This study deals with age estimation based on colour changes of human tissue from the intervertebral discs, Achilles tendon and rib cartilage. The investigated colour changes are the result of the accumulation of non-enzymatic browning products in the tissue. Samples of excised tissues were photographed with a digital camera and the pictures were evaluated using the image analysis processor Lucia G 4.11 processor. The values of the intensities of the RGB channels (MeanRed, Mean Green, MeanBlue) and parameters from the IHS system (MeanSaturation, HueTypical, HueVariation, BrightVariation and MeanBrightness) were evaluated. The results confirm that colour changes of some tissues depend on ageing and are a good tool for age estimation.

• MeSH
• Achillova šlacha patologie   MeSH
• barva MeSH
• biologické markery MeSH
• chrupavka patologie   MeSH
• dospělí MeSH
• financování organizované MeSH
• fotografování MeSH
• lidé středního věku MeSH
• lidé MeSH
• meziobratlová ploténka patologie   MeSH
• mladiství MeSH
• počítačové zpracování obrazu MeSH
• produkty pokročilé glykace metabolismus   MeSH
• reprodukovatelnost výsledků MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• soudní vědy metody   přístrojové vybavení   MeSH
• stárnutí MeSH
• žebra MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH

Despite recent advances in targeted therapeutics, administration of 5-fluorouracil (5-FU) remains a common clinical strategy for post-surgical treatment of solid tumors. Although it has been proposed that RNA metabolism is disturbed by 5-FU treatment, the key cytotoxic response is believed to be enzymatic inhibition of thymidylate synthase resulting in nucleotide pool disproportions. An operating p53 tumor suppressor signaling network is in many cases essential for the efficiency of chemotherapy, and malfunctions within this system remain a clinical obstacle. Since the fate of chemotherapy-insensitive tumor cells is rarely described, we performed a comparative analysis of 5-FU toxicity in p53-deficient cells and conclude that p53 acts as a facilitator rather than a gatekeeper of cell death. Although p53 can act as a regulator of several cellular stress responses, no rerouting of cell death mode was observed in absence of the tumor suppressor. Thus, the final death outcome of 5-FU-treated p53-/- cells is demonstrated to be caspase-dependent, but due to a slow pace, accumulation of mitochondrial reactive oxygen species contributes to necrotic characteristics. The oligomerization status of the p53 target gene DR5 is determined as a significant limiting factor for the initiation of caspase activity in an intracellular TRAIL-dependent manner. Using several experimental approaches, we further conclude that RNA-rather than DNA-related stress follows by caspase activation irrespectively of p53 status. A distinct 5-FU-induced stress mechanism is thereby functionally connected to a successive and discrete cell death signaling pathway. Finally, we provide evidence that silencing of PARP-1 function may be an approach to specifically target p53-deficient cells in 5-FU combinatorial treatment strategies. Together, our results disclose details of impaired cell death signaling engaged as a consequence of 5-FU chemotherapy. Obtained data will contribute to the comprehension of factors restraining 5-FU efficiency, and by excluding DNA as the main stress target in some cell types they propose alternatives to currently used and suggested synergistic treatment regimens.

• MeSH
• apoptóza účinky léků   MeSH
• fluorescenční protilátková technika MeSH
• fluoruracil farmakologie   MeSH
• imunoblotting MeSH
• imunoprecipitace MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• nádorový supresorový protein p53 metabolismus   MeSH
• polymerázová řetězová reakce MeSH
• protein TRAIL metabolismus   MeSH
• protinádorové antimetabolity farmakologie   MeSH
• RNA účinky léků   MeSH
• signální transdukce účinky léků   fyziologie   MeSH
• transdukce genetická MeSH
• transmisní elektronová mikroskopie MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The aim of this study was to determine if cytokeratins (CKs) 8 and 18--typical epithelial cell markers--are constitutively expressed in adult human corneal endothelium. Cryosections, paraffin-embedded sections and corneal endothelial imprints obtained from eleven adult human corneal discs not suitable for transplantation were used. Different fixative solutions were applied before indirect immunofluorescent or enzymatic staining was performed with antibodies against CK8 (Chemicon), CK18 (Dako and Sigma) and CK8/18 (Novocastra). Semi-quantitative RT-PCR and Western blotting (mRNA or proteins were isolated from Millicell membranes) were used to determine cytokeratin mRNA and protein levels. Approximately 50% of the corneal endothelial cells were positive for CK8 (Chemicon), CK18 (Sigma) and the CK pair 8/18 (Novocastra) in the endothelium when acetone was used for fixation. Four and 52% CK18-positive cells were observed using immunofluorescent and enzymatic immunohistochemistry, respectively, when the CK18 antibody from Dako was used. No signal was detected when 4% formalin or 10% paraformaldehyde was used as a fixative, irrespective of the antibody used. CK8 and CK18 proteins and mRNAs were detected in the endothelium of all tested corneas by Western blotting or semi-quantitative RT-PCR, respectively. We detected both CK8 and CK18 in the endothelium of all specimens at both the protein and mRNA levels. These results clearly demonstrate that cells of the corneal endothelium express CKs 8 and 18 and share some features with simple epithelia.

• MeSH
• exprese genu MeSH
• fluorescenční protilátková technika nepřímá metody   MeSH
• keratin-18 genetika   metabolismus   MeSH
• keratin-8 genetika   metabolismus   MeSH
• lidé středního věku MeSH
• lidé MeSH
• messenger RNA genetika   MeSH
• oční proteiny genetika   metabolismus   MeSH
• polymerázová řetězová reakce s reverzní transkripcí metody   MeSH
• rohovkový endotel metabolismus   MeSH
• senioři MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• senioři MeSH
• Publikační typ
• práce podpořená grantem MeSH

• MeSH
• antikoagulancia terapeutické užití   MeSH
• časové faktory MeSH
• extrakce katarakty MeSH
• ischemie komplikace   MeSH
• lidé středního věku MeSH
• lidé MeSH
• lokální anestezie škodlivé účinky   MeSH
• městnavá papila etiologie   MeSH
• nervus opticus krevní zásobení   MeSH
• nitrooční tlak MeSH
• pooperační komplikace MeSH
• poranění nervus opticus MeSH
• prednison terapeutické užití   MeSH
• senioři MeSH
• skotom etiologie   MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH

The aim of this study was to find out whether protease inhibitor 9 (PI-9) and granzyme B (GrB) molecules that contribute to immune response and the immunological privilege of various tissues are expressed in healthy and pathological human corneas. Using cryosections, cell imprints of control corneoscleral discs, we showed that PI-9 was expressed particularly in the endothelium, the superficial and suprabasal epithelium of healthy corneas, limbus, and conjunctiva. GrB was localized in healthy corneal and conjunctival epithelium, while the endothelium showed weak immunostaining. The expression of PI-6 and GrB was confirmed by qRT-PCR. Increased expression levels of the PI-9 and GrB genes were determined when the corneas were cultured with proinflammatory cytokines. Fluorescent and enzymatic immunohistochemistry of pathological corneal explants (corneal melting and herpes virus keratitis) showed pronounced PI-9, GrB, human leucocyte antigen (HLA)-DR, and leukocyte-common antigen (CD45) signals localized in multicellular stromal infiltrates and inflammatory cells scattered in the corneal stroma. We conclude that increased expression of the PI-9 and GrB proteins under pathological conditions and their upregulation in an inflammatory environment indicate their participation in immune response of the cornea during the inflammatory process.

• Publikační typ
• časopisecké články MeSH