[Use of fluorescence microspectral method for studying bone marrow EGFP+ cells 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+ cells 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+ cells 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.

[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 the 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 the 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.

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

The experimental results on syngeneic and allogeneic transplantation of whole fraction of mice bone marrow cells without irradiation have been presented. Data on the dynamics of the donor cell colonization of bone marrow, spleen, thymus and blood of the recipient mice were obtained. The degree of immunogenicity of donor cells with syngeneic and allogeneic bone marrow transplantation based on the microspectral fluorescence method was evaluated. Within the framework of the experiment a low degree of immunogenicity of donor cells with syngeneic and allogeneic transplantation is shown. Importantly, allogeneic bone marrow transplantation did not cause any reduction in the mean life span of mice. These data and the results of our previous studies, demonstrating the mean life span increased by 34% with syngeneic transplantation in line C57BL/6 EGFP+, allow for developing different methods of cell therapy with no risk of fatal consequences of the immunological incompatibility between donor and recipient.

[On the possibility of determination of stereological characteristics of interactions of biological micro objects with nanoparticles based on analysis of 2-dimensional images of cytological and histological specimens].

In this article the method for studying 3-dimensional (stereological) characteristics of interactions of biological micro objects with nanoparticles on a basis of morphodensitometry analysis of standard 2-dimensional images of cytological and histological specimens is proposed. The performance of the task of determining the distance from registered nanoparticle to the center of the nucleus of the cell is described in detail. It is shown that using specific nanoparticles the results obtained may find application in science and diagnostics. Furthermore, it is possible to employ these results in nanotoxicology, in particular for determination of quantity characteristics of translocation of nanoparticles of different types in biological structures.

[About a new biophysical approach for total vaccination of humans and animals].

The new method of total nonspecific vaccination of humans and animals is presented based on the experience of practical use of a powerful light pulse for air disinfection in laboratory facilities.

[Serapeo Temple in Pozzuoli, Italy--the unique gauge for the sea (world ocean) level and the Earth surface temperature for over 2100 years].

The changes in the sea level relative to the position of the Serapeo Temple in Pozzuoly (Italia) over a period of 2100 years are discussed in the context of the well known periods of climate cooling off (Neoglacial, Little Ace Period) and climate warming (Middle Ages Optimum, Modern climate warming). It is noted that the rate of sea level lifting relative to the position of the Serapeo Temple in the modern phase of climate warming, which began the end of the 18th Century is approximately two times higher than in the previous phase of climate warming in the period from the fifth to the mid-tenth century A.D. This indicates that not only the natural cyclic component contributes to the mechanisms of Modern Climate warming but also the anthropogenic component of approximately equivalent power, which results from the waste of CO2 caused by the burning of fossilized fuels.

[PCR technique for prenatal diagnosis of fetal rhesus factor from peripheral maternal blood].

A procedure is proposed for the noninvasive determination of fetal rhesus factor in rhesus-negative pregnant women, which is based on the detection of the RHD gene in peripheral maternal blood by a PCR technique. The studies have shown the high sensitivity and specificity of determination of rhesus factor in a fetus at more than 15 weeks gestation. The technique does not require the use of such invasive and pregnancy-threatening procedures, as amniocentesis, cordocentesis or chorion biopsy. Furthermore, determination of fetal blood rhesus factor in rhesus-negative patients makes it possible to reduce expenses on the management of pregnancy, to avoid multiple determination of antibody rhesus and prevention of rhesus immunization, and to timely initiate therapeutic and preventive measures.

[Analysis of the temporal organization of the synthesis of RNA and protein in blood lymphocytes during the immune response of the body].

The temporal organization of the synthesis of RNA and antibodies in immunocompetent cells has been investigated by fluorecsent spectroscopy. Two fluorochromes: acridine orange and 1-anilino-8-naphthalenesulfonate were used. Temporal changes in the mean values of the characteristic parameters alpha' and beta in blood lymphocytes stained with acridine orange and 1-anilino-8-naphthalenesulfonate in the immune reaction to the introduction of ovalbumin were obtained. It was shown that the maximal increase in RNA and protein synthesis in blood lymphocytes by a factor of 6-8 occurs after the fourth immunization. The simultaneous increase in RNA and protein synthesis in cells was indicative of cell activation. A general response of immunocompetent blood cells to the introduction of the antigen was observed. The results confirm the idea about the cooperative interaction between different lymphocyte subpopulations during the synthesis of antibodies.

[The radioprotective effect of hypothermia on the immune and hematopoietic systems in mammals].

The functional activity of the synthetic apparatus (parameter alpha) in blood lymphocytes, bone marrow hemopoietic cells, and thymus cells, as well as the total number of blood and bone marrow cells in rats after y-irradiation at a dose of 8 Gy in the conditions of normothermia and hypothermia (16-18 degrees C) with hypoxia-hypercapnia were investigated after 2 h and on days 1 and 4. The recovery processes in blood in both groups of rats after acute X-irradiation at a dose of 7 Gy for 36 days were analyzed too. Under hypothermia, on days 1-4 after acute gamma-irradiation, a decrease in the synthetic activity in remaining cells and devastation in the hemopoietic system were pronounced to a lesser degree. After X-irradiation, the restoration of synthetic activity in blood lymphocytes was shown to begin earlier and to finish faster in "hypothermic" rats as compared with the animals irradiated in the state of normothermia. The survival of "hypothermic" rats was 100% as compared with 30% in "normothermic" animals. Thus, the data show that hypothermia exerts a radioprotective effect on the cells of the immune and hemopoietic systems, thus enhancing the resistance of the organism to radiation.

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

The functional (synthetic) activity of blood lymphocytes and bone marrow haemopoietic cells in ground squirrels during the annual cycle as well as in hibernating and awaken animals in winter have been studied by fluorescent microspectrometry. The effect of ionizing radiation on animals in different functional states of the hibernation-arousal bout was investigated too. It was shown that the synthetic activity (parameter alpha) in blood lymphocytes was minimal in hibernating state in winter and maximal in active euthermic spring animals, then slightly decreased in June and more considerably decreased in the prehibernating autumn period. In awake animals in winter, the values of parameter alpha reached the same values as in summer. The changes of parameter alpha in bone marrow haemopoietic cells were essentially the same: the minimal values were observed in the prehibernation autumn period and in awake animals in winter the alpha values were slightly higher than in active euthermic animals in summer. The maximal synthetic activity in bone marrow haemopoietic cells in active euthermic spring animals is due mainly to cells in G1-G2 phases of the cell cycle. The decrease of the synthetic activity in summer is a result of the cell transition from G2 to mitosis and transition of a part of cells to G0 When investigating the hibernation-arousal bout in ground squirrels in winter, during arousal, we found two stages considerably differing in both the values of parameter alpha in bone marrow haemopoietic cells and the number of blood cells. The synthetic activity and the total number of blood and bone marrow cells in ground squirrels irradiated in the state of deep hibernation did not differ significantly from the state of non-irradiated hibernating animals. The negative effect of radiation appeared upon the arousal of these animals but it was expressed to a lesser degree in comparison with the animals irradiated in the active state. It was found that the acute irradiation of animals during arousal from hibernation in the second stage caused the most pronounced functional inactivation and cell death. The physiological state of ground squirrels subjected to ionizing irradiation at different phases of the hibernation-arousal bout plays a determining role in the changes of the qualitative and quantitative characteristics of blood system cells. Thus, the hypometabolic state of ground squirrels in hibernation is a factor of protection from the action of ionizing radiation on the organism and the immune system.

[A study of the correlation between the functional activity of blood lymphocytes in different animals and the intensity of neutron radiation near the Earth surface].

A correlation between the functional activity of the synthesis apparatus of blood lymphocytes in different animals and the variations in the neutron count rate near the Earth surface has been studied. From 1999 to 2002 for rats and from 2002 to 2003 for ground squirrels, a reliable positive correlation between the synthetic activity of animal blood lymphocytes and the neutron count rate was found. The correlation was observed for both the neutron flux directed towards the Earth and the total neutron flux (the neutron field of the Earth). There was no a correlation for the neutron flux directed away from the Earth in this period. In 2005-2006, which is characterized by a minor variations in the neutron count rate, no correlation was observed. Thus, the biotropic effect of thermal neutrons makes its self-evident as changes in the functional activity of blood lymphocytes in periods of significant variations of neutron fluxes.

[Study of radioprotective effect of "Soma" preparate].

Changes in the functional activity of the synthesis apparatus of rat blood lymphocytes under different scheme in application of bioadditive Soma after acute X-irradiation by fluorescent microspectrometry. Some metabolic indices in animals were investigated too. It has been shown the bioadditive Soma using in norm reliably increased the synthetic activity on days 13 and 20. Preliminary Soma using during a month followed by the same interval increased the animal radioresistance (scheme 1), while the Soma using immediately after irradiation (scheme 2) revealed no pronounced radioprotective effect. It was found the Soma increased metabolism that may be important to recover homeostasis. The results show the expediency of further investigation of the Soma radioprotective properties with different concentrations and schemes as well as the necessity of monitoring the immune system during Soma using.

[Survival of amphibian embryos after continuous ultrasound treatment].

The influence of continuous ultrasound on the embryonic development of grass frog Rana temporaria has been investigated. Intact embryos at the blastula stage were treated by ultrasound of different frequency (0.88 and 2.64 MHz), intensity (0.05-1.0 W/cm2), and duration (1-15 min). The treatment with ultrasound of frequency 0.88 MHz and intensity 0.05 W/cm2 for 1-5 min tended to increase the proportion of normally developing embryos up to hatch (10-25% of control). Increasing the intensity of ultrasound (0.88 MHz) to 0.7-1.0 W/cm2 and the duration of its action to 5-15 min induced the death of almost all of treated embryos. No significant differences were found between the development of control embryos and embryos treated with ultrasound of middle intensity (0.2-0.7 W/cm2) for 1-5 min. The exposure of amphibian embryos to ultrasound of frequency 2.64 MHz and intensity 0.05-0.7 W/cm2 for 1-5 min did not change their survival. Increasing the intensity of ultrasound (2.64 MHz) to 1.0 W/cm2 and the duration of its action to 5 min decreased the number of normal developing embryos (by 35%).

[Changes in the permeability of amphibian embryo envelope for fluorochromes under the action of ultrasound].

It was determined whether high-frequency ultra sound can change the permeability of gray toad Bufo bufo and grass frog Rana temporaria under the action of high-frequency ultrasound. The changes in the permeability of embryonic envelope were assessed by using slowly penetrating fluorochromes ANS, FDA, and fluorescein. It was found that the ultrasound of 0.88 MHz and 0.4 - 0.7 W/cm2 intensity increased the permeability of amphibian embryonic envelope for ANS and FDA, whereas the ultrasound of 2.64 MHz and the same intensity increased that for fluorescein with the retention of low permeability for FDA. Embryos continued the normal development after treatment with ultrasound under these conditions.

[Changes in fluorescence spectral characteristics of blood nuclear cells by the action of physical factors associated with 11- and 22-year cyclic variations in solar activity]. / Izmenenie fluorestsentnykh spektral'nykh kharakteristik iadernykh kletok krovi pod vozdeistviem fizicheskikh faktorov, sviazannykh s 11- i 22-letnimi tsiklicheskimi variatsiami solnechnoi aktivnosti.

A correlation between the functional (synthetic) activity of animal blood lymphocytes and the intensity of radiation of the Sun at a frequency of 2800 MHz in 2000 and 2002 was studied. The results were compared with the data obtained in 1993 and 1994. These data suggest that there are seasonal regularities in the interrelationship of the processes studied. A change in the sign of the correlation coefficients at the maximum of the 23rd solar cycle was shown. Possible mechanisms of the phenomena are discussed.

[Microspectral studies of neuroendocrine regulation of gametogenesis in mollusks]. / Mikrospektral'nye issledovaniia neiéndokrinnoi reguliatsii gametogeneza u molliuskov.

An associated culture of nervous ganglia and gonads was used to study the gametogenesis neuroendocrine regulation in mollusks (Mytilus edulis L.). The state of oocytes was controlled by microspectral fluorescent analysis on slides stained with acridine orange. The object of investigations was Mytilus edulis L. collected in La Manch in April-May. A microspectrofluorimetrical investigation of samples was carried out in Pushchino. As a criterion of cell synthetic activity, the parameter alpha was used. Parameter alpha is the ratio of fluorescence intensities in the red (I640) and green (I530) regions of the visible spectrum: alpha = I640/I530. The results obtained showed that the nervous system realized the endocrine regulation of gametogenesis in mollusks. There was no normal development of oocytes in an isolated mantle culture. The development of oocytes in a tissue culture proceeds in a similar manner as in the gonads of the native organism in normal natural conditions only if the mantle tissue is associated either with all nervous ganglia or with the cerebral ganglion alone. It was found that the cerebral ganglion plays a key role in the neuroendocrine regulation of gametogenesis.

[Spectral studies of a single cell of marine microplankton. History and perspectives]. / Spektral'nye issledovaniia odinochnykh kletok morskogo mikroplanktona. Istoriia i perspektivy.

Methods and instruments for the spectral analysis of single cells of sea microplankton under the conditions of expeditions to the Mediterranean Sea and tropical zone of the Atlantic Ocean were proposed. Special emphasis was given to the ways of adaptation of sea microplankton communities to unfavorable environment with the formation of autotrophic and heterotrophic symbiont organisms (Radiolarian, Foraminifera, etc.) having closed cycles of phosphate metabolism and variotrophic microorganisms (Cyanophyceae and Dinoflagellata class).

[Reactive oxygen forms and luminescence of intact microspore cells]. / Aktivnye formy kisloroda i liuminestsentsiia intaktnykh kletok mikrospor.

The participation of reactive oxygen species (ROS) in luminescence (chemiluminescence and autofluorescence induced by ultraviolet light of 360-380 nm) was analyzed. Microspores, the pollen (male gametophyte) of Hippeastrum hybridum, Philadelphus grandiflorus, and Betula verrucosa and vegetative microspores of the spore-breeding plant Equisetum arvense served as models. It was found that the addition of the chemiluminescent probe lucigenin, which luminesces in the presence of superoxide anionradicals, leads to intensive chemiluminescence of microspores. No emission was observed in the absence of lucigenin and in the presence of the dye luminol as a chemiluminescent probe. The emission decreased significantly if superoxide dismutase, an enzyme of the superoxide anionradical dismutation during which this radical disappeared, was added before the dye addition. The autofluorescence intensity of microspores decreased in the presence of both superoxide dismutase and peroxidase, an enzyme destroying hydrogen peroxide and organic peroxides. The most significant effect was noted after the addition of peroxidase, which indicates a greater contribution of peroxides to this type of emission. The fumigation with ozone, which increases the amount of ROS on the cell surface, enhanced the intensity of the chemiluminescence of microspores with lucigenin, but decreased the intensity of the autofluorescence of microspores. Exogenous peroxides (hydrogen peroxide and tert-butylhydroperoxide) stimulated the autofluorescence of pollen and vegetative spores in a concentration-dependent manner. It was shown that the formation of ROS contributes to the luminescence of plant microspores, which reflects their functional state.

[Functional activity of ganglion neurons from the pond snail upon temperature variations]. / Issledovanie funktsional'noi aktivnosti neironov gangliev bol'shogo prudovika pri izmenenii temperatury.

Changes in the functional state of the synthetic apparatus of neurons of the peripharyngeal ring of the mollusk Lymnaea stagnalis upon elevating the temperature of the habitat from 4 to 18-22 degrees C were studied. In the first series of experiments, it was found that the synthetic activity of neurons of the great and small parietal, visceral, and pleural ganglia increases on the average by 70%. In the second series of experiments, changes in the average synthetic activity for all ganglia at 4.0, 6.5, 13.0, 16.8, 20.5, 21.5, 21.9, and 22.0 degrees C were studied. The increase in the synthetic activity of neuron cytoplasm (35% higher than the control values at 4.0 degrees C) was maximal at 13.0 degrees C; then the synthetic activity decreased to reach the level 20% higher than the control. The physiological range of temperature-dependent changes in the functional state of neurons was determined.