PURPOSE: To document the expression of apical-basal polarity (ABP) determinants in the mouse corneal epithelium (CE) and elucidate the functions of Pard3 in establishment and maintenance of ABP, stratification, homeostasis, and barrier function in the CE. METHODS: Pard3Δ/ΔC mice (Pard3LoxP/LoxP; Aldh3A1-Cre/+) with cornea-specific Pard3 ablation were generated by breeding Aldh3A1-Cre/+ with Pard3LoxP/LoxP mice. The control (Aldh3A1-Cre/+ or Pard3LoxP/LoxP alone) and Pard3Δ/ΔC corneal histology, ocular surface properties, barrier function, and actin cytoskeleton were assessed by Haematoxylin and Eosin staining of paraformaldehyde-fixed, paraffin-embedded tissues, scanning electron microscopy, fluorescein staining, and phalloidin staining, respectively. The expression of specific markers of interest was evaluated by qRT-PCR, immunoblots and immunofluorescent staining. RESULTS: Dynamic changes were observed in the expression and localization of ABP determinants as the CE stratified and matured between post-natal day 5 (PN5) and PN52. Adult Pard3Δ/ΔC CE contained fewer cell layers with rounded basal cells, and loosely adherent superficial cells lacking microplicae. Adult Pard3Δ/ΔC CE also displayed impaired barrier function with decreased expression of tight junction, adherens junction, and desmosome components, disrupted actin cytoskeletal organization, increased proliferation, and upregulation of transcription factors that drive epithelial-mesenchymal transition (EMT). CONCLUSIONS: Disruption of ABP in Pard3Δ/ΔC CE, altered expression of cell junction complex components and disorganized actin cytoskeleton, increased cell proliferation, and upregulated EMT transcription factors suggest that the ABP-determinant Pard3 promotes CE features while suppressing mesenchymal cell fate. Collectively, these results elucidate that Pard3-mediated ABP is essential for CE stratification, homeostasis and barrier function.

• MeSH
• adaptorové proteiny signální transdukční * MeSH
• cytoskelet * metabolismus   MeSH
• homeostáza fyziologie   MeSH
• mikroskopie elektronová rastrovací MeSH
• myši MeSH
• polarita buněk * fyziologie   MeSH
• rohovkový epitel * metabolismus   ultrastruktura   MeSH
• těsný spoj * metabolismus   fyziologie   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Hydrogen sulfide (H2S) is a gas neurotransmitter that is synthesized in various mammalian tissues including vascular tissues and regulates vascular tone. The aim of this study is to investigate whether the endogenous L-cysteine/H2S pathway is impaired due to aging and endothelial denudation in mouse isolated thoracic aorta. For this purpose, young (3-4 months) and old (23-25 months) mice were used in the experiments. The effects of aging and endothelium on endogenous and exogenous H2S-induced vasorelaxation were investigated by cumulative L-cysteine-(1 microM-10 mM) and NaHS-(1 microM-3 mM) induced vasorelaxations, respectively. The L-cysteine-induced relaxations were reduced in old mice aorta compared to the young mice. Also, vasorelaxant responses to L-cysteine (1 microM-10 mM) were reduced on aorta rings with denuded-endothelium of young and old mice. However, the relaxation responses to NaHS were not altered by age or endothelium denudation. The loss of staining of CSE in the endothelial layer was observed in old thoracic aorta. Ach-induced (1-30 microM) relaxation almost abolished in endothelium-denuded rings from both mice group. Also, relaxation Ach reduced in intact endothelium tissue of old mice aorta. In conclusion, the vasorelaxant responses to L-cysteine but not NaHS decreased and the protein expression of CSE reduced in old thoracic aorta rings consistent with a decrease in H2S concentration with aging and endothelium damage, suggesting that aging may be lead to decrease in enzyme expression and H2S signaling system due to endothelium damage in mouse thoracic aorta. Key words Aging, Hydrogen sulfide, L-cysteine, Endothelium, Thoracic aorta.

• MeSH
• aorta thoracica * účinky léků   metabolismus   fyziologie   MeSH
• cévní endotel * metabolismus   účinky léků   MeSH
• cystein metabolismus   farmakologie   MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• signální transdukce MeSH
• stárnutí * metabolismus   MeSH
• sulfan * metabolismus   MeSH
• vazodilatace * účinky léků   fyziologie   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

For the treatment of bilateral limbal stem cell deficiency (LSCD), cell therapy with transplantation of cultivated oral mucosa epithelial cells (COMET) is a promising alternative. Although not yet established, current protocols on the cultivation of oral mucosal epithelial cell (OMECs) sheets are based mainly on substrates and xenobiotic additives that may lead to variable outcomes and undesirable immune responses by the patient. The aim of this study was to characterize OMECs cultivated in xenobiotic-free media (XF) seeded on fibrin gel, in comparison to conventional complex (COM) medium. Oral mucosal biopsies were retrieved from 31 donors. After cultivation in COM or XF medium, OMECs were compared based on growth kinetics, morphology, cell size and viability. Using immunofluorescence and gene expression analyses, the degree of stemness, proliferation and differentiation was evaluated in OMEC cultures. Our findings showed that although OMECs showed a similar morphology and viability, and comparable growth kinetics, immunofluorescence revealed the preservation of stemness (p63 + p40 positivity in cells ≤11 μm) and proliferation in both COM and XF. Gene expression analyses showed that keratin (K)13 and K15 expression levels were significantly higher in XF (adj. p < 0.001), but otherwise COM and XF-treated OMECs had comparable transcriptional profiles in a panel of stemness, proliferation and differentiation genes. These results demonstrate the feasibility of culturing OMECs on fibrin gel without xenogeneic additives, while maintaining their undifferentiated state and preserving stemness. In conclusion, both in terms of results and methodology, the procedures presented here are suitable for implementation in clinical practice.

• MeSH
• buněčná diferenciace MeSH
• buněčné kultury * MeSH
• deficit limbálních kmenových buněk MeSH
• dospělí MeSH
• epitelové buňky * metabolismus   účinky léků   MeSH
• fibrin * MeSH
• gely MeSH
• kmenové buňky * metabolismus   cytologie   MeSH
• kultivační média MeSH
• kultivované buňky MeSH
• lidé středního věku MeSH
• lidé MeSH
• limbus corneae * cytologie   patologie   metabolismus   MeSH
• nemoci rohovky patologie   farmakoterapie   metabolismus   MeSH
• proliferace buněk účinky léků   MeSH
• rohovkový epitel metabolismus   cytologie   účinky léků   patologie   MeSH
• senioři MeSH
• transplantace kmenových buněk metody   MeSH
• ústní sliznice * cytologie   MeSH
• viabilita buněk MeSH
• xenobiotika farmakologie   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

BackgroundSulodexide is a glycosaminoglycan-based drug prescribed to patients with angiopathy. We performed a pilot study to investigate whether sulodexide positively modulates the endothelial glycocalyx (EG) layer and the microcirculation in a porcine model of EG enzymatic damage. The EG is a sugar-based endothelial lining that is involved in the physiology of the capillary wall and the pathogenesis of many diseases.MethodsEG damage was induced in eight piglets by hyaluronidase III and heparanase I given intravenously. Four animals received sulodexide 600 IU intravenously before the enzymes and four animals after the enzymes were administered. Four animals constituted a control group. Sublingual microcirculation by side-stream dark field imaging and plasmatic concentration of syndecan-1 by ELISA were measured at baseline, 20 min after intervention, and at the 40th, and 60th minute onwards. The statistics were performed with a one-way ANOVA test with Turkey's correction for multiple comparisons testing. Timepoint comparison was performed by Student t-test or Mann-Whitney test.ResultsAt baseline, there were no statistically significant differences between the animal groups. After the intervention, the levels of syndecan-1 were significantly lower in the control group. While there were no differences between the two intervention groups. The sublingual microcirculation analysis showed that the DeBacker score was significantly higher in the control group. At 60 min, there was also a statistically significant difference in DeBacker score between the groups (8.1 ± 1.6 mm-1 in the group with enzymes given first and 11 ± 0.92 mm-1 in the group with sulodexide given first, p = 0.03). The analysis of the proportion of perused vessels did not show any statistically significant differences.ConclusionThe results of the study demonstrated a working model of EG damage but no specific action of sulodexide on EG modulation. In the sublingual microcirculation analysis, the sulodexide reduced the fall in absolute tissue perfusion in 60 min.

• MeSH
• cévní endotel * účinky léků   MeSH
• glykokalyx * účinky léků   metabolismus   MeSH
• glykosaminoglykany * farmakologie   MeSH
• hyaluronoglukosaminidasa MeSH
• mikrocirkulace účinky léků   MeSH
• modely nemocí na zvířatech MeSH
• pilotní projekty MeSH
• prasata MeSH
• syndekan-1 krev   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Kontext: Endotelová dysfunkce (ED) je jedním z hlavních mechanismů rozvoje různých kardiovaskulárních onemocnění. Endotelová dysfunkce je spojena s obezitou a hypertenzí, což se odráží v nízkých hodnotách průtokem stimulované dilatace tepny (flow-mediated dilatation, FMD) a vysokých hodnotách endotelinu-1 (ET-1) u dospělých osob a starších. Méně se ví o hodnotách uvedených dvou parametrů u adolescentů. Cílem této studie bylo srovnat hodnoty FMD a ET-1 obézních adolescentů s hypertenzí, obézních adolescentů bez hypertenze a u adolescentů s normální tělesnou hmotností. Metoda: Celkem 72 indonéských adolescentů průměrného věku 195 (178–217) měsíců, převážně chlapců (61 %), bylo zařazeno do tří následujících skupin: obezita s hypertenzí (n = 21), obezita bez hypertenze (n = 19) a bez obezity a bez hypertenze (n = 32). Hodnota FMD se měřila na pažní tepně po vyvinutí zvýšeného systolického tlaku na předloktí po dobu pěti minut. Hodnoty ET-1 se měřily metodou ELISA v sendvičovém uspořádání. Výsledek: Medián hodnoty ET-1 byl 42 (2,4–384) pg/ml. Mezi skupinami však nebyly nalezeny žádné významné rozdíly v hodnotách ET-1 (p 0,269). I když byly hodnoty FMD podle deskriptivní analýzy vyšší u zdravých adolescentů, nebyly zjištěny žádné statisticky významné rozdíly (p 0,159) v hodnotách FMD ve skupinách obezita s hypertenzí (9,1 ± 5 %), obezita bez hypertenze (8,3 ± 3,1 %) a bez obezity a bez hypertenze (10,9 ± 6,2 %). Srovnání hodnot ET-1 mezi skupinami s ED a bez ED (mezní hodnota FMD 7,1 %) nevykázalo žádné rozdíly (p 0,77). Navíc hodnoty ET-1 nekorelovaly statisticky významně s hodnotami FMD (p 0,66, r –0.053). Závěr: U indonéských adolescentů s obezitou a hypertenzí, obezitou bez hypertenze a s normální tělesnou hmotností a bez hypertenze nebyly zjištěny žádné rozdíly v hodnotách ET-1 a FMD. Korelace mezi ET-1 a FMD není u adolescentů statisticky významná.

Background: Endothelial dysfunction (ED) is one of the major mechanisms in various cardiovascular dis- eases. ED is associated with obesity and hypertension, which is reflected by low flow-mediated dilatation (FMD) value and high endothelin-1 (ET-1) level in adults or older. However, their value in adolescents is less known. This study aims to compare the FMD and ET-1 between obesity with hypertension, obesity without hypertension, and normal adolescents. Method: A total of 72 Indonesian adolescents with a mean age of 195 (178-217) months and dominated by males (61%) were classified into the three following groups obesity with hypertension (n = 21), obesity with- out hypertension (n = 19), and normal (n = 32) adolescents. FMD was measured using the brachial artery by previously applied supra systolic pressure in the forearm in 5 minutes. ET-1 was measured by Sandwich ELISA. Results: The median ET-1 level was 42 (2.4-384) pg/mL. However, there are no significant differences in ET-1 levels between all groups (p 0.269). Although FMD values are descriptively higher in normal adolescents, however, there are no statistical differences (p 0.159) in FMD values between obese with hypertension (9.1 ± 5%), obese without hypertension (8.3 ± 3.1%), and normal (10.9 ± 6.2%) groups. Comparison of ET-1 levels between groups with ED and without ED (FMD cut-off 7.1%) showed no differences (p 0.77) among them. In addition, ET-1 was not significantly correlated with FMD (p 0.66, r -0.053). Conclusion: There are no differences in ET-1 and FMD values between obesity with hypertension, obesity without hypertension, and normal Indonesian adolescents. The correlation between ET-1 and FMD is not significant in adolescents.

• MeSH
• arteria brachialis patologie   MeSH
• dilatace patologická etiologie   krev   MeSH
• endotel patofyziologie   MeSH
• endotelin-1 * krev   MeSH
• hypertenze komplikace   krev   MeSH
• lidé MeSH
• mladiství MeSH
• obezita dětí a dospívajících * komplikace   krev   MeSH
• statistika jako téma MeSH
• Check Tag
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• klinická studie MeSH
• práce podpořená grantem MeSH

• MeSH
• chronická lymfatická leukemie genetika   MeSH
• DNA-dependentní DNA-polymerasy genetika   metabolismus   MeSH
• DNA-primasa genetika   metabolismus   MeSH
• extracelulární matrix metabolismus   patologie   MeSH
• geny p53 genetika   MeSH
• glioblastom metabolismus   patologie   MeSH
• pericyty metabolismus   patologie   MeSH
• protein BRCA1 genetika   metabolismus   MeSH
• Publikační typ
• souhrny MeSH

Since the 1960s, more than 350,000 new chemicals have been introduced into the lives of humans and domestic animals. Many of them have become part of modern life and some are affecting nature as pollutants. Yet, our comprehension of their potential health risks for both humans and animals remains partial. The "epithelial barrier theory" suggests that genetic predisposition and exposure to diverse factors damaging the epithelial barriers contribute to the emergence of allergic and autoimmune conditions. Impaired epithelial barriers, microbial dysbiosis, and tissue inflammation have been observed in a high number of mucosal inflammatory, autoimmune and neuropsychiatric diseases, many of which showed increased prevalence in the last decades. Pets, especially cats and dogs, share living spaces with humans and are exposed to household cleaners, personal care products, air pollutants, and microplastics. The utilisation of cosmetic products and food additives for pets is on the rise, unfortunately, accompanied by less rigorous safety regulations than those governing human products. In this review, we explore the implications of disruptions in epithelial barriers on the well-being of companion animals, drawing comparisons with humans, and endeavour to elucidate the spectrum of diseases that afflict them. In addition, future research areas with the interconnectedness of human, animal, and environmental well-being are highlighted in line with the "One Health" concept.

• MeSH
• domácí zvířata * imunologie   MeSH
• epitel imunologie   MeSH
• kočky MeSH
• lidé MeSH
• psi MeSH
• vystavení vlivu životního prostředí škodlivé účinky   MeSH
• zvířata MeSH
• Check Tag
• kočky MeSH
• lidé MeSH
• psi MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

• Klíčová slova
• buněčná denzita, hexagonalita, index variačního koeficientu,
• MeSH
• denzitometrie MeSH
• klinické zkoušky jako téma statistika a číselné údaje   MeSH
• kouření * škodlivé účinky   MeSH
• lidé MeSH
• oční nemoci * etiologie   MeSH
• poruchy zraku * etiologie   MeSH
• rohovkový endotel patologie   MeSH
• úbytek endoteliálních buněk rohovky etiologie   patologie   MeSH
• Check Tag
• lidé MeSH

• Klíčová slova
• ortokeratologické kontaktní čočky,
• MeSH
• kontaktní čočky * MeSH
• lidé MeSH
• myopie prevence a kontrola   terapie   MeSH
• ortokeratologické metody * dějiny   metody   MeSH
• refrakce oka fyziologie   MeSH
• refrakční vady prevence a kontrola   terapie   MeSH
• rohovkové aberace terapie   MeSH
• rohovkový epitel MeSH
• Check Tag
• lidé MeSH

Ciel: Zistiť efekt opakovaných intravitreálnych aplikácii afliberceptu na endotel rohovky u pacientov s diabetickým makulárnym edémom (DME) a edémom makuly pri oklúzii retinálnej vény (RVO). Metodika: Do prospektívneho sledovania v období od januára 2021 do novembra 2023 bolo zaradených 87 naivných očí s diagnózou DME a RVO. Exklúzne kritérium pre zaradenie bol operačný alebo laserový zákrok počas sledovacieho obdobia, nosenie kontaktných šošoviek, ope rácia katarakty pred menej ako 6 mesiacmi, dystrofie alebo iné ochorenia rohovky, ktoré môžu spôsobiť zmeny endotelu. Okrem rutinných vyšetrení sme v deň 1., 4. a 8. injekcie vyšetrovali aj endotel rohovky pomocou bezkontaktnej endotelovej mikroskopie. Vyhodnocovali sme 4 parametre: hustotu endotelových buniek (CD), hexagonalitu (HEX), koeficient variability (CV) a centrálnu hrúbku rohovky (CCT). Najskôr sme vyhodnocovali celý súbor očí a následne sme ho rozdelili podľa 2 kritérií; podľa diagnózy na DME/RVO a podľa šošovky na fakické/pseu dofakické oči. Výsledky: Vyhodnotených bolo 87 očí (68 pacientov). Priemerný vek pacientov v čase diagnózy bol 66,8 ±9,3 rokov. Z celkového počtu 87 bolo 51 (59 %) fakických a 36 (41 %) pseudofakických očí. S diagnózou DME bolo liečených 61 (70 %) očí a s diagnózou RVO 26 (30 %). V prie behu sledovania nedošlo vplyvom liečby k štatisticky signifikantnej zmene priemerných hodnôt CD, HEX, CV, CCT či už v súbore všetkých očí alebo v rozdelení do podskupín podľa diagnózy či stavu šošovky. Záver: Zistili sme, že intravitreálne aplikácie afliberceptu u pacientov s DME a RVO neovplyvňujú parametre rohovkového endotelu ako CCT, HEX, CD, CV hodnotené pomocou endotelovej mikroskopie pri sledovaní po 8 injekciu.

Aim: To determine the effect of repeated intravitreal injections of aflibercept on the corneal endothelium in patients with diabetic macular edema (DME) and macular edema due to retinal vein occlusion (RVO). Methods: In a prospective study conducted between January 2021 and November 2023, a total of 87 treatment-naive eyes with DME and RVO were evaluated. The exclusion criteria were surgery or laser intervention during the follow-up period, contact lens wear, cataract surgery in the last 6 months, dystrophy, or other corneal condition that may cause endothelial damage. In addition to routine examinations on the day of application, we also measured the corneal endothelium using specular microscopy on the 1st, 4th and 8th day of injection. We evaluated 4 parameters: endothelial cell density (CD), hexagonality (HEX), coefficient of variability (CV) and central corneal thickness (CCT). First of all, we evaluated the entire cohort of eyes, and then divided it according to 2 criteria; the diagnosis into DME/RVO and according to the lens status into phakic/pseudophakic eyes. Results: A total of 87 eyes of 68 patients were evaluated. The average age of the patients at the time of diagnosis was 66.8 ±9.3 years. Within the cohort 51 (59%) eyes were phakic and 36 (41%) pseudophakic. A total of 61 (70%) eyes with a diagnosis of DME were treated, and 26 (30%) with RVO. During the follow-up, there were no significant changes in the average values of CD, HEX, CV, CCT due to aflibercept treatment, either in the whole group or in subgroups according to diagnosis or lens condition. Conclusions: The results of this study suggest that intravitreal administration of aflibercept in patients with DME and RVO did not have an impact on corneal endothelial parameters, including CCT, HEX, CD and CV. These parameters were measured using endothelial microscopy during an 8-injection observation period.

• Klíčová slova
• aflibercept,
• MeSH
• bevacizumab aplikace a dávkování   farmakologie   terapeutické užití   MeSH
• endoteliální buňky účinky léků   MeSH
• humanizované monoklonální protilátky farmakologie   klasifikace   terapeutické užití   MeSH
• inhibitory angiogeneze * aplikace a dávkování   farmakologie   klasifikace   terapeutické užití   MeSH
• injekce intravitreální * klasifikace   metody   MeSH
• lidé MeSH
• makulární edém chemicky indukované   etiologie   MeSH
• mikroskopie klasifikace   metody   MeSH
• počet buněk MeSH
• prospektivní studie MeSH
• rohovkový endotel diagnostické zobrazování   účinky léků   MeSH
• vaskulární endoteliální růstový faktor A antagonisté a inhibitory   MeSH
• Check Tag
• lidé MeSH