Matvey Anatol'EVich Vadyukhin

Background/Objectives: Serous ovarian neoplasms comprise biologically distinct entities with substantial differences in clinical behavior and molecular characteristics. Comparative data on the quantitative and spatial distribution of T-cell populations across the spectrum of serous ovarian neoplasms, particularly borderline ovarian tumors (BOT) and serous cystadenomas (CA), remain limited. The aim of this study was to compare the quantitative and spatial distribution of CD3+, CD4+, CD8+, and FOXP3+ cells across serous ovarian neoplasms and to assess their associations with selected molecular characteristics. Methods: This retrospective, single-center observational study included 227 patients with high-grade serous carcinoma (HGSC; n = 108), low-grade serous carcinoma (LGSC; n = 49), BOT (n = 16), and CA (n = 54). The primary endpoint was the density of CD3+, CD4+, CD8+ and FOXP3+ immunopositive cells per mm², evaluated by immunohistochemistry and digital morphometry. Spatial distribution, differences across early pT substages, the CD8+/FOXP3+ ratio, and associations with selected molecular characteristics were additionally assessed. Age-adjusted generalized linear models were used to evaluate whether differences among histological groups were independent of patient age. Results: Compared with CA, HGSC showed an 18.58-fold higher CD3+ cell density and a 9.07-fold higher FOXP3+ cell density (both p < 0.001). Histological type remained significantly associated with CD3+, CD4+, CD8+, and FOXP3+ cell densities after adjustment for patient age (all p < 0.001), whereas age itself was not independently associated with any of the four markers (all p > 0.05). CD3+ cells were predominantly distributed in peritumoral areas, whereas HGSC also demonstrated diffuse intratumoral and stromal infiltration. Comparatively high T-cell densities were observed in BRCA1/2-mutated HGSC and HRD-positive subgroups. No statistically significant differences in immunopositive cell densities were detected across the analyzed pT1 substages. HGSC showed the lowest CD8+/FOXP3+ ratio (Me 1.18 [1.06–1.34]). Conclusions: Serous ovarian neoplasms demonstrate distinct histology-associated patterns of T-cell infiltration and spatial distribution, with additional heterogeneity across selected molecular subgroups. These differences remained evident after adjustment for patient age. Their functional and prognostic significance requires further validation using expanded immunophenotypic panels, multiplex approaches, and longitudinal clinical outcomes.

Background and Objectives: Oral squamous cell carcinoma (OSCC) belongs to a broad group of malignant neoplasms of the head and neck. It is known that components of the tumor microenvironment (TME) influence tumor progression. Similar patterns are being actively investigated in OSCC. In particular, a high density of CD68⁺ macrophages is associated with longer survival, whereas an abundance of M2 macrophages (CD163⁺) correlates with metastasis and an unfavorable prognosis. The present study compares the density and spatial distribution of CD68⁺ and CD163⁺ macrophages between intratumoral and peritumoral compartments in OSCC of the tongue and lip. Materials and Methods: Data from 112 patients with a verified OSCC diagnosis were included in this study. For the comparative morphological analysis, two core groups were formed: (a) Group I (n = 84), patients with Squamous Cell Carcinoma (pT1) of the tongue; (b) Group II (n = 28), patients with Squamous Cell Carcinoma (pT1) of the lip. Immunohistochemistry was performed according to standard protocols. Results: Tongue OSCC was characterized by the predominant accumulation of both CD68⁺ and CD163⁺ macrophages in the peritumoral stroma compared with the intratumoral zones (34.2 ± 8.1 vs. 48.1 ± 6.4 cells/mm²). In contrast, in lip OSCC, a higher density of CD68⁺ macrophages was observed in the intratumoral compartment, whereas the number of CD163⁺ macrophages remained considerably lower in both the intratumoral (51.1 ± 4.3 vs. 9.4 ± 2.1 cells/mm²) and peritumoral stroma of the tumor. Conclusions: These findings demonstrate that tongue and lip OSCC harbor distinct patterns of macrophage distribution and polarization, underscoring the role of the immune microenvironment in the biological heterogeneity of OSCC and warranting further investigation into macrophage subtype-specific prognostic markers.

Aging profoundly modifies neuronal responses to ischemia. We aimed to define age-dependent features of neuronal metabolism and cell death after ischemic stroke by assessing NeuN, NSE, and Caspase-3 in human cortical neurons and by comparing transcriptional activity within PI3K/Akt/mTOR and PI3K/Akt/FOXO3a pathways across age groups. The aim of this study was to determine age-dependent features of neuronal metabolism and cellular degradation in ischemic stroke based on immunohistochemical assessment of NeuN, NSE, and Caspase-3 markers in human cerebral cortex neurons, as well as to conduct a comparative analysis of gene expression in the PI3K/Akt/mTOR and PI3K/Akt/FOXO3a signaling pathways involved in the regulation of neuronal survival and apoptosis. For the investigation, frontal cortex autopsies from patients with ischemic stroke (n = 154; “young”, “middle” and “elderly”; death ≤7 days post-onset) were examined. Histology (hematoxylin–eosin) and Nissl staining were used for morphology and neuron counts. Multiplex immunofluorescence (NeuN, NSE, Caspase-3) quantified metabolically active and apoptotic neurons, and the percentage of Caspase-3⁺ among NeuN⁺ cells was calculated. qRT-PCR measured PIK3CA, AKT2, MTOR, and FOXO3A expression in the infarct border zone. Based on our results, neuronal density and NeuN/NSE expression declined with aging, and the fraction of Caspase-3⁺ among NeuN⁺ neurons in the penumbra rose (young 42%, middle 82%, elderly 89%). Morphologically “intact” penumbral neurons frequently lacked NeuN/NSE, revealing covert dysfunction. Young brains showed balanced activation of PI3K/Akt/mTOR and PI3K/Akt/FOXO3a, whereas elderly brains exhibited reduced Akt/mTOR activity with FOXO3A predominance, consistent with pro-apoptotic, inflammatory, and dysregulated autophagic signaling. Thus, aging markedly reduces neuronal metabolic activity and increases apoptotic death in the infarct border zone after ischemic stroke. In older patients, there is an almost complete loss of NeuN and NSE expression in penumbral neurons with robust activation of the caspase cascade, whereas younger patients retain a pool of metabolically active neurons. Age-dependent dysregulation of PI3K/Akt signaling—characterized by FOXO3a hyperactivation and mTOR suppression—further promotes apoptosis and dysregulated autophagy. These changes likely underlie the limited efficacy of standard neuroprotection in ischemic stroke and support the need for age-tailored neurotropic therapy aimed at enhancing pro-survival pathways within the infarct border zone.

Purpose of the study . To evaluate the roles of the growth factors IGF‑1, TGF-β, and VEGF-A in regulating delayed regenerative mechanisms induced by N-acetylcysteine (NAC) administered prior to local pancreatic irradiation with electrons at a total dose of 30 Gy. Materials and methods . An experimental study was conducted on 60 male Wistar rats, which were divided into four groups: Group 1 (control, n = 10): rats received an intraperitoneal injection of NaCl solution without local electron irradiation; Group 2 (n = 20): intraperitoneal administration of 0.9 % NaCl solution prior to local electron irradiation in a fractionated mode with a total focal dose (TFD) of 30 Gy; Group 3 (n = 20): intraperitoneal administration of N-AC at a dose of 120 mg/kg prior to local electron irradiation in a fractionated mode with a TFD of 30 Gy; Group 4 (n = 10): intraperitoneal administration of N-AC at a dose of 120 mg/kg without local electron irradiation. Thirty days after irradiation, blood levels of glucose, insulin, and amylase were measured. Histological analysis of pancreatic islet architecture, the degree of perivascular fibrosis (Masson’s trichrome staining), and the expression of IGF‑1, TGF-β, and VEGF-A were assessed by immunohistochemistry through counting positively stained cells. Results . Irradiation resulted in elevated blood glucose and amylase levels and decreased insulin levels. Signs of radiation-induced pancreatic injury were accompanied by reduced expression of IGF‑1 and VEGF-A in the islets by 39 % and 56 %, respectively, while the proportion of TGF-β-positive cells exceeded the control values by 3.4‑fold. Pre-irradiation administration of NAC partially preserved biochemical parameters within the normal range, suppressed TGF-β expression, and maintained the numbers of IGF‑1- and VEGF-A-positive endocrine cells at levels 2.5 and 3 times higher than those in the irradiation-only group, respectively. Conclusion . Pre-irradiation administration of NAC exerts a modulatory effect on the expression of growth factors IGF‑1, TGF-β, and VEGF-A in the late period following local electron irradiation of the pancreas, promoting restoration of the organ’s structural integrity and endocrine function. The obtained data suggest that N-AC is a promising agent for the prevention of delayed post-radiation complications, including fibrosis as well as metabolic and secretory insufficiency of the pancreatic islets.

Ischemic stroke remains a leading cause of death and disability worldwide, with neuroinflammation playing a central role in its pathogenesis. This study aimed to investigate age-related differences in neuroinflammatory responses in the human cerebral cortex following ischemic stroke. Using autopsy-derived brain tissue from 184 patients histological, histochemical, multiplex immunofluorescence, ELISA, and qRT-PCR analyses were conducted to assess neuronal damage, immune cell infiltration and cytokine expression. Morphological examination revealed pannecrosis in infarct cores and moderate inflammatory infiltration in penumbral regions. Multiplex immunofluorescence demonstrated active migration of NK (CD45⁺CD56⁺CD3⁻) and NKT (CD45⁺CD56⁺CD3⁺) cells in young patients, whereas elderly individuals showed a predominance of T lymphocytes (CD45⁺CD56⁻CD3⁺) and a decline in NK/NKT activity. Molecular assays indicated elevated NKG2D receptor expression and higher IFN-γ and proinflammatory cytokine (TNF-α, IL-1β, IL-6) levels in young patients, contrasting with dysregulated cytokine balance and reduced NK cytotoxicity in the elderly. These findings highlight distinct age-dependent immune mechanisms underlying ischemic injury, demonstrating that NK and NKT cells play a crucial role in early neuroinflammation, while aging shifts the balance toward adaptive immune dominance. The study underscores the need to consider patient age when designing neuroprotective and anti-inflammatory therapeutic strategies for ischemic stroke.

Ischemic stroke remains one of the leading causes of death and disability, especially among elderly patients. One of the key adaptive mechanisms in the post-ischemic period is neoangiogenesis, which promotes revascularization and restoration of the metabolism of neurons in the penumbra zone. The aim of the study was an immunohistochemical assessment of the age — related features of neoangiogenesis in the acute period of cerebral cortex infarction. Methods. The study was conducted on archived autopsy material — fragments of the frontal lobe of the cerebral cortex of patients with verified ischemic stroke (n=154) and the control group (n=30). The patients of both groups were divided by age into subgroups: young (18–44 years old), middle-aged (45–59 years old) and elderly (60–74 years old). The samples were examined histologically and immunohistochemically using antibodies to CD31 and VEGF-A. Results. The most pronounced neoangiogenesis (high number of vessels and VEGF-positive cells) was observed in young patients with ischemic stroke already in the early stages. VEGF-A expression was observed mainly in small-caliber blood vessels and neurons of the penumbra zone, especially in the group of young patients. On the contrary, in elderly patients with a large number of risk factors, the development of cerebral infarction was accompanied by a significant decrease in these indicators, especially in penumbra, which correlated with early high mortality. Conclusion. Increased VEGF expression combined with an increase in the number of small-caliber blood vessels is one of the key initiating factors of compensatory neoangiogenesis and activation of collateral blood supply and neurogenesis, collectively aimed at maintaining the functional activity of neurons in the penumbra zone. The high mortality of elderly patients in the first days after an ischemic stroke may be due to dysregulation of proangiogenic factors and a decrease in the activity of these adaptive reactions with age.

Ischemic stroke remains the leading cause of mortality and disability, with patient age significantly influencing risk factor structure, disease severity, and outcome. Despite numerous studies, comparative analysis of clinical characteristics in different age groups and their relationship to early mortality remain debatable. Purpose of the study: to conduct an age-stratified analysis of clinical characteristics in patients with ischemic stroke, identify key predisposing factors associated with neurological deficit severity, hospitalization time, and early mortality, and propose a simple prognostic scale for early stratification of adverse outcomes within the first week of illness. Materials. The study analyzed clinical data from 154 patients with ischemic stroke aged 18–74 years, divided into three age groups: young (n=38), middle-aged (n=51), and elderly (n=65). Anamnestic and clinical neurological indicators, comorbidities, hospitalization timing, and outcomes within the first seven days were evaluated. Univariate and multivariate analyses were applied, calculating odds ratios and standardized mean differences. Results. Among risk factors, older patients more frequently had hyperlipidemia, chronic obstructive pulmonary disease, anemia, dementia, and prolonged immobilization; younger patients had cardiomyopathies and atrial fibrillation. Stroke severity, delay before hospitalization, and early mortality increased proportionally with age (40% among elderly vs. 15.8% among young). Correlation analysis confirmed the contribution of hyperlipidemia, COPD, dementia, NIHSS score, and delayed hospitalization to early mortality risks. Conclusion. Clinical characteristics of patients with ischemic stroke demonstrate significant age-related differences in risk factor profile, disease severity, delay before hospitalization, and early mortality; based on multivariable analysis, an age-stratified predictive scale incorporating major clinical parameters was developed and proposed.

Background: Mast cells, key effectors of the innate immune system, are known to participate in various stages of tumor progression, including inflammation, angiogenesis, and extracellular matrix remodeling. Their role in melanoma, particularly in relation to Breslow thickness, pT stage, and AJCC staging, remains unclear. This study aims to quantitatively and phenotypically assess mast cell infiltration in cutaneous melanoma at different stages of progression, focusing on Tryptase- and Chymase-positive subtypes. Methods: This retrospective multicenter study included 124 patients with cutaneous melanoma (AJCC 8th edition, stages IA–IIIC). Histological sections were stained with hematoxylin and eosin, and mast cells were visualized using toluidine blue and immunohistochemistry with anti-Tryptase and anti-Chymase antibodies. Mast cells were counted manually in intratumoral and peritumoral regions by two independent observers. Quantitative data were analyzed using non-parametric tests and presented as median [Q1–Q3]. Results: Histological examination of 124 melanoma samples confirmed typical features of cutaneous melanoma, with nodular melanoma being the most common subtype (68 cases, 54.8%) and the lower extremities identified as the predominant tumor location (47 cases, 37.9%). Toluidine blue staining verified the presence of mast cells in both intratumoral and peritumoral compartments, with the highest density observed in early-stage melanomas. Immunohistochemical analysis identified both Tryptase+ and Chymase+ mast cells. The intratumoral number of Tryptase+ cells declined from 17 [14–19] per HPF at AJCC stage IA to 6 [5–7] per HPF at stage IIIC, while Chymase+ mast cells decreased from 14 [11–16] per HPF to 2 [1–3] per HPF over the same stages. Peritumoral counts also showed a downward trend, although less pronounced. Overall, the most significant reduction was observed in Chymase+ mast cells, suggesting their potential role as markers of melanoma progression. Conclusions: This study highlights the dynamic changes in mast cell populations in cutaneous melanoma, with a pronounced decrease in Chymase⁺ mast cells as the tumor progresses. Further research is needed to explore the mechanistic role of mast cells and their phenotypic shifts in melanoma progression.

Background: Macrophages are key components of the tumor microenvironment and play a critical role in melanoma progression. However, the dynamics of different macrophage subsets, particularly CD68⁺ and CD163⁺ populations, in relation to tumor thickness and stage remain insufficiently characterized. Objective: This study aimed to quantitatively assess intratumoral and peritumoral CD68⁺ and CD163⁺ macrophages in cutaneous melanoma and to investigate their associations with Breslow thickness, pT stage, and AJCC stage. Methods: We conducted a retrospective cohort study of 126 patients with cutaneous melanoma (AJCC stages IA–IIID). Tumor samples were examined histologically and immunohistochemically for CD68⁺ and CD163⁺ macrophages, and quantitative analysis was performed in intratumoral and peritumoral regions. Results: This study included 126 patients with cutaneous melanoma, ranging in stage from IA to IIID. Histopathological analysis revealed that melanoma tissues were primarily composed of irregular clusters of atypical melanocytic cells infiltrating the dermis and subcutaneous fat. Immunohistochemical staining identified CD68⁺ and CD163⁺ macrophages both within the tumor and in the surrounding stroma. Macrophage infiltration significantly increased with tumor thickness (Breslow) and progression to more advanced stages. Specifically, at Breslow thickness 4.0 mm. CD163⁺ macrophages showed similar trends, with a rise from 15.6 ± 2.8 cells at 4.0 mm in the intratumoral zone. Additionally, macrophage density was higher in tumors with ulceration, and both CD68⁺ and CD163⁺ macrophage numbers increased progressively with tumor stage, particularly in advanced stages. The number of CD68⁺ macrophages at stage IA in the intratumoral zone was 21.6 ± 3.1 cells and increased to 56.4 ± 6.8 cells at stage IIID, while CD163⁺ macrophages rose from 13.8 ± 3.2 cells at stage IA to 36.4 ± 4.6 cells at stage IIID. This suggests that macrophage infiltration, particularly CD163⁺ cells, correlates with melanoma progression. Conclusions: Our findings highlight distinct spatial and phenotypic patterns of macrophage infiltration in melanoma progression. The prominent increase in CD68⁺ and CD163⁺macrophages suggests their important role in tumor-associated immunomodulation. Further studies are warranted to elucidate macrophage polarization states and their prognostic and therapeutic implications in melanoma.

Background: A critical-size bone defect is defined as damage that exceeds the body’s intrinsic regenerative capacity for repair. The restoration of critical-size bone defects remains a major challenge in regenerative medicine and tissue engineering. Recent research emphasizes the importance of combining structural scaffolds with cellular and biologically active components to enhance bone regeneration. Objective: to develop and evaluate the structural and functional effects on bone repair and neoangiogenesis of a collagen-based scaffold implanted into a critical-size bone defect with platelet-rich plasma (PRP) in combination with stromal vascular fraction (SVF) cells. Methods: Forty Wistar rats were divided into groups: Group I (n = 10) received a collagen sponge (CS); Group II (n = 15) received a CS + SVF; Group III (n = 15) received a CS/SVF scaffold + PRP. Foreign material was received locally at the site of a critical-size bone defect. Micro-CT, haematological (clinical blood analyses), and histological (haematoxylin and eosin staining) analyses were performed. All animals were euthanized three months after implantation. Results: Micro-CT and histological evaluation revealed enhanced scaffold osteointegration, increased bone formation, and vascularisation in the CS+SVF+PRP group compared to controls. PRP significantly improved bone volume, surface area, and material density in the defect zone. Histological scoring confirmed more pronounced osteogenesis and neovascularisation in experimental groups. Haematological data indicated increased hemoglobin without signs of systemic inflammation. Local inflammatory response was minimal in the PRP group. Conclusion: The combination of collagen matrix, SVF, and PRP exhibits high biocompatibility and pronounced regenerative potential, providing a synergistic effect that promotes osteogenesis and angiogenesis. This approach shows promise for future applications in bone tissue engineering and warrants further molecular-level investigation.