Surface expression
Dotaz
Zobrazit nápovědu
Acta pathologica, microbiologica et immunologica Scandinavica, ISSN 0108-0172 section C, 1985, suppl. no. 286, vol. 93
35 s. : tab.
- MeSH
- antigeny povrchové MeSH
- interferony MeSH
- leukocyty mononukleární MeSH
- Publikační typ
- vysokoškolské kvalifikační práce MeSH
- Konspekt
- Patologie. Klinická medicína
- NLK Obory
- alergologie a imunologie
- MeSH
- buněčné linie mikrobiologie ultrastruktura MeSH
- cytologické techniky metody MeSH
- lidé MeSH
- membránové proteiny MeSH
- radionuklidy MeSH
- techniky in vitro MeSH
- virus Epsteinův-Barrové MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- srovnávací studie MeSH
Sublethal concentrations (sub-MICs) of certain disinfectants are no longer effective in removing biofilms from abiotic surfaces and can even promote the formation of biofilms. Bacterial cells can probably adapt to these low concentrations of disinfectants and defend themselves by way of biofilm formation. In this paper, we report on three Staphylococcus aureus biofilm formers (strong B+++, moderate B++, and weak B+) that were cultivated with sub-MICs of commonly used disinfectants, ethanol or chloramine T, and quantified using Syto9 green fluorogenic nucleic acid stain. We demonstrate that 1.25-2.5% ethanol and 2500 μg/mL chloramine T significantly enhanced S. aureus biofilm formation. To visualize differences in biofilm compactness between S. aureus biofilms in control medium, 1.25% ethanol, or 2500 μg/mL chloramine T, scanning electron microscopy was used. To describe changes in abundance of surface-exposed proteins in ethanol- or chloramine T-treated biofilms, surface proteins were prepared using a novel trypsin shaving approach and quantified after dimethyl labeling by LC-LTQ/Orbitrap MS. Our data show that some proteins with adhesive functions and others with cell maintenance functions and virulence factor EsxA were significantly upregulated by both treatments. In contrast, immunoglobulin-binding protein A was significantly downregulated for both disinfectants. Significant differences were observed in the effect of the two disinfectants on the expression of surface proteins including some adhesins, foldase protein PrsA, and two virulence factors.
- MeSH
- bakteriální proteiny metabolismus MeSH
- biofilmy účinky léků růst a vývoj MeSH
- dezinficiencia aplikace a dávkování MeSH
- druhová specificita MeSH
- membránové proteiny metabolismus MeSH
- potravinářská mikrobiologie * MeSH
- regulace genové exprese u bakterií účinky léků fyziologie MeSH
- Staphylococcus aureus klasifikace účinky léků metabolismus MeSH
- viabilita buněk účinky léků MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- Publikační typ
- časopisecké články MeSH
Východisko. Perspektivní léčba žen s karcinomy mléčné žlázy preparátem Herceptin zaměřeným proti membránovému proteinu nádorových buněk, erbB-2 (Her2/neu), zvyšuje nároky na přesné stanovení intenzity exprese erbB-2 proteinu. Zvýšená exprese erbB-2 proteinu je spojena se zvýšenou expresí genu c-erbB-2, nejčastěji s jeho amplifikací. Tento vztah není dostatečně ověřen pro klinicky aplikovanou diagnostiku. Imunohistochemická (IHC) detekce membránové exprese erbB-2 proteinu není vždy jednoznačně interpretovatelná. Cílem práce bylo vymezit vztah mezi expresí proteinu a počtem kopií c-erbB-2 genu a zjistit, zda ve sporných případech IHC detekce erbB-2 proteinu by vyšetření počtu kopií genu přineslo řešení pro indikaci k léčbě Herceptinem. Metody a výsledky. V pilotní studii jsme vyšetřili karcinomy mléčné žlázy u 34 pacientek v průměrném věku 59 let (medián 57 let), s rozpětím od 33 do 88 let. Ve všech vyšetřeních jsme získali tkáň z primární operace karcinomu bez předchozí onkologické léčby. Počet kopií genu c-erbB-2 jsme stanovili metodou fluorescenční in situ hybridizace (FISH). Membránovou expresi erbB-2 proteinu jsme zjišťovali imunohistochemicky (IHC). Výsledky jsme porovnali s gradem karcinomu, se stavem hormonálních receptorů, a s nálezem v resekovaných axilárních lymfatických uzlinách. Karcinomy jsme rozdělili podle počtu kopií c-erbB-2 genu do tří skupin. První skupina (7 případů) byla charakterizována více jak 10 kopiemi genu c-erbB-2 v jádrech nádorových buněk. Druhá skupina (9 případů) byla charakterizována stupněm amplifikace pod 10 kopií genu c-erbB-2. Třetí skupinu (18 případů) tvořily karcinomy bez amplifikace. Nejlépe koreloval stav genu c-erbB-2 s expresí erbB-2 proteinu u první skupiny karcinomů - silná amplifikace genu, silná exprese proteinu. Morfologický nález odpovídal u všech invazivnímu duktálnímu karcinomu, u 6 ze 7 nemocných grade 3. Receptory pro estrogen a progesteron byly negativní u většiny karcinomů této skupiny. U pěti ze sedmi nemocných v první skupině jsme prokázali metastázy do axilárních lymfatických uzlin. Ve druhé a třetí skupině karcinomů se slabou amplifikací genu byla exprese erbB-2 proteinu střední, slabá nebo negativní. Grade karcinomů a exprese hormonálních receptorů byly variabilní a nezjistili jsme signifikantní vztah k metastatickému postižení lymfatických uzlin v době diagnózy. Závěr. Invazivní duktální karcinomy mléčné žlázy se silnou amplifikací genu c-erbB-2 mají silnou expresi membránového proteinu erbB-2. Mezi amplifikací genu c-erbB-2 s počtem kopií genu nad 10 a silnou overexpresí proteinu erbB-2 je přímý vztah. Z pilotní studie vyplývá, že tyto karcinomy mají agresivnější charakter než karcinomy s nižším počtem kopií genu c-erbB-2 (méně jak 10 kopií), nebo karcinomy se dvěma signály pro gen c-erbB-2. Exprese membránového proteinu erbB-2 je u karcinomů se slabou amplifikací nebo bez amplifikace genu c-erbB-2 slabá nebo negativní. Slabá pozitivita nebo negativní IHC výsledek však může být způsoben technickými problémy (fixace tkáně a další). Proto stanovení počtu kopií genu c-erbB-2 je vhodné v případech slabšího nebo nejistého výsledku IHC stanovení intenzity exprese erbB-2 proteinu.
Background. Perspective therapy of women with breast carcinoma using Herceptin, a monoclonal antibody against a membrane protein of the tumor cells, erbB-2 (Her2/neu), requires a precise evaluation of the intensity of erbB-2 protein expression. Overexpression of the erbB-2 protein is associated with an overexpression of the c-erbB-2 gene, most frequently with the gene amplification. This relation is not sufficiently verified to be used in clinically applied diagnostics. Immunohistochemical (IHC) detection of the membrane expression of the erbB-2 protein is not always reliable. The aim of the study was to determine the relation of the protein expression and the c-erbB-2 gene copy number and to ascertain whether in cases with controversial IHC of erbB-2 protein investigation of the gene copy numbers would help to decide if the Herceptin treatment is indicated. Methods and Results. We investigated 34 patients with breast carcinoma in a pilot study. The average age of the patients was 59 years (median 57 years), with the range from 33 years to 88 years. In all cases we investigated the tissue from primary breast carcinomas prior to oncology treatment. The number of c-erbB-2 gene copies was evaluated using fluorescence in situ hybridization (FISH). The membrane expression of the erbB-2 protein was expression of hormonal receptors, and with the findings in the axillary lymph nodes. The carcinomas were divided according to the number of c-erbB-2 gene copies in three groups. The first group (7 patients) was characterized by more than 10 copies of c-erbB-2 gene in the tumor cell nuclei (strong amplification). In the second group (9 patients) there were carcinomas with less than 10 copies of the c-erbB-2 gene. The third group (18 patients) formed carcinomas without c-erbB-2 gene amplification. Best correlation of the c-erbB-2 gene status with the erbB-2 protein expression was found for the group 1 (strong gene amplification, strong protein expression). The morphology was that of invasive duct carcinoma, in 6 of 7 patients grade 3. Estrogen and progesterone receptors were negative in a majority of the patients. In five of the seven patients axillary lymph node metastases were detected. In group 2 and 3 with low c-erbB-2 gene copy numbers the erbB-2 protein expression was moderate, weak or negative. The patients had carcinomas of variable grades (predominantly grade 2) and had a variable expression of the hormonal receptors. No significant association with lymph node metastases was found. Conclusions. Invasive duct carcinoma of the breast with a strong amplification of the c-erbB-2 gene have a strong expression of the membrane protein erbB-2. There is a direct relation between amplification of the c-erbB-2 gene with more than 10 copy numbers and a strong overexpression of the erbB-2 protein. Our results show that breast carcinomas with a strong c-erbB-2 gene amplification have more aggressive features than carcinomas with a lower number of c-erbB-2 gene copies or carcinomas with two signals of c-erbB-2 gene. Expression of the membrane protein erbB-2 in a carcinoma with a low amplification or without the c-erbB-2 gene amplification is weak or negative. However, a weak positivity or negative IHC result may be caused by technical problems (e.g. fixation of the tissue). Therefore, the assessment of the copy numbers of the c-erbB-2 gene is a matter of choice in cases of weak and difficult to interpret results of erbB-2 protein detection.
BACKGROUND: Yeast infections are often connected with formation of biofilms that are extremely difficult to eradicate. An excellent model system for deciphering multifactorial determinants of yeast biofilm development is the colony biofilm, composed of surface ("aerial") and invasive ("root") cells. While surface cells have been partially analyzed before, we know little about invasive root cells. In particular, information on the metabolic, chemical and morphogenetic properties of invasive versus surface cells is lacking. In this study, we used a new strategy to isolate invasive cells from agar and extracellular matrix, and employed it to perform genome wide expression profiling and biochemical analyses of surface and invasive cells. RESULTS: RNA sequencing revealed expression differences in 1245 genes with high statistical significance, indicating large genetically regulated metabolic differences between surface and invasive cells. Functional annotation analyses implicated genes involved in stress defense, peroxisomal fatty acid β-oxidation, autophagy, protein degradation, storage compound metabolism and meiosis as being important in surface cells. In contrast, numerous genes with functions in nutrient transport and diverse synthetic metabolic reactions, including genes involved in ribosome biogenesis, biosynthesis and translation, were found to be important in invasive cells. Variation in gene expression correlated significantly with cell-type specific processes such as autophagy and storage compound accumulation as identified by microscopic and biochemical analyses. Expression profiling also provided indications of cell-specific regulations. Subsequent knockout strain analyses identified Gip2p, a regulatory subunit of type 1 protein phosphatase Glc7p, to be essential for glycogen accumulation in surface cells. CONCLUSIONS: This is the first study reporting genome wide differences between surface and invasive cells of yeast colony biofilms. New findings show that surface and invasive cells display very different physiology, adapting to different conditions in different colony areas and contributing to development and survival of the colony biofilm as a whole. Notably, surface and invasive cells of colony biofilms differ significantly from upper and lower cells of smooth colonies adapted to plentiful laboratory conditions.
Background:The intratumoural heterogeneity, often driven by epithelial-to-mesenchymal transition (EMT), significantly contributes to chemoresistance and disease progression in adenocarcinomas. Methods:We introduced a high-throughput screening platform to identify surface antigens that associate with epithelial–mesenchymal plasticity in well-defined pairs of epithelial cell lines and their mesenchymal counterparts. Using multicolour flow cytometry, we then analysed the expression of 10 most robustly changed antigens and identified a 10-molecule surface signature, in pan-cytokeratin-positive/EpCAM-positive and -negative fractions of dissociated breast tumours. Results:We found that surface CD9, CD29, CD49c, and integrin ß5 are lost in breast cancer cells that underwent EMT in vivo. The tetraspanin family member CD9 was concordantly downregulated both in vitro and in vivo and associated with epithelial phenotype and favourable prognosis. Conclusions:We propose that overall landscape of 10-molecule surface signature expression reflects the epithelial–mesenchymal plasticity in breast cancer.
- MeSH
- antigeny CD9 biosyntéza imunologie MeSH
- antigeny nádorové biosyntéza imunologie MeSH
- antigeny povrchové biosyntéza imunologie MeSH
- epitelo-mezenchymální tranzice imunologie MeSH
- genetická transkripce MeSH
- lidé MeSH
- metastázy nádorů MeSH
- nádorové biomarkery MeSH
- nádorové buněčné linie MeSH
- nádory prsu genetika imunologie patologie MeSH
- plasticita buňky imunologie MeSH
- přeprogramování buněk fyziologie MeSH
- průtoková cytometrie MeSH
- rychlé screeningové testy MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Class C G-protein coupled receptors form obligatory dimers. Metabotropic glutamate receptors (mGluRs) are found commonly as homodimers. Alternative splicing of mGluR1 gene results in vivo in the expression of a long variant mGluR1a and at least two short variants mGluR1b and d. The amino acid sequences diverge within their carboxyl-termini six amino acid residues following RRKK motif. This four basic residue sequence was shown to have pronounced impact on function and trafficking of the short variants, while for mGluR1a the long C-terminus reduces the effects caused by presence of the RRKK motif. Here we investigated consequences of interactions between long mGluR1a and short mGluR1b variants. Our results show that mGluR1a interferes with mGluR1b trafficking to the cell surface in HEK293 transfected cells. Expression of a mGlu1a mutant incapable of activating G-proteins with mGluR1b mutated in the glutamate binding site led to the formation of a functional heterodimer. Moreover, we show that swapping long mGluR1a and/or short mGluR1b C-termini with corresponding regions in chimerical GB1 and GB2 gamma-amino butyric acid b (GABAb) receptor subunits do not exclude heterodimerization. These data reveal that the C-terminal ends of mGluR1 do not control subunit association, such that mGluR1 dimers with two distinct C-termini can form and function properly.
- MeSH
- alternativní sestřih genetika MeSH
- exprese genu fyziologie MeSH
- fosfatasy metabolismus MeSH
- imunoprecipitace metody MeSH
- lidé MeSH
- mutageneze fyziologie MeSH
- receptory metabotropního glutamátu genetika klasifikace metabolismus MeSH
- terciární struktura proteinů fyziologie MeSH
- transfekce metody MeSH
- transformované buněčné linie fyziologie MeSH
- transport proteinů fyziologie MeSH
- vápník metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
