Comparison of Direct Counts and Conductometric Measurements on a Hematology Analyzer Abacus Junior 5 Vet Myelokaryocyte Content in the Red Bone Marrow of Mice.

We studied the possibility of conductometric measurement of myelokaryocyte content in the red bone marrow of mice using a hematological Abacus Junior 5 Vet analyzer (Diatron). Statistical, correlation, and regression analyses were performed to assess of the results of myelokaryocyte counting in the suspensions of mouse red bone marrow by a direct method in cytometers and by using Abacus Junior 5 Vet analyzer. It was shown that in both intact mice and animals with modelled red bone marrow hypoplasia, irrespectively of the state of hematopoiesis in representative samples, conductometric measurements of myelokaryocyte content on the Abacus analyzer with high confidence reproduced direct counting results (in different tests p=0.64-0.82, p=0.83-0.98). This indicates that myelokaryocyte counting on the Abacus Junior 5 Vet analyzer can be an acceptable alternative to counting chamber measurements in mouse samplings. However, the variability of single measurements with the Abacus Junior 5 Vet in red bone marrow suspensions is high (5%) and this has to be considered in small samples.

Experimental Study of the Efficacy of Sodium Deoxyribonucleate Used in Combination with Co-Transplantation of Mesenchymal and Hematopoietic Stem Cells after Exposure to γ-Radiation.

We studied the possibility of using sodium deoxyribonucleate (Derinat) for improving the efficiency of co-transplantation of mesenchymal (MSC) and hematopoietic stem cells (HSC) to female F1(CBA×C57BL/6) mice with bone marrow aplasia caused by exposure to γ-radiation. It was found that immunomodulator Derinat enhanced the effect of co-transplantation, in particular, triple post-irradiation administration of Derinat accelerated hematopoiesis recovery judging from the parameters of peripheral blood, total cellularity of the bone marrow and spleen, and animal survival. Single or double administration of Derinat prior to irradiation was ineffective. The optimal result was obtained when the following scheme was applied: MSC→HSC with an interval of 48 h starting during the first hours after irradiation and triple administration of Derinat (in 10-15 min, 3 and 7 days after irradiation) in a dose of 3 mg/mouse.

Effect of Rat Exposure to Moderate Pressure Oxygen Atmosphere on Resistance of Erythrocytes against Hemolysis.

Exposure of male Wistar rats to oxygen atmosphere at moderate pressure (1.10-1.15 atm) for 4 h resulted in significant, transient, and reversible decrease in hemolytic resistance of peripheral blood erythrocytes. In 2 days after oxygen exposure, the bone marrow released young erythrocytes with enhanced hemolytic resistance as a compensation for oxygen injury. These findings suggest that normobaric oxygen load can be viewed as a stress factor of moderate intensity.

Effect of Different Regimens of Co-Transplantation of Hemopoietic and Mesenchymal Hemopoietic Stem Cells on the Rates of Hemopoietic Recovery in Laboratory Mice Treated with Cyclophosphamide in Sublethal Doses.

We studied the effect of mesenchymal stem cells and hemopoietic stem cells co-transplanted in different regimens (sequence and intervals between the injections within 0-48 h) on the rate of the bone marrow hemopoiesis recovery in 468 inbred mice with cytostatic aplasia caused by a single injection of cyclophosphamide in a dose of 500 mg/kg. The efficacy of stem cells was evaluated by animal survival and general physical condition, body weight dynamics, peripheral blood counts (leucocytes, platelets, and reticulocytes), total cellularity and the presence of hemopoietic stem cells in the bone marrow on day 9 after cyclophosphamide injection. The relative content of hemopoietic stem cells in the total myelokaryocyte pool was assessed by flow cytofluorometry using specific monoclonal antibodies to CD117 and CD184. The stimulating effect of co-transplantation of mesenchymal stem cells and hemopoietic stem cells on hemopoiesis was demonstrated by the indices of total cellularity and the relative content of CD117+ hemopoietic stem cells. The schemes with injection of mesenchymal stem cells 24-48 h prior to injection of hemopoietic stem cells were most effective.

Effects of Exposure of Animals to Oxygen Atmosphere at Low Pressure on Lipid Peroxidation and Antioxidant Defense.

Relatively short-term (2.5 or 5 h) exposure of Wistar rats to oxygen atmosphere at moderate pressure (1.10-1.15 atm) resulted in an increase in LPO level and reduction of antioxidant activity in the blood serum. An increase in malondialdehyde concentration 1 day after termination of the exposure was followed by a decrease in the inhibiting activity of free radical oxidation of liposomal phospholipids induced by Fe(II) ions (100 µm). Malondialdehyde concentration increased by 1.29 times already after 2.5-h exposure and did not changed when the duration of the exposure to oxygen atmosphere was prolonged to 5 h. These data confirm the necessity of using substances potentiating antioxidant defense of the body during exposure to normobaric oxygenation.

[Study of the influence of low-dose γ-irradiation on the functional state of peripheral blood erythrocytes of rats].

Low-intensity radiation at the absorbed dose of 4 µGy/min is a stressor of medium strength. In male Wistar rats, a pronounced and long-lasting response occurs in the system of red blood cells at the accumulated dose of 4.8 mGy. Functional deficiency of circulating cells was evaluated by the resistance of erythrocytes to acid lyse and the activity of the main antioxidant enzymes--superoxide dismutase (SOD) and catalase (CAT). The minimum "threshold" doses of radiation that cause systemic reactions occur in the range of units of miligrey.

[Increase in the number of cancer stem cells after exposure to low-LET radiation].

Radioresistance of cancer stem cells (CSCs) is regarded as one of the possible causes of cancer recurrence after radiotherapy. Since the regularities and mechanisms of radiation effects on this population of cells have not been sufficiently studied, the aim of this work is to elucidate the changes in the CSC number after γ-irradiation in stable cultures of tumor cells in vitro and tumor tissue in vivo (in the course of radiation therapy of patients with cancers of the upper respiratory tract). CSCs were identified in the cell lines B16, MCF-7, HeLa by the ability to exclude the fluorescent dye Hoechst 33342 (SP method) 48-72 h after irradiation at the doses of 1-20 Gy and in biopsy material by immunophenotype CD44+CD24(-/low) before and 24 h after irradiation at the total dose of 10 Gy. The essential differences in the response of CSCs and other cancer cells were found after exposure to low-LET radiation. The absolute number of CSCs increased after a single exposure at the doses ranging from 1 to 5-10 Gy in different cell cultures, but a further dose increase maintained the current number of CSCs or decreased it. At the same time, the number of non CSCs significantly decreased with increasing doses of radiation exposure, as expected. Fractionated irradiation in vivo at a total dose of 10 Gy increased the relative amount of CSCs in most patients. The registered changes are an integral indicator of cell death, cell division delay immediately after irradiation, proliferation at a later time, possible dedifferentiation of non CSCs, etc. The exact contribution of each of them to the radiation-induced increase of the CSCs number is of considerable interest and requires further research.

[Experimental justification of possible mechanisms of action of low intensity electromagnetic radiation (EMR) on animals' behavior].

Effects of EMR on the behavior of Wistar rats (196 males, 180-240 g of mass) under the conflict of opposed motivations: strong positive, drinking, motivation, and strong negative, pain, motivation were studied. The animals were exposed to low intensity EMR (40 microW/cm2) produced by two independent sources, 475 MHz (Albatross) with two orthogonal E vectors, and synchronization of rhythm modulation in the range of electroencephalography (EEG) frequency. The effect on behavior was observed during 10 min: 1) following the 5-minute exposure to EMR and 2) during the 10-minute exposure. Low intensity EMR of the above mentioned parameters and pulse modulation of 4, 8, 10 and 13 Hz was found to inhibit development of phobia to pain, increase the number of punishable contacts. It testifies to the existence of a weak anxiolytic effect which is similar to the effect of tranquilizers. If animals were exposed to EMR following administration of phenazepam, the radiation was shown to produce potentiation of the anxiolytic effect ofphenazepam. Effect of phenazepam is associated with activation ofbenzdiazipine receptors in the structure ofGABA-ergic receptor complex, which regulates neural membrane chloride channel conductance. We can suggest that anxiolytic and neurodepressive effects of EMR are realized to some extent at the level of ionophore and regulatory receptor complexes.

[Effect of synthetic tripeptides on hemopoietic stem cells in norm and after gamma-irradiation].

Our work was aimed at researching into the influence of dipeptide (gamma-dGlu-dTrp) "Timodepressin" and this dipeptide-based tripeptides on the colony-forming ability of the irradiated in vitro bone marrow and hemopoietic stem cells of the normal organism. Also studied was the effect of various doses (1-1000 microg/kg) of one oftripeptides (dAla-gammadGlu-dTrp) on the output of exogenous splenic colonies in the case of its introduction 48 hours before irradiation. It is shown that the mode of influence of the preparations produced on the basis ofdipeptides dGlu-dTrp and gamma-dGlu-dTrp on the initial stages ofa hemopoiesis in the normal and irradiated organism depends on the nature of the additional amino-acid residue and its optical orientation.

Hemostatic system in Wistar rats in different types of oxidative stress.

We studied parameters of hemostatic homeostasis in isolated and combined exposure to low-intensity γ-irradiation in a low dose, hyperbaric oxygenation, and antiorthostatic hypokinesia. Complex effects of the above stress factors are accompanied by a pronounced hypercoagulable shift with signs of thrombinemia against the background of depressed fibrinolysis, which indicates the risk of intravascular blood coagulation.

[Experimental assessment of embryotoxic action of low intensity ionizing irradiation in different periods of intrauterine development].

The prolonged ionizing radiation exposure at the absorbed doze rate of 2.6 mGy/hour and the total accumulated doze of about 20 cGy during different periods of the intrauterine development caused disorders of a coordinated development of organs and the decrease of endurance in physical activity in the postnatal ontogenesis of the irradiated posterity. The results obtained are comparable to the consequences of irradiation exposure of the fetus at the accumulated doze of 125 cGy during all stages of pregnancy and give grounds for considering radiation exposure at the above-mentioned rate (20 cGy) as a biologically significant factor.

[Influence of peptide derivatives of diketopiperazines on hemopoietic stem cells of an intact organism and irradiated bone marrow cells in vitro].

We have performed structural and functional studies of the hemotropic activity for a number of novel 2,5-diketopiperazine peptidomimetic derivatives. We employed a mouse model of hemopoietic stem cells cloning in the spleen of lethally irradiated animals. Biologic activity of synthetic products was studied in two experimental models: 1) in vitro irradiated bone marrow SFU-S was used for studying the radio modifying activity; 2) the biological effect of peptidomimetics on the intact non-irradiated bone marrow was evaluated in vivo. Various ways of administration of the peptidomimetics studied were used in the in vivo experiments: intravenous, intraperitoneal or subcutaneous injections and oral administration in the dose range of 10-10000 microg/kg. As a result of our work, we have discovered 2,5-diketopiperazine peptidomimetic derivatives with the dual activity: stimulation of intact committed SFU-S and radiomodifying activity.

[Psychophysiological development of two generations of female rats chronically exposed to gamma-irradiation during pregnancy in the total dose of 1 Gy and its modification by melanin].

The purpose of the work was to study the embryotoxic action of chronic gamma-irradiation of pregnant female rats (F0) during the first 10 days of pregnancy in the total dose of 1 Gy (mean dose rate of 5.31 mGy/hour) on psychophysiological development of posterity of the first (F1) and the second (F2) generations and its modification by natural pigment melanin (peroral 10 mg/kg once per day during the irradiation). 54 pregnant female Wistar rats were the objects of research were their 180 descendants of the first generation and about 400 descendants of the second generation of maternal and of paternal lines. Psychophysiological development and its correction by melanin estimated on ability to learning with the test of training a conditioned avoidance reflex in the shuttle box. Precise negative action of gamma-irradiation in the aforesaid dose on psychophysiological development of posterity of the first generation is established. At rats of the second generation the inferiority is shown mainly at descendants of maternal line. Application of melanin of natural origin in most cases diminished negative consequences of the irradiation.

[Experimental study of the radioprotective action of melanin on the somatic development during irradiation in the antenatal period of ontogenesis].

Daily introduction per os of the exogenous melanin in a doze of weight of 10 mg/kg pregnant female rats Wistar eliminated the functional deficiency of somatic development revealed at posterity at chronic gamma-irradiation in a doze 1.00-1.25 Gy for all period of pregnancy. The irradiation or introduction melanin antenatal to a phase ontogenesis resulted in stimulation of the immune answer, which was determined at offspring on 3rd week after birth. On the basis of the received data it is concluded presence radioembryoprotective actions melanin in the relation embryotoxic effects of small dozes of ionizing radiation and its participation in regulation immunogenes.

[Experimental study of the neuroprotective properties of the melanin in embryos irradiated during antenatal development].

Daily introduction per os of the exogenous melanin in a doze of weight of 10 mg/kg pregnant female rats of line Wistar on a background continuous irradiations (dose rate of 2.6 mGy/h within 20 days of pregnancy) eliminated deficiency cognitive functions at posterity. On the basis of the received data it is concluded presence radioembryoprotective actions of melanin in the relation neuro embryotoxic effects of small dozes ionizing radiation. Taking into account small toxicity of melanin, the preparation can be perspective for practical application.

Evaluation of psychoemotional status of rats after brain injury and systemic transplantation of mesenchymal stem cells.

We studied the effects of systemic transplantation of mesenchymal stem cells obtained after culturing of autologous bone marrow on psychophysiological status of Wistar rats after diffuse brain trauma. Two months after systemic injection of mesenchymal stem cells we observed a decrease in manifestations of emotional behavioral reactions (anxiety and excitability) and shortening of the time of realization of drinking behavior in a T-maze. A significant positive effect of systemic transfusion of mesenchymal stem cells on avoidance conditioning in a shuttle box was observed 3 months after brain injury.

[The effect of pulsed cyclical microware radiation on the conditioned behavior of rats]. / Vliianie tsiklicheskogo impul'snogo mikrovolnovogo oblucheniia na uslovno-reflektornuiu deiatel'nost' krys.

Research has been carried out to investigate the effects of pulsed cyclical microware exposure (7 GHz, 400 pps, 100 mcs, 10-20 mW/cm2, 10 or 20 cycles of "5 min exposure--4 min pause") on avoidance learning of rats. It was shown that reductions in conditioned behavior after cyclical pulsed microware exposure occurred at an SAR of 2.1 W/kg (10 mW/cm2). It was found the cumulation of the effects of the cycles at prolonged cyclical microwave exposures.

[Radiation-induced and embryotoxic effects in experimental hyperthyroidism]. / Eksperimental'no indutsirovannyi gipertireoz: ego radioembriotoksicheskoe deistvie.

The purpose of this article was investigation the development of the posterity rat (line Wistar) in the postnatal ontogenesis after combined and separate action of gamma-radiation (1.25 Gy at 20 days) and hyperthyroidism, provoked by toxic thyroid doses of 100 mg thyroidin per rat. It was shown that hyperthyroidism did not affect physical development of the irradiated posterity, but in a long-time period (7 month) it was revealed a decrease in ability to the training of posterity rats after the combined action of the factors.

[The influence of exogenous melanin on the development of female Wistar rats' posterity of first generation exposed to low doses at different dose rates during embryogenesis]. / Vliianie ékzogennogo melanina na razvitie potomstva pervogo pokoleniia krys Wistar, obluchennogo v émbriogeneze v maloi doze raznoi moshchnosti.

To research a radio-protective effect of melanin on the reproductive system during its creation the development of first generation posterity (900 descendants) of 125 pubertal female Wistar rats was examined after the single gamma-irradiation with a dose of 1 Gy (mean dose rate of 0.03 mGy/h), which they have been exposed to on the day ten of pregnancy, and the chronic gamma-irradiation with a total dose of 1 Gy (mean dose rate of 5.31 mGy/h) which they have been being exposed to during the first 10 days of pregnancy. The melanin of natural animal origin has the anti-radial effectiveness with the single irradiation increasing the number of alive newborn descendants in a litter and doubling their survival during the first 30 days after the birth. The melanin's effect is less expressed with the chronic irradiation.