Aleksandra S. Rogozhina

Tumor-associated macrophages (TAMs), key effectors of the innate immune system, actively participate in extracellular matrix remodeling, angiogenesis, and tumor growth. However, their age-associated quantitative and phenotypic distribution in uterine leiomyoma microenvironment remains insufficiently understood. This study aims to evaluate the density and spatial distribution of CD68⁺ and CD163⁺ macrophage immunophenotypes in uterine leiomyoma stroma across distinct patient age groups. Methods: A total of 318 patients with histologically confirmed uterine leiomyoma were stratified into three chronological age cohorts: Group I (≤30 years, n = 42), Group II (31–45 years, n = 216), and Group III (≥46 years, n = 60). Archival paraffin-embedded tumor tissue sections were analyzed histologically with hematoxylin and eosin, and immunohistochemically using antibodies against CD68 and CD163. Digital morphometric analysis was performed using QuPath software on whole-slide images, evaluating median immunopositive cell densities (cells/mm²) across five standardized regions of interest per case. Results: Histological examination confirmed typical benign leiomyoma morphology, with secondary degenerative alterations (hyalinosis, sclerosis, calcification, and aseptic necrosis) being most pronounced in Group III. Immunohistochemical profiling demonstrated an age-associated increase in macrophage infiltration within the tumor stroma. Median CD68⁺ macrophage density was higher in older cohorts, ranging from 11.4 cells/mm² [IQR: 8.6–13.8] in Group I to 16.7 cells/mm² [IQR: 13.9–19.8] in Group II, and 22.4 cells/mm² [IQR: 18.8–26.2] in Group III (p < 0.001). Concurrently, CD163⁺ macrophage density was likewise higher across older cohorts, measured at 9.8 cells/mm² [IQR: 7.4–12.1] in Group I, 19.4 cells/mm² [IQR: 16.1–22.9] in Group II, and 28.6 cells/mm² [IQR: 23.8–33.4] in Group III (p < 0.001). The observed numerical predominance of CD163⁺ cells over CD68⁺ cells in Group III topographically correlated with areas of secondary tissue remodeling and ischemic injury. Conclusions: This study reveals significant unadjusted differences in separately quantified CD68-positive and CD163-positive cell densities within uterine leiomyoma stroma across distinct patient age groups. Higher CD163⁺ cell densities in older cohorts topographically co-distribute with areas of secondary tissue remodeling and ischemic alteration. Because markers were evaluated on separate sequential sections and multivariable adjustments were unfeasible due to subgroup collinearity, these parameters reflect descriptive, marker-specific stromal variations among real-world clinical cohorts rather than independent causal effects of biological aging.

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. A comprehensive understanding of the relationship between uterine scars and the blood flow in adjacent myometrial tissue is crucial for the early identification of risk groups, which can significantly reduce the incidence of complications, facilitate differential diagnosis, and inform effective treatment strategies for patients with uterine scar defects (USD). Aim. To study the histoarchitectural features, area and percentage of collagen fibers in excised scar tissue from the uterus, contingent upon the blood flow status in the adjacent myometrium. Materials and methods. Laparoscopic metroplasty was performed on patients diagnosed with USD, categorized into two groups based on blood flow assessment via functional magnetic resonance imaging (fMRI): preserved blood flow (n = 6) and impaired blood flow (n = 6). A histochemical examination was performed on samples of excised fibrous tissue from the scars, focusing on morphometric analysis, area, and the percentage of collagen fibers contingent upon the blood flow characteristics in the adjacent myometrium. Results. The mean age of the participants was 35.17 ± 3.35 years. The predominant complaints included postmenstrual spotting in 8 of 12 patients (66.7%), heavy menstruation in 3 patients (25%), pain in 2 patients (16.7%), and infertility in 3 patients (25%). According to fMRI, the average scar thickness was 2.16 ± 0.74 mm. Scar samples with impaired blood flow demonstrated a statistically significant reduction in both the area and percentage of collagen fibers by 2.2 times (p = 0.03) and 1.9 times (p = 0.01), respectively, when compared to samples without circulatory disorders. Conclusions. The state of blood flow in the adjacent myometrium significantly influences the development of the connective tissue component in scar tissue. Assessment of perfusion blood flow in the adjacent myometrium using fMRI could serve as an indicator of the morphological integrity of the uterine scar following cesarean section. Consequently, fMRI may serve as an additional diagnostic tool and assist in formulating management strategies for patients with USD.

Backgraund. In order to analyze the diversity of somatic mutations in the MED12 gene in the myomatous nodes of patients with multiple fibroids, to compare the occurrence of various types of such mutations in multiple nodes with data on the frequency of mutations in fibroids in general and the results of studies on the detection of such mutations in the myometrium. Materials and methods. samples of 62 patients with multiple uterine fibroids (from 2 to 14 operatively removed myomatous nodes, 264 nodes in total) were studied. After DNA isolation, PCR and amplicon sequencing, the obtained nucleotide sequences were analyzed for the presence of single nucleotide substitutions, as well as deletions and insertions of nucleotides in exon 2 of the MED12 gene. Results. The total frequency of detected somatic mutations in the MED12 gene in the studied sample was 56%. The obtained mutation rates in multiple nodes were compared with data on large samples of fibroids and data on the presence of such mutations in intact myometrial cells. The correspondence of the frequencies of different mutation variants was revealed. The data obtained on the distribution of somatic mutations in exon 2 of the MED12 gene in multiple uterine fibroids indicate frequent spontaneous mutagenesis in myometrial cells and a high probability of developing myomatous nodules with various mutations in multiple fibroids. The distribution of mutation frequencies in the MED12 gene of different types in the nodes of multiple fibroids practically does not differ from the distribution in the general samples obtained both in our work and taken from published data. The data on the detection of such mutations in the myometrium also generally repeat the distribution by type of mutations, which allows us to conclude that deletions occur with the same frequency in myometrial cells. Conclusions. Mutations in the MED12 gene are the most common trigger that causes myometrial cell degeneration. In the Russian population, the prevalence of such mutations in uterine leiomyoma nodules is 50—60%, while with multiple fibroids, patients have nodes with various mutations (both single-nucleotide and deletions), along with nodes that do not have such mutations.

The presence of nervous tissue is a key characteristic of mature and immature ovarian teratomas, therefore, differential diagnosis between ependymal tubules and neuroectodermal rosettes is of fundamental diagnostic and prognostic importance. Ependymal tubules are differentiated glial tissue characteristic of mature teratomas and indicate a favorable prognosis. In contrast, neuroectodermal rosettes reflect primitive neuroepithelial differentiation and are a pathognomonic sign of an immature teratoma. Their number directly determines the degree of malignancy of the tumor and the patient’s treatment strategy. Thus, a thorough morphological analysis combined with immunohistochemical examination underlies accurate diagnosis and optimal management of patients with ovarian teratomas.

Background/Objectives: Low-grade serous ovarian carcinoma (LGSOC) is known as an uncommon subtype of cancer with poor response to standard chemotherapy, so novel targets are required. The current study aims to highlight the Human Epidermal Growth Factor Receptor-2 (HER2/neu) immunohistochemical (IHC) expression in LGSOCs. Methods: The study was conducted using 33 cases of LGSOCs from the calendar years 2017–2024. IHC staining was performed using antibody anti-HER2/neu (clone 4B5). HER2/neu scoring was performed based on the American Society of Clinical Oncology and the College of American Pathologists (ASCO/CAP) criteria for breast carcinoma. Results: The mean age of the 33 patients was 46.5 years. International Federation of Gynecology and Obstetrics (FIGO) stage data for patients revealed a predominance of advanced disease: 82.7% (24/29) were in advanced stages. Early stages comprised 17.3% (5/29) of cases. The study did not find HER2/neu overexpression in all cases. In 3.0% of samples (1/33), HER2/neu IHC staining was scored as 1+, and in 6.1% (2/33) of all LGSOCs, ultralow phenotype was observed. Of 23 cases in the HER2-negative group, 6 patients were alive with progressive disease, 1 patient died in 5 months, and 16 were alive with no evidence of disease. Of two patients with the HER2-ultralow phenotype, one was alive with no evidence of disease at 16 months follow-up. Conclusions: The results support the idea that HER2/neu overexpression is exceptionally rare in LGSOC; nevertheless, future trials are essential to fully characterize the spectrum of HER2/neu alterations in LGSOC and to determine definitively whether the rare cases with mutations or ultralow expression could represent a small subgroup that might benefit from specific targeted agents.

High-grade serous ovarian carcinoma (HGSOC) is the most aggressive and the commonest histological subtype of epithelial ovarian cancer and is characterized by a high recurrence rate associated with the development of primary or acquired chemoresistance. Modern molecular classifications have identified four major HGSOC subtypes. This review aimed to analyze the morphological and molecular mechanisms underlying chemoresistance in different molecular subtypes and to evaluate their diagnostic and prognostic significance. HGSOC molecular subtypes demonstrated distinct morphological patterns, including differences in tumor architecture, stromal desmoplasia, lymphoid infiltration, mitotic activity, and genomic instability. The integration of morphological, immunohistochemical, and molecular data is essential for improving patient stratification, predicting therapeutic response, and developing personalized treatment strategies. Employing digital pathology and artificial intelligence provides additional opportunities for identifying molecular HGSOC subtypes and detecting tumors with a high risk of chemoresistance. Keywords: high-grade serous ovarian carcinoma, HGSOC, chemoresistance, molecular subtypes, tumor microenvironment, platinum resistance, digital pathology

Background/Objectives: Atypical cytological findings in cervical screening, such as ASC-US, ASC-H, and AGC, present a clinical challenge due to their variable risk of underlying high-grade lesions. The precise stratification of this risk is crucial for effective patient management. This study aimed to correlate Bethesda cytology categories with HPV genotyping, including viral load, and histological follow-up to improve risk prediction for cervical intraepithelial neoplasia grade 2 or worse (CIN2+). Materials and Methods: In this retrospective single-center study, we analyzed 407 patients with cytological reports of ASC-US, ASC-H, or AGC. All patients underwent HPV DNA testing with genotyping for 21 types, with viral load quantification for HPV16/18, and subsequent histological verification. Statistical analyses included non-parametric tests, correlation analysis, and multivariate logistic regression to identify independent predictors of CIN2+. Results: The prevalence of CIN2+ differed significantly among the cytological categories: 23.2% in ASC-US, 47.3% in ASC-H, and 19.5% in AGC. ASC-H and a high HPV16 viral load were identified as independent predictors of CIN2+ in the multivariate analysis. An ASC-H result increased the probability of CIN2+ by 2.5 times (aOR = 2.51; 95% CI: 1.28–4.94). For each 1 log10 increase in HPV16 viral load, the risk of CIN2+ increased by 30% (aOR = 1.30; 95% CI: 1.16–1.46). Stratification of ASC-US cases by HPV16 status revealed a dramatically higher positive predictive value (PPV) for CIN2+ in HPV16-positive patients (66%) compared to HPV16-negative patients (12.6%). The AGC category showed the strongest association with glandular pathology, including adenocarcinoma in situ. Conclusions: The combination of cytological findings and HPV16 viral load provides a powerful model for risk stratification. An ASC-H result is a strong independent risk marker, while the clinical significance of ASC-US is fundamentally determined by HPV16 status. These findings advocate for a risk-based management algorithm that integrates liquid-based cytology with extended HPV genotyping and viral load assessments to optimize patient triage and follow-up.

Background/Objectives: Fumarate hydratase-deficient leiomyomas (dFH-LMs) are a rare subtype of uterine smooth muscle tumors (USMTs) with implications for hereditary leiomatosis and renal cell carcinoma (HLRCC). Although several morphologic clues have been proposed, their diagnostic reproducibility is poorly defined. We aimed to determine the diagnostic significance of histopathologic features associated with fumarase deficiency and the reproducibility of key morphologic criteria for defining dFH-LMs. Methods: A retrospective analysis was performed on 45 USMTs that were initially classified as atypical leiomyomas (ALMs). The cohort comprised patients aged 21 to 75 years who had surgery at one tertiary medical care center from 2016 to 2025. Hematoxylin–eosin (H&E) slides were independently examined by three pathologists for features associated with FH deficiency, including eosinophilic globules, staghorn-like vessels, diffuse nuclear atypia, “bizarre” nuclei, and prominent nucleoli with halos. Molecular status was determined by immunohistochemistry (IHC) for fumarate hydratase (FH) and S-(2-succino)-cysteine (2SC). Interobserver agreement was quantified using Fleiss’s κ and Cohen’s κ. Results: Loss of FH expression was detected in 26/45 tumors (57.7%). Eosinophilic globules occurred in 88.5% of dFH-LMs, but only in 15.8% of ALMs (p < 0.001). By majority consensus (≥2 of 3 reviewers), the eosinophilic globules predicted FH deficiency with a sensitivity of 88.0% and a specificity of 85.0%; interobserver reproducibility was substantial (κ = 0.63). Staghorn-like vessels occurred in 73.1% of dFH-LMs vs. 26.3% of ALMs (p = 0.02) and diffuse nuclear atypia (84.6%) was also more frequent in dFH-LMs (p = 0.01). Patients with dFH-LMs were significantly younger than those with ALMs (Median, 34 vs. 41 years). Conclusions: Eosinophilic globules, staghorn-like vessels and diffuse nuclear atypia have been shown to have high diagnostic value and could be considered morphologic indicators of dFH-LMs. The substantial interobserver reproducibility of eosinophilic globules makes this feature particularly promising for routine clinical practice.