Institute of Cytology

ctive: The endometrium, the inner lining of the uterine cavity, plays a key role in women’s reproductive health due to its capacity for cyclic regeneration, decidualization, and embryo implantation. Investigating the potential contribution of endometrial stromal cells ( EnSCs) to the pathogenesis of various reproductive disorders requires long-term cultivation of these cells, which is hindered by the replicative senescence of primary cultures. The aim of this work was to establish an immortalized EnSC line. Methods: An EnSC line with constitutive expression of the human telomerase catalytic subunit (hTERT) was generated via lentiviral transduction. The established line was characterized for proliferative activity, cell size, accumulation of lipofuscin granules, senescence-associated activity of β-galactosidase, expression of total histone H3, the cyclin-dependent kinase inhibitor p21, and the secretory profile component PAI-1. Furthermore, the immunophenotype of immortalized EnSCs was assessed using specific antibodies against CD73, CD90, and CD105. The dynamics of stress-induced apoptosis in hTERT-expressing EnSCs were analyzed using Annexin V and DAPI double staining. Results: hTERT expression in EnSCs significantly increased the replicative potential of the cells and delayed the onset of replicative senescence. Importantly, hTERT-expressing EnSCs maintained their immunophenotype and capacity for stress-induced apoptosis throughout the entire culture period. Conclusions: The established immortalized EnSC line represents a valuable tool for fundamental research in reproductive biology.

Palladium(II) cyclometalated complexes combine the properties of aromatic compounds and platinum group metals in terms of the ability to bind DNA and even double-stranded RNA. In this study, the activity of palladium(II) ethylenediamine complexes with Nile red (PdEnNR), coumarine 6 (PdEnC6) and 7,8-benzoquinoline (PdEnBhq) was tested against human cancerous and non-cancerous cells, as well as against mycoplasma pathogens of humans, poultry and plants. It was found that only PdEnC6 transiently reduced A549 cell proliferation after 72h, and cell counts were normalized after 96h, while PdEnNR and PdEnBhq did not demonstrate any significant influence on cancer cells. In relation to non-cancerous FLEH-1097/30 cells, only PdEnBhq had a limited toxic effect. At the same time, PdEnNR showed weak activity against mycoplasmas, but the minimum inhibitory concentrations of PdEnC6 and PdEnBhq against poultry pathogen Mycoplasma gallisepticum and human pathogen Mycoplasma arginini correspond to that of a number of commercial antibacterial drugs. We assume that these complexes or their derivatives can be promising agents for treating mycoplasma-infected cell cultures used in science or biotechnology or perspective drugs for controlling mycoplasma infections.

For patients with glioblastoma, the LMR serves as a diagnostically significant inflammatory biomarker independent of glucocorticoid administration. In patients with metastatic brain lesions, the most significant diagnostic value was demonstrated by monocyte counts, PIV and LMR.

Lampbrush chromosomes (LBCs) are a feature of amphibian oocytes and are typically associated with high levels of transcription during active oocyte growth. However, their state during winter hibernation has not been studied. Here, we investigated LBCs in early vitellogenic oocytes (early stage 4) of the grass frog Rana temporaria during winter hibernation. We found that the chromosomes retained their lampbrush morphology, and the phosphorylated form of RNA polymerase II resided on the lateral loops. Transcription on the lateral loops was reduced but detectable at cold conditions and significantly increased when the oocytes were transferred at room temperature. Satellite S1a transcripts were detected at the lateral loops of the chromosomes by RNA FISH. The possible significance of maintaining chromosomes in the lampbrush form during hibernation is discussed.

Sexual reproduction is a hallmark of most eukaryotes, yet hybridization can lead to alternative reproductive strategies, such as hybridogenesis. During hybridogenesis, found in the European water frog (Pelophylax esculentus) complex, one of the parental genomes is eliminated during gametogenesis, and the other one propagates clonally to gametes. We analyzed diploid hybrid males from populations common for hybrids and Pelophylax lessonae (L-E systems) from the eastern part of their distribution range. All but one hybrid produced spermatocytes and spermatids with Pelophylax ridibundus chromosomes, suggesting premeiotic elimination of P. lessonae genome and endoreplication of the P. ridibundus genome. We also observed spermatocytes with 13 or 26 univalents, indicating alterations in genome elimination and endoreplication. By comparing spermatocytes with germ cell genome composition, we suggest that genome elimination and endoreplication do not occur in adult males. Further, we examined introgressions using mitochondrial and nuclear genetic markers and comparative genomic hybridization. Most hybrids carried P. ridibundus and P. lessonae haplogroups, but five had Pelophylax cf. bedriagae mitochondrial DNA. In one hybrid, we revealed introgression of P. ridibundus chromosomal segments on P. lessonae chromosome. Although L-E systems are generally considered as stable, our results indicate variability in hybrid male gametogenesis, which may be related to introgressive hybridization.

Abstract Background We aimed to develop and validate a transcriptomic signature that correlates with clinical disease activity and therapy response using the Inflammatory Bowel Disease (IBD) Plexus® real-world data platform (Crohn’s & Colitis Foundation), CytoReason Disease Model based on published ulcerative colitis (UC) molecular studies, and a large prospective randomized clinical trial (VARSITY1). Methods A molecular disease activity score was built by identifying shared differential expression signals in transcriptomics data from inflamed v healthy control UC samples using an elastic net regression model trained against disease activity scores.2 The resulting model assigned a continuous molecular activity score per sample based on expression of the selected genes, with interpretability supported by feature importance and downstream biological evaluation. Signature module involvement was assessed using disease model data from 1985 mucosal biopsies from 18 UC clinical studies, including pts treated with golimumab, infliximab or vedolizumab. Pathway differential expression analysis was performed using gene set variation analysis (GSVA) with validation using molecular data from VARSITY.1 Results An 85 gene signature was identified, which represents enrichment of endothelial cells, alpha-beta T cells, epithelial, fibroblasts, and plasma cells (Figure). In contrast, published molecular inflammation gene sets (400–600 genes) 3, 4 are primarily enriched for immune and epithelial cells.3, 4 Our molecular activity signature also captured tertiary lymphoid structure, vascular, connective tissue, and hematopoiesis signaling—processes implicated in severe UC—thus demonstrating broader representation of disease mechanisms. Our molecular signature correlated with Mayo total/endoscopic scores across multiple IBD cohorts, and real-world registries eg., IBD Plexus. Assessment of our gene signature in VARSITY data revealed a significant correlation with therapy response: ie.,change in endoscopic and histological activity at week 52 (Table). Conclusion We developed and validated a refined molecular signature of UC disease activity encompassing diverse cellular populations which correlated with change in endoscopic and histologic disease severity in a prospective centrally read clinical trial. Validation across multiple real-world registries and clinical trials highlights its potential as a biomarker for clinical evaluation and the gene panel size relative to prior work offers enhanced feasibility for use to define residual disease burden in pts with endoscopic and histologic improvements in disease activity. This is an important step towards precision medicine in IBD using a molecular endotype-based classification of UC.

Cancer therapy resistance emerges from highly integrated molecular systems that enable tumor cells to evade cell death and survive cytotoxic therapeutic stress. High Mobility Group Box 1 (HMGB1) is increasingly gaining recognition as a central coordinator of these resistance programs. This review delineates how HMGB1 functions as a molecular switch that dynamically redistributes between cellular compartments in response to stress, with each localization enabling a distinct layer of resistance. In the nucleus, HMGB1 enhances chromatin accessibility and facilitates the recruitment of DNA repair machinery, strengthening resistance to radio- and chemotherapeutic damage. Cytosolic HMGB1 drives pro-survival autophagy, maintains redox stability, and modulates multiple regulated cell death pathways, including apoptosis, ferroptosis, and necroptosis, thereby predominantly shifting cell-fate decisions toward survival under therapeutic pressure. Once released into the extracellular space, HMGB1 acts as a damage-associated molecular pattern (DAMP) that activates key pro-survival and inflammatory signaling pathways, establishing microenvironmental circuits that reinforce malignant progression and therapy escape. HMGB1 further intensifies resistance through upregulation of multidrug resistance transporters, amplifying drug efflux. Together, these compartmentalized functions position HMGB1 as a central node in the networks of cancer therapy resistance. Emerging HMGB1-targeted agents, ranging from peptides and small molecules to receptor antagonists and nanoformulations, show promise in reversing resistance, but clinical translation will require precise, context- and redox-informed HMGB1 targeting to overcome multifactorial resistance program in refractory cancers.

The clinical efficacy of first-line oxaliplatin-based treatments in metastatic colorectal cancer (CRC) is universally limited by acquired resistance, a process driven by metabolic plasticity that allows tumors to dynamically reprogram their energy metabolism for survival. To exploit this vulnerability, we developed a novel organometallic scaffold based on palladium(II) complexes bearing acyclic diaminocarbene ligands. The complexes exhibit potent, low-micromolar antiproliferative activity against glycolytic CRC models and, crucially, display undiminished efficacy against oxaliplatin-resistant cells. We establish that their antiproliferative action hinges on the formation of stable H-bonded lipophilic cations under physiological conditions, which selectively target mitochondria of metabolically dysregulated cancer cells. This triggers a critical surge in mitochondrial lipid peroxidation, culminating in a regulated cell death that adapts to the cellular context, manifesting as either apoptosis or non-apoptotic death depending on the metabolic and redox status of the tumor cell.