The Role of Immunomodulator Betaleukin in Recovery of Hepatocytic Ploidy Profile in Delayed Terms after Irradiation.

We studied the radioprotective effect of betaleukin administered to (CBA×C57Bl/6)F1 mice in single doses of 50 and 3 µg/kg 2 and 22 h, respectively, prior to long-term (21 h) whole-body low-intensity (10 mGy/min) γ-radiation (137Сs; total dose 12.65 Gy). Hepatocyte ploidy, a biomarker of metabolic disorders of the liver, was evaluated, and nuclearity and ploidy indices were calculated. In 10 months after irradiation, a significant decrease in the ploidy index was revealed in the group of irradiated mice, while in animal receiving 3 or 50 µg/kg betaleukin, this parameter did not differ and even surpassed the control level, respectively. Thus, in vivo assessment of hepatocytic ploidy profile in mice revealed negative delayed effects of γ-irradiation in a dose of 12.65 Gy and a protective effect a single injection of immunomodulator betaleukin.

Changes in the Activity of Genes Involved in the Regulation of Hematopoiesis during Tumorigenesis in Irradiated Mice.

We used 183 F1 CBA×C57Bl hybrid mice to study the delayed effects of low-power long-term γ-irradiation at a dose of 12.6 Gy (10 mGy/min) 8 and 10 months after the treatment. Eight months after the treatment we found the increased expression of the transcription factor NFκB and its target genes iNOS and G-SCF in the bone marrow (BM). Ten months after the treatment malignant lymphomas were revealed in 14 of 94 mice in the liver, abdominal cavity, and subcutaneously. In the BM of these mice, the transcription of the PTEN, NFκB, and iNOS genes was inhibited and the contents of long non-coding RNAs (lncRNAs) lnc p21, NEAT1, and microRNA miR-125b were decreased. The expression of the NFκB(p65) gene and miR-125b was inhibited in the BM of irradiated mice without tumors ten months after the treatment. These data show the deregulation of the P53 system supporting the genome stability in the BM of irradiated mice. These indices will be studied as potential markers of risk of development of irradiation-induced tumors.

The Dependence of the Mutagenic Effect on the Dose of X-Ray Irradiation in an In Vivo Experiment on Female (CBA×C57Bl/6)F1 Mice.

We studied the mutagenic effect of X-ray irradiation in doses of 0.5, 1, and 1.5 Gy on female (CBA×C57Bl/6)F1 mice. The mutagenic effect (assessed by the parameter "frequency of bone marrow polychromatophilic erythrocytes with micronuclei") linearly depended on the dose of X-ray irradiation in the range of up to 1 Gy and reached the plateau at 1.5 Gy. The fraction of polychromatophilic erythrocytes was 45, 45, and 46% under control conditions (without exposure) and exposure to the irradiation in the doses of 0.5 and 1 Gy, respectively. Irradiation in a dose of 1.5 Gy induced a slight inhibition of erythropoiesis. These data confirm the hypothesis on possible death of highly aberrant erythrocyte precursors after irradiation in high doses.

[Evaluation of the treatment effectiveness of domestic G-SCF preparations in experiments on irradiated dogs].

We have evaluated the treatment effectiveness of Leucostim and Neupomax in dogs exposed to radiation at lethal doses of 3 and 3.5 Gy, correspondingly, by testing the dynamics of the blood cell number, first of all, leucocytes and neutrophiles, and the 45-day survival. Supportive therapy for all the dogs, including the control ones, consisted in antibiotic treatment during the acute period of 7-24 days. It was shown that both pre-parations administered consecutively for about 17-21 days after irradiation positively influenced the dynamics of all blood cells but predominantly impacted the neutrophile number dynamics. The latter ones manifested a higher nadir level and an earlier onset of restoration in the G-SCF treated dogs in comparison with the control ones. The tendency to a positive influence on the survival has been shown in Neupomax-treated dogs exposed to 3.5 Gy of radiation (plus about 40%). The results of the experiments were in good accordance with the data by foreign authors who used Neupogen. This allows a conclusion that home-produced G-SCF preparations can replace their foreign analogues.

[Actual problems of searching and studying radiation countermeasures].

The state of radiation counterdrug elaboration has been analyzed. The main criterion of estimation is how various possible radiation incidents are provided with radiation countermeasures. The latter are differentiated in 3 principal groups: radioprotectors, radiomodificators (these are able to have a positive effect when administered preliminary, before the exposure, or provide a delayed nonspecific protection after the exposure--urgent therapy) and hemopoietic growth factors demanding course administration. It should be underlined that the list ofofficinal radiation countermeasures is rather short. The most dynamic now are investigations aimed at developing a home preparation of recombinant human interleukine-1beta named betaleukine, and the preparation CBLB502, a modified microbe polypeptide elaborated in the USA. Also elaborated is a scheme of emergency exposure treatment. It includes urgent administration of the cytokine combination (betaleukine and thrombopoietin) with subsequent supportive therapy and a hemopoietic growth factors course. In the case of medical radiation- and chemotherapy the preparations betaleukine and thiol compound amifostine are used rather seldom. Official countermeasures for protection against low dose rate prolonged exposure are still absent. The problem of an indicator/marker of the radioresistance induced by a radioprotector or radiomodificator still remains unsolved. Reliable indicators/markers are needed to provide the 2nd stage of clinical trials of radioprotectors/modificators.

[Recombinant thrombopoietin antiradiation therapeutic effectiveness evaluation on dogs according to hemopoiesis and survival criteria].

Recombinant human thrombopoietin (rh TPO) has been investigated as a means of acute radiation disease urgent treatment in the experiments on 24 mongrel dogs. The animals were exposed to total acute gamma-irradiation at the doses of 3.5 Gy (exceeding LD50/45 under our conditions) and 3 Gy. All the dogs including control ones received antibiotics Ampicillin and Gentamicin twice a day during the acute period from the 7th to the 21st day. TPO was injected one time s/c or i/v at the doses of5 or 10 mkg/kg 1.5-2 h after exposure. TPO at a dose of 5 mkg/kg was ineffective. TPO at a dose of 10 mkg/kg had a positive effect on the kinetics of blood forming units, especially platelets (nadir, restoration rate) in terms of the 45-day survival. As a result, in TPO groups, nadir averaged at both exposure doses on leucocytes (1.3-1.4) x 10(9)/l vs (0.70-0.75) x 10(9)/l in control groups and on thrombocytes (102-112) x 10(9)/l vs (44-33) x 10(9)/l in control ones. Despite the low number of animals in experimental groups, the results permit to regard rhTPO as a worth-while urgent therapeutic means for the acute radiation damage treatment and preventing thrombopenia.

[Comparison of different G-CSF treatment effectiveness in experiments on irradiated mice].

In the experiments on F1 (CBA x C57BL) and BALB mice irradiated by 137Cs gamma-rays, preparations of unglycosilated G-SCF such as Neupogen and their domestic analogs Leucostim and Neupomax were investigated. The tests such as 9-day bone marrow cellularity (BMC) and endogenous CFUs, the neutrophile number restoration, the 30-day survival index have shown that all three preparations have an approximately equal effectiveness relating to acute radiation disease treatment and granulopoiesis stimulation after a 5-10 day consecutive administration following irradiation of mice at lethal and sublethal doses. We have come to the conclusion that Leucostim and Neupomax can be regarded as adequate substitutes for Neupogen.

[The threshold for radiation stochastic effects: arguments "pro" and "contra". Applied realization].

This study represents the analysis of the data available in the literature and the author's findings concerning the issue of a shape of the dose stochastic effect curve in the range of low levels of radiation (LLR). The data obtained from radioepidemiological and experimental investigations are used. Also considered are the arguments "pro" and "contra" regarding approximation of these curves by means of a linear function (linear non-threshold conception) or as a quasi-plateau (threshold conception). The above analysis allows us to conclude that the threshold conception is more reliable than the non-threshold one from the standpoint of the analysis of postulate bases, theoretical paradigms, the mechanisms for radiobiological effects, epidemiological and experimental data. It is suggested that a separate radiogenic cancer risk estimation should be used in case of LLR and high level radiation instead of one overall estimation by means of the linear non-threshold model.

[Radiobiological analysis of cancerogenic risk values in radioepidemiological investigations].

The aim of the present article consisted in critical analysis of the epidemiological approach to radiocancerogenic risk estimation in region of low level radiation (LLR). The estimation is making by means of mathematician models that ignore a principal difference in biological action of LLR and high level radiation (HLR). The main formal characteristic of LLR action is the presence of a plateau in beginning of a dose-effect curve of radiogenic risk. It may be argued by the following positions: repeating the plateau-phenomenon on various radiobiological effects, in different tests and bioobjects, first; a paradoxical trend of reciprocal ERR/Sv increasing regarding dose decreasing in region of plateau, second, and third, the increasing of the curvature in dose-effect curve beginning. The presence of a plateau is associated with the presence of a real radiogenic risk threshold. Besides, the analysis of processes influencing significantly the dynamics of initial radiation injury of biologically important macromolecules showed the preference in region of LLR those, decreasing/eliminating genome damages. There is follows from mentioned above a necessity to evaluate radiogenic risks in LLR region separately from HLR region.

[Recombinant human interleukine-1beta (betaleukine) usage for acute radiation sickness of severe degree treatment at canines].

Recombinant human interleukine-1beta (betaleukune) of Institute of especially pure biopreparation production had been examined as a treatment means at acute radiation disease of severe degree at dogs. Dogs were irradiated totally in doses above the LD95/45. Betaleukine had been administered s/c twice in day in 15 min - 2 h after irradiation. All the dogs, including control ones, received in acute period 8-24/26 days after irradiation antibiotics ampicillin and gentamycin i/m. Betaleukine increased 45 day-survival by 37% at 4 Gy and by 25% at 4.4 Gy. The effect correlated with more high level of nadir and more early beginning the leucocyte number restoration. We observed the regularity at all the dogs that received betaleukine as survived as died, but in the latter case in a lesser degree certainly. Besides the noticed character of leucocytes kinetics had been repeated at all the blood cell types but in the different degree. It had been concluded on the base of these observations that betaleukine acts on hemopoietic stem cells preferentially. The effect is in preventing death of a stem cells part, or in stimulating survived stem cells proliferation, or in both together. Betaleukin can be regarded as a suitable means of urgent pathogenic therapy at radiation accidents.

[The analytical review of the schemes of the acute radiation disease treatment used in experiment and in clinic].

The analysis of the original sources showed that there are some differences in approaches to radiation injuries treatment used mainly by Russian and foreign researches in their experiments. In Russian radiobiology the main way was the single usage of high molecular substances of different chemical structure and of natural origin soon after radiation exposure. These substances were designated then as cytokine inductors. Foreign researchers investigated in the beginning analogous substances too, but then--recombinant cytokines exclusively. In native radiobiology the urgent (in limits of 2 h) and early (in limits of 6-24 h) pathogenic therapy develops already for a long time. There are preparations, betaleukine (recombinant human interleukine 1 beta) and dezoksinat (degraded DNA), as results of developing the directions. Both have licenses on the administration at radiation accidents. But the issues of various medicaments optimal combinations in a common scheme of early (in first 24 h) medical help remain undefined accurately. The consensus protocol of radiation injuries treatment in clinic consists of the supportive therapy (aseptic conditions, wide antimicrobic antibiotics, thrombocytes and erythrocytes transfusions) and the myelostimulation therapy by cytokine combinations. The hemopoietic stem cells transplantation in cases of severe radiation damages treatment remains under question in the reason of immunological conflict between a host and a graft.

[Analysis of the epidemiological data concerning radiation carcinogenic effects and approaches to the low doses' upper limits determination in the aspect of a threshold of the unhealthy influences of ionizing radiation]. / Analiz dannykh épidemiologicheskikh issledovanii radiokantserogennogo éffekta i podkhodov k opredeleniiu granitsy malykh doz v aspekte porogovosti biologicheski brednogo deistviia ioniziruiushchei radiatsii.

The analysis of the epidemiological data regarding cancer mortality in cohorts of Japanese A-bomb survivors and Chermobyl liquidators exposed to different doses suggests that there are good reasons for recognizing the threshold of the radiocarcinogenic effect in the region of about 200 Gy (mSv). The analysis of solid cancer mortality in Japanese cohort, which exceeded the expected one in a dose diapason of 5-200 mSv, revealed a (quasi) plateau in a dose-effect curve and led to the conclusion that the nature of the overshoot is non-radiogenic. The analysis of supposedly dose dependent leucosis incidence in the limited low dose diapason in the Chernobyl cohort showed that the real coefficient of the excess absolute or relative radiation risk could not be received in the case because the larger part curve was placed under the control level. In supporting the principle of single hit in a cell nucleus as a base of microdosimetric determination of low radiation doses, the approach to objective delimitation between low, intermediate and high doses regions has been proposed. The low doses upper limit of sparse ionizing radiation for cell nucleus of 8 microns in diameter has been evaluated as 0.65 mGy. It can serve for evaluation of the dose rate threshold regarding the safe chronic radiation levels in the environment.

[Study of mechanisms of anti-irradiation effects of interleukin-1beta in long-term bone marrow cultures]. / Issledovanie mekhanizmov protivoluchevogo deistviia interleikina-1beta na modeli dlitel'nykh kul'tur kostnogo mozga.

The influence of betaleukin (human recombinant interleukin-1 beta) on the processes of postirradiation recovery of haemopoietic precursors (GM-CFC) and the level of granulocyte-macrophag colony-stimulating factor (GM-CSF) were studied in long-term bone marrow cultures after gamma-irradiation with a dose 2 Gy. Then the betaleukin action on the contents of GM-CFC and induction of GM-CSF in the non irradiated cultures was studied. It was shown that betaleukin increased the induction of GM-CSF and raised the contents of GM-CFC in long-term bone marrow cultures, and the maximal increase of a GM-CSF level and GM-CFC amount was marked in 20 hours after introduction. At an irradiation of long-term bone marrow cultures in conditions of betaleukin introduction 20 hours prior to influence of radiation the smaller degree of damage and faster recovery of GM-CFC was observed. The data in this report suggest that one of the mechanisms of antiirradiation action of betaleukin apparently is connected to the action of the preparation on hematopoietic microenvironment cellular elements, that causes the release of a colony-stimulating factor and stimulation of recovery of haemopoietic precursors.

[Dependence of the therapeutic effectiveness of interleukin-1beta on the time of the drug administration after irradiation of mice]. / Zavisimost' lechebnoi éffektivnosti interleikina-1beta ot sroka vvedeniia preparata posle oblucheniia myshei.

In experiments on mice F1(CBA x C57BL/6) the dependence of 30th days survival on the time of betaleukin (medicine form of interleukin-1 beta) administering after exposure to 7.5 Gy whole body gamma-irradiation from 137Cs (approximately DL80/30) was studied. Betaleukin was injected subcutaneously in dose 25 mcg/kg 0.2, 1, 3, 6 and 24 h after the exposure. The highest therapeutic effect took place in case of 0.2 h interval, then it dropped but was slightly expressed at 1 h. The hemopoiesis condition was studied in 7 or 9 days after the mice exposure to 6 Gy and betaleukin administering 1 h later in dose 25 or 50 mcg/kg. The positive effect on granulopoiesis beginning from the level of CFU-GM was observed. There were analyzed the reasons of weaker betaleukin effect and shorter period of its effectiveness after exposure in comparison with literature data regarding IL-1.

[Pro and contra regarding the threshold/non-threshold mutagenic (carcinogenic) action of low-level ionizing radiation]. / Pro i contra porogovosti/besporogovosti mutagennogo (kantserogennogo) deistviia ioniziruiushchego izlucheniia nizkogo urovnia.

According to the analysis of epidemiological and experimental data there are evidences in threshold on criterion of mutagenic (in broad sense) ionising radiation action. A threshold takes place in the region of about 20-200 mGy concerning a dose in case of an acute exposure and in the region of the third order above a background level regarding a dose rate in case of chronic or prolonged exposure. Evidences in favour of linear non-threshold conception are based on primitive mathematical extrapolations and approximations of the study results, mainly, in the region of intermediate doses/dose rates. Besides real data in region of low doses are ignored. The latter has a reason in paradigm of stochastic damage action of ionising radiation and in underestimation of cell and organism protection-reparation systems action. Regarding axiomatic bases of ionising radiation action it is possible to conclude more reliable substantiation of the dual (positive-negative) nature and the threshold conception concerning ionising radiation action in comparison with the non-threshold one.

[Biological effects of low level ionizing radiation (the threshold of effects and radiosensitivity/radioreactivity of biological structures with different level of organization)]. / Kontseptsiia biologicheskogo deistviia ioniziruiushchei radiatsii nizkogo urovnia (analiz problemy v aspektakh porogovosti éffektov i radiochuvstvitel'nosti/radioreaktivnosti biostruktur razlichnogo urovnia organizatsii).

There have been analysed the data as own as taken from the literature concerning the action of low level radiation (LLR) on different structure level in a mammalian organism. There have been compared the types of various structures early reactions regarding an acute or prolonged exposure depending on the test type and the studied structure organization level. The data analysis has shown it should differentiate the damaging and irritating action of LLR. The former reveals the radiosensitivity and the latter does the radioreactivity of the structure studied. The damaging radiation action has an threshold and the latter increases in the way from molecular to whole body level. It is supposed the irritating LLR action is realized through the involvement of organism and cell regulative systems. The existence of a threshold in the case is unknown so far.

[The estimation of the level of mutations at the glycophorin A locus in subjects exposed to the chronic action of ionizing radiation]. / Otsenka urovnia mutatsii v lokuse glikoforina A u lits, podvergavshikhsia khronicheskomu vozdeistviiu ioniziruiushchei radiatsii.

11 persons, who had been irradiated chronically at low dose rate under occupational conditions in 1950s in doses 220-581 cGy according data of individual film dosimeters, and 5 control persons were examined regarding the level of glycophorin A (GPA) mutation type NO and NN in blood erythrocytes. Significantly higher level of GPA mutations type NO was registered in average in the group of exposed persons (23.2 +/- 4.6 x 10(-6)) compared with the control group (10.2 +/- 2.1 x 10(-6)) through the dose dependence was expressed slightly. The coefficient of the linear regression has equaled (2.3 +/- 1.2 x 10(-6)) Gy. The outlook on GPA assay usage in retrospective biodosimetry is discussed.

[Cytokines in the aspect of pathogenesis and therapy of acute radiation sickness]. / Tsitokiny v aspekte patogeneza i terapii ostrogo luchevogo porazheniia.

The statement about dominating role of stem cell failure (exhaustion) in acute radiation bone marrow syndrome outcome needs to be changed. Cytokine-producing cells lacking reactivity that prevent usage of stem cell compartment reserves can play important role in immunohemopoiesis radiation damage.

[Radiobiological, clinical, experimental and methodological aspects of postradiation repair of hematopoietic stem cells]. / Postluchevaia reparatsiia stvolovykh krovetvornykh kletok v obshcheradiobiologicheskom, klinicheskom, éksperimental'nom i metodicheskom aspektakh.

On the basis of literature and author's data the topics dealt with revealing and evaluation of post-radiation repair as well as possibility of its acceleration in hemopoietic stem (colony-forming) cells are presented and discussed. Some conceptions formed in this field of science cannot be regarded as final ones and should be checked by means of different independent methods. Evaluation of stem cell repair pronouncing and its acceleration possibility is rather indefinite.