values in lymphocytes were minimal in hibernating animals (January–March), reached a peak upon their complete awakening (April), slightly decreased in the summer activity period, and decreased further in the prehibernation autumn period (November). During winter arousals between torpor bouts, this parameter reached the same values as in summer. The dynamics of parameter α in bone marrow hematopoietic cells were generally similar: minimal values in November and higher between torpor bouts than in summer. The peak of synthetic activity of proliferating hematopoietic cells recorded upon awakening from hibernation in April was mainly due to the accumulation of cells in the G1 and G2 phases of the cell cycle, and its decrease in summer reflected prevalent transition from G2 to mitosis and then partly to G0. In the torpor-arousal-euthermia cycle, two stages of awakening were distinguished, differing considerably in most of the test parameters. The synthetic activity and the total number of blood and bone marrow cells in ground squirrels irradiated in the state of torpor did not differ significantly from those in nonirradiated torpid animals. The adverse effect of radiation in animals irradiated at the initial stage of awakening was lesser than in animals irradiated in the active state, whereas animals at the second stage of awakening proved more vulnerable to acute irradiation. The physiological state of ground squirrels exposed to ionizing radiation at different phases of the torpor-arousal-euthermia cycle plays a key role in the dynamics of qualitative and quantitative characteristics of blood system cells. The results of this study indicate that the hypometabolic state of ground squirrels during hibernation is a factor of protection from the impact of ionizing radiation on the whole body and on the immune system in particular.

Informational Theory of Aging: The Life Extension Method Based on the Bone Marrow Transplantation.

The method of lifespan extension that is a practical application of the informational theory of aging is proposed. In this theory, the degradation (error accumulation) of the genetic information in cells is considered a main cause of aging. According to it, our method is based on the transplantation of genetically identical (or similar) stem cells with the lower number of genomic errors to the old recipients. For humans and large mammals, this method can be realized by cryopreservation of their own stem cells, taken in a young age, for the later autologous transplantation in old age. To test this method experimentally, we chose laboratory animals of relatively short lifespan (mouse). Because it is difficult to isolate the required amount of the stem cells (e.g., bone marrow) without significant damage for animals, we used the bone marrow transplantation from sacrificed inbred young donors. It is shown that the lifespan extension of recipients depends on level of their genetic similarity (syngeneity) with donors. We have achieved the lifespan increase of the experimental mice by 34% when the transplantation of the bone marrow with high level of genetic similarity was used.

Comparative analysis of efficiency of syngeneic and allogeneic transplantation without irradiation of the EGFP+ mice bone marrow cells with the microspectral fluorescence method

The results of experiments on syngeneic and allogeneic transplantation of the whole fraction of mice bone marrow cells without irradiation are presented. The data were obtained on the dynamic of donor cell colonization of bone marrow, spleen, thymus, and blood of the recipient mice. The degree of immunogenicity of the donor cells for both syngeneic and allogeneic transplantation of bone marrow was determined based on the method of microspectral fluorescence analysis. The low immunogenicity degree of donor cells was demonstrated for both transplantation variants within the framework of the conducted experiment. It is important to note that the allogeneic transplantation did not cause any reduction in the mean lifespan of the mice. These results, in combination with the results of our previous studies in which an increase of average lifespan by 34% was shown with syngeneic transplantation within the C57BL/6 EGFP+ strain allow one to anticipate the development of various methods of cellular therapy without risk of fatal consequences of immunological incompatibility between donor and recipient.

Use of fluorescence microspectral method for studying bone marrow EGFP+ cell transplantation in 5-fluorouracil-treated mice

In this paper the experimental results of bone marrow transplantation from C57BL/6-Tg(ACTB-EGFP)1Osb/J transgenic mice into C57BL/6 mice subjected to 5-fluorouracil treatment are represented. It has been shown that EGFP+ cell engraftment in bone marrow, spleen and thymus of host mice after 5-Fu treatment significantly increased. More long-term engraftment was recorded after transplantation between closely related donors and 5-fluorouracil treatment hosts. We have also obtained data on differences in the dynamics of EGFP+ cell engraftment in host investigated organs. To assess the effect of the donor’s bone marrow cells on the host immune system, functional activity of the synthetic apparatus (synthetic activity) of cells in bone marrow, spleen, thymus and blood have been investigated with fluorescence microspectral method. The results obtained allow of improving techniques for bone marrow transplantation without host irradiation in order to minimize the adverse effects.

A Study of the Regenerative Potential of Bone Marrow Cells of Donor Mice that Carry the egfp Gene in Irradiated Mice

A study of the regenerative potential of bone marrow cells of donor mice that express the enhanced green fluorescent protein was conducted in mice irradiated at a dose of 7 Gy. Expression of this protein allowed us to carry out monitoring of the presence of donor cells in recipient blood over the entire lifespan of the recipient. The lifespan of young recipients increased by 93% after transplantation; for old recipients it increased by 15%. Total acceptance of the bone marrow, spleen, thymus, and blood of the recipient with donor bone marrow cells was demonstrated over the entire life of the recipient. Only the donor colonies were detected with the studied irradiation dose and number of transplanted cells (11.7 ± 0.4) · 106 on the spleen surface. The percentage of bone marrow and spleen cells that expressed the CD117 and CD34 stem cell markers in the recipient mice was above the control level for a long period of time after the irradiation. More than half of the cells with CD117, CD34, CD90.2, and CD45R/B220 phenotypes in the studied organs were donor cells. Further detailed study of the peculiarities of the engraftment of bone marrow cells, both without preliminary treatment of recipients and after the effects of extreme factors, will allow improvement of the methods of cell therapy.

The information theory of aging: The major factors that determine lifespan

The information theory of aging assumes that decreased functionality of the whole organism and its systems is a result of decreased functionality of cellular repair mechanisms caused by the accumulation of genomic damages in the cells. A four-level cause-effect model of the interaction between the key processes during aging of highly organized multicellular organisms is proposed. Based on this model, some mathematical laws of changes in the viability of an organism are examined. The main fundamental factors that influence individual life expectancy, such as the initial amount of genomic damages at birth, the rate of its accumulation, and the pressure of the environment, are described. Additive and multiplicative interactions of the processes in different complex systems of a multicellular organism and their effects on its overall functionality (viability) are analyzed and the quantitative patterns of aging in a population of model organism are studied. The simulation provided the survival curves for species with different types of aging and a description of a compensation effect for species with aging that follows the Gompertz–Makeham law.

Fluorescent analysis in cell monitoring of environment and human health

One of the important methods to study the intracellular regulation of metabolism is the fluorescent spectroscopy. Fluorochrome acridine orange (AO) is widely used to investigate the nucleic acids in living as well as fixed cells. The objective of this research was to study the functional activity of the cell synthesizing apparatus with spectral fluorescent method. The cell functional (synthetic) activity is described by the parameter α representing the RNA/DNA ratio in fixed cells stained with AO. The possibilities of fluorescent cytodiagnosis have been studied to analyze the blood system cells in some diseases, under ionizing and electromagnetic irradiations, in natural and artificial hypomethabolic states as well as nervous and reproductive systems in the mollusks.

Dynamics of ribosomal activity and protein production in peripheral blood lymphocytes during an immune response

Microscopic spectrofluorimetry with acridine orange and 1-anilino-8-naphthalene sulfonate was used to assess the levels of translationally active rRNA (parameter α’, red/green emission ratio) and protein synthesis (parameter β, blue/green) in peripheral blood lymphocytes during rabbit immunization with ovalbumin. The averaged α’ and β changed synchronously and increased maximally after the third booster injection. At the height of the immune response, the distributions of lymphocytes in either parameter were broad and polymodal, suggesting differential activation of particular subpopulations. This two-dye, three-color assay holds much promise for monitoring the functional state of various cells.

Correlation between the synthetic activity of lymphocytes and the intensity of neutron radiation near the Earth surface

The synthetic activity of lymphocytes from rat and ground squirrel was assessed for correlation with the rate of neutron radiation about the Earth surface. In the periods 1999–2002 for rats and 2002–2003 for ground squirrels, significant positive correlation was obtained with the neutron flux incident on Earth and the overall “terrestrial neutron field” but not with the neutron efflux from Earth. No correlation was observed in 2005–2006 when the solar activity and the variation in neutron count were minimal. It is concluded that low-energy neutrons may be bioeffective in periods of substantial variation in flux intensity.