Many respiratory pathogens compromise epithelial barrier function during lung infection by disrupting intercellular junctions, such as adherens junctions and tight junctions, that maintain intercellular integrity. This includes Streptococcus pneumoniae, a leading cause of pneumonia, which can successfully breach the epithelial barrier and cause severe infections such as septicemia and meningitis. Fluorescence microscopy analysis on intercellular junction protein manipulation by respiratory pathogens has yielded major advances in our understanding of their pathogenesis. Unfortunately, a lack of automated image analysis tools that can tolerate variability in sample-sample staining has limited the accuracy in evaluating intercellular junction organization quantitatively. We have created an open source, automated Python computer script called "Intercellular Junction Organization Quantification" or IJOQ that can handle a high degree of sample-sample staining variability and robustly measure intercellular junction integrity. In silico validation of IJOQ was successful in analyzing computer generated images containing varying degrees of simulated intercellular junction disruption. Accurate IJOQ analysis was further confirmed using images generated from in vitro and in vivo bacterial infection models. When compared in parallel to a previously published, semi-automated script used to measure intercellular junction organization, IJOQ demonstrated superior analysis for all in vitro and in vivo experiments described herein. These data indicate that IJOQ is an unbiased, easy-to-use tool for fluorescence microscopy analysis and will serve as a valuable, automated resource to rapidly quantify intercellular junction disruption under diverse experimental conditions.

• MeSH
• adhezní spoje MeSH
• mezibuněčné spoje metabolismus   MeSH
• respirační sliznice MeSH
• Streptococcus pneumoniae * MeSH
• těsný spoj * metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH

The skin is known to be the largest organ in human organism creating interface with outer environment. The skin provides protective barrier against pathogens, physical and chemical insults, and against uncontrolled loss of water. The barrier function was primarily attributed to the stratum corneum (SC) but recent studies confirmed that epidermal tight junctions (TJs) also play important role in maintaining barrier properties of the skin. Independent observations indicate that barrier function and its recovery is impaired in aged skin. However, trans-epidermal water loss (TEWL) values remains rather unchanged in elderly population. UV radiation as major factor of photoageing impairs TJ proteins, but TJs have great self-regenerative potential. Since it may be possible that TJs can compensate TEWL in elderly due to its regenerative and compensatory capabilities, important question remains to be answered: how are TJs regulated during skin ageing? This review provides an insight into TJs functioning as epidermal barrier and summarizes current knowledge about the impact of ageing on the barrier function of the skin and epidermal TJs.

• MeSH
• epidermis metabolismus   patologie   účinky záření   MeSH
• kožní absorpce MeSH
• lidé MeSH
• permeabilita MeSH
• perspiratio insensibilis MeSH
• stárnutí kůže * účinky záření   MeSH
• stárnutí metabolismus   patologie   MeSH
• těsný spoj metabolismus   patologie   účinky záření   MeSH
• ultrafialové záření škodlivé účinky   MeSH
• věkové faktory MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH

Tumor cells interact with blood constituents and these interactions promote metastasis. Selectins are vascular receptors facilitating interactions of tumor cells with platelets, leukocytes, and endothelium, but the role of endothelial E-selectin remains unclear. Here we show that E-selectin is a major receptor for monocyte recruitment to tumor cell-activated endothelium. Experimental and spontaneous lung metastasis using murine tumor cells, without E-selectin ligands, were attenuated in E-selectin-deficient mice. Tumor cell-derived CCL2 promoted endothelial activation, resulting in enhanced endothelial E-selectin expression. The recruitment of inflammatory monocytes to metastasizing tumor cells was dependent on the local endothelial activation and the presence of E-selectin. Monocytes promoted transendothelial migration of tumor cells through the induction of E-selectin-dependent endothelial retractions and a subsequent modulation of tight junctions through dephosphorylation of VE-cadherin. Thus, endothelial E-selectin shapes the tumor microenvironment through the recruitment, adhesion, and activation of monocytes that facilitate tumor cell extravasation and thereby metastasis. These findings provide evidence that endothelial E-selectin is a novel factor contributing to endothelial retraction required for efficient lung metastasis. Cancer Res; 76(18); 5302-12. ©2016 AACR.

• MeSH
• CD antigeny metabolismus   MeSH
• cévní endotel metabolismus   MeSH
• E-selektin metabolismus   MeSH
• endoteliální buňky metabolismus   MeSH
• imunoblotting MeSH
• imunoprecipitace MeSH
• invazivní růst nádoru patologie   MeSH
• kadheriny metabolismus   MeSH
• modely nemocí na zvířatech MeSH
• monocyty metabolismus   MeSH
• mutantní kmeny myší MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• polymerázová řetězová reakce MeSH
• průtoková cytometrie MeSH
• těsný spoj metabolismus   patologie   MeSH
• transendoteliální a transepiteliální migrace fyziologie   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

The Caco-2 cell monolayer model is widely used as a standard screening tool for studying the mechanisms of cellular drug transport. Caffeine was chosen as a model drug and is supposed to be class I of the Biopharmaceutics Classification System (BCS). Our study was conducted 1) to characterize the mechanisms of caffeine transport across the intestinal barrier, 2) to classify caffeine according to BCS, 3) to predict drugs intestinal absorption in humans. METHODS: Caffeine transport (0.1, 0.3, 1 and 10 mmol/l) was studied in Caco-2 cell monolayer in apical to basolateral (AP-BL) and basolateral to apical (BL-AP) direction, under iso-pH 7.4 and pH-gradient (6/7.4) conditions. The relative contribution of the paracellular route was estimated using Ca2+- free transport medium (opening tight junctions). RESULTS: The caffeine transport was linear with time, transport direction and pH independent, displaying non-saturable (first-order) kinetics, with high permeability coefficient (Papp): in AP-BL direction Papp = 46.3-53.5 x 10-6 cm/s; in BL-AP direction Papp = 45.6-49.4 x 10-6 cm/s. Thus, the transport seems to be transcellular mediated by passive diffusion. Using Ca2+- free transport medium tight junctions were opened (confirmed by increased Papp of mannitol) but the caffeine Papp was not changed. Thus, the paracellular route is only a minor way of caffeine transport. CONCLUSION: High solubility and high permeability of caffeine rank it among class I of BCS and well absorbed compounds

• MeSH
• biologický transport fyziologie   účinky léků   MeSH
• buněčné kultury MeSH
• Caco-2 buňky MeSH
• difuze MeSH
• financování organizované MeSH
• intestinální absorpce MeSH
• kinetika MeSH
• kofein farmakokinetika   farmakologie   MeSH
• koncentrace vodíkových iontů MeSH
• lidé MeSH
• lineární modely MeSH
• mannitol metabolismus   MeSH
• permeabilita účinky léků   MeSH
• střevní sliznice metabolismus   MeSH
• těsný spoj metabolismus   účinky léků   MeSH
• viabilita buněk účinky léků   MeSH
• xenobiotika farmakokinetika   farmakologie   MeSH
• Check Tag
• lidé MeSH

OBJECTIVES: The Caco-2 cell monolayer model is widely used as a standard screening tool for studying the mechanisms of cellular drug transport. Caffeine was chosen as a model drug and is supposed to be class I of the Biopharmaceutics Classification System (BCS). Our study was conducted 1) to characterize the mechanisms of caffeine transport across the intestinal barrier, 2) to classify caffeine according to BCS, 3) to predict drugs intestinal absorption in humans. METHODS: Caffeine transport (0.1, 0.3, 1 and 10 mmol/l) was studied in Caco-2 cell monolayer in apical to basolateral (AP-BL) and basolateral to apical (BL-AP) direction, under iso-pH 7.4 and pH-gradient (6/7.4) conditions. The relative contribution of the paracellular route was estimated using Ca2+- free transport medium (opening tight junctions). RESULTS: The caffeine transport was linear with time, transport direction and pH independent, displaying non-saturable (first-order) kinetics, with high permeability coefficient (Papp): in AP-BL direction Papp = 46.3-53.5 x 10-6 cm/s; in BL-AP direction Papp = 45.6-49.4 x 10-6 cm/s. Thus, the transport seems to be transcellular mediated by passive diffusion. Using Ca2+- free transport medium tight junctions were opened (confirmed by increased Papp of mannitol) but the caffeine Papp was not changed. Thus, the paracellular route is only a minor way of caffeine transport. CONCLUSION: High solubility and high permeability of caffeine rank it among class I of BCS and well absorbed compounds.

• MeSH
• biologický transport účinky léků   fyziologie   MeSH
• buněčné kultury MeSH
• Caco-2 buňky MeSH
• difuze MeSH
• intestinální absorpce MeSH
• kinetika MeSH
• kofein farmakokinetika   farmakologie   MeSH
• koncentrace vodíkových iontů MeSH
• lidé MeSH
• lineární modely MeSH
• mannitol metabolismus   MeSH
• permeabilita účinky léků   MeSH
• střevní sliznice metabolismus   MeSH
• těsný spoj účinky léků   metabolismus   MeSH
• viabilita buněk účinky léků   MeSH
• xenobiotika farmakokinetika   farmakologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Hypertension-induced myocardial metabolic, structural and electrophysiological remodeling deteriorates with aging and contributes to both heart failure and occurrence of malignant arrhythmias. It has been shown in clinical trials that n-3 polyunsaturated fatty acids (n-3 PUFA) reduce the incidence of cardiovascular diseases and sudden cardiac death. We investigated the cardioprotective effects of n-3 PUFA in aged spontaneously hypertensive rats (SHR) and possible cellular mechanisms involved. Male and female 14-moth-old SHR were fed with n-3 PUFA (Vesteralens, Norway, 20 mg/day for two months) and compared with untreated SHR. Results showed that n-3 PUFA supplementation led to 1) significant decline of blood pressure; 2) suppression of inducible ventricular fibrillation (VF) by 57 % (male) and 67 % (female), although the arrhythmogenic substrates, like fibrosis, hypertrophy and abnormal gap junctions distribution were not eliminated; 3) preservation of the cardiomyocytes and the integrity of their junctions; 4) enhancement of energetic metabolism enzyme activity; 5) augmentation of capillary density associated with increased alkaline phosphatase and decreased dipeptidyl peptidase-4 (DPP4) activity and 6/ increase in gap junction channel connexin-43 expression. Thus, aged male as well as female SHR benefit from n-3 PUFA supplementation that results in decrease in VF susceptibility, partly due to an improvement of myocardial metabolic state, cardiomyocyte and cell-to-cell junctions integrity and Cx43 up-regulation.

• MeSH
• energetický metabolismus účinky léků   MeSH
• fibrilace komor etiologie   metabolismus   patofyziologie   prevence a kontrola   MeSH
• hypertenze komplikace   farmakoterapie   metabolismus   patofyziologie   MeSH
• kapiláry účinky léků   MeSH
• kardiomyocyty účinky léků   metabolismus   MeSH
• konexin 43 metabolismus   MeSH
• krevní tlak účinky léků   MeSH
• krysa rodu rattus MeSH
• kyseliny mastné omega-3 farmakologie   MeSH
• modely nemocí na zvířatech MeSH
• myokard enzymologie   metabolismus   patologie   MeSH
• potkani inbrední SHR MeSH
• potravní doplňky * MeSH
• těsný spoj účinky léků   metabolismus   MeSH
• věkové faktory MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

BACKGROUND AND AIM: Melibiose/rhamnose permeability test is used for noninvasive intestinal mucosa barrier testing. However, the possible escape route of the absorbed saccharides through either intact or impaired blood-biliary barriers has not so far been explored. The objective of the present study was therefore two-fold: First, to describe in detail the biliary pharmacokinetics of melibiose and rhamnose in rats; second, to evaluate the changes of both sugars' pharmacokinetics upon impairment of the blood-biliary barrier by acute extrahepatic cholestasis in rats.

• MeSH
• akutní nemoc MeSH
• cholestáza diagnóza   metabolismus   MeSH
• diagnostické techniky gastrointestinální MeSH
• financování organizované MeSH
• injekce intravenózní MeSH
• krysa rodu rattus MeSH
• melibiosa aplikace a dávkování   diagnostické užití   farmakokinetika   moč   MeSH
• modely nemocí na zvířatech MeSH
• permeabilita MeSH
• potkani Wistar MeSH
• rhamnosa aplikace a dávkování   diagnostické užití   farmakokinetika   moč   MeSH
• těsný spoj metabolismus   MeSH
• upregulace MeSH
• vysokoúčinná kapalinová chromatografie MeSH
• žluč metabolismus   MeSH
• žlučové kanálky metabolismus   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH