Breast cancer is the most prevalent cancer type in women worldwide. It proliferates rapidly and can metastasize into farther tissues at any stage due to the gradual invasiveness and motility of the tumor cells. These crucial properties are the outcome of the weakened intercellular adhesion, regulated by small guanosine triphosphatases (GTPases), which hydrolyze to the guanosine diphosphate (GDP)-bound conformation. We investigated the inactivating effect of ARHGAP1 on Rho GTPases involved signaling pathways after treatment with a high dose of doxorubicin. Label-free quantitative proteomic analysis of the proteome isolated from the MCF-7 breast cancer cell line, treated with 1 μM of doxorubicin, identified RAC1, CDC42, and RHOA GTPases that were inactivated by the ARHGAP1 protein. Upregulation of the GTPases involved in the transforming growth factor-beta (TGF-beta) signaling pathway initiated epithelial-mesenchymal transitions. These findings demonstrate a key role of the ARHGAP1 protein in the disruption of the cell adhesion and simultaneously allow for a better understanding of the molecular mechanism of the reduced cell adhesion leading to the subsequent metastasis. The conclusions of this study corroborate the hypothesis that chemotherapy with doxorubicin may increase the risk of metastases in drug-resistant breast cancer cells.

• MeSH
• cdc42 protein vázající GTP metabolismus   MeSH
• doxorubicin farmakologie   MeSH
• lidé MeSH
• MFC-7 buňky MeSH
• nádory prsu * farmakoterapie   MeSH
• proteiny aktivující GTPasu * metabolismus   MeSH
• proteomika MeSH
• rac1 protein vázající GTP metabolismus   MeSH
• rho proteiny vázající GTP * metabolismus   MeSH
• rhoA protein vázající GTP metabolismus   MeSH
• Check Tag
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

Protection against water- and air-borne bacteria as well as their detection at very low levels is a big challenge for the health care profession. The study’s main goal was to prepare bacterial filters with a tunable trapping effectivity. We revealed that the trapping efficiency of Escherichia coli estimated from the optical density of bacteria passed through the filter was exponentially dependent on the surface density of the polyacrylonitrile nanofibre membranes. This log/linear regression profile was proven for bacterial trapping efficiency higher than 99.9% which opens a door for easy and tunable constructions of ultrasensitive filters and/or nanosensors as well as for the standardization and quality control of nanofibre membranes.

The incisional hernia is one of the most frequent complications affecting around 20% of all laparotomies. Around half of them suffer a recurrence when operated and one forth when surgical mesh is used. Unfortunately, even the surgical mesh can cause a wide range of complications and poses a lifelong risk for the patient. The biological mesh has brought a certain change to this concept. However, due to its biological origin this material triggers an immunological response of the organism and its manufacturing is extremely expensive. The biodegradable polycaprolactone nanofibres mimicking the extracellular matrix with their 3D structure could become an alternative as they are biocompatible and support fibroplasia. They have been tested as a material suitable for abdominal wall reconstruction already. Nanofibres will be further functionalized with platelet derivatives as a source of natural growth factors. Final samples will be tested on small animal model (rabbit).

Kýla v jizvě patří k nejčastějším pooperačním komplikacím postihujících zhruba 20% všech pacientů po operaci břicha. Zhruba polovina pacientů po plastice kýly v jizvě recidivuje do 5 let, při případě použití kýlní síťky zhruba čtvrtina. Bohužel i kýlní síťka je zatížena řadou nežádoucích vedlejších účinků s celoživotním rizikem. Určitou změnu přinesl koncept biologických kýlních sítěk. Díky svému původu však vedou k imunologické odpovědi organismu a jejich příprava je velmi ekonomicky nákladná. Alternativou biologických materiálů by se mohla stát biokompatibilní nanovlákna polykaprolaktonu, které svou strukturou napodobují extracelulární matrix, což výrazně zlepšuje biokompatibilitu materiálu a vede k urychlení regenerace tkáně. V experimentech již byla ověřena jako materiál využitelný pro regeneraci břišní stěny. Nanovlákna budou dále funkcionalizována trombocytárními deriváty, jako zdroj růstových faktorů pro urychlení hojení tkání. Nosiče budou testovány in vitro a in vivo na malém zvířecím modelu (králik).

• MeSH
• abdominální hernie komplikace   MeSH
• biokompatibilní materiály terapeutické užití   MeSH
• hojení ran MeSH
• incizní kýla prevence a kontrola   MeSH
• králíci MeSH
• lidé MeSH
• modely nemocí na zvířatech MeSH
• nanovlákna terapeutické užití   MeSH
• testování materiálů MeSH
• trombocyty MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• lidé MeSH
• zvířata MeSH
• Publikační typ
• hodnotící studie MeSH

• Konspekt
• Patologie. Klinická medicína
• NLK Obory
• preventivní medicína
• NLK Publikační typ
• závěrečné zprávy o řešení grantu AZV MZ ČR

Úvod a cieľ práce: Exozómy, ktoré sú generované endozomálnou cestou, sa exocytózou uvoľňujú do extracelulárneho priestoru vrátane slín. Obsahujú nukleové kyseliny, proteíny a lipidy, ktoré transportujú do rôznych častí tela. Podieľajú sa na medzibunkovej komunikácii, či už podporujú alebo narúšajú rôzne fyziologické procesy. Cieľom práce bolo overiť efektívnosť ultracentrifugačnej metódy izolácie exozómov zo slín, izolovať a identifikovať proteíny v nich obsiahnuté, s dôrazom na exozomálne proteíny súvisiace s ochoreniami ústnej dutiny alebo systematickými ochoreniami s prejavmi v ústnej dutine. Metódy: Exozómy boli z plných nestimulovaných ľudských slín izolované opakovanými centrifugačnými krokmi s premývaním vo fosfátovom tlmivom roztoku s následnou metanolovo/chloroformovou precipitáciou proteínov. Proteíny boli identifikované bottom-up prístupom pomocou hmotnostnej spektrometrie s predradenou separáciou na kvapalinovom chromatografe. Výsledky: Identifikované proteíny boli klasifikované podľa proteínových tried, molekulových funkcií a biologických procesov. Najviac identifikovaných proteínov bolo z proteínových tried: cytoskeletálne proteíny, obranné/imunitné proteíny a štrukturálne proteíny, a proteínov zodpovedajúcich za katalytickú aktivitu a štrukturálnu molekulovú aktivitu. Závery: Proteíny boli klasifikované do skupín na základe ich molekulovej funkcie a biologických procesov, na ktorých sa v ľudskom organizme zúčastňujú. Boli identifikované niektoré proteíny/skupiny proteínov, ktoré môžu byť zaujímavé z hľadiska skúmania etiológie niektorých orálnych ochorení, napr. anexín A1, zymogén granulárny proteín 16 homológ B, mucín-5B, oblasti Ig lambda-3 reťazca C, oblasť Ig kapa reťazca C, oblasť Ig alfa-2 reťazca C a oblasť Ig alfa-1 reťazca C.

Introduction, aim: Exosomes generated by the endosomal pathway are released by the exocytosis into the extracellular space, including saliva. They contain nucleic acids, proteins and lipids that are transported to different parts of the body. They participate in intercellular communication, whether they support or disrupt various physiological processes. The aim of this work was to verify the efficiency of the ultracentrifugation method of isolation of exosomes from saliva, to exctract and identify proteins contained therein, with an emphasis on exosomal proteins related to diseases of the oral cavity or systemic diseases with manifestations in the oral cavity. Methods: Exosomes were isolated from full unstimulated human saliva by repeated centrifugation steps with washing in phosphate buffer followed by methanol/chloroform protein precipitation. Proteins were identified by a bottom-up approach using mass spectrometry with pre-separation by liquid chromatography. Results: The identified proteins were classified according to protein classes, molecular functions and biological processes. The most identified proteins were of the protein classes: cytoskeletal proteins, defense/immune proteins and structural proteins and proteins responsible for catalytic activity and structural molecular activity. Conclusions: Proteins have been classified into groups based on their molecular function and biological processes in which they participate in the human body. Some proteins/family of proteins have been identified that may be of interest in investigating the etiology of some oral diseases e.g. annexin A1, zymogen granular protein 16 homologue B, mucin-5B, Ig lambda-3 chain C region, Ig kappa chain C region, Ig alpha-2 chain C region and Ig alpha-1 chain C region.

• MeSH
• exozómy MeSH
• klinická studie jako téma MeSH
• lidé MeSH
• nemoci úst * diagnóza   MeSH
• proteomika MeSH
• sliny chemie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH

The authors present their contribution to the improvement of methods suitable for the detection of the freezing and thawing damage of cells of cryopreserved venous grafts used for lower limb revascularization procedures. They studied the post-thaw viability of cells of the wall of cryopreserved venous grafts (CVG) immediately after thawing and after 24 and 48 h culture at +37 °C in two groups of six CVG selected randomly for slow thawing in the refrigerator and rapid thawing in a water bath at +37 °C. The grafts were collected from multi-organ and tissue brain-dead donors, cryopreserved, and stored in a liquid nitrogen vapor phase for five years. The viability was assessed from tissue slices obtained by perpendicular and longitudinal cuts of the thawed graft samples using in situ staining with fluorescence vital dyes. The mean and median immediate post-thaw viability values above 70% were found in using both thawing protocols and both types of cutting. The statistically significant decline in viability after the 48-h culture was observed only when using the slow thawing protocol and perpendicular cutting. The possible explanation might be the "solution effect damage" during slow thawing, which caused a gentle reduction in the graft cellularity. The possible influence of this phenomenon on the immunogenicity of CVG should be the subject of further investigations.

• MeSH
• alografty diagnostické zobrazování   účinky léků   MeSH
• apoptóza účinky léků   MeSH
• dárci tkání MeSH
• dimethylsulfoxid farmakologie   MeSH
• fluorescenční barviva * MeSH
• konfokální mikroskopie metody   MeSH
• kryoprezervace metody   MeSH
• kryoprotektivní látky farmakologie   MeSH
• lidé MeSH
• optické zobrazování metody   MeSH
• transplantace cév metody   MeSH
• vena femoralis diagnostické zobrazování   účinky léků   MeSH
• vena saphena diagnostické zobrazování   účinky léků   MeSH
• viabilita buněk účinky léků   MeSH
• zmrazování * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Cancer cell invasion through tissue barriers is the intrinsic feature of metastasis, the most life-threatening aspect of cancer. Detailed observation and analysis of cancer cell behaviour in a 3D environment is essential for a full understanding of the mechanisms of cancer cell invasion. The inherent limits of optical microscopy resolution do not allow to for in-depth observation of intracellular structures, such as invadopodia of invading cancer cells. The required resolution can be achieved using electron microscopy techniques such as FIB-SEM. However, visualising cells in a 3D matrix using FIB-SEM is challenging due to difficulties with localisation of a specific cell deep within the resin block. We have developed a new protocol based on the near-infrared branding (NIRB) procedure that extends the pattern from the surface grid deep inside the resin. This 3D burned pattern allows for precise trimming followed by targeted 3D FIB-SEM. Here we present detailed 3D CLEM results combining confocal and FIB-SEM imaging of cancer cell invadopodia that extend deep into the collagen meshwork.

• MeSH
• blízká infračervená spektroskopie metody   MeSH
• fibrosarkom patologie   MeSH
• invazivní růst nádoru MeSH
• lidé MeSH
• mikroskopie elektronová rastrovací metody   MeSH
• nádorové buňky kultivované MeSH
• nádory prsu patologie   MeSH
• počítačové zpracování obrazu MeSH
• podozomy patologie   MeSH
• zobrazování trojrozměrné metody   MeSH
• Check Tag
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

Úvod a cíl: Kompozitné materiály používané v zubnom lekárstve sa stávajú čoraz populárnejšími z viacerých dôvodov – estetiky, precitlivenosti niektorých jedincov na amalgám a obáv z možného uvoľňovania reaktívnych iónov kovov z amalgámu a zároveň aj obmedzovania jeho používania kvôli negatívnemu vplyvu na životné prostredie jednak počas výroby ako aj pri jeho likvidácii. Kompozitné materiály sú zložené z monomérov, plniva, iniciačného systému polymerizácie a z ďalších látok zlepšujúcich vlastnosti výplňového materiálu. Po aplikácii do kavity výplňový materiál tuhne polymerizačným procesom, ktorý je iniciovaný chemicky alebo svetlom, prípadne duálne. Konverzia monomérov používaných v stomatológii nikdy nedosiahne sto percent, čo predstavuje jednu zo slabín týchto materiálov. Pri styku kompozitných materiálov s vodou, alkoholom ako aj kyslým prostredím sa zistilo, že niektoré komponenty ako napríklad nespolymerizované monoméry sa môžu z kompozitu postupne uvoľňovať. V ústnej dutine sú kompozitné výplne vystavené neustálemu pôsobeniu slín a zároveň aj potravín a nápojov, ktoré vplývajú na uvoľňovanie zložiek kompozitov. Tieto môžu spôsobovať lokálne alergické reakcie, ale prehĺtaním slín s obsahom monomérov ovplyvňovať aj celý ľudský organizmus. Metodika: V tejto práci bolo stanovené množstvo nespolymerizovaných monomérov uvoľnených z vybraných kompozitných materiálov pripravených v tvare valčeka s výškou približne 2 mm s priemerom 3 mm, s hmotnosťou približne 0,05 g a vytvrdených – spolymerizovaných podľa postupu udávaného výrobcom. Následne boli kompozity ponorené do nestimulovane odobratých slín pri teplote 37 °C a množstvo uvoľnených monomérov stanovené v niekoľkých časových intervaloch. Na analýzu slín bola použitá technika vysokoúčinnej kvapalinovej chromatografie s UV detektorom. Experimentálne bolo sledované, aké množstvo monomérov sa vyplaví do slín s postupujúcim časom od vytvrdnutia kompozitu. Výsledky: V priebehu uvoľňovania monomérov z kompozitných materiálov do slín postupne vzrastá ich množstvo, rôzne monoméry sa vyplavujú v rôznych koncentračných hladinách. Najväčšie množstvo uvoľnených monomérov bolo namerané z kompozitných materiálov obsahujúcich trietylénglykol-dimetakrylát, najmenej sa vyplavil nespolymerizovaný bisfenol-A-glycidyl-metakrylát. Závěr: Vzhľadom na pomerne vysoké hladiny uvoľnených monomérov je pri vývoji kompozitných materiálov dôležité monitorovať ich hladiny v slinách v prvých hodinách po aplikácii ako aj v dlhodobom časovom odstupe za účelom výroby biologicky kompatibilnejšieho kompozitu.

Introduction, aim: Composite materials used in dentistry are increasingly used for several reasons – aesthetics, hypersensitivity of some individuals to amalgam and concerns about the possible release of amalgam metals, and at the same time limiting its use due to the negative impact on the environment both during production and disposal. Composite materials consist of monomers, filler, polymerization initiation system and other substances that improve the properties of the filler. After application to the cavity, the filler material solidifies by a polymerization process which is initiated chemically, by light or dual. The conversion of monomers used in dentistry will never reach 100%, which is one of the weaknesses of these materials. Upon contact of the composite materials with water, alcohol as well as an acidic environment, it has been found that some components, such as non-polymerized monomers, can be gradually released from the composite. In the oral cavity, composite fillings are exposed to the constant action of saliva as well as food and beverages, which affect the release of composite components. They can cause local allergic reactions, but they can also affect the whole human body by swallowing saliva-containing monomers. Methods: In this work, the amount of unpolymerized monomers released from selected composite materials, prepared in the shape of a cylinder with a height of approx. 2 mm and diameter of 3 mm, weighing approximately 0.05 g and cured – polymerised according to the procedure specified by the manufacturer, was determined. Subsequently, the composites were immersed in unstimulated saliva at 37 °C and the amount of monomers released was determined at several time intervals. A high performance liquid chromatography with a UV detector was used for saliva analysis. The amount of monomers released into saliva over time from the curing of the composite was monitored. Results: During the elution of monomers from composite fillers into saliva, their amount gradually increases, different monomers leach out at different concentration levels. The largest amount of released monomers was measured from composite materials containing triethylene glycol dimethacrylate, while bisphenol A-glycidyl methacrylate was eluted in the smallest quantity. Conclusion: Due to the relatively high levels of released monomers, it is important in the development of composite materials to monitor their levels in saliva in the first hours after application as well as in the long term in order to produce a more biologically compatible composite.

• MeSH
• chromatografie metody   MeSH
• klinická studie jako téma MeSH
• lidé MeSH
• sliny MeSH
• složené pryskyřice škodlivé účinky   MeSH
• zubní materiály * škodlivé účinky   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH

Tissue homeostasis mainly depends on the activity of stem cells to replace damaged elements and restore tissue functions. Within this context, mesenchymal stem cells and fibroblasts are essential for maintaining tissue homeostasis in skin, in particular in the dermis. Modifications in collagen fibers are able to affect stem cell features. Skin properties can be significantly reduced after injuries or with aging, and stem cell niches, mainly comprising extracellular matrix (ECM), may be compromised. To this end, specific molecules can be administrated to prevent the aging process induced by UV exposure in the attempt to maintain a youngness phenotype. NanoPCL-M is a novel nanodevice able to control delivery of Mediterranean plant myrtle (Myrtus communis L.) extracts. In particular, we previously described that myrtle extracts, rich in bioactive molecules and nutraceuticals, were able to counteract senescence in adipose derived stem cells. In this study, we analyzed the effect of NanoPCL-M on skin stem cells (SSCs) and dermal fibroblasts in a dynamic cell culture model in order to prevent the effects of UV-induced senescence on proliferation and collagen depot. The BrdU assay results highlight the significantly positive effect of NanoPCL-M on the proliferation of both fibroblasts and SSCs. Our results demonstrate that-M is able to preserve SSCs features and collagen depot after UV-induced senescence, suggesting their capability to retain a young phenotype.

• MeSH
• fibroblasty metabolismus   MeSH
• fytonutrienty * chemie   farmakologie   MeSH
• kmenové buňky metabolismus   MeSH
• lidé MeSH
• Myrtus chemie   MeSH
• nanovlákna chemie   MeSH
• rostlinné extrakty * chemie   farmakologie   MeSH
• stárnutí buněk účinky léků   MeSH
• tuková tkáň metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Endometrial cancer is one of the most frequent gynecological malignancies present in more than 95 % of all uterine cancers. In spite of that, screening of such disease is not commonly performed in clinical practice due to enormous costs and relatively low sensitivity. Therefore, developing an effective screening test to diagnose endometrial cancer at early stages is of great importance for the clinical area of investigation. In this work, we applied urinary proteomics (i.e., bottom-up proteomic approach followed by nano HPLC-ESI-MS/MS) in patients with endometrial cancer, with respect to find proteins aimed for the early diagnostics and screening. According to the results, the significant semi-quantitative changes were observed in urinary proteome of treated patients. The proteins that may be pivotal in pathogenesis of endometrial cancer, like cadherin-1 (CDH1), vitronectin (VTN) and basement membrane specific-heparan sulphate proteoglycan core protein (HSPG2) were down-regulated, when compared to the control group. Ultimately, it can be stated that urinary proteomics has a potential for the searching of cancer protein biomarkers based on their altered concentration.

• MeSH
• biologické markery moč   MeSH
• endometroidní karcinom moč   MeSH
• lidé středního věku MeSH
• lidé MeSH
• nádory endometria moč   MeSH
• proteom * MeSH
• studie případů a kontrol MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

The laser radiation absorbed by cells induces production of reactive oxygen species (ROS), followed by the development of oxidative stress. Proteins are major targets for ROS due to their abundance in biological systems. The aim of the present pilot study was to examine the effects of transcutaneous laser blood irradiation (TLBI), i.e., low-level laser therapy (LLLT) at 830 nm on plasma proteome in Wistar rats. Rats were irradiated in the heart area (i.e. coronary arteries) daily (i.e., for 9-day period), by commercially available GaAsAl diode laser (Maestro/CCM, Medicom Prague, Czech Republic, lambda=830 nm, power density 450mW/cm(2), daily dose 60,3 J/ cm(2), irradiation time 134 sec). The comparison of blood plasma proteome from irradiated and non-irradiated rats was performed utilizing 2D electrophoresis followed by MALDI TOF/TOF mass spectrometry. LLLT led to a quantitative change in the acute phase proteins with antioxidant protection i.e., haptoglobin (log(2) fold change (FC)=3.5), hemopexin (log(2) FC=0.5), fibrinogen gamma (log2 FC=1.4), alpha-1-antitrypsin (log(2) FC=-2.2), fetuin A (log2 FC=-0.6) and fetuin B (log2 FC=-2.3). In comparison to conventional biochemical methods, the changes in protein levels in blood plasma induced by LLLT offer a deeper insight into the oxidative stress response.

• MeSH
• fetuiny metabolismus   MeSH
• krev účinky záření   MeSH
• laserová terapie s nízkou intenzitou světla * MeSH
• pilotní projekty MeSH
• potkani Wistar MeSH
• proteiny akutní fáze metabolismus   MeSH
• proteom účinky záření   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH