Evgeniya B. Logashenko

Background. The influence of nanoparticles of industrial aerosols on phenotypes of occupational chronic obstructive pulmonary disease (COPD) is not studied enough. In this connection the system signs and symptoms of the disease including sarcopenia is of a great interest for investigation, because there associations with quality of life and lifespan. Aim. To establish the features of sarcopenia in patients with COPD due to aerosols containing nanoparticles. Materials and methods. A prospective observational study was performed. Previously, chemical and hygienic investigation of nanoparticles in the workplaces air on the machine building enterprise was done. The groups under investigation were occupational COPD patients, who were employed at the workplaces that has been investigated and contacting with aerosols containing metal (n = 48) or silica (n = 55) nanoparticles. Groups were matched by sex, age, COPD duration. Study procedures were Sarcopenia Fast questionnaire, ultrasound measurement of quadriceps femoris, bioelectrical impedance analysis, hand grip strength by dynamometry, sit-to-stand test, short physical performance battery, pulmonary function tests, serum molecular markers measured by enzyme-linked immunosorbent assay or by kinetic method, serum fibrinogen by Klauss method. COPD was diagnosed when postbronchodilator forced expiratory volume in one second divided by forced vital capacity was less than 0.7. Sarcopenia was diagnosed by EWGSOP2 criteria (European Working Group on Sarcopenia in Older People). Statistical analysis included descriptive methods and liner regression. Results. Sarcopenia rate was 26 (47.2%) in COPD due to aerosols containing silica nanoparticles, 17 (35.4%) in COPD due to aerosols containing metal nanoparticles and 15 (30.0%) in control group; p = 0.015. The differences of sarcopenia severity between COPD due to different environmental conditions were seen – Ме (Q2–Q3). COPD due to aerosols containing silica nanoparticles was characterized by minimal values of quadriceps cross-sectional area: 4.9 (4.0–5.5) sm2 compared with 9.4 (8.4–9.9) sm2 in COPD due to aerosols containing metal nanoparticles patients and with 9,0 (7.8; 9.2) sm2 in control group respectivel; р = 0.010, quadriceps thickness, appendicular fat-free mass index: 7.1 (5.5–7.4), 7.5 (6.2–8.3) and 8.2 (6.8–9.1) kg/m2; р = 0.009, sit-to-stand test 15.8 (13.2–16.7), 12.1 (10.5–13.0) and 11.5 (9.4–13.8) s; р = 0.009, by elevated muscle echogenicity. COPD due to aerosols containing metal nanoparticles patients had the mid values. After 12 months these relationships were preserved. The associations of silica nanoparticles with quadriceps cross-sectional area (В = -0.95; р = 0.009), echogenicity (В = 1.02; р = 0.001), appendicular fat-free mass index (В = -1.05; р = 0.001) and sit-to-stand test (В = 0.91; р = 0.011) were explored. Conclusion. Occuational COPD is characterized by sarcopenia rate, less values of muscle quantity and quality, especially pronounced in COPD due to aerosols containing silica nanoparticles.

Cervical cancer (CC) is the fourth most common cancer among women worldwide and a leading cause of cancer-related death. While immortalized cell lines like HeLa are widely used, they often lose characteristic features of the original tumor due to long-term cultivation. Primary cell cultures offer a more representative model, as they retain key features of the native tumor microenvironment. Here, we establish and characterize two novel human CC cell lines, AdMer35 and AdMer43, uniquely derived from paired tumor biopsies of the same patient before (AdMer35) and after (AdMer43) chemoradiotherapy for stage IIIB squamous cell carcinoma. Both lines exhibited stable epithelial morphology (cobblestone pattern), adhesion, were positive by cytokeratin staining, and have HPV-59 DNA genomes. They displayed distinct karyotypes: 80% of AdMer35 cells were near-tetraploid (modal number 95), while 98% of AdMer43 cells were near-triploid (modal number 77). Functional assays revealed significant differences: AdMer43 proliferated faster (Cell Index ~ 3.8 vs. ~2.1 at 100 h), but AdMer35 migrated more rapidly. Crucially, AdMer43 demonstrated increased resistance in comparison with AdMer35 to both radiotherapy (minimal apoptosis and G2/M arrest post-5 Gy irradiation) and chemotherapy (IC₅₀ for carboplatin: 87.8 µM vs. 54.3 µM; for paclitaxel: 7.3 nM vs. 3.0 nM). Both lines were tumorigenic in nude mice, forming histologically distinct tumors. Thus, we present a novel paired in vitro model of acquired chemoradioresistance in cervical cancer. The AdMer35/AdMer43 cell line pair provides a unique tool for mechanistic studies of therapy resistance and preclinical drug screening.

Cervical cancer remains a significant global health burden, with chemoradioresistance representing a major obstacle to successful treatment. To elucidate the mechanisms underlying this resistance, we established a unique pair of isogenic primary cervical cancer cell lines, AdMer35 and AdMer43, obtained from a patient with squamous cell carcinoma of the cervix before and after radiation therapy. The aim of our study was to characterize the transcriptomic and cellular heterogeneity of these cells. We conducted an in-depth comparative analysis using single-cell RNA sequencing. Analysis of this paired, patient-derived isogenic model suggests that chemoradioresistance can arise through coordinated multilevel cellular adaptations. Resistant AdMer43 cells demonstrated transcriptional reprogramming, with the upregulation of embryonic stemness factors (HOX, POU5F1, SOX2), a shift in extracellular matrix from fibrillar to non-fibrillar collagens, and activation of inflammatory pathways. We identified and characterized critical cell-state dynamics: resistant cells exhibited a remodeled ecosystem with a metabolically reprogrammed senescent-like cell population showing an enhanced pro-tumorigenic communication via EREG, SEMA3C, BMP, and WNT pathways. Furthermore, we identified a progenitor-like cell population with a minimal CNV burden, potentially serving as a reservoir for tumor persistence. These findings offer novel insights for developing targeted strategies to eliminate resistant cell pools and improve cervical cancer outcomes.

BACKGROUND: The characteristics of vascular pathology in different phenotypes of occupational chronic obstructive pulmonary disease (COPD), as well as the causal role of various industrial aerosol components, especially nanoparticles, are poorly understood. AIM: To determine the characteristics of arterial remodeling and endothelial function in patients with occupational COPD caused by exposure to aerosol nanoparticles. METHODS: An observational, cohort cross-sectional study was performed in patients with occupational COPD caused by exposure to aerosols containing metal (n = 48) or silicon (n = 55) nanoparticles, compared with COPD caused by tobacco smoking (n = 50). Scanning electron microscopy and inductively coupled plasma atomic emission spectroscopy were used to measure the size and chemical composition of particles, respectively. The procedures used in the study included color-flow duplex scanning of the brachiocephalic arteries, brachial arteries, and aorta; flow-mediated dilation test of the brachial arteries; and enzyme-linked immunosorbent assay of molecular markers. Linear regression was used to determine relationships. RESULTS: The group of occupational COPD caused by aerosols with silicon nanoparticles had the highest values of carotid intima-media thickness: 1.2 [0.9; 1.5] mm; in the group of occupational COPD caused by aerosols with metal nanoparticles and the control group, these values were 0.9 [0.7; 1.0] mm and 0.8 [0.7; 0.9] mm, respectively (p = 0.009). Moreover, the group of occupational COPD caused by aerosols with silicon nanoparticles had the highest incidence of atherosclerosis compared to the group of occupational COPD caused by aerosols with metal nanoparticles and the control group (41.8% vs. 22.9% and 18.0%, respectively; р = 0.003). The aortic pulse wave velocity in the three groups was 12.6 [11.2; 14.1], 9.3 [8.9; 10.7], and 7.2 [6.9; 8.4] m/s, respectively (р = 0.001). The minimum flow-mediated dilation of the brachial arteries was 2.5 [2.1; 3.4], 3.8 [3.3; 4.6], and 4.7 [4.5; 5.3]%, respectively (р = 0.001). Occupational COPD caused by aerosols with silicon nanoparticles was associated with the highest serum levels of vascular cell adhesion molecule 1, von Willebrand factor, transforming growth factor β1, procollagen III N-terminal propeptide, and fibroblast growth factor 2. Regression relationships were found between the intima-media thickness and the concentration of metal (adjusted R-squared [R2]: 0.36) and silicon (adjusted R2: 0.47) nanoparticles, as well as the length of employment (adjusted R2: 0.27). Moreover, regression relationships were found between the flow-mediated dilation of the brachial arteries and the concentration of metal (adjusted R2: 0.51) and silicon (adjusted R2: 0.71) nanoparticles, the length of employment (adjusted R2: 0.68), and the total concentration of silicon-containing dust (adjusted R2: 0.55). CONCLUSION: Occupational COPD caused by exposure to aerosol nanoparticles (especially silicon-containing ones) is associated with significant vascular remodeling and endothelial dysfunction, which must be considered during follow-up care.

The significant prevalence of industrial aerosols with unintentional nanoparticles and the increasing use of engineering nanoparticles in modern production determine the relevance of research on the patterns of interaction with the human bronchopulmonary system. Unintentional nanoparticles are formed during various production processes: melting and welding of metals, combustion, crushing, grinding of minerals, plasma processing of materials. Engineering nanoparticles are released into the air of the work area during their production or use in any technological processes. The respiratory system, as a barrier organ, is most vulnerable to the effects of adverse environmental factors. At the same time, nanoparticles are the least studied component of industrial aerosols. In order to systematize data on the role of nanoparticles in the development of occupational bronchopulmonary pathology, the authors conducted a literature review. The size determines the features of the physical, chemical and biological properties of nanoparticles. They have high values of the ratio of surface area to volume, total surface area, which leads to an increase in reactogenicity and penetrating power. The pathogenic properties of nanoparticles depend on the chemical composition, shape, curvature of the surface, structure, charge, mass concentration, total surface area of the particles, and exposure time. The multidimensionality of hygienic characteristics determines the complexity of hygienic rationing and monitoring of nanoparticles. To date, safe exposure levels have not been established. When interacting with cells of the respiratory tract, nanoparticles induce the following cellular and molecular mechanisms: oxidative stress, inflammation, violation of the epithelial barrier, autophagy and dysfunction of lysosomes, stress of the endoplasmic reticulum, apoptosis, senescence, fibrosis, endothelial dysfunction, DNA damage. This leads to the development of interstitial pneumonitis, pulmonary fibrosis and obstructive disorders, and an increase in the activity of allergic inflammation. All these mechanisms are present in the pathogenesis of occupational lung diseases. Under the influence of nanoparticles of various chemical compositions, separate phenotypes of occupational chronic obstructive pulmonary disease are formed. There is a lack of epidemiological studies of the possible etiological role of nanoparticles. Nanoparticles of industrial aerosols are a significant factor in the development of occupational diseases of the bronchopulmonary system and have a significant impact on the formation of phenotypes.

Acute lung injury is a complex cascade process that develops in response to various damaging factors, which can lead to acute respiratory distress syndrome. Within this study, based on bioinformatics reanalysis of available full-transcriptome data of acute lung injury induced in mice and humans by various factors, we selected a set of genes that could serve as good targets for suppressing inflammation in the lung tissue, evaluated their expression in the cells of different origins during LPS-induced inflammation, and chose the tissue inhibitor of metalloproteinase Timp1 as a promising target for suppressing inflammation. We designed an effective chemically modified anti-TIMP1 siRNA and showed that Timp1 silencing correlates with a decrease in the pro-inflammatory cytokine IL6 secretion in cultured macrophage cells and reduces the severity of LPS-induced acute lung injury in a mouse model.

Introduction. Influence of incidental nanoparticles of industrial aerosols on occupational lung diseases development is not studied enough. As nanoparticles has properties to induce inflammation and fibrosis, it is hypothesized that they affect occupational chronic obstructive pulmonary disease (COPD) phenotype. The aim was to establish monocyte subsets, airway inflammation, clinical and functional features in occupational COPD due to aerosols containing nanoparticles exposure. Materials and methods. Study design was a single center prospective cohort observational. Subjects with occupational COPD (GOLD 2011-2021 criteria) exposed to aerosols containing nanoparticles (n=50) enrolled. Comparison group - COPD in tobacco smokers (n=50), control group - healthy people (n=50). Groups were matched by demographics and COPD duration. Nanoparticles at workplaces air were measured by inductively coupled plasma atomic emission spectrometry and by scanning electron microscopy. Of participants 26 were exposed to maximal concentrations of metal nanoparticles and 24 - of silica nanoparticles. Spirography, body pletysmography, lung diffusing capacity (DLco/Va), Doppler-ehocardiography, induced sputum cytology, COPD exacerbations assessment were done. Monocyte subsets were determined by flow cytometry. Linear regression model was used to explore relationships. Results. COPD due to aerosols containing metal nanoparticles was characterized by most severe airflow limitation, lung hyperinflation, pulmonary hypertension, most prominent decrease in DLco/Va, frequent and severe COPD exacerbations, eosinophilic inflammation. The largest proportion of «classical» CD14+CD16- monocytes subset, 96,4% (90,3%; 97,2%), high level of CCR5 expression were seen in this group. The features of COPD due to aerosols containing silica nanoparticles were substantial decrease in DLco/Va, the least airflow limitation, mild lung hyperinflation, rare COPD exacerbations, paucigranulocytic inflammation. The largest proportion of «non-classical» CD14DimCD16+ monocytes with high level of CCR2 expression revealed. Mass concentration of metal nanoparticles was associated with «classical» monocytes, (B=1,5), silica nanoparticles - with «non-classical» monocytes (B=1,4). On their turn, «classical» monocytes were associated with DLco (B=-1,6), functional residual volume (B=1,2), mean pulmonary artery pressure (B=-1,4), eosinophilic inflammation. «Non-classical» monocytes were associated with DLco (B=-1,5) and paucigranulocytic inflammation (B=1,2), p<0,015. Conclusions. Exposure of incidental nanoparticles was associated with circulated monocyte subsets, airway inflammation and occupaitonal CODP phenotype.

Plant-extracted triterpenoids belong to a class of bioactive compounds with pleotropic functions, including antioxidant, anti-cancer, and anti-inflammatory effects. In this work, we investigated the anti-inflammatory and anti-oxidative activities of a semisynthetic derivative of 18βH-glycyrrhetinic acid (18βH-GA), soloxolone methyl (methyl 2-cyano-3,12-dioxo-18βH-olean-9(11),1(2)-dien-30-oate, or SM) in vitro on lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages and in vivo in models of acute inflammation: LPS-induced endotoxemia and carrageenan-induced peritonitis. SM used at non-cytotoxic concentrations was found to attenuate the production of reactive oxygen species and nitric oxide (II) and increase the level of reduced glutathione production by LPS-stimulated RAW264.7 cells. Moreover, SM strongly suppressed the phagocytic and migration activity of activated macrophages. These effects were found to be associated with the stimulation of heme oxigenase-1 (HO-1) expression, as well as with the inhibition of nuclear factor-κB (NF-κB) and Akt phosphorylation. Surprisingly, it was found that SM significantly enhanced LPS-induced expression of the pro-inflammatory cytokines interleukin-6 (IL-6), tumour necrosis factor-α (TNF-α), and interleukin-1β (IL-1β) in RAW264.7 cells via activation of the c-Jun/Toll-like receptor 4 (TLR4) signaling axis. In vivo pre-exposure treatment with SM effectively inhibited the development of carrageenan-induced acute inflammation in the peritoneal cavity, but it did not improve LPS-induced inflammation in the endotoxemia model.

Semi-synthetic triterpenoids, bearing cyano enone functionality in ring A, are considered to be novel promising therapeutic agents with complex inhibitory effects on tissue damage, inflammation and tumor growth. Previously, we showed that the cyano enone-containing 18βH-glycyrrhetinic acid derivative soloxolone methyl (SM) effectively suppressed the inflammatory response of macrophages in vitro and the development of influenza A-induced pneumonia and phlogogen-stimulated paw edema in vivo. In this work, we reported the synthesis of a novel 18βH-glycyrrhetinic acid derivative trioxolone methyl (TM), bearing a 2-cyano-3-oxo-1(2)-en moiety in ring A and a 12,19-dioxo-9(11),13(18)-dien moiety in rings C, D, and E. TM exhibited a high inhibitory effect on nitric oxide (II) production by lipopolysaccharide-stimulated J774 macrophages in vitro and dextran sulfate sodium (DSS)-induced colitis in mice, displaying higher anti-inflammatory activity in comparison with SM. TM effectively suppressed the DSS-induced epithelial damage and inflammatory infiltration of colon tissue, the hyperproduction of colonic neutral mucin and TNFα and increased glutathione synthesis. Our in silico analysis showed that Akt1, STAT3 and dopamine receptor D2 can be considered as mediators of the anti-colitic activity of TM. Our findings provided valuable information for a better understanding of the anti-inflammatory activity of cyano enone-bearing triterpenoids and revealed TM as a promising anti-inflammatory candidate.