Bulletin of RSMU

ISSN Print 2500–1094 ISSN Online 2542–1204

Bulletin of RSMU

BIOMEDICAL JOURNAL OF PIROGOV UNIVERSITY (MOSCOW, RUSSIA)

Scopus

SCImago Journal & Country Rank

Log In Registration RU

Adrianov MA, Gautier MS, Degtyareva AV, Marey MV, Manukhova LA, Rastorguev SM, Simonov VS, Ushakova LV, Vyssokikh MYu

The most common biochemical defect in Leigh syndrome is aberrations in proteins involved in the assembly of the electron transport chain (ETC) complex IV subunits — cytochrome C oxidase (COX). Among these, mutations in the SURF1 gene are the most common. The SURF1 protein is embedded in the inner mitochondrial membrane and plays a crucial role in the COX complex assembly. All mutations in the SURF1 gene result in the truncated protein biosynthesis and damage to the COX complex. Adeno-associated viral vectors (AAV9), which carry the not mutated SURF1 gene (AAV9-SURF1), are being investigated for the treatment of this disease. The aim of this study was to evaluate the expression levels of SURF1 and MTCO1 proteins in whole blood from patients with Leigh syndrome compared to reference values obtained for a pool of patients without mutations, as well as to evaluate the expression of the MTCO1 cytochrome c oxidase subunit in skin fibroblast cultures from patients with Leigh syndrome treated with AAV9. To investigate the gene therapy efficacy, AAV9-SURF1 was added to fibroblasts derived from the skin of a patient with a mutation in the Surf1 gene and to control skin fibroblasts at an optimal dose that did not impair cell viability in the MTT assay. We used Western blot analysis and quantitative PCR to evaluate changes in the relative amounts of the studied proteins after the addition of AAV9-SURF1 to control cells and cells obtained from the patient and identified significant compensatory changes in skin fibroblasts from a patient with a SURF1 mutation.

Received: 2025-12-12

Published online: 2025-12-29

DOI: 10.24075/brsmu.2025.083

Comments 0

VIEWS 170

Zhygmitova EB, Kosykh AV, Gurskaya NG

Diseases associated with limbal stem cell deficiency, such as chronic epithelial erosion and corneal scarring, require new therapeutic approaches rooted in regenerative medicine. This study aimed to develop a protocol for obtaining progenitor cells of the epithelium of the limb and cornea from induced pluripotent stem cells (iPSCs). We differentiated iPSCs toward eye organoids to obtain three-dimensional heterogeneous structures within three weeks. The resulting organoids contain corneal epithelial progenitor cells expressing keratin 3 and collagen 7, which confirms the possibility of generating functional epithelium in vitro. The protocol enables the generation of isogenic, patient-specific cell lines for treating limbal insufficiency and dystrophic epidermolysis bullosa, including applications following preliminary genome editing of iPSCs.

Received: 2025-11-25

Published online: 2025-12-27

DOI: 10.24075/brsmu.2025.070

Comments 0

VIEWS 156

Plotnikova MA, Oleynik VA, Klotchenko SA

Monoclonal antibody therapy is one of the most promising approaches for effective influenza control. In this study, we evaluated the antiviral activity of exogenous mRNA-encoded single-chain variable fragment (scFv) antibodies, which are capable of binding viral antigens inside the cell with high affinity. Two influenza virus proteins, hemagglutinin (antibody FI6) and nucleoprotein (antibody 2/3), were chosen as targets. Each scFv encoded by mRNA was produced in two variants: one containing a signal peptide (SP) to direct secretion into the extracellular space (scFv-SP) and one lacking the signal peptide (scFv-WO) for cytosolic localization and function. These variants showed distinct intracellular localization patterns: scFv-SP localized to regions characteristic of the endoplasmic reticulum and the Golgi complex, whereas scFv-WO was distributed diffusely throughout the cytoplasm. mRNAs encoding scFv-FI6-SP, scFv-2/3-SP, and scFv-2/3-WO exhibited antiviral activity against influenza A virus in vitro. The scFv-FI6-SP mRNA showed the strongest antiviral effect, reducing viral load by approximately tenfold compared to the control. For influenza B virus, both scFv-2/3 mRNA variants, with and without the signal peptide, reduced viral load by an average of 50%. These findings highlight the antiviral potential of intracellular antibodies and point to new opportunities for targeting viral components that are not accessible to conventional antiviral therapies.

Received: 2025-11-27

Published online: 2025-12-26

DOI: 10.24075/brsmu.2025.077

Comments 0

VIEWS 286

Polyakova VA, Davydova EE, Luparev AR, Matsvay AD, Gnusareva NI, Gordukova MA, Galeeva EV, Tolokonceva AA, Shipulin GA

The diversity and succession of epidemiologically significant enteroviruses (EV) lead to constant changes in the clinical presentation and morbidity levels. The aim of the study is to investigate cases of EV infection in hospitalized children during the resurgence of the epidemic process following the COVID-19 pandemic. We collected clinical samples from 156 patients with EV infection across a range of ages. Virus genotyping was performed using the Sanger sequencing of the 5’UTR-VP2 and VP1 genome fragments. Sixteen types of enteroviruses were identified, with one additional case identified only to the species level (EV-C). The dominant EV type was Coxsackie CV-A6, with a share of 80.6% (95% CI: 66.7–95.5) in 2021 and 36.1% (95% CI: 27.5–44.6) in 2022. Most commonly, CV-A6 caused skin lesions (exanthema or HFMD) and respiratory manifestations. In 2022, the proportion of CV-A10 cases increased considerably to 27.0% (95% CI: 19.2–34.9) compared with 6.4% (95% CI: 0–15.1) in 2021. The most common clinical manifestation of CV-A10 was herpangina. The most severe EV infection cases were associated with ECHO 6 — four out of 11 patients were diagnosed with meningitis, while the remaining patients exhibited neurological symptoms (meningism, intense headache, vomiting) accompanied by fever. We observed a large number of EV cases accompanied by the presence of other infectious agents in biological samples, which may result from immune suppression during EV infection development. The most common of these agents was human herpesvirus 6 (HHV-6). The nucleotide sequences of the characterized enteroviruses have been deposited in the NCBI database to enable subsequent epidemiological analysis of enterovirus circulation in the Russian Federation.

Received: 2025-11-24

Published online: 2025-12-25

DOI: 10.24075/brsmu.2025.082

Comments 0

VIEWS 316

Semikina EV, Azarova YuE, Panichev SA, Basareva OI, Dzhanchatova NV, Alferova EJ, Polonikov AV

Diabetic nephropathy (DNP) is a serious complication of type 2 diabetes mellitus (T2D), leading to early disability and mortality from end-stage renal failure. Experimental and clinical studies have shown the leading role of oxidative stress-induced damage to macromolecules, including DNA, in the development and progression of DNF against the background of hyperglycemia. On the contrary, repair of these DNA lesions serves as a signal to end ongoing oxidative stress. The key DNA repair enzyme is 8-oxoguanine DNA glycosylase, encoded by the OGG1 gene. The aim of this study was to analyze the associations of five polymorphic variants (rs2072668, rs1052133, rs293795, rs2304277, and rs6443265) of the OGG1 gene with the risk of developing DNF in patients with type 2 diabetes. The study included 1461 patients with type 2 diabetes, 577 of whom were diagnosed with DNF. DNA genotyping was performed by real-time polymerase chain reaction using allele-specific fluorescently labeled probes. Associations were established between the rs293795-G/G genotype (OR = 1.97, 95% CI = 1.23-3.16, p = 0.007) and the rs2072668C-rs1052133C-rs293795G-rs2304277G-rs6443265C haplotype (OR = 1.30, 95% CI = 1.06-1.60, p = 0.012) of the OGG1 gene with a predisposition to DNF in the background of T2D. Moreover, six OGG1 diplotypes associated with an increased risk of DNF and one diplotype associated with a reduced risk of DNF in patients with T2D were identified. Thus, in our study, we presented for the first time data on the association of the OGG1 gene polymorphism with DNF, which creates a scientific foundation for further research on the contribution of disturbances in the DNA oxidative damage repair system to the development of microvascular complications of T2D.

Received: 2025-12-01

Published online: 2025-12-24

DOI: 10.24075/brsmu.2025.084

Comments 0

VIEWS 304

Grigorova IL

Follicular helper (Tfh) and follicular regulatory (Tfr) T cells play critical roles in inducing and controlling B cell responses, including the generation of high-affinity humoral immunity, the antibody class-switching, and the prevention of autoreactivity. Successful Tfh responses are linked to robust vaccine-induced neutralizing antibody production and efficient clearance of various pathogens. Conversely, dysregulation of follicular T cells is often linked to autoimmune diseases and allergic reactions. Furthermore, these cells are implicated in the formation of ectopic lymphoid structures (ELS), contribute to certain vascular pathologies, and hold prognostic value in several cancers. Consequently, the analysis of follicular T cell subpopulations in human peripheral blood is increasingly utilized to investigate the mechanisms underlying various diseases. In this opinion article, the current understanding of follicular T cell subsets, their functions, and the evolving methods for analyzing their circulating counterparts in human blood are discussed. In the author's opinion, the central unresolved questions remaining in the field are the precise phenotypic definition of circulating Tfr cell subpopulations, the elucidation of their developmental trajectory from precursors cells to mature regulatory forms, and the identification of their anatomical differentiation niches. The collection and translation of these essential data into reliable cellular signatures for peripheral blood analysis are critical for advancing personalized patient prognosis and developing tailored therapies.

Received: 2025-12-10

Published online: 2025-12-23

DOI: 10.24075/brsmu.2025.076

Comments 0

VIEWS 338

Shamovskaya DV, Gordukova MA, Smertina MA, Galeeva EV, Oscorbin IP, Khrapov EA, Boyarskikh UA, Filipenko ML

Salmonellosis remains one of the leading causes of bacterial gastrointestinal infections in humans and animals. Molecular diagnostics has dramatically reshaped the diagnostic landscape for Salmonella infection; however, it remains time- and resource-intensive. Isothermal DNA amplification, for example loop isothermal amplification (LAMP), performed at a constant temperature, is the basis for the development of rapid diagnostic tests that can be adapted to the point-of-care (PoC) formats and implemented in resource-limited settings or remote from centralized laboratories. The aim of this study was to develop and validate a novel LAMP-based method for detecting Salmonella enterica in human stool samples, wherein amplification results are monitored using a loop primer labeled with a fluorophore and an internal quencher. The proposed method achieves a limit of detection (LoD95) of 250 copies per reaction, with a sensitivity of 86.84% (95% CI: 71.91–95.59%) and specificity of 96.49% (95% CI: 87.89–99.57%) relative to qPCR, and demonstrates increased robustness against DNA amplification inhibitors present in fecal samples. Incorporation of distinct fluorophores into loop primers for FLP-LAMP targeting different genes could potentially enable multiplexing and simultaneous detection of multiple pathogens, thereby expanding the diagnostic utility of isothermal amplification.

Received: 2025-11-23

Published online: 2025-12-22

DOI: 10.24075/brsmu.2025.066

Comments 0

VIEWS 314

Zhulai GА, Kurbatova IV, Dudaniva OP

Experimental studies have demonstrated the protective role of ectonucleotidases — particularly CD39 and CD73 — in limiting inflammation connected to a liver pathology. However, their expression in metabolic-associated fatty liver disease (MAFLD) has not been thoroughly investigated. This study aimed to evaluate the mRNA levels of the ENTPD1 and NT5E genes, which encode CD39 and CD73, respectively, in patients with different forms of MAFLD (liver steatosis (LS) and metabolic-associated steatohepatitis (MASH)), and to assess the expression of CD39- and CD73-positive cells following immune cell activation in vitro. The sample included 29 healthy donors and 56 MAFLD patients. We measured the mRNA levels of the ENTPD1 and NT5E genes, pro-inflammatory cytokines (IL-6, TNFα), fragmented cytokeratin-18, and the blood content of CD39+ cells. Another parameter measured in vitro was the effect of immune cell activation on the proportion of CD39+ and CD73+ cells in patients with MASH and healthy donors. The expression of the ENTPD1 gene (p = 0.007 vs. control group; p = 0.010 vs. LS group) and the proportion of CD39+ cells among monocytes (p = 0.004 vs. control group; p = 0.003 vs. LS group) and lymphocytes (p = 0.034 vs. control group) were lower in the MASH group compared with both the control and LS groups. Activation of cells from MASH patients increased the proportion of CD39+ lymphocytes, but not that of CD14+ monocytes. It also increased the proportion of CD73+ cells among both lymphocytes and CD14+ monocytes. Thus, further investigation into the roles of CD39 and CD73 in the context of MAFLD progression represents a promising avenue for future research.

Received: 2025-11-11

Published online: 2025-12-21

DOI: 10.24075/brsmu.2025.079

Comments 0

VIEWS 295