Folia Biologica最新文献

Breast milk, as the optimal food for infants and young children, contains all the components necessary for proper growth and development. It is a rich source of both essential nutrients and biologically active factors, making breast milk a unique food with scientifically proven health-promoting properties. Among the entire range of biologically active factors, breast milk microorganisms and prebiotic factors, in the form of breast milk oligosaccharides, occupy an important place. The aim of our research was to determine the occurrence of bacteria with probiotic potential, belonging to the Lactobacillaceae family, in the environment of breast milk and breast milk oligosaccharides. The study included 63 human milk samples from breastfeeding women at various stages of lactation. Microorganism identification based on culture tests and MALDI TOF/MS, macronutrient analysis using the MIRIS human milk analyser, as well as analysis of human milk oligosaccharides using ultra-high-performance liquid chromatography coupled with high-resolution mass spectrometry were performed. The results have shown that breast milk from different breastfeeding women is characterized by great diversity in terms of the presence of Lacto-bacillaceae bacteria in its microbiological composition. These bacteria were present in 22.2 % of the tested breast milk samples. Analysis of the human milk oligosaccharide profile revealed a slightly higher content of prebiotic factors in breast milk samples containing Lactobacillaceae, including 2'-fucosyllactose, oligosaccharide occurring in the highest amount in breast milk.

Trypsin (TRY) combined with ethylenediaminetetraacetic acid (EDTA) is a widely used dissociation agent due to its efficiency and cost-effectiveness. However, its impact on preserving stem cell marker expression, such as C-X-C chemokine receptor type 4 (CXCR4) (critical for cell migration and homing) and cluster of differentiation 146 (CD146) (involved in pluripotency and angiogenesis), may be suboptimal compared to alternatives such as Accuta-se (ACC) and Accumax (ACMX), as shown previous-ly in bone marrow-derived stem cells (BM-MSCs). Limited data exist on these agents' effects on dental pulp stem cells (DPSCs). This study aims to investigate the influence of TRY, ACC, and ACMX on the expression of CXCR4 and CD146 in DPSCs. Seven characterized DPSC lines were cultured under standardized conditions and detached using TRY-EDTA, ACC, or ACMX. The expression of CXCR4 and CD146 was quantified via multicolour flow cytometry using an innovative DURAClone SC panel with supplementary anti-CXCR4 antibody. Comprehen-sive statistical analyses were performed to evaluate differences in marker preservation. No statistically significant differences in CXCR4 or CD146 expression were observed across the detachment methods (P > 0.05). ACMX consistently demonstrated margi-nally higher mean expression levels for both markers (CXCR4: 84.77 %; CD146: 93.91 %) compared to ACC (CXCR4: 83.45 %; CD146: 93.41 %) and TRY (CXCR4: 83.95 %; CD146: 92.99 %). While differences were not statistically significant, ACMX consistently yielded higher mean expression of both CXCR4 and CD146, indicating a potential advantage in preserving marker integrity during the cell detachment.

Prostate cancer (PCa) represents the most common malignancy among men in many countries, including the Czech Republic. While most cases are sporadic, approximately 10 % are attributed to hereditary factors, particularly germline pathogenic variants in genes such as BRCA1 and BRCA2, ATM, and CHEK2. These variants are associated with higher disease susceptibility, aggressive tumour behaviour and earlier onset of the disease. In parallel, modifiable and non-modifiable risk factors - including age, ethnicity, lifestyle, obesity, exposure to environmental carcinogens - also play an important role in the development and progression of PCa. This review summarizes the current state of knowledge on both genetic and non-genetic risk factors for PCa and emphasizes their relevance for clinical risk stratification, early detection strategies and prevention. The paper also provides an overview of the implications of hereditary PCa in the context of targeted therapy, particularly PARP inhibitors and immunotherapy, and discusses how somatic tumour profiling may refine the therapeutic decision-making. By integrating epidemiological data, molecular diagnostic and recent clinical advances, this review enhances understanding of the genetic underpinnings of PCa and advocates for a comprehensive, precision medicine approach in PCa management. The review emphasizes the importance of early identification of high-risk individuals through germline genetic testing and polygenic risk assessment, which can guide screening and personalized treatment and ultimately improve patient outcomes.

Nerol (NRL), a monoterpene in essential oils of various plants, exhibits anticancer activity, albeit with contrasting effects on normal cells. Because anaemia is induced by anticancer drugs, this study was initiated to profile the cytotoxic mechanisms of NRL in human erythrocytes. Cells were treated with NRL (0.01-0.1 %) for 24 h at 37 °C in Ringer buffers. Lytic cell death was measured photometrically and eryptosis was examined by flow cytometry using forward scatter, annexin V-affinity assay, H2DCFDA, and Fluo4/AM to evaluate cell volume, phosphatidylserine (PS) externalization, oxidative stress, and Ca2+, respectively. Small molecule inhibitors were used to probe the molecular mechanisms governing NRL-induced cytotoxicity. NRL led to eryptotic volume loss associa-ted with increased PS externalization and lysis through oxidative stress and Ca2+ nucleation. Ca2+ deprivation and K+ gradient dissipation, along with glucose, guanosine and sucrose, significantly blunted NRL-induced eryptosis and lytic death. Notably, both forms of cell death were inhibited by staurosporine, necrostatin 2 and myriocin, whereas urea, Z-VAD-FMK, SB205830 and D4476 only attenuated eryptosis. Furthermore, while lytic death was inhibited by polyethylene glycol, it was potentiated by Ca2+ deprivation and heparin, whereas eryptosis was aggravated by uric acid. In conclusion, NRL triggers cation channel- and redox-mediated eryptosis and lytic death through energy deprivation and activation of protein kinase C, receptor-interacting protein 1, serine palmitoyltransferase, caspase, p38 MAPK and casein kinase 1α. Altogether, these findings underscore the differential mechanisms by which NRL modulates divergent erythrocyte injury pathways and collectively advance the current knowledge of the extent of its cellular effects.

Cardiovascular diseases due to atherosclerosis remain a dominant medical problem. Macro-phages play a crucial role in both atherosclerosis progression and recycling of body iron. Epidemio-logical data point to elevation of body iron stores as one of non-classical cardiovascular risk factors, and we know that iron must be contained within macrophages to be atherogenic. Presumably, iron already contained within circulating monocytes turns cells to a more pro-inflammatory and hence atherogenic phenotype, but experimental evidence for such relationship remains limited. In this study, human monocytic THP-1 cells were loaded with micromolar iron in the forms of transferrin, ferric-ammonium citrate (FAC) and haemin for 2 and 24 hours. Only haemin was cytotoxic. All kinds of iron elevated the labile iron pool at 2 hours, as well as ferritin expression at 24 hours. Expression of scavenger receptors A and B, pro-inflammatory as well as anti-inflammatory cytokines, haem oxygenase and haptoglobin receptor were measured by quantitative PCR. Very few changes, none pro-inflammatory, were observed in response to transferrin or FAC. Haemin suppressed the expression of scavenger receptors, increased the expression of pro-inflammatory, and variably, anti-inflammatory cytokines, and dramatically induced haem oxygenase. The effects of haemin were not prevented by apotransferrin. We conclude that iron must be in the haemin form to act as a pro-inflammatory stimulus in THP-1 monocytes. Non-haem iron might require the presence of other factors to be atherogenic. Haemin treatment of THP-1 cells may represent a convenient experimental model to study the pro-inflammatory effects of haem that are observed in late stages of atherosclerosis.

Ovarian cancer remains one of the most lethal gynaecological malignancies, with paclitaxel resistance being a major therapeutic challenge that limits treatment efficacy and patient survival. We found that although the BARD1 level was not signi-ficantly altered in patients with ovarian cancer (OC), patients with higher BARD1 levels had increased survival time, suggesting that the down-regulation of BARD1 may be related to the paclitaxel sensitivity. Through examining the expression of BARD1 in tumour samples from paclitaxel responders and non-responders, we observed that the BARD1 level was significantly reduced in non-responders. CYP2C8 was up-regulated in non-responders. Also, the BARD1 level was negatively correlated with the level of CYP2C8. BARD1 over-expression in OC cells could repress the CYP2C8 expression, while knockdown of BARD1 could up-regulate CYP2C8 expression, which could be rescued by H2A-Ub. Results from gain and loss of functional experiments indicated that BARD1 functions as a tumour suppressor during paclitaxel treatment, and BARD1 down-regulation increased the IC50 of paclitaxel from 2.46 nM to 5.33 nM in SK-OV-3 cells and from 3.11 nM to 7.51 nM in CaoV-3 cells. We are the first to demonstrate that the down-regulation of BARD1 contributes to paclitaxel resistance via up-regulating CYP2C8 in patients with OC, which provides a potent target for clinical OC treatment.

Numerous studies have reported that increased interleukin 6 (IL-6) and soluble IL-6 receptor (sIL-6) levels induce inflammatory conditions. However, the exact mechanisms by which IL-6 drives inflammatory conditions remain unclear. Therefore, we investigated the potential role of IL-6/sIL-6R in inducing energy metabolism, including glycolysis, oxidative phosphorylation, lactate secretion and Akt/mTOR phosphorylation, in Jurkat cells, and whether IL-6 would increase the risk of developing inflammatory conditions due to the high metabolic profile of the T cells. Jurkat CD4 T-cell lines were stimulated with IL-6/sIL-6R for 24 h prior to 48-h stimulation with anti-CD3/CD28. Lactate secretion, glycolysis and oxidative phosphorylation levels were characterized using the Seahorse XF analyser. The Akt and mTOR phosphorylation status was detected using Western blotting. IL-6/sIL-6R significantly induced glycolysis and oxidative phosphorylation and their related parameters, including glycolytic capacity and maximal respiration, followed by significantly increased lactate secretion. Akt and mTOR phosphorylation were increased, which could have resulted from energy metabolism. Here we show that IL-6 enhanced the metabolic profile of Jurkat cells. This effect could have consequences for the metabolism-related signalling pathways, including Akt and mTOR, suggesting that IL-6 might promote T-cell energy metabolism, where T-cell hyperactivity might increase the inflammatory disease risk. The findings should be validated using studies on primary cells isolated from humans.

The poor prognosis of colorectal cancer (CRC) contributes to a yearly increase in CRC mortality, while microRNAs (miRNAs) were found to play a regulatory function in diversiform cancers, including CRC. The objective of this research was to evaluate the clinical value and possible regulatory mechanisms of miR-767-5p in CRC. The expression level of miR-767-5p in CRC tissues and cells was examined. The Kaplan-Meier curve was utilized to analyse the function of miR-767-5p in CRC prognosis. The independent prognostic factors in CRC were assessed by a multivariate COX regression analysis. Additionally, the regulatory mechanism of miR-767-5p in CRC was determined through an in vitro cell experiment. The miR-767-5p expression was down-regulated in CRC tumour tissues and CRC cells. Indicators such as tumour differentiation, TNM, LNM and miR-767-5p were identified as independent prognostic factors for a poor CRC prognosis. The regulatory relationship between miR-767-5p and nuclear factor I A (NFIA) was verified by the dual-luciferase reporter assay, and the NFIA expression level was significantly suppressed by over-expressed miR-767-5p. The proliferation, migration and invasion of CRC cells were inhibited by over-expressing miR-767-5p, while the inhibition effect could be reversed by over-expressing NFIA. The over-expressed miR-767-5p could serve as a tumour suppressor to inhibit the progression of CRC by suppressing the expression level of NFIA.

Fragility fractures have been a cause for concern because of their high incidence. For the prevention and treatment of osteoporotic fractures, it is important to understand how to promote bone formation and increase bone mass. This study investigated miR-296-3p expression and function in fragility fracture. The study enrolled 98 patients with hip fractures, 90 patients with wrist fractures and 35 healthy controls. RT-qPCR was used to detect the miR-296-3p level changes before and after surgery in fracture patients and during the differentiation of human bone mesenchymal stem cells (BMSCs). The starBase bioinformatics database was used for prediction of the miR-296-3p target gene, and dual luciferase report was used for verification of the target relationship. Our results demonstrated that miR-296-3p levels are up-regulated in fracture patients, while they gradually decrease during human BMSC differentiation. The up-regulation of miR-296-3p inhibited the proliferation and differentiation ability of human BMSCs, while inhibition of its expression had the opposite effects. miR-296-3p negatively regulates osteogenic differentiation, and over-expression of inhibitor of β-catenin and TCF (ICAT) could counteract the negative regulatory effect. miR-296-3p targets ICAT and affects the expression of key proteins in the Wnt/β-catenin signalling pathway. In conclusion, miR-296-3p can regulate the division and differentiation of osteoblasts by affecting the expression of ICAT and participate in fracture healing.

Osteoarthritis (OA) is a degenerative arthritis associated with aging. It is recognized that telomere attrition is a hallmark of aging. However, the transcriptional dynamics of synovial telomere-related genes (TRGs) in OA has not yet been elucidated. OA synovium microarray profiles were sourced from GEO and TRGs from TelNet. GO, KEGG, DO and GSVA enrichment analyses were employed to explore the underlying mechanisms. WGCNA and machine learning methods were utilized to screen hub differentially expressed TRGs (TRDEGs) that highly correlated with OA traits (hub OA-TRDEGs). Nomograms and receiver operating characteristic (ROC) curves were used to evaluate the diagnostic performance of hub OA-TRDEGs. An RNA-binding protein (RBP) network was developed to predict potential RBP target genes.​ The CIBERSORT analysis was performed to assess associations between hub OA-TRDEGs and immune infiltration. We identified 77 TRDEGs in normal and OA synovium samples. Functional enrichment analysis implicated these ge-nes primarily in metabolism regulation, DNA repair and inflammatory response. LTA4H, HNMT, ANKMY2, UFSP2, HLTF and RPA3 were established as hub OA-TRDEGs, demonstrating strong diagnostic performance for OA. Wilcox testing confirmed significant up-regulation of all hub OA-TRDEGs in OA synovium - a finding validated through independent datasets and qRT-PCR assays. Immune infiltration analysis further indicated that ​​resting mast cells, CD4+ memory resting T cells and activated mast cells are implicated in OA pathogenesis and exhibit significant correlations with hub OA-TRDEGs. These results nominate hub OA-TRDEGs as potential dia-gnostic biomarkers and underscore immune cell infiltration as a critical driver of OA progression.