Medical Radiology and Radiation Safety. 2024. Vol. 69. № 1

DOI:10.33266/1024-6177-2024-69-1-15-19

A.K. Chigasova^{1, 2, 3}, M.V. Pustovalova^{1, 4}, A.A. Osipov², S.A. Korneva⁵,
P.S. Eremin⁶, E.I. Yashkina^{1, 2}, M.A. Ignatov^{1, 2},Yu.A. Fedotov^{1, 2},
N.Yu. Vorobyeva^{1, 2}, A.N. Osipov^{1, 2}

Post-Radiation Changes in The Number of Phosphorylated H2ax
and Atm Protein Foci in Low Dose X-Ray Irradiated Human Mesenchymal Stem Cells

¹ A.I. Burnazyan Federal Medical Biophysical Center, Moscow, Russia

² N.N. Semenov Federal Research Center for Chemical Physics, Moscow, Russia

³ Institute of Biochemical Physics, Moscow, Russia

⁴ Moscow Institute of Physics and Technology, Moscow region, Dolgoprudny, Russia

⁵ M.V. Lomonosov Moscow State University, Moscow, Russia 

⁶ National Medical Research Center of Rehabilitation and Balneology, Moscow, Russia

Contact person: N.Yu. Vorobyeva, e-mail: This email address is being protected from spambots. You need JavaScript enabled to view it.

ABSTRACT

Aim: To study the patterns of changes in the number of foci of phosphorylated DNA double-strand break repair proteins H2AX (γH2AX) and ATM (pATM) in cultured human mesenchymal stem cells (MSCs) 1‒48 hours after exposure to X-ray radiation at doses of 40, 80, 160 and 250 mGy. 

Material and methods: We used the primary culture of human MSCs, obtained from the collection of LLC “BioloT” (Russia). Cells were irradiated using a RUB RUST-M1 X-ray biological unit (Diagnostika-M LLC, Moscow, Russia) equipped with two X-ray emitters at a dose rate of 40 mGy/min (voltage of 100 kV, an anode current of 8 mA, and a 1.5 mm Al filter) and 4 °C temperature. To quantify the yield of γH2AX and pATM foci immunocytochemical staining was carried out with the use of γH2AX and pATM antibody respectively. Statistical analysis of the obtained data was carried out using the statistical software package Statistica 8.0 (StatSoft). To assess the significance of differences between samples, Student’s t-test was used.

Results: It was shown that the kinetics of changes in the number of γH2AX foci after irradiation at doses of 160 and 250 mGy and low (40‒80 mGy) doses are significantly different. In contrast to the significant (50‒60 %) decrease in the number of γH2AX foci observed
6 hours after irradiation at doses of 160 and 250 mGy, after irradiation at low doses, no significant decrease in γH2AX foci was observed at this time point. Analysis of the colocalization of γH2AX foci with pATM foci indicates that the mechanisms for maintaining a high number of γH2AX foci 24‒48 hours after low-dose irradiation are ATM independent. A hypothesis has been put forward to explain the phenomenon of maintaining the number of γH2AX foci 24‒48 hours after irradiation in low doses by replicative stress caused by stimulation of proliferation against the background of hyperproduction of free radicals, resulting in additional formation of DNA double-strand breaks and phosphorylation of H2AX by ATR kinase.

Keywords: mesenchymal stem cells, γH2AX, pATM, DNA double-strand breaks, X-ray radiation, low doses

For citation: Chigasova AK, Pustovalova MV, Osipov AA, Korneva SA, Eremin PS, Yashkina EI, Ignatov MA, Fedotov YuA, Vorobyeva NYu, Osipov AN. Post-Radiation Changes in The Number of Phosphorylated H2ax and Atm Protein Foci in Low Dose X-Ray Irradiated Human Mesenchymal Stem Cells. Medical Radiology and Radiation Safety. 2024;69(1):15–19. (In Russian). DOI:10.33266/1024-6177-2024-69-1-15-19

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 PDF (RUS) Full-text article (in Russian)

Conflict of interest. The authors declare no conflict of interest.

Financing. The research was carried out with the support of the RNF (project No. 23-14-00078).

Contribution. Article was prepared with equal participation of the authors.

Article received: 20.10.2023. Accepted for publication: 27.11.2023.

Medical Radiology and Radiation Safety. 2024. Vol. 69. № 1

DOI:10.33266/1024-6177-2024-69-1-28-32

L.A. Romodin¹, E.I. Yashkina¹, A.A. Moskovskij²

Fluorimetric Evaluation of the Effect of Malic, Succinic and Ascorbic Acids on the Growth Properties of A549 Cells in Culture

¹ A.I. Burnazyan Federal Medical Biophysical Center, Moscow, Russia

² Russian Biotechnological University, Moscow, Russia

Contact person: L.A. Romodin, e-mail: This email address is being protected from spambots. You need JavaScript enabled to view it.

ABSTRACT

Relevance: A number of researchers consider the study of the radioprotective properties of non-toxic or low-toxic natural substances to be a promising direction. A special place among them is occupied by antioxidants and participants in the basic reactions of metabolism. In order to avoid methodological errors when performing these studies, it is necessary to conduct a number of additional experiments. For example, in order to study the properties of various substances on cell cultures using tablet readers, it is first necessary to make sure that these substances do not affect the ability of cells to adsorb to the bottom of the wells of the tablet and do not interfere with cell proliferation. And if such an influence is detected, further experiments with these substances should be planned taking into account the information received.

Purpose: To search the effect of ascorbic, malic and succinic acids on the ability of lung adenocarcinoma cells (A549) to adhere in a 96-well plate, followed by the onset of proliferation by fluorescence registration method using Hoechst-33342 fluorophore.

Methodology: The experiment was carried out in a 96-well tablet. The working concentration of Hoechst-33342 was 1 μg/ml (1.62 μM). Fluorescence was recorded at a wavelength of 460 nm when the samples were excited by light with a wavelength of 355 nm. In an experiment to study the effect of ascorbate, malate and succinate on cell adhesion and proliferation, 20,000 cells and a solution of one of these substances in a working concentration of 2 mM were introduced into the cells of the tablet. The number of cells in the wells was estimated based on the fluorescence of Hoechst-33342 after a day of incubation.

Result: In samples containing 2 mM succinic acid and ascorbic acid, a statistically significant decrease in the intensity of fluorescence was observed compared with a sample that did not contain the drug. This suggests that these compounds negatively affect the growth properties of the A549 culture: they inhibit cell adhesion or slow down their proliferation.

Scope of the results and conclusions:The results obtained are necessary for the methodologically correct planning of the most detailed studies on the A549 cell line model using fluorescent methods, including studies on the radioprotective properties of ascorbate, malate and succinate under the influence of rare ionizing and neutron radiation.

Keywords: cell culture, A549, ascorbic acid, succinate, malic acid, Hoechst-33342 flatbed fluorimeter, influence estimation

For citation: Romodin LA, Yashkina EI, Moskovskij AA. Fluorimetric Evaluation of the Effect of Malic, Succinic and Ascorbic Acids on the Growth Properties of A549 Cells in Culture. Medical Radiology and Radiation Safety. 2024;69(1):28–32. (In Russian). DOI:10.33266/1024-6177-2024-69-1-28-32

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 PDF (RUS) Full-text article (in Russian)

Conflict of interest. The authors declare no conflict of interest.

Financing. The work was carried out within the framework of the research project “Technology-3” (registration number of the research project in the EGISU R&D system: 1230113001053).

Contribution. Article was prepared with equal participation of the authors.

Article received: 20.10.2023. Accepted for publication: 27.11.2023.

Medical Radiology and Radiation Safety. 2024. Vol. 69. № 1

DOI:10.33266/1024-6177-2024-69-1-20-27

V.A. Anikina ¹, S.S. Sorokina ¹, A.E. Shemyakov ^1,2, E.A. Zamyatina ¹,
N.R. Popova ¹

Comparative Assessment of the Effect of Local Proton Radiation
with a Dose of 30 Gy in BALB/c and C57BL/6 Mice

¹ Institute of Theoretical and Experimental Biophysics, Moscow Region, Pushchino, Russia

² Branch “Physio-Technical Center” of the P.N. Lebedev Physical Institute, Moscow Region, Protvino, Russia

Contact person: N.R. Popova, e-mail: This email address is being protected from spambots. You need JavaScript enabled to view it.

ABSTRACT

Purpose: To evaluate the effect of local proton irradiation at a dose of 30 Gy on Balb/c and C57BL/6 mice in terms of the degree and dynamics of radiation-induced skin damage formation, changes in body weight and peripheral blood elements count.

Material and methods: Experiments were performed on non-depilated male mice aged 7‒8 weeks from two strains: Balb/c and C57BL/6 (n=15). Local irradiation of the skin was carried out on the dorsal side of the animals using a scanning proton beam at an extended Bragg peak in the proton therapy complex «Prometheus» of the LPI Physico-technical Centre (Protvino) at a dose of 30 Gy with a proton energy of 87.8 MeV. During the irradiation session, animals were subjected to intraperitoneal anesthesia using a combination of Zoletil 100 (Virbac, France) and Xyla (Interchemie, Netherlands) in a previously determined ratio 1:3 (20‒40 mg/kg). Photographic documentation of radiation-induced skin damage was performed weekly for 70 days. Animals were examined daily for clinical manifestations of radiation-induced skin damage formation according to the RTOG international scale for 21 days following irradiation. The body weight dynamics of mice were evaluated one day before irradiation and then weekly for 70 days. Blood samples were collected from the tail vein by cutting the tip of the tail and analyzed using a DH36 Vet hematology analyzer (Dymind, China) one day before irradiation, one day and three days after irradiation, and weekly thereafter for 70 days. Experimental data were presented as mean ± standard deviation (M ± SD).

Results: In this study, the impact of a single local exposure to proton radiation at a dose of 30 Gy on the degree and dynamics of radiation-induced skin damage formation was evaluated. It was demonstrated that Balb/c mice exhibited a higher frequency and degree of radiation-induced skin damage formation compared to the C57BL/6 mice. Analysis of body weight in mice after radiation exposure revealed no significant decrease in either mouse strain. A comparative analysis of the number of platelets, erythrocytes and hemoglobin concentration in both mouse strains did not reveal any changes, while a tendency towards a decrease in the number of leukocytes, lymphocytes, and granulocytes was observed in the irradiated Balb/c mice group compared to the control group. Conversely, in irradiated C57BL/6 mice, the number of lymphocytes was higher compared to control animals.

Conclusion: In this study, Balb/c mice exhibited higher radiosensitivity compared to C57BL mice in response to a single local proton irradiation at a dose of 30 Gy.

Keywords: proton radiation, radiation dermatitis, radiation burn, hematological analysis, BALB/c and C57BL/6 mice

For citation: Anikina VA, Sorokina SS, Shemyakov AE, Zamyatina EA, Popova NR. Comparative Assessment of the Effect of Local Proton Radiation with a Dose of 30 Gy in BALB/c and C57BL/6 Mice. Medical Radiology and Radiation Safety. 2024;69(1):20–27. (In Russian). DOI:10.33266/1024-6177-2024-69-1-20-27

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 PDF (RUS) Full-text article (in Russian)

Conflict of interest. The authors declare no conflict of interest.

Financing. The study was done with the support of the RNF grant No. 22-63-00082.

Contribution. Development of the research concept: Popova N.R., Sorokina S.S.; development of the research design: Popova N.R., Sorokina S.S., Anikina V.A.; conducting experiments: Anikina V.A., Zamyatina E.A., Shemyakov A.E., development and modification of research methods: Anikina V.A., Shemyakov A.E.; collection and analysis of literary material: Anikina V.A., Sorokina S.S., Popova N.R.; statistical data processing: Anikina V.A.; writing and scientific editing of the text: Sorokina S.S., Popova N.R.

Article received: 20.10.2023. Accepted for publication: 27.11.2023.

Medical Radiology and Radiation Safety. 2024. Vol. 69. № 1

DOI:10.33266/1024-6177-2024-69-1-33-40

L.I. Baranov, A.N.Tsarev, F.S. Torubarov, A.S. Kretov, V.V. Petrova, E.Vasilyev,
S.M. Dumansky, O.A. Tikhonova, T.M. Bulanova, M.V. Kalinina, P.A. Shulepov,
I. Dibirgadzhiyev, A.S. Samoilov

Digital Twin of Worker of Nuclear Facility at the Stage of Pre-Shift Control

A.I. Burnazyan Federal Medical Biophysical Center, Moscow, Russia

Contact person: L.I. Baranov, e-mail: This email address is being protected from spambots. You need JavaScript enabled to view it.

ABSTRACT

Introduction. Digital twin. Digital twin in medicine using the example of Philips. Digital twin as an object of medical information space. Digital twin as an abstraction. Digital twin of worker of nuclear facility at the stage of pre-shift control. Conclusion.

Keywords: worker of nuclear facility, digital twin, pre-shift control, medical information space, abstraction

For citation: Baranov LI, Tsarev AN, Torubarov FS, Kretov AS, Petrova VV, Vasilyev EV, Dumansky SM, Tikhonova OA, Bulano-
va TM, Kalinina MV, Shulepov PA, Dibirgadzhiyev I, Samoilov AS. Digital Twin of Worker of  Nuclear Facility at the Stage of Pre-Shift Control. Medical Radiology and Radiation Safety. 2024;69(1):33–40. (In Russian). DOI:10.33266/1024-6177-2024-69-1-33-40

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 PDF (RUS) Full-text article (in Russian)

Conflict of interest. The authors declare no conflict of interest.

Financing. The study had no sponsorship.

Contribution. Article was prepared with equal participation of the authors.

Article received: 20.10.2023. Accepted for publication: 27.11.2023.