A novel series of proflavine ureas, derivatives 11a-11i, were synthesized on the basis of molecular modeling design studies. The structure of the novel ureas was obtained from the pharmacological model, the parameters of which were determined from studies of the structure-activity relationship of previously prepared proflavine ureas bearing n-alkyl chains. The lipophilicity (LogP) and the changes in the standard entropy (ΔS°) of the urea models, the input parameters of the pharmacological model, were determined using quantum mechanics and cheminformatics. The anticancer activity of the synthesized derivatives was evaluated against NCI-60 human cancer cell lines. The urea derivatives azepyl 11b, phenyl 11c and phenylethyl 11f displayed the highest levels of anticancer activity, although the results were only a slight improvement over the hexyl urea, derivative 11j, which was reported in a previous publication. Several of the novel urea derivatives displayed GI50 values against the HCT-116 cancer cell line, which suggest the cytostatic effect of the compounds azepyl 11b-0.44 μM, phenyl 11c-0.23 μM, phenylethyl 11f-0.35 μM and hexyl 11j-0.36 μM. In contrast, the novel urea derivatives 11b, 11c and 11f exhibited levels of cytotoxicity three orders of magnitude lower than that of hexyl urea 11j or amsacrine.
- MeSH
- chemické jevy MeSH
- entropie * MeSH
- fibroblasty cytologie účinky léků MeSH
- inhibiční koncentrace 50 MeSH
- kinetika MeSH
- lidé MeSH
- močovina chemická syntéza chemie farmakologie MeSH
- molekulární modely MeSH
- proflavin chemická syntéza chemie farmakologie MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
Although some metallic nanoparticles (NPs) are commonly used in the food processing plants as nanomaterials for food packaging, or as coatings on the food handling equipment, little is known about antimicrobial properties of palladium (PdNPs) and platinum (PtNPs) nanoparticles and their potential use in the food industry. In this study, common food-borne pathogens Salmonella enterica Infantis, Escherichia coli, Listeria monocytogenes and Staphylococcus aureus were tested. Both NPs reduced viable cells with the log10 CFU reduction of 0.3-2.4 (PdNPs) and 0.8-2.0 (PtNPs), average inhibitory rates of 55.2-99% for PdNPs and of 83.8-99% for PtNPs. However, both NPs seemed to be less effective for biofilm formation and its reduction. The most effective concentrations were evaluated to be 22.25-44.5 mg/L for PdNPs and 50.5-101 mg/L for PtNPs. Furthermore, the interactions of tested NPs with bacterial cell were visualized by transmission electron microscopy (TEM). TEM visualization confirmed that NPs entered bacteria and caused direct damage of the cell walls, which resulted in bacterial disruption. The in vitro cytotoxicity of individual NPs was determined in primary human renal tubular epithelial cells (HRTECs), human keratinocytes (HaCat), human dermal fibroblasts (HDFs), human epithelial kidney cells (HEK 293), and primary human coronary artery endothelial cells (HCAECs). Due to their antimicrobial properties on bacterial cells and no acute cytotoxicity, both types of NPs could potentially fight food-borne pathogens.
- MeSH
- antibakteriální látky chemie farmakologie MeSH
- Bacteria klasifikace účinky léků růst a vývoj MeSH
- fibroblasty cytologie účinky léků MeSH
- kovové nanočástice aplikace a dávkování chemie MeSH
- kultivované buňky MeSH
- ledviny cytologie účinky léků MeSH
- lidé MeSH
- nemoci přenášené potravou prevence a kontrola MeSH
- palladium chemie MeSH
- platina chemie MeSH
- potravinářská mikrobiologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Diamond-like carbon (DLC) is a biocompatible material that has many potential biomedical applications, including in orthopaedics. DLC layers doped with Cr at atomic percent (at.%) of 0, 0.9, 1.8, 7.3, and 7.7 at.% were evaluated with reference to their osteoinductivity with human bone marrow mesenchymal stromal cells (hMSCs), immune activation potential with RAW 264.7 macrophage-like cells, and their effect on apoptosis in Saos-2 human osteoblast-like cells and neonatal human dermal fibroblasts (NHDFs). At mRNA level, hMSCs on DLC doped with 0.9 and 7.7 at.% of Cr reached higher maximum values of both RUNX2 and alkaline phosphatase. An earlier onset of mRNA production of type I collagen and osteocalcin was also observed on these samples; they also supported the production of both type I collagen and osteocalcin. RAW 264.7 macrophages were screened using a RayBio™ Human Cytokine Array for cytokine production. 10 cytokines were at a concentration more than 2 × as high as the concentration of a positive control, but the values for the DLC samples were only moderately higher than the values on glass. NHDF cells, but not Saos-2 cells, had a higher expression of pro-apoptotic markers Bax and Bim and a lower expression of anti-apoptotic factor BCL-XL in proportion to the Cr content. Increased apoptosis was also proven by annexin V staining. These results show that a Cr-doped DLC layer with a lower Cr content can act as an osteoinductive material with relatively low immunogenicity, but that a higher Cr content can induce cell apoptosis.
- MeSH
- aktiny metabolismus MeSH
- alkalická fosfatasa genetika metabolismus MeSH
- apoptóza účinky léků imunologie MeSH
- buněčná adheze účinky léků MeSH
- buněčná diferenciace účinky léků imunologie MeSH
- buněčné linie MeSH
- chrom farmakologie MeSH
- cytokiny metabolismus MeSH
- diamant farmakologie MeSH
- fibroblasty cytologie účinky léků MeSH
- kolagen typu I genetika metabolismus MeSH
- lidé MeSH
- makrofágy účinky léků metabolismus MeSH
- mezenchymální kmenové buňky cytologie účinky léků imunologie metabolismus MeSH
- myši MeSH
- osteogeneze účinky léků MeSH
- osteokalcin genetika metabolismus MeSH
- počet buněk MeSH
- proliferace buněk účinky léků MeSH
- protein PEBP2alfaA genetika metabolismus MeSH
- regulace genové exprese účinky léků MeSH
- RNA metabolismus MeSH
- tvar buňky účinky léků MeSH
- vápník metabolismus MeSH
- vinkulin metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Cells attaching to the extracellular matrix spontaneously acquire front-rear polarity. This self-organization process comprises spatial activation of polarity signaling networks and the establishment of a protruding cell front and a non-protruding cell rear. Cell polarization also involves the reorganization of cell mass, notably the nucleus that is positioned at the cell rear. It remains unclear, however, how these processes are regulated. Here, using coherence-controlled holographic microscopy (CCHM) for non-invasive live-cell quantitative phase imaging (QPI), we examined the role of the focal adhesion kinase (FAK) and its interacting partner Rack1 in dry mass distribution in spreading Rat2 fibroblasts. We found that FAK-depleted cells adopt an elongated, bipolar phenotype with a high central body mass that gradually decreases toward the ends of the elongated processes. Further characterization of spreading cells showed that FAK-depleted cells are incapable of forming a stable rear; rather, they form two distally positioned protruding regions. Continuous protrusions at opposite sides results in an elongated cell shape. In contrast, Rack1-depleted cells are round and large with the cell mass sharply dropping from the nuclear area towards the basal side. We propose that FAK and Rack1 act differently yet coordinately to establish front-rear polarity in spreading cells.
- MeSH
- buněčná adheze genetika fyziologie MeSH
- buněčné linie MeSH
- fibroblasty cytologie metabolismus MeSH
- fokální adhezní tyrosinkinasy genetika metabolismus MeSH
- krysa rodu rattus MeSH
- mikroskopie fázově kontrastní MeSH
- pohyb buněk genetika fyziologie MeSH
- polarita buněk genetika fyziologie MeSH
- receptory pro aktivovanou kinasu C genetika metabolismus MeSH
- RNA interference MeSH
- tvar buňky genetika fyziologie MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Chronic wounds and their associated bacterial infections are major issues in modern health care systems. Therefore, antimicrobial resistance (AMR), treatment costs, and number of disability-adjusted life-years have gained more interest. Recently, photodynamic therapy emerged as an effective approach against resistant and naïve bacterial strains with a low probability of creating AMR. In this study, needleless electrospinning was used to produce an indocyanine green (ICG) loaded poly(d,l-lactide) nanofibrous mesh as a photoresponsive wound dressing. The non-woven mesh had a homogeneous nanofibrous structure and showed long-term hydrolytic stability at different pH values. The antimicrobial activity was tested against several bacterial strains, namely Staphylococcus saprophyticus subsp. bovis, Escherichia coli DH5 alpha, and Staphylococcus aureus subsp. aureus. Upon irradiation with a laser of a specific wavelength (λ = 810 nm), the bacterial viability was significantly reduced by 99.978% (3.66 log10), 99.699% (2.52 log10), and 99.977% (3.64 log10), respectively. The nanofibrous mesh showed good biocompatibility, which was confirmed by the proliferation of mouse fibroblasts (L929) on the surface and into deeper parts of the mesh. Furthermore, a favorable proangiogenic effect was observed in ovo using the chorioallantoic membrane assay. In general, it can be concluded that ICG loaded nanofibers as an innovative wound dressing represent a promising strategy against chronic wounds associated with skin infections.
- MeSH
- biokompatibilní materiály chemie farmakologie MeSH
- buněčné linie MeSH
- chorioalantoická membrána krevní zásobení účinky léků MeSH
- Escherichia coli účinky léků růst a vývoj MeSH
- fibroblasty cytologie účinky léků MeSH
- fotochemoterapie MeSH
- indokyanová zeleň chemie farmakologie MeSH
- mikrobiální viabilita účinky léků MeSH
- myši MeSH
- nanovlákna MeSH
- obvazy MeSH
- polyestery chemie MeSH
- proliferace buněk účinky léků MeSH
- Staphylococcus aureus účinky léků růst a vývoj MeSH
- Staphylococcus saprophyticus účinky léků růst a vývoj MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
We recently developed a new light source that allows for the continuous monitoring of light-induced changes using common spectrophotometric devices adapted for microplate analyses. This source was designed primarily to induce photodynamic processes in cell models. Modern light components, such as LED chips, were used to improve the irradiance homogeneity. In addition, this source forms a small hermetic chamber and thus allows for the regulation of the surrounding atmosphere, which plays a significant role in these light-dependent reactions. The efficacy of the new light source was proven via kinetic measurements of reactive oxygen species generated during the photodynamic reaction of chloroaluminium phthalocyanine disulfonate (ClAlPcS2) in three cell lines: human melanoma cells (G361), human breast adenocarcinoma cells (MCF7), and human fibroblasts (BJ).
Little attention was given to the interaction between tumor and stromal cells in urothelial bladder carcinoma (UBC). While recent studies point towards the existence of different fibroblast subsets, no comprehensive analyses linking different fibroblast markers to UBC patient survival have been performed so far. Through immunohistochemical analysis of five selected fibroblast markers, namely alpha smooth muscle actin (ASMA), CD90/Thy-1, fibroblast activation protein (FAP), platelet derived growth factor receptor-alpha and -beta (PDGFRa,-b), this study investigates their association with survival and histopathological characteristics in a cohort of 344 UBC patients, involving both, muscle-invasive and non-muscle-invasive cases. The data indicates that combinations of stromal markers are more suited to identify prognostic patient subgroups than single marker analysis. Refined stroma-marker-based patient stratification was achieved through cluster analysis and identified a FAP-dominant patient cluster as independent marker for shorter 5-year-survival (HR(95% CI)2.25(1.08-4.67), p = 0.030). Analyses of interactions between fibroblast and CD8a-status identified a potential minority of cases with CD90-defined stroma and high CD8a infiltration showing a good prognosis of more than 80% 5-year-survival. Presented analyses point towards the existence of different stroma-cell subgroups with distinct tumor-modulatory properties and motivate further studies aiming to better understand the molecular tumor-stroma crosstalk in UBC.
- MeSH
- aktiny metabolismus MeSH
- antigeny Thy-1 metabolismus MeSH
- CD8-pozitivní T-lymfocyty cytologie imunologie metabolismus MeSH
- fenotyp MeSH
- fibroblasty cytologie metabolismus MeSH
- Kaplanův-Meierův odhad MeSH
- lidé MeSH
- membránové proteiny metabolismus MeSH
- nádorové biomarkery metabolismus MeSH
- nádory močového měchýře metabolismus mortalita patologie MeSH
- prognóza MeSH
- proporcionální rizikové modely MeSH
- růstový faktor odvozený z trombocytů - receptor alfa metabolismus MeSH
- senioři MeSH
- serinové endopeptidasy metabolismus MeSH
- shluková analýza MeSH
- želatinasy metabolismus MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Fibroblast growth factor (FGF) signaling is crucial for mammary gland development. Although multiple roles for FGF signaling in the epithelium have been described, the function of FGF signaling in mammary stroma has not been elucidated. In this study, we investigated FGF signaling in mammary fibroblasts. We found that murine mammary fibroblasts express FGF receptors FGFR1 and FGFR2 and respond to FGF ligands. In particular, FGF2 and FGF9 induce sustained ERK1/2 signaling and promote fibroblast proliferation and migration in 2D cultures. Intriguingly, only FGF2 induces fibroblast migration in 3D extracellular matrix (ECM) through regulation of actomyosin cytoskeleton and promotes force-mediated collagen remodeling by mammary fibroblasts. Moreover, FGF2 regulates production of ECM proteins by mammary fibroblasts, including collagens, fibronectin, osteopontin and matrix metalloproteinases. Finally, using organotypic 3D co-cultures we show that FGF2 and FGF9 signaling in mammary fibroblasts enhances fibroblast-induced branching of mammary epithelium by modulating paracrine signaling, and that knockdown of Fgfr1 and Fgfr2 in mammary fibroblasts reduces branching of mammary epithelium. Our results demonstrate a pleiotropic role for FGF signaling in mammary fibroblasts, with implications for regulation of mammary stromal functions and epithelial branching morphogenesis.
- MeSH
- fibroblastový růstový faktor 2 metabolismus MeSH
- fibroblastový růstový faktor 9 metabolismus MeSH
- fibroblasty cytologie metabolismus MeSH
- MAP kinasový signální systém * MeSH
- mléčné žlázy zvířat cytologie embryologie MeSH
- myši inbrední ICR MeSH
- myši MeSH
- parakrinní signalizace * MeSH
- receptor fibroblastových růstových faktorů, typ 1 metabolismus MeSH
- receptor fibroblastových růstových faktorů, typ 2 metabolismus MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Oct4-mediated reprogramming has recently become a novel tool for the generation of various cell types from differentiated somatic cells. Although molecular mechanisms underlying this process are unknown, it is well documented that cells over-expressing Oct4 undergo transition from differentiated state into plastic state. This transition is associated with the acquisition of stem cells properties leading to epigenetically "open" state that is permissive to cell fate switch upon external stimuli. In order to contribute to our understanding of molecular mechanisms driving this process, we characterised human fibroblasts over-expressing Oct4 and performed comprehensive small-RNAseq analysis. Our analyses revealed new interesting aspects of Oct4-mediated cell plasticity induction. Cells over-expressing Oct4 lose their cell identity demonstrated by down-regulation of fibroblast-specific genes and up-regulation of epithelial genes. Interestingly, this process is associated with microRNA expression profile that is similar to microRNA profiles typically found in pluripotent stem cells. We also provide extensive network of microRNA families and clusters allowing us to precisely determine the miRNAome associated with the acquisition of Oct4-induced transient plastic state. Our data expands current knowledge of microRNA and their implications in cell fate alterations and contributing to understanding molecular mechanisms underlying it.
- MeSH
- buněčné linie MeSH
- embryo savčí * MeSH
- fibroblasty cytologie metabolismus MeSH
- lidé MeSH
- mikro RNA * biosyntéza genetika MeSH
- oktamerní transkripční faktor 3 * biosyntéza genetika MeSH
- regulace genové exprese * MeSH
- techniky buněčného přeprogramování * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Cancer-associated fibroblasts (CAFs) represent a crucial component of cancer microenvironment. CAFs significantly influence biological properties of various types of cancer in terms of local aggressiveness, recurrence, and metastatic behaviour. This chapter summarizes a simple protocol for isolation of normal fibroblasts and their cancer-associated counterparts from normal human skin and mucosa, respectively, as well as from samples of human tumours such as basal/squamous carcinoma, melanoma, and breast cancer, and employment of this procedure for other types of cancer is possible. Isolated fibroblasts can be expanded in vitro and employed for further analysis of, e.g., DNA, RNA, protein, etc.
