Formation of peritoneal adhesions (PA) is one of the major complications following intra-abdominal surgery. It is primarily caused by activation of the mesothelial layer and underlying tissues in the peritoneal membrane resulting in the transition of mesothelial cells (MCs) and fibroblasts to a pro-fibrotic phenotype. Pro-fibrotic transition of MCs-mesothelial-to-mesenchymal transition (MMT), and fibroblasts activation to myofibroblasts are interconnected to changes in cellular metabolism and culminate in the deposition of extracellular matrix (ECM) in the form of fibrotic tissue between injured sides in the abdominal cavity. However, ECM is not only a mechanical scaffold of the newly synthetized tissue but reciprocally affects fibrosis development. Hyaluronan (HA), an important component of ECM, is a non-sulfated glycosaminoglycan consisting of N-acetyl-D-glucosamine (GlcNAc) and D-glucuronic acid (GlcUA) that can affect the majority of processes involved in PA formation. This review considers the role of endogenously produced HA in the context of different fibrosis-related pathologies and its overlap in the development of PA.

• Klíčová slova
• fibrosis, hyaluronan, inflammation, mesothelial cell, mesothelial-to-mesenchymal transition, metabolism, peritoneal adhesion,
• MeSH
• epitel MeSH
• fibroblasty fyziologie   MeSH
• kyselina hyaluronová * metabolismus   MeSH
• myofibroblasty metabolismus   MeSH
• peritoneum * metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH
• Názvy látek
• kyselina hyaluronová * MeSH

ETHNOPHARMACOLOGICAL RELEVANCE: The prevalence of different types of chronic wounds, due to the ageing population and increase incidence of diseases, is becoming a worldwide problem. Various medicinal plants used in folk medicine have demonstrated wound healing and antimicrobial properties, and some of these species are currently used in commercial preparations. Despite the well-documented and rich tradition of the use of local herbs for the treatment of skin injuries in Samoan folk medicine, their wound healing potential has not yet been systematically studied. AIM OF THE STUDY: Investigation into the in vitro antibacterial activity of ethanol extracts from 14 medicinal plants used in Samoan traditional medicine for the healing of wounds, burns and sores, and their effects on the proliferation and migration of human fibroblasts. MATERIALS AND METHODS: The antibacterial activity of these extracts was tested against pathogens associated with infected skin injuries, using the broth microdilution method. The effect on migration, proliferation and viability of human dermal fibroblasts was evaluated using wound healing scratch assay, cell proliferation assay, and thiazolyl blue tetrazolium bromide cytotoxicity test. RESULTS: The extracts from Cerbera manghas, Commelina diffusa, Kleinhovia hospita, Mikania micrantha, Omalanthus nutans, Peperomia pellucida, Phymatosorus scolopendria, Piper graeffei, Psychotria insularum, and Schizostachyum glaucifolium inhibited the growth of Staphylococcus aureus at the minimum inhibitory concentration (MIC) of ≥4 μg/mL, whereas C. manghas and P. pellucida produced the same MIC against both Escherichia coli and Pseudomonas aeruginosa. Among the antibacterially active species, C. diffusa, K. hospita, P. scolopendria, P. insularum, and S. glaucifolium did not produce toxicity towards the standard line of normal adult human dermal fibroblasts (IC80 > 128 μg/mL). In addition, extracts from Barringtonia asiatica, C. manghas, M. micrantha, O. nutans, P. insularum, and Piper graeffei stimulated significant migration of dermal fibroblasts, while M. micrantha, O. nutans, and P. insularum did not affect cell proliferation at a concentration of 32 μg/mL. CONCLUSIONS: The results suggest that the above-mentioned species of Samoan medicinal plants can be used for the development of new wound healing agents. However, further phytochemical and pharmacological research is needed regarding the isolation and identification of their active constituents.

• Klíčová slova
• Antibacterial activity, Medicinal plant, Samoa, Wound healing,
• MeSH
• antibakteriální látky izolace a purifikace   farmakologie   MeSH
• fibroblasty účinky léků   fyziologie   MeSH
• léčivé rostliny * MeSH
• lidé MeSH
• mikrobiální testy citlivosti metody   MeSH
• nadzemní části rostlin MeSH
• pohyb buněk účinky léků   fyziologie   MeSH
• proliferace buněk účinky léků   fyziologie   MeSH
• rostlinné extrakty izolace a purifikace   farmakologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Samoa etnologie   MeSH
• Názvy látek
• antibakteriální látky MeSH
• rostlinné extrakty MeSH

Despite widespread and prolonged use of adult novelties, their health safety is not regularly tested or legally regulated. In the EU, adult novelties are subjected to the General Product Safety Directive, placing the burden of proof regarding safe products onto the manufacturers. The aim of our pilot study was to expand knowledge on potential application of in vitro methods for hazard prediction of extracts from final products. We subjected extracts of 20 adult novelties, purchased on the Czech market to toxicological tests including NRU cytotoxicity assay, sensitization tests DPRA and LuSens and the YES/YAS endocrine assay. Four samples produced cytotoxicity. Sensitization potential was recorded by DPRA (three samples) while the LuSens reported ten samples. Regarding endocrine disruption, three samples produced antiestrogen and antiandrogen effects. Six samples exhibited androgenic potential and one sample showed estrogenic potential. Positive results with possible health effects were recorded repeatedly for samples made of ABS, PVC and latex. The study has confirmed promising usefulness of our test methods combination with regard to safety testing of this type of consumer products. The results should be evaluated with care, however, the data bring added-value to the limited knowledge of mixture toxicology and are indicative for further testing.

• Klíčová slova
• Chemical mixtures safety, Cytotoxicity, Endocrine disruption, In vitro toxicology, Public health, Risk assessment, Sex toy industry, Skin sensitization,
• MeSH
• buňky BALB 3T3 MeSH
• endokrinní disruptory toxicita   MeSH
• fibroblasty účinky léků   fyziologie   MeSH
• hra a hračky * MeSH
• lidé MeSH
• myši MeSH
• pilotní projekty MeSH
• plastické hmoty toxicita   MeSH
• Saccharomyces cerevisiae účinky léků   fyziologie   MeSH
• sexuální chování účinky léků   fyziologie   MeSH
• spotřebitelská bezpečnost produktů normy   MeSH
• techniky in vitro metody   MeSH
• testy akutní toxicity metody   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• endokrinní disruptory MeSH
• plastické hmoty MeSH

Effects of low-frequency electromagnetic fields (LF EMF) on the activation of different tissue recovery processes have already been fully understood. Preliminary recent data demonstrated that a special case of sinusoidal electromagnetic fields, known as amplitude-modulated currents (AMC) could have a potential to accelerate the cell metabolism or cell migration. An AMC generator was designed to generate sinusoidal induced electric currents with the amplitude modulation and the harmonic carrier frequency of 5,000 Hz was modulated by frequencies of 1 to 100 Hz. The magnetic field peak was 6 mT, electric field intensity 2 V/m and the current density of induced electrical currents was approximately 1 A/m(2). The coil of the generator was adapted to easy handling and safe integration into the shelf of the CO(2) incubator. The shelf with the coil was prepared for the introduction of cells in standard plastic in vitro chambers. The tests focused on cells with migratory capacity after injury or during immunological processes and thus, mesenchymal stromal cells (MSC), dendritic cells (DC), and fibroblasts were chosen. The tests involved exposures of the cells to LF EMF (180 min/day) every day, for a period of three days, before examining them for cell death, morphology changes, and CD markers. The samples were tested by using MTT assay and the effects on the intracellular concentration of reactive oxygen species were quantified. The cell migration was finally measured with the help of the transwell migration assay. None of the cell types showed any decrease in the cell viability after the LF EMF application and the cells displayed minimum changes in reactive oxygen species. Functional changes (acceleration of cell migration) after AMC exposure were statistically significant for the MSC samples only. The acceleration of MSCs is associated with the production of MMP by these cells. The EMF has a potential to be a safe, clinically applicable selective activator of MSC homing, MSC paracrine production, and subsequent regeneration processes.

• MeSH
• buňky 3T3 MeSH
• dendritické buňky fyziologie   MeSH
• elektromagnetická pole * MeSH
• fibroblasty fyziologie   MeSH
• lidé MeSH
• matrixová metaloproteinasa 2 metabolismus   MeSH
• mezenchymální kmenové buňky fyziologie   MeSH
• myši MeSH
• parakrinní signalizace MeSH
• pohyb buněk * MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• matrixová metaloproteinasa 2 MeSH

BACKGROUND: Several techniques for cell volume measurement using fluorescence microscopy have been established to date. In this study, we compare the performance of three different approaches which allow for estimations of the cell volume changes in biological samples containing individual fluorescently labeled cells either in culture or in the tissue context. The specific requirements, limitations and advantages of individual approaches are discussed. NEW METHOD: Global morphometric data are quantitatively compared with local information about the overall cell volume, represented by the concentration of a mobile fluorophore accumulated within the monitored cell. RESULTS: Volume changes induced by variations in the extracellular osmolarity in murine fibroblasts and astrocytes either in the culture or in the acute brain slices were registered by the three- and two-dimensional morphometries and by local fluorescence intensity measurements. The performance of the latter approach was verified using FRAP assessment of the fluorophore mobility. Significantly lower amplitudes of the cortical astrocytes swelling were detected by three-dimensional morphometry, when compared to the other two approaches. Consequently, it failed to detect temperature-induced cell volume changes. COMPARISON WITH EXISTING METHOD(S): The three most popular methods of cell volume measurement are compared to each other in this study. CONCLUSIONS: We show that the effectivity of global morphometry-based volumetric approaches drops with the increasing cell shape complexity or in the tissue context. In contrast to this, the performance of local fluorescence intensity monitoring, which is also fully capable of reflecting the instant cell volume variations remains stable, independent of the system used and application.

• Klíčová slova
• Calcein, Cell volume changes, Fluorescence microscopy, Morphometry,
• MeSH
• astrocyty cytologie   fyziologie   MeSH
• buňky 3T3 MeSH
• fibroblasty cytologie   fyziologie   MeSH
• fluorescenční barviva MeSH
• fluorescenční mikroskopie metody   MeSH
• hypertonické roztoky MeSH
• hypotonické roztoky MeSH
• isotonické roztoky MeSH
• konfokální mikroskopie metody   MeSH
• kultivované buňky MeSH
• mozková kůra cytologie   fyziologie   MeSH
• myši MeSH
• velikost buňky * MeSH
• zobrazování trojrozměrné metody   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• fluorescenční barviva MeSH
• hypertonické roztoky MeSH
• hypotonické roztoky MeSH
• isotonické roztoky MeSH

Cellular senescence is a hallmark of normal aging and aging-related syndromes, including the premature aging disorder Hutchinson-Gilford Progeria Syndrome (HGPS), a rare genetic disorder caused by a single mutation in the LMNA gene that results in the constitutive expression of a truncated splicing mutant of lamin A known as progerin. Progerin accumulation leads to increased cellular stresses including unrepaired DNA damage, activation of the p53 signaling pathway and accelerated senescence. We previously established that the p53 isoforms ∆133p53 and p53β regulate senescence in normal human cells. However, their role in premature aging is unknown. Here we report that p53 isoforms are expressed in primary fibroblasts derived from HGPS patients, are associated with their accelerated senescence and that their manipulation can restore the replication capacity of HGPS fibroblasts. We found that in near-senescent HGPS fibroblasts, which exhibit low levels of ∆133p53 and high levels of p53β, restoration of Δ133p53 expression was sufficient to extend replicative lifespan and delay senescence, despite progerin levels and abnormal nuclear morphology remaining unchanged. Conversely, Δ133p53 depletion or p53β overexpression accelerated the onset of senescence in otherwise proliferative HGPS fibroblasts. Our data indicate that Δ133p53 exerts its role by modulating full-length p53 (FLp53) signaling to extend the replicative lifespan and promotes the repair of spontaneous progerin-induced DNA double-strand breaks (DSBs). We showed that Δ133p53 dominant-negative inhibition of FLp53 occurs directly at the p21/CDKN1A and miR-34a promoters, two p53 senescence-associated genes. In addition, Δ133p53 expression increased the expression of DNA repair RAD51, likely through upregulation of E2F1, a transcription factor that activates RAD51, to promote repair of DSBs. In summary, our data indicate that Δ133p53 modulates p53 signaling to repress progerin-induced early onset of senescence in HGPS cells. Therefore, restoration of ∆133p53 expression may be a novel therapeutic strategy to treat aging-associated phenotypes of HGPS in vivo.

• MeSH
• časové faktory MeSH
• fibroblasty patologie   fyziologie   MeSH
• kultivované buňky MeSH
• lidé MeSH
• nádorový supresorový protein p53 genetika   fyziologie   MeSH
• poškození DNA genetika   MeSH
• předčasné stárnutí genetika   patologie   MeSH
• progerie genetika   patologie   MeSH
• protein - isoformy fyziologie   MeSH
• stárnutí buněk genetika   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH
• Názvy látek
• nádorový supresorový protein p53 MeSH
• protein - isoformy MeSH
• TP53 protein, human MeSH Prohlížeč

Dermal fibroblasts, which make up the major cell type in the dermis, have, historically, been considered to be relatively 'passive' cells which are responsible for the synthesis and remodeling of extracellular matrix proteins. However, the dermal fibroblast population is composed of heterogeneous and distinct cell types, and it has been established that, under the stress conditions of healing wound environments, dermal fibroblasts participate in the regulation of ongoing inflammation and cell proliferation by secreting a variety of signaling molecules that modulate the functions of immune cells, keratinocyte, endothelial cells and mast cells via both direct cell to cell communication and autocrine and paracrine interactions. This review describes the capacity of dermal fibroblasts to sense and respond to signals from the micro-environment and to communicate with surrounding cells during cutaneous wound healing. The review further emphasizes the, to date, poorly understood roles of heterogeneous dermal fibroblast populations in the wound healing process.

• Klíčová slova
• Dermal fibroblast, Intercellular communication, Papillary, Reticular, Wound healing,
• MeSH
• endoteliální buňky fyziologie   MeSH
• extracelulární matrix - proteiny fyziologie   MeSH
• fibroblasty fyziologie   MeSH
• hojení ran * MeSH
• keratinocyty fyziologie   MeSH
• lidé MeSH
• mezibuněčná komunikace * MeSH
• proliferace buněk MeSH
• signální transdukce MeSH
• škára cytologie   MeSH
• zánět imunologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH
• Názvy látek
• extracelulární matrix - proteiny MeSH

Each cell types or tissues contain certain "physiological" levels of R-2-hydroxyglutarate (2HG), as well as enzymes for its synthesis and degradation. 2HG accumulates in certain tumors, possessing heterozygous point mutations of isocitrate dehydrogenases IDH1 (cytosolic) or IDH2 (mitochondrial) and contributes to strengthening their malignancy by inhibiting 2-oxoglutarate-dependent dioxygenases. By blocking histone de-methylation and 5-methyl-cytosine hydroxylation, 2HG maintains cancer cells de-differentiated and promotes their proliferation. However, physiological 2HG formation and formation by non-mutant IDH1/2 in cancer cells were neglected. Consequently, low levels of 2HG might play certain physiological roles. We aimed to elucidate this issue and found that compared to highest 2HG levels in hepatocellular carcinoma HepG2 cells and moderate levels in neuroblastoma SH-SY5Y cells, rat primary fibroblast contained low basal 2HG levels at early passages. These levels increased at late passage and likewise 2HG/2OG ratios dropped without growth factors and enormously increased at hypoxia, reaching levels compared to cancer HepG2 cells. Responses in SH-SY5Y cells were opposite. Moreover, external 2HG supplementation enhanced fibroblast growth. Hence, we conclude that low 2HG levels facilitate cell proliferation in primary fibroblasts, acting via hypoxia-induced factor regulations and epigenetic changes.

• MeSH
• buňky Hep G2 MeSH
• experimentální nádory patologie   patofyziologie   MeSH
• fibroblasty cytologie   fyziologie   MeSH
• glutaráty metabolismus   MeSH
• krysa rodu Rattus MeSH
• kultivované buňky MeSH
• lidé MeSH
• mutace MeSH
• potkani Wistar MeSH
• proliferace buněk fyziologie   MeSH
• vývojová regulace genové exprese fyziologie   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• lidé MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH
• Názvy látek
• alpha-hydroxyglutarate MeSH Prohlížeč
• glutaráty MeSH

Patch clamp recordings carried out in the inside-out configuration revealed activity of three kinds of channels: nonselective cation channels, small-conductance K(+) channels, and large-conductance anion channels. The nonselective cation channels did not distinguish between Na(+) and K(+). The unitary conductance of these channels reached 28 pS in a symmetrical concentration of 200 mM NaCl. A lower value of this parameter was recorded for the small-conductance K(+) channels and in a 50-fold gradient of K(+) (200 mM/4 mM) it reached 8 pS. The high selectivity of these channels to potassium was confirmed by the reversal potential (-97 mV), whose value was close to the equilibrium potential for potassium (-100 mV). One of the features of the largeconductance anion channels was high conductance amounting to 493 pS in a symmetrical concentration of 200 mM NaCl. The channels exhibited three subconductance levels. Moreover, an increase in the open probability of the channels at voltages close to zero was observed. The anion selectivity of the channels was low, because the channels were permeable to both Cl(-) and gluconate - a large anion. Research on the calcium dependence revealed that internal calcium activates nonselective cation channels and small-conductance K(+) channels, but not largeconductance anion channels.

• MeSH
• buněčná membrána fyziologie   MeSH
• buněčné linie MeSH
• fibroblasty fyziologie   MeSH
• iontové kanály fyziologie   MeSH
• myši MeSH
• napětím ovládané aniontové kanály fyziologie   MeSH
• nízkovodivostní draslíkové kanály aktivované vápníkem fyziologie   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• iontové kanály MeSH
• napětím ovládané aniontové kanály MeSH
• nízkovodivostní draslíkové kanály aktivované vápníkem MeSH
• nonselective cation channel protein, mouse MeSH Prohlížeč

Live-cell imaging of focal adhesions requires a sufficiently high temporal resolution, which remains a challenge for super-resolution microscopy. Here we address this important issue by combining photoactivated localization microscopy (PALM) with super-resolution optical fluctuation imaging (SOFI). Using simulations and fixed-cell focal adhesion images, we investigate the complementarity between PALM and SOFI in terms of spatial and temporal resolution. This PALM-SOFI framework is used to image focal adhesions in living cells, while obtaining a temporal resolution below 10 s. We visualize the dynamics of focal adhesions, and reveal local mean velocities around 190 nm min-1. The complementarity of PALM and SOFI is assessed in detail with a methodology that integrates a resolution and signal-to-noise metric. This PALM and SOFI concept provides an enlarged quantitative imaging framework, allowing unprecedented functional exploration of focal adhesions through the estimation of molecular parameters such as fluorophore densities and photoactivation or photoswitching kinetics.

• MeSH
• barvení a značení MeSH
• buněčná adheze fyziologie   MeSH
• časové faktory MeSH
• fibroblasty fyziologie   MeSH
• krysa rodu Rattus MeSH
• mikroskopie metody   MeSH
• myši MeSH
• paxilin chemie   genetika   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• paxilin MeSH