Analysis of the Effect of ß-Endorphin-Like Peptide Pentarphin on Intracellular Ca²âº Concentration in Early Mouse Embryos Using Fluorescent Probes.

At the early stages of mammalian embryo development, Ca(2+) influences the processes of cleavage and differentiation. An important role in these processes is played by peptide growth factors. One of them, pentarphin, is a peptide growth factor similar to the central part of ß-endorphin molecule and a fragment 369-373 of IgG heavy chain. We used fluorescent Ca(2+) indicators Fluo-3 AM and Fura Red AM. Pentarphin added to embryos in a concentration of 0.1 µM 1.5-2 fold increased intercellular Ca(2+) level in comparison with the initial level.

[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.

[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.

[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.

[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.

[Changes in functional activity of the synthetic apparatus of rat thymocytes under acute and chronic gamma-irradiation]. / Izmenenie funktsional'noi aktivnosti sinteticheskogo apparata timotsitov krys pod deistviem ostrogo i khronicheskogo gamma-izlucheniia.

The changes in functional activity of rat thymocyte synthetic apparatus (synthetic activity) under acute (7.5 Gy) and continuous (dose rates 14.4 and 0.43 cGy/day) gamma-irradiation were studied by the fluorescent microspectral analysis. It has been shown that after the acute irradiation the changes in synthetic activity occurred in three main stages. The stages reflect the depression and activation of synthetic processes that is due to interphase and reproductive cell death and urgent recovery of thymus cellularity and secondary repopulating. Under continuous irradiation with a dose rate 14.4 cGy/day in long-term period both the decrease of thymocyte synthetic activity (in most animals) and activation (in the animals with pronounced symptoms of radiation damage) were observed. This reflects the depression processes in immune system and augmentation of immunoreactivity due to mass antigen influence of transformed cells and infectious agents on thymocytes. Under low dose ionizing irradiation (dose rate 0.43 cGy/day) the undulating changes in synthetic processes in thymus cells were observed. This depends on the recurrence of depression and recovery processes in the blood-forming tissue.

[Fluorescent evaluation of blood nuclear cells in experimental myocardial infarction]. / Fliuorestsentnaia otsenka iadernykh kletok krovi pri éksperimental'nom infarkte miokarda.

Functional (synthetic) activity of acridine orange-stained blood nuclear cells was studied by microspectral fluorescent analysis in experimental myocardial infarction. A specific time course of changes in the synthetic activity of blood nuclears, described by alpha parameter, was detected in uneventful myocardial infarction and infarction complicated by suppurative inflammation of the suture. This time course reflected the reparative changes in the myocardium. Hence, alpha parameter can be regarded as an indicator of reparative processes in the myocardium, and the above method recommended for fluorescent cytodiagnosis of reparation processes after acute myocardial infarction and for predicting its course.

[Correlation between change in solar activity and functional activity of animal blood lymphocytes in norm and pathology]. / Korrleiatsiia mezhdu izmeneniem solnechnoi aktivnosti i funktsional'noi aktivnost'iu limfotsitov krovi zhivotnykh v norme i pri patologii.

It is shown that the correlation between solar activity variations and the state of lymphocyte in norm is depressed under influence of stronger internal (gall-stone disease) and external (continuous gamma-irradiation) effects in pathology.

[Study of the functional status of a synthetic apparatus of blood lymphocytes under the action of weak low-frequency magnetic fields]. / Issledovanie funktsional'nogo sostoianiia sinteticheskogo apparata limfotsitov krovi pri deistvii slabykh nizkochastotnykh magnitnykh polei.

The functional state of the synthetic apparatus of animal blood lymphocytes under weak low-frequency magnetic fields was studied. The changes in blood cell synthetic activity and leukocyte formula, a signal index of general nonspecific adaptation reactions, were shown. In the majority of cases examined, the variable magnetic fields increase the functional activity of the lymphocyte synthetic apparatus, improve the type of adaptation reaction, and enhance the level of immune resistance of the organism.

[Correlations between functional activity of animal blood lymphocytes and change in solar activity]. / Korreliatsiia mezhdu funktsional'noi aktivnost'iu limfotsitov krovi zhivotnykh i izmeniem solnechnoi aktivnosti.

It is shown that increase of Solar activity as measurement of the intensity of solar radio emissions at frequency of 2804 MHz leads to the reducing of the functional activity of immunocompetent cells in animal blood defining by parameter alpha.

[Quantity and quality of animal immunocompetent cells in relation with variations in solar activity]. / Kolichestvo i kachestvo immunokompetentnykh kletok zhivotnykh v sopostavlenii s variatsiiami solnechnoi aktivnosti.

It is shown that the number and quality (synthetic activity) of immunocompetent cells in animal peripheral blood correlate with the parameters of solar activity (number of spots on the Sun and solar fluctuations).

[Change in the fluorescent characteristics of rat lymphocytes stained with acridine orange after acute gamma-irradiation]. / Izmenenie fluorestsentnykh kharakteristik okrashennykh akridinovym oranzhevym limfotsitov krovi krys posle ostrogo gamma-oblucheniia.

Changes of parameter alpha of rat blood lymphocytes after acute gamma-irradiation have been investigated by luminescence microspectral analysis. Parameter alpha reflects the cell synthetic activity and represents the ratio of intensities of the red (640 nm) and green (530 nm) components the luminescence spectrum of acridine orange fluorochromed cells. It has been demonstrated that changes of parameter alpha occur in three stages: a) sharp decrease at the 1st or 2nd day after irradiation: b) increase with the maximum at the 4th-15th day; c) fluctuations on the level higher then the control values. At each stage there exist typical specific shapes of parameter alpha distribution histograms. These results indicate that this method may be used for control of the immune system state during gamma-radiation sickness or in the process of human cancer radiation therapy.