Bulletin of RSMU

ISSN Print 2500–1094 ISSN Online 2542–1204

Bulletin of RSMU

BIOMEDICAL JOURNAL OF PIROGOV UNIVERSITY (MOSCOW, RUSSIA)

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Bilan DS, Shokhina AG, Panova AS, Belousov VV

The zebrafish Danio rerio is a popular model species for biomedical research focusing on modeling human diseases and screening of therapeutic drugs. Genetically encoded biosensors based on fluorescent proteins are widely used in many laboratories worldwide to study the biology of living systems of various complexity in vivo. The main advantage of these molecular tools is that they allow observing biological processes in intact systems in real time. In the present work we study the dynamics of the intracellular ratio of NAD+ to NADP in the yolk sac and tissues of D. rerio larvae using the SoNar biosensor following the injection of glycolisis end products, lactate and pyruvate.

Received: 2017-12-05

Published online: 2018-03-14

DOI: 10.24075/brsmu.2018.005

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VIEWS 5880

Osipova ZM, Shcheglov AS, Yampolsky IV

Bioluminescence is chemical oxidation of a small luciferin molecule by air catalyzed by luciferase and accompanied by the emission of photons in the visible spectrum. This reaction is used in bioluminescent bioimaging, the method for the visualization of organism’s interior. Bioimaging is a popular tool used in medical research. However, it has an unfortunate drawback: it requires introduction of external luciferin to the system before every experiment. In this work we discuss a possibility of developing an autonomous luminescent system in eukaryotes based on the bioluminescent system of higher fungi.

Received: 2017-12-15

Published online: 2018-03-04

DOI: 10.24075/brsmu.2018.004

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VIEWS 5428

Pinto TCA, Banerjee A, Nazarov PA

There is an urgent need for new antimicrobial and therapeutic strategies to deal with the ever evolving antimicrobial resistance among the most prevalent bacterial pathogens. Infections due to virulent bacteria remain significant causes of morbidity and mortality despite progress in antimicrobial therapy, primarily because of the increasing of antimicrobial resistance levels among such group of bacteria. Despite significant advances in the understanding of the pathogenesis of infection due to these organisms, there are only limited strategies to prevent infection. Recently it was reported that SkQ1, triphenyl phosphonium-based mitochondria-targeted antioxidant and antibiotic, effectively kills all tested Gram-positive laboratory strains including of Bacillus subtilis, Staphylococcus aureus and Mycobacterium sp. Moreover, SkQ1 demonstrated effectiveness towards Gram-negative strains too, except Escherichia coli. The mechanism of the bactericidal action of TPP-based antibiotics could be also described by its ability to suppress bacterial bioenergetics by collapsing membrane potential through activation of protonophorous uncoupling. To this date, there are no reports of resistance to SkQ1 among Gram-positive strains; therefore, triphenyl phosphonium-based antibacterial agents would be effective towards planktonic and sessile cells of clinical resistant strains.

Received: 2018-01-23

Published online: 2018-03-04

DOI: 10.24075/brsmu.2018.003

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VIEWS 6296

Nazarov PA

Multiple drug resistance of nosocomial bacterial strains provoked by the unwise and uncontrolled use of antibiotics in medicine and agriculture seriously threatens modern healthcare: minor and trivial infections are about to kill again. A solution may lie in the development of new alternatives to antibiotics. This review highlights the most interesting approaches to the development of antibacterial drugs focusing on the most promising ones, such as phage therapy and phage lytic enzymes (lysins).

Received: 2018-01-23

Published online: 2018-03-04

DOI: 10.24075/brsmu.2018.002

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VIEWS 5829

Komech EA, Lebedev YuB, Koshenkova AV, Syrko DS, Musatkina EA, Lukyanov SA, Chudakov DM, Zvyagin IV

Recent studies of T-cell clonal repertoires of patients with ankylosing spondylitis (AS) have led to the discovery of AS-associated T-cell clones with a highly homologous T-cell receptor structure. The role of T-lymphocytes in the disease progression cannot be elucidated without analyzing the diversity and abundance of functionally different T-cell clones found in patients with AS. Using a state-of-the-art technique for T-cell repertoire profiling based on massively parallel sequencing, we, for the first time, studied the T-cell receptor repertoire of activated T-cells from the peripheral blood of a patient with AS. We have demonstrated that a subpopulation of CD38+HLA-DR+ T-lymphocytes is highly diverse both in terms of clonal diversity and abundance of the identified clonotypes, suggesting diverse antigen specificity of the activated peripheral blood T-cells. Most of the activated T-cell clonotypes had low abundance in total population of peripheral blood T-cells. In the repertoire of activated T-cells we have found the clonotype TRBV9_CASSVGVYSTDTQYF_TRBJ2-3, previously discovered in AS and reactive arthritis, and a few other clonotypes of cytotoxic and helper T-cells that may have a role in promoting inflammation in AS patients. Presence of the AS-associated clonotype in activated T-cell subset suggests that the T-cells might play an active role in ongoing inflammation during the disease progression. This provides rationale for further research of their antigen specificity and role in triggering or maintaining AS.

Received: 2017-12-15

Published online: 2018-02-16

DOI: 10.24075/brsmu.2018.001

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VIEWS 5449

Pozdniakova NV, Turobov VI, Garanina EE, Ryabaya OA, Biryukova YuK, Minkevich NI, Trubnikova EV, Shevelev AB, Kuznetsova TV, Belyakova AV, Udovichenko IP

In this work we explore the temporal dynamics of cytokines in Dark Agouti rats with experimentally induced autoimmune encephalomyelitis (EAE). The main group consisted of 11 animals who were injected with 100 μl (per leg) of spinal cord homogenate obtained from random-bred rats and combined with incomplete Freund’s adjuvant to the hind footpads. The control group included 7 animals who received 100 μl of normal saline mixed with incomplete Freund’s adjuvant. Blood samples (500 μl) were collected daily, starting from day 1 through day 7. We ran a Bio-Plex-based multiplex cytokine assay on the samples using the Bio-Plex Pro Rat Cytokine 24-plex Assay kit. EAE in rats was shown to simulate progression of multiple sclerosis in humans in terms of temporal dynamics of lymphoproliferative and hematopoietic factors IL-1b, IL-2, IL-4, IL-5, IL-6, and IL-7. The studied model satisfactory imitates the dynamics of factors stimulating migration of lymphocytes, monocytes and other immune cells, including IL-17, RANTES (CCL-5) and MCP-1 (CCL-2) but excluding GRO/KC (CXCL1), which shows a different dynamics. The model also resembles patterns of human multiple sclerosis in terms of factors affecting cytotoxic and apoptotic reactions, including IFNγ, IL-6 and IL-17, but excluding TNFα.

Received: 2017-12-15

Published online: 2018-01-25

DOI: 10.24075/brsmu.2017-06-12

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VIEWS 5364

Bilan DS, Kelmanson IV, Belousov VV

Studies of ischemic brain injury are an important area of modern biomedical research. So far, a lot of ischemic stroke models have been proposed, along with different imaging and staining modalities aimed to visualize the damaged tissue. In this work we use a rat model to investigate how the experimental setup affects the interpretation of experimental data obtained in the acute phase of ischemic stroke (5 hours after the occlusion of the middle cerebral artery). We show the association between the choice of the type of anesthesia and the severity of ischemic injury: in our experiments brain damage was the most pronounced in the animals anesthetized with a combination of chloral hydrate and Rometar; the least damage was observed for isoflurane. Staining was performed using the popular dye 2,3,5-triphenyltetrazolium chloride (TTC). We demonstrate that parameters of brain slices incubation in TTC also need to be accounted for when interpreting the results obtained during the acute phase of stroke, the optimum incubation time being 30 min and temperature 37 °С.

Received: 2017-11-29

Published online: 2018-01-25

DOI: 10.24075/brsmu.2017-06-11

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VIEWS 4970

Grabovsky EV, Oleynik GM, Krastelev EG, Smirnov VP, Khmelevsky EV, Bozhenko VK, Shishkin AM, Ivanov AV, Kulinich TM

Relative biological effectiveness of ionizing radiation is determined by a number of factors, including a dose rate. Radiotherapy equipment employs low dose rates of up to a few Gy per minute. But very little is known about the biological effect of high and ultrahigh (≥ 10<sup>8</sup> Gy/min) dose rate radiation. Our study aimed to investigate the apoptotic effect of ultrahigh gamma dose rates on human peripheral blood lymphocytes. Blood samples were collected from seemingly healthy donors. Lymphocytes were isolated by density gradient separation. Lymphocyte suspensions were irradiated with low-rate doses on the Rokus- AM gamma-ray machine for clinical use (Russia) and with 10<sup>8</sup> Gy/s doses on the experimental pulse generators Angara-5-1 and Mir-M (Russia). Apoptosis was measured by flow cytometry using annexin V and propidium iodide double staining. We established that in comparison with low dose rates, ultrahigh gamma dose rates (with doses ranging from 1 to 6 Gy) induced significantly more pronounced apoptosis in peripheral blood lymphocytes (p < 0.05) with fewer necrotic cells. Total radiation-induced cell death did not differ significantly between the therapeutic gamma machine and the experimental pulse generators. Further research is needed to assess biological and medical significance of our findings.

Received: 2017-11-22

Published online: 2018-01-24

DOI: 10.24075/brsmu.2017-06-10

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VIEWS 5329