This study was focused on characterizing the differentiation of bone marrow-derived mesenchymal stem cells (MSCs) into corneal-like cells. Mouse MSCs were isolated from the bone marrow, grown in cell culture for 3 weeks, and purified using a magnetic activated cell sorter. Purified MSCs were cultured with an extract prepared from excised corneas and in the presence or absence of insulin-like growth factor-I (IGF-I). Analysis by quantitative real-time polymerase chain reaction showed that the expression of corneal specific markers, such as cytokeratin 12 (K12), keratocan, and lumican, was already induced after a 3-day cultivation and gradually increased during the 10-day incubation of MSCs with the extract. The presence of IGF-I significantly increased differentiation. Immunofluorescence analysis of differentiated MSCs showed positive results for the K12 protein. The morphology of the differentiated cells and the expression of cell surface markers CD45, CD11b, CD73, CD44, and CD105 were comparable in the control and differentiated MSCs. Proliferative activity was even higher in differentiated cells than in untreated MSCs. Both untreated and differentiated MSCs inhibited the production of interleukin-2 and interferon-γ in spleen cells stimulated with Concanavalin A. The results thus show that MSCs cultured in the presence of corneal extract and IGF-I efficiently differentiate into corneal-like cells. The differentiated cells possess characteristics of corneal epithelial cells and keratocytes, while at the same time maintaining MSC properties.

• MeSH
• biologické markery metabolismus   MeSH
• buněčná diferenciace účinky léků   genetika   MeSH
• imunosupresivní léčba MeSH
• insulinu podobný růstový faktor I farmakologie   MeSH
• mezenchymální kmenové buňky cytologie   účinky léků   metabolismus   MeSH
• myši inbrední BALB C MeSH
• proliferace buněk účinky léků   genetika   MeSH
• regulace genové exprese účinky léků   MeSH
• rohovka cytologie   MeSH
• tvar buňky účinky léků   genetika   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• biologické markery MeSH
• insulinu podobný růstový faktor I MeSH

To vitrify human amniotic membrane specimens so that the maximum of epithelial cells survives in order to develop a procedure for the eventual vitrification of posterior corneal lamellae without using cryoprotective agents. To assess different methods of tissue sample preparation preceding vitrification. In group 1, the amniotic membrane specimens were stretched on nitrocellulose support. In group 2, mechanical pressure was used to remove the excess culture medium between the support and the membrane. The samples were frozen in liquid ethane (-183 °C) and stored in liquid nitrogen. The specimens in the control group were not vitrified. Re-warming was performed at 40 °C. The epithelial cell survival rate was assessed after 1, 3 and 7 days of storage following re-warming using calcein and ethidium homodimer-1 fluorescence. A wide range of values was observed among the different groups and among individual specimens within the groups. Resulting average survival rate was 41 % for group 1 and 53 % for group 2; in several samples the cell survival rate exceeded 70 %. The storage period did not significantly affect the survival rates. The results of the rapid cooling of amniotic membranes in liquid ethane indicate that significant percentage of epithelial cells remain viable after the re-warming.

• MeSH
• amnion cytologie   fyziologie   MeSH
• epitelové buňky cytologie   fyziologie   MeSH
• kryoprezervace metody   MeSH
• kryoprotektivní látky MeSH
• lidé MeSH
• placenta cytologie   MeSH
• rohovka cytologie   fyziologie   MeSH
• těhotenství MeSH
• viabilita buněk MeSH
• vitrifikace * MeSH
• zmrazování MeSH
• Check Tag
• lidé MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• kryoprotektivní látky MeSH

To assess the quantitative and qualitative parameters of pre-cut posterior corneal lamellae for Descemet membrane endothelial keratoplasty with a stromal rim (DMEK-S) prepared manually in the Ocular Tissue Bank Prague. All 65 successfully prepared pre-cut posterior corneal lamellae provided for grafting during a 2-year period were analyzed retrospectively. The lamellae, consisting of a central zone of endothelium-Descemet membrane surrounded by a supporting peripheral stromal rim, were prepared manually from corneoscleral buttons having an endothelial cell density higher than 2,500 cells/mm(2). The live endothelial cell density, the percentage of dead cells, the hexagonality and the coefficient of variation were assessed before and immediately after preparation as well as after 2 days of organ culture storage at 31 °C. Altogether, the endothelium of 57 lamellae was assessed. Immediately after preparation, the mean live endothelial cell density was 2,835 cells/mm(2) and, on average, 1.8 % of dead cells were found. After 2 days of storage, the cell density decreased significantly to 2,757 cells/mm(2) and the percentage of dead cells to 1.0 %. There was a significant change in the mean hexagonality and the coefficient of variation after lamellar preparation and subsequent storage. The amount of tissue wasted during the preparation was 23 %. The endothelial cell density of posterior corneal lamellae sent for DMEK-S was higher than 2,700 cells/mm(2) in average with a low percentage of dead cells; 65 pre-cut tissues were used for grafting during a 2-year period.

• MeSH
• apoptóza MeSH
• Descemetova membrána cytologie   chirurgie   MeSH
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• nemoci rohovky chirurgie   MeSH
• oční banky MeSH
• počet buněk MeSH
• retrospektivní studie MeSH
• rohovka cytologie   chirurgie   MeSH
• rohovkový endotel cytologie   chirurgie   MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• stroma rohovky cytologie   chirurgie   MeSH
• techniky tkáňových kultur metody   MeSH
• zadní lamelární keratoplastika metody   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• práce podpořená grantem MeSH

OBJECTIVES: Malathion is generally not classified as toxic. However, the toxicity seems to be species-dependent. Local and systemic toxicity data for birds are rare, but a decrease of wild bird densities in areas where malathion was applied was reported. Aim of the study was to extend knowledge on malathion toxicity on cellular and organ level and to evaluate embryotoxicity and genotoxicity for birds using the chick embryo model HET-CAM. METHODS: Skin and eye irritation was determined using reconstructed skin and eye cornea tissues and the chorioallantoic membrane of chick embryo to simulate conjunctiva. Cytotoxicity in 3T3 Balb/c fibroblast culture was determined to estimate acute systemic toxicity. Chick embryo model was further employed to evaluate acute embryotoxicity for birds (mortality and genotoxicity). Data were analysed by means of general linear models. RESULTS: Malathion is not a skin and eye irritant. Cytotoxicity in vitro test provided LD50 value of 616 mg/kg suggesting higher toxic potential than is generally published based on in vivo tests on laboratory rodents. Embryotoxicity studies revealed dose and age dependent mortality of chick embryos. Genotoxicity was identified by means of micronucleus test in erythroid cells isolated from chorioallantois vascular system of chick embryos. CONCLUSIONS: Using in vitro alternative toxicological methods, a higher toxic potential of malathion was demonstrated than is generally declared. An increased health and environmental hazard may occur in areas with intensive agricultural production. The environmental consequences of delayed effects and embryotoxicity for bird populations in areas exposed to organophosphate insecticides, such as malathion, are obvious.

• MeSH
• biologické modely MeSH
• buňky BALB 3T3 cytologie   účinky léků   MeSH
• chorioalantoická membrána cytologie   účinky léků   MeSH
• dráždivé látky toxicita   MeSH
• druhová specificita MeSH
• embryo nesavčí cytologie   účinky léků   MeSH
• insekticidy toxicita   MeSH
• kur domácí MeSH
• kuřecí embryo MeSH
• lineární modely MeSH
• malathion toxicita   MeSH
• mitóza účinky léků   MeSH
• myši MeSH
• rohovka cytologie   účinky léků   MeSH
• techniky in vitro MeSH
• testy akutní toxicity MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• zvířata MeSH
• Check Tag
• kuřecí embryo MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• dráždivé látky MeSH
• insekticidy MeSH
• malathion MeSH

AIMS: To compare the number of antigen-presenting cells (APC) at various locations in fresh human corneoscleral disks and in those that are stored for grafting under hypothermic conditions or in organ culture (OC) with the aim of determining the conditions under which the decline in APC numbers is most substantial. METHODS: Cryosections obtained from fresh corneoscleral disks and disks stored under hypothermic (Optisol-GS) or OC conditions were used. The density of HLA-DR-positive cells was determined on cross sections using enzyme immunohistochemistry (alkaline phosphatase-antialkaline phosphatase technique). Longitudinal sections were used for detecting ATPase activity. RESULTS: The densities of HLA-DR-positive cells in both the epithelium and stroma increased from the central (3.79 and 0.61/mm(2)) to the peripheral cornea (5.56 and 1.35/mm(2)) as well as to the limbus and conjunctiva. A marked decrease in the number of HLA-DR-positive cells occurred after 7 days of storage in all corneal areas, the limbus and conjunctiva, compared to fresh tissue. No positive cells were found in the epithelium of corneas after 14 days in OC and after 21 days in hypothermic storage. Twenty-eight days of storage in OC led to the complete absence of HLA-DR-positive cells in the epithelium of the limbus and conjunctiva, and to a significant reduction in the stroma. CONCLUSION: Corneas stored in OC longer than 14 days are likely to be less immunogenic than corneas stored under hypothermic conditions, thus resulting in a possible positive effect on prolonging graft survival.

• MeSH
• adenosintrifosfatasy metabolismus   MeSH
• chondroitin sulfáty MeSH
• dextrany MeSH
• gentamiciny MeSH
• HLA-DR antigeny metabolismus   MeSH
• imunoenzymatické techniky MeSH
• komplexní směsi MeSH
• kryoprezervace * MeSH
• lidé MeSH
• orgánové kultury - kultivační techniky MeSH
• počet buněk MeSH
• rohovka cytologie   metabolismus   MeSH
• uchovávání orgánů * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• adenosintrifosfatasy MeSH
• chondroitin sulfáty MeSH
• dextrany MeSH
• gentamiciny MeSH
• HLA-DR antigeny MeSH
• komplexní směsi MeSH
• Optisol MeSH Prohlížeč

The genus Acanthamoeba includes more than 20 morphological species, but classification is problematical. Recently, the discovery of substantial interstrain differences in ribosomal DNA (rDNA) sequences has prompted questions about the relatedness of strains of the same species. In this study, therefore, we have investigated relationships between two isolates of A. polyphaga, CCAP 1501/3c and ATCC 30871, using morphological, biochemical, physiological, molecular and cytotoxicity assays. We observed that A. polyphaga ATCC 30871 exhibited up to six arms in endocyst while A. polyphaga CCAP 1501/3c exhibited a maximum of 5 arms thus indicating their position in group 2 and 3, respectively. Acanthamoeba polyphaga ATCC 30871 exhibited growth at 37 degrees C and growth on 1M mannitol plates while A. polyphaga CCAP 1501/3c did not. In addition, both isolates exhibited differences in isoenzyme banding patterns and rDNA restriction fragment polymorphisms. More importantly, A. polyphaga ATCC 30871 produced cytotoxicity on corneal epithelial cells while A. polyphaga CCAP 1501/3c had no effects, suggesting differences in pathogenicity. Thus, all the results provide evidence for significant differences between the strains and further provided the basis for reclassification of the isolates. Implications of these results in the clinical diagnosis of pathogenic Acanthamoeba are discussed.

• MeSH
• Acanthamoeba klasifikace   genetika   patogenita   fyziologie   MeSH
• akantamébová keratitida parazitologie   MeSH
• epitelové buňky parazitologie   patologie   MeSH
• fenotyp MeSH
• genotyp MeSH
• izoenzymy metabolismus   MeSH
• králíci MeSH
• lidé MeSH
• protozoální DNA analýza   MeSH
• restrikční mapování MeSH
• ribozomální DNA analýza   MeSH
• rohovka cytologie   parazitologie   patologie   MeSH
• virulence MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• izoenzymy MeSH
• protozoální DNA MeSH
• ribozomální DNA MeSH

This paper presents results of experiments on the effects of electromagnetic radiation in the millimeter range (frequency 34.0 +/- 0.1 GHz, power density 20 muW/cm2) emitted by a police radar device. Considering the physical properties of the radiation in millimeter range (skin effects), the experiments were carried out on hairless mice. The main physiological parameters tested were body mass, body temperature, peripheral blood, and mass and cellularity of several important organs. Critical organs, the skin, and cornea were examined by electron microscopy. Differentiation ability of hematopoietic cells, progenitors of granulocytes and macrophages, and DNA synthesis in the cornea were compared in irradiated and nonirradiated animals. None of the parameters tested was affected to an extent that would indicate the start of a pathological process or the risk of damage to genetic material.

• MeSH
• elektromagnetické jevy * MeSH
• femur cytologie   účinky záření   MeSH
• krev účinky záření   MeSH
• kůže účinky záření   MeSH
• myši bezsrsté MeSH
• myši MeSH
• policie * MeSH
• radar * MeSH
• rohovka cytologie   účinky záření   MeSH
• tělesná hmotnost účinky záření   MeSH
• tělesná teplota účinky záření   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

• MeSH
• dítě MeSH
• dospělí MeSH
• endotel cytologie   patologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mikroskopie metody   MeSH
• mladiství MeSH
• nemoci rohovky patologie   MeSH
• rohovka cytologie   patologie   MeSH
• senioři MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• anglický abstrakt MeSH
• časopisecké články MeSH

• MeSH
• endotel cytologie   MeSH
• králíci MeSH
• manganistan draselný škodlivé účinky   MeSH
• rohovka cytologie   účinky léků   MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• zvířata MeSH
• Publikační typ
• anglický abstrakt MeSH
• časopisecké články MeSH
• Názvy látek
• manganistan draselný MeSH

• MeSH
• hojení ran * MeSH
• jizva patofyziologie   MeSH
• králíci MeSH
• poranění oka patofyziologie   MeSH
• poranění rohovky MeSH
• rohovka cytologie   MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• zvířata MeSH
• Publikační typ
• anglický abstrakt MeSH
• časopisecké články MeSH