Background: Hyperbaric oxygen (HBO2) therapy can have a positive effect on wound healing, angiogenesis and blood flow. No prior study has described the effects of HBO2 therapy and gene expression of this process. The goal of our research was to show the effects of HBO2 and its impact at the molecular level on angiogenesis, proliferation, differentiation, oxidative stress, inflammation, and extracellular matrix formation. Live animal subjects were used for simulating the process of wound healing under standard conditions and under the influence of HBO2. Methods: Two experimental groups were created using injured rabbits (N=24), one group (N=12) treated with hyperbaric therapy twice a day and one (N=12) with standard wound care management. Wounds were surgical, uninfected, and in healthy animal test subjects. We compared the whole genomic analysis of the transcriptome with the use of microarray technology at three intervals during treatment. Results: The induction of the wounds in rabbit skin increased expression of hundreds of genes in both treatment groups. The numbers of elevated and decreased genes gradually reduced as the wound healed. Gene expression analysis showed elevated expression of several genes associated with inflammation in both groups of injured animals. Genes connected to the process of angiogenesis, proliferation, differentiation, oxidative stress and extracellular matrix formation were without statistically significant changes. Conclusion: The evidence did not support that HBO2 had any significant effect on gene expression during wound healing. Additionally, there was no evidence to support that there were changes in gene expression in either treatment group.
- MeSH
- chirurgická rána genetika terapie MeSH
- čipová analýza tkání metody MeSH
- exprese genu * MeSH
- hojení ran genetika MeSH
- hyperbarická oxygenace * MeSH
- králíci MeSH
- kůže zranění MeSH
- messenger RNA analýza MeSH
- zvířata MeSH
- Check Tag
- králíci MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
The urokinase-type plasminogen activator (uPA) and PA inhibitor 1 (PAI-1) play important roles in breast cancer metastasis through cell migration and invasion. They are clinically applicable prognostic and predictive markers. High levels of uPA and PAI-1 are associated with high risk of recurrence and adjuvant chemotherapy provides substantial benefit for this breast cancer population. The current sole validated method for quantifying uPA level in breast tumour tissue is ELISA assay. It requires 50–300 mg of fresh or frozen tissue, which is the main limitation for routine use. In this study, we evaluated the performances of customized antibody microarray to quantify uPA concentration from reduced extraction solution of breast tumour tissue and compared it with standard ELISA kit. We firstly optimized the elaboration of customized antibody microarray in order to sensitively detect and quantify uPA standard solutions. In the best conditions, we analysed uPA concentration in 16 cytosolic extracts from breast tumour tissue. Results showed that our customized antibody microarray could correctly quantify uPA concentration while consuming 100 times less volume of tumour tissue extraction solution than ELISA. Our antibody microarray is a powerful and promising tool for the miniaturization of the immunoassay quantification of uPA from breast tumour tissue extracts.
- MeSH
- aktivátor plazminogenu urokinázového typu * analýza imunologie škodlivé účinky MeSH
- biologické markery MeSH
- čipová analýza tkání * metody přístrojové vybavení MeSH
- ELISA MeSH
- imunoanalýza metody MeSH
- lidé MeSH
- metastázy nádorů MeSH
- nádory prsu * diagnóza imunologie MeSH
- prognóza MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
- srovnávací studie MeSH
The successful development and characterization of human induced pluripotent stem cells (iPSCs) provides a powerful tool to study the molecular mechanisms that control cell fate decisions and differentiation toward distinct lineages. Here we focus on the ability of donors derived iPSCs to differentiate toward hematopoietic progenitor cells and on the analysis of their telomere length. The ability to screen telomere length in individual donors is important for defining cellular senescence, which correlates with their differentiation potential toward hematopoietic lineages. We have modified iPSC culture protocol and telomere length analysis to suit for high throughput screening of telomere length in large number of individual donors. This approach can be used to demonstrate the heterogeneity or changes of telomere length and its shortening as an exclusion criterion for selection of suitable donors for future stem cell therapies.
- MeSH
- biologické markery * MeSH
- buněčné kultury * MeSH
- čipová analýza tkání metody MeSH
- hematopoetické kmenové buňky cytologie metabolismus MeSH
- indukované pluripotentní kmenové buňky cytologie metabolismus MeSH
- lidé MeSH
- rychlé screeningové testy * MeSH
- stárnutí buněk * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
AIM: To identify tissue biomarkers that are predictive of the therapeutic effect of sunitinib in treatment of metastatic clear cell renal cell carcinoma (mCRCC). MATERIALS AND METHODS: Our study included 39 patients with mCRCC treated with sunitinib. Patients were stratified into two groups based on their response to sunitinib treatment: non-responders (progression), and responders (stable disease, regression). The effect of treatment was measured by comparing imaging studies before the initiation treatment with those performed at between 3rd and 7th months of treatment, depending on the patient. Histological samples of tumor tissue and healthy renal parenchyma, acquired during surgery of the primary tumor, were examined with immunohistochemistry to detect tissue targets involved in the signaling pathways of tumor growth and neoangiogenesis. We selected mammalian target of rapamycine, p53, vascular endothelial growth factor, hypoxia-inducible factor 1 and 2 and carbonic anhydrase IX. We compared the average levels of biomarker expression in both, tumor tissue, as well as in healthy renal parenchyma. Results were evaluated using the Student's t-test. RESULTS: For responders, statistically significant differences in marker expression in tumor tissue versus healthy parenchyma were found for mTOR (4%/16.7%; p=0.01031), p53 (4%/12.7%; p=0.042019), VEGF (62.7%/45%; p=0.019836) and CAIX (45%/15.33%; p=0.001624). A further significant difference was found in the frequency of high expression (more than 60%) between tumor tissue and healthy parenchyma in VEGF (65%/35%; p=0.026487) and CAIX (42%/8%; p=0.003328). CAIX was expressed at high levels in the tumor tissue in both evaluated groups. CONCLUSION: A significantly higher expression of VEGF in CRCC in comparison to healthy parenchyma can predict a better response to sunitinib.
- MeSH
- čipová analýza tkání metody MeSH
- dospělí MeSH
- imunoenzymatické techniky MeSH
- indoly terapeutické užití MeSH
- karcinom z renálních buněk farmakoterapie mortalita sekundární MeSH
- ledviny metabolismus patologie MeSH
- lidé středního věku MeSH
- lidé MeSH
- lymfatické metastázy MeSH
- míra přežití MeSH
- nádorové biomarkery metabolismus MeSH
- nádory ledvin farmakoterapie mortalita patologie MeSH
- následné studie MeSH
- prognóza MeSH
- protinádorové látky terapeutické užití MeSH
- pyrroly terapeutické užití MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- staging nádorů MeSH
- studie případů a kontrol MeSH
- stupeň nádoru 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
- práce podpořená grantem MeSH
- srovnávací studie MeSH
Interactions between a micro-magnet array and living cells may guide the establishment of cell networks due to the cellular response to a magnetic field. To manipulate mesenchymal stem cells free of magnetic nanoparticles by a high magnetic field gradient, we used high quality micro-patterned NdFeB films around which the stray field's value and direction drastically change across the cell body. Such micro-magnet arrays coated with parylene produce high magnetic field gradients that affect the cells in two main ways: i) causing cell migration and adherence to a covered magnetic surface and ii) elongating the cells in the directions parallel to the edges of the micro-magnet. To explain these effects, three putative mechanisms that incorporate both physical and biological factors influencing the cells are suggested. It is shown that the static high magnetic field gradient generated by the micro-magnet arrays are capable of assisting cell migration to those areas with the strongest magnetic field gradient, thereby allowing the build up of tunable interconnected stem cell networks, which is an elegant route for tissue engineering and regenerative medicine.
- MeSH
- buněčná adheze MeSH
- časové faktory MeSH
- čipová analýza tkání metody MeSH
- krysa rodu rattus MeSH
- kultivační média chemie MeSH
- magnetické pole MeSH
- magnety * MeSH
- mezenchymální kmenové buňky cytologie MeSH
- nanočástice MeSH
- pohyb buněk MeSH
- potkani Wistar MeSH
- viabilita buněk MeSH
- železité sloučeniny chemie MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Drug resistance is one of the reasons for chemotherapy failure in non-small-cell lung carcinoma (NSCLC). One of the major mechanisms of drug resistance is the inhibition of chemotherapy-induced apoptosis. Therefore, the study of novel cell death pathways could possibly enable us to overcome resistance to apoptosis in NSCLC. One of the non-caspase types of cell death is autophagy. BNIP3 protein, a Bcl-2 family member, highly expressed in some tumours, plays a key role in the induction of autophagy. In the present study, we investigated the immunohistochemical expression and subcellular localization of BNIP3 in a series of early- and late-stage non-small-cell lung carcinomas and normal bronchial tissues, and correlated this expression with the occurrence of metastasis and survival. BNIP3 was strongly expressed in the nucleus of cancer cells in 16/79 (20.3%) cases. This BNIP3 positivity did not correlate with histological grade, stage, histology type, metastatic potential, or expression of BNIP3 according to median values. No significant correlation was observed between the expression of BNIP3 and the overall survival of NSCLC patients (p = 0.55). Nor did we find any significant correlation between BNIP3 expression and the occurrence of site-specific metastasis (p = 0.85).
- MeSH
- buněčné jádro chemie MeSH
- čipová analýza tkání metody MeSH
- imunohistochemie MeSH
- lidé středního věku MeSH
- lidé MeSH
- membránové proteiny analýza MeSH
- nádory plic chemie mortalita MeSH
- nemalobuněčný karcinom plic chemie mortalita MeSH
- protoonkogenní proteiny analýza MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Úvod: I když je vrozený defekt femuru (PFFD) dobře charakterizován morfologicky, změny na molekulární úrovni nejsou v literatuře popsány. Tento informační nedostatek společně s neznámou etiologií vady nás vedl k myšlence analýzy procesů angiogeneze a osteogeneze v tkáni pakloubu pacientů s PFFD v porovnání s fyziologickou kostí. Očekávali jsme rozdíly v genové expresi, zejména kvantitativní. Materiál a metodika: Během plánovaných ortopedických výkonů jsme odebrali kostní bloček, vložili do prezervačního média (RNA stabilization reagent), které brání degradaci RNA, a hluboce zmrazili. Poté jsme izolovali RNA a transkripční profil studovali biočipovou analýzou (SuperArray Bioscience Corporation). Celkem je tímto způsobem možno detekovat 113 genů osteogeneze a 113 genů angiogeneze. Od října roku 2005 do konce roku 2008 jsme vyšetřili vzorky od 7 pacientů s PFFD a 3 kontrolní vzorky. Některé analýzy jsme pro ověření výsledků opakovali, celkem jsme použili 13 čipů pro osteogenezi a 11 pro angiogenezi. Výsledky: Zaznamenali jsme rozdíly v kvantitě i typech exprimovaných genů. Některé geny byly exprimovány u PFFD více proti kontrolnímu vzorku (gen pro kalcitoninový receptor, kolagen XII, kolagen I alfa2, kolagen II, kolagen IX, FGFR2, fibronektin, integrin), jiné naopak vykazovaly sníženou expresi (gen pro annexin A5, kolagen XVIII alfa 1, kolagen I alfa1, katepsin K, FGFR1, FGFR3, IGF2, VEGFB). Závěr: Naše původní předpoklady se potvrdily rozdílnou genovou expresí. Prozatím získané výsledky nedovolují zobecnění, jsou pouze prvním krokem k dalším experimentům, kterými je třeba potvrdit avizované informace a propojit je s klinickými nálezy, jako je alternativní cévní zásobení postižené končetiny u některých pacientů.
