The detection of HPV infection and microbial colonization in cervical lesions is currently done through PCR-based viral or bacterial DNA amplification. Our objective was to develop a methodology to expand the metaproteomic landscape of cervical disease and determine if protein biomarkers from both human and microbes could be detected in distinct cervical samples. This would lead to the development of multi-species proteomics, which includes protein-based lateral flow diagnostics that can define patterns of microbes and/or human proteins relevant to disease status. In this study, we collected both non-frozen tissue biopsy and exfoliative non-fixed cytology samples to assess the consistency of detecting human proteomic signatures between the cytology and biopsy samples. Our results show that proteomics using biopsies or cytologies can detect both human and microbial organisms. Across patients, Lumican and Galectin-1 were most highly expressed human proteins in the tissue biopsy, whilst IL-36 and IL-1RA were most highly expressed human proteins in the cytology. We also used mass spectrometry to assess microbial proteomes known to reside based on prior 16S rRNA gene signatures. Lactobacillus spp. was the most highly expressed proteome in patient samples and specific abundant Lactobacillus proteins were identified. These methodological approaches can be used in future metaproteomic clinical studies to interrogate the vaginal human and microbiome structure and metabolic diversity in cytologies or biopsies from the same patients who have pre-invasive cervical intraepithelial neoplasia, invasive cervical cancer, as well as in healthy controls to assess how human and pathogenic proteins may correlate with disease presence and severity.
- MeSH
- biologické markery * analýza metabolismus MeSH
- biopsie MeSH
- cervix uteri * mikrobiologie patologie MeSH
- dospělí MeSH
- galektin 1 metabolismus analýza genetika MeSH
- Lactobacillus MeSH
- lidé MeSH
- lumican MeSH
- mikrobiota MeSH
- nádory děložního čípku patologie mikrobiologie MeSH
- proteomika * metody MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
N-Acetyllactosamine (LacNAc; Galβ4GlcNAc) is a typical disaccharide ligand of galectins. The most abundant members of these human lectins, galectin-1 (Gal-1) and galectin-3 (Gal-3), participate in a number of pathologies including cancerogenesis and metastatic formation. In this study, we synthesized a series of fifteen N-(2-hydroxypropyl)methacrylamide (HPMA)-based glycopolymers with varying LacNAc amounts and presentations and evaluated the impact of their architecture on the binding affinity to Gal-1 and Gal-3. The controlled radical reversible addition-fragmentation chain transfer copolymerization technique afforded linear polymer precursors with comparable molecular weight (Mn ≈ 22,000 g mol-1) and narrow dispersity (D̵ ≈ 1.1). The precursors were conjugated with the functionalized LacNAc disaccharide (4-22 mol % content in glycopolymer) prepared by enzymatic synthesis under catalysis by β-galactosidase from Bacillus circulans. The structure-affinity relationship study based on the enzyme-linked immunosorbent assay revealed that the type of LacNAc presentation, individual or clustered on bi- or trivalent linkers, brings a clear discrimination (almost 300-fold) between Gal-1 and Gal-3, reaching avidity to Gal-1 in the nanomolar range. Whereas Gal-1 strongly preferred a dense presentation of individually distributed LacNAc epitopes, Gal-3 preferred a clustered LacNAc presentation. Such a strong galectin preference based just on the structure of a multivalent glycopolymer type is exceptional. The prepared nontoxic, nonimmunogenic, and biocompatible glycopolymers are prospective for therapeutic applications requiring selectivity for one particular galectin.
- MeSH
- akrylamidy chemie MeSH
- aminocukry chemie MeSH
- Bacillus enzymologie MeSH
- beta-galaktosidasa metabolismus MeSH
- disacharidy chemická syntéza MeSH
- ELISA MeSH
- epitopy MeSH
- galektin 1 analýza metabolismus MeSH
- galektiny analýza metabolismus MeSH
- katalýza MeSH
- krevní proteiny analýza metabolismus MeSH
- magnetická rezonanční spektroskopie MeSH
- polymerizace MeSH
- polymery chemie metabolismus farmakologie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
BACKGROUND/AIM: Having previously initiated genome-wide expression profiling in head and neck squamous cell carcinoma (HNSCC) for regions of the tumor, the margin of surgical resecate (MSR) and normal mucosa (NM), we here proceed with respective analysis of cases after stratification according to the expression status of tenascin (Ten). MATERIALS AND METHODS: Tissue specimens of each anatomical site were analyzed by immunofluorescent detection of Ten, fibronectin (Fn) and galectin-1 (Gal-1) as well as by microarrays. RESULTS: Histopathological examination demonstrated that Ten+Fn+Gal-1+co-expression occurs more frequently in samples of HNSCC (55%) than in NM (9%; p<0.01). Contrary, the Ten-Fn+Gal-1-(45%) and Ten-Fn-Gal-1-(39%) status occurred with significantly (p<0.01) higher frequency than in HNSCC (3% and 4%, respectively). In MSRs, different immunophenotypes were distributed rather equally (Ten+Fn+Gal-1+=24%; Ten-Fn+Gal-1-=36%; Ten-Fn-Gal-1-=33%), differing to the results in tumors (p<0.05). Absence/presence of Ten was used for stratification of patients into cohorts without a difference in prognosis, to comparatively examine gene-activity signatures. Microarray analysis revealed i) expression of several tumor progression-associated genes in Ten+HNSCC tumors and ii) a strong up-regulation of gene expression assigned to lipid metabolism in MSRs of Ten-tumors, while NM profiles remained similar. CONCLUSION: The presented data reveal marked and specific changes in tumors and MSR specimens of HNSCC without a separation based on prognosis.
- MeSH
- fibronektiny genetika metabolismus MeSH
- galektin 1 genetika metabolismus MeSH
- genová ontologie MeSH
- lidé MeSH
- nádorové biomarkery genetika MeSH
- nádory hlavy a krku genetika metabolismus chirurgie MeSH
- přežití bez známek nemoci MeSH
- regulace genové exprese u nádorů * MeSH
- resekční okraje MeSH
- sliznice metabolismus MeSH
- spinocelulární karcinom genetika metabolismus chirurgie MeSH
- stanovení celkové genové exprese metody MeSH
- tenascin genetika metabolismus MeSH
- transkriptom * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
PURPOSE: To assess the impact of Acanthamoeba keratitis (AK) and amniotic membrane transplantation (AMT) in corneal explants on presence of two multifunctional endogenous lectins, i.e. galectins-1 and -7. METHODS: Ten corneal explants from AK patients (five with previous AMT and five controls without this treatment) and seven specimens of disease-free control cornea were processed by indirect fluorescent immunohistochemistry. RESULTS: Immunostaining for both galectins was obtained in the epithelium, stroma and the endothelial layer of all controls, with the strongest positivity in the epithelium. Significantly decreased intensity for galectin-1 was recorded in the epithelium of corneal explants from patients with AK and AMT. The signal for galectin-7 was significantly decreased in the epithelium of AK patients and normalized after AMT. CONCLUSIONS: AMT has a marked impact on presence of the two galectins in opposite directions, encouraging complete profiling for this family of endogenous effectors.
- MeSH
- akantamébová keratitida metabolismus chirurgie MeSH
- amnion transplantace MeSH
- biologické krytí * MeSH
- biologické markery metabolismus MeSH
- dospělí MeSH
- galektin 1 MeSH
- galektiny metabolismus MeSH
- imunohistochemie MeSH
- keratoplastika perforující metody MeSH
- lidé středního věku MeSH
- lidé MeSH
- oční infekce parazitární diagnóza metabolismus chirurgie MeSH
- rohovka metabolismus chirurgie MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- multicentrická studie MeSH
Galektíny patria medzi endogénne lektíny – bielkoviny špecificky rozpoznávajúce cukorné motívy. Galektíny hrajú významné úlohy v procesoch bunkovej proliferácie, diferenciácie, migrácie a tvorby medzibunkovej hmoty. Navyše sú schopné prenášať bunkové signály a podieľať sa na medzibunkových interakciách. Podobne bolo dokázané, že galektíny zohrávajú dôležitú úlohu pri tvorbe mikroprostredia nádoru a/alebo hojacej sa rany. Táto práca poskytuje prehľad experimentálnych a klinických štúdií zaoberajúcich sa biologickými úlohami galektínov v tkanivovej reparácii a jej paralele k raste nádoru.
Galectins are representatives of endogenous lectins – molecules specifically recognizing distinct sugar motifs. They play an important role in the processes of cell proliferation, differentiation, migration and extracellular matrix formation. Furthermore, galectins are able to transfer cellular signals and to participate in intercellular interaction. It has been proven that galectins play an important role in the formation of tumor and/or wound healing microenvironment. This review contains an overview of experimental and clinical studies dealing with biological roles of galectins in tissue repair and in its parallel – the tumor growth.
- MeSH
- exprese genu fyziologie MeSH
- galektin 1 fyziologie metabolismus MeSH
- galektin 3 fyziologie metabolismus MeSH
- galektiny * fyziologie klasifikace metabolismus MeSH
- hojení ran fyziologie MeSH
- lidé MeSH
- modely u zvířat MeSH
- nádory patofyziologie MeSH
- tkáně fyziologie metabolismus MeSH
- výzkum MeSH
- zánět patofyziologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- přehledy MeSH
Cancer-associated fibroblasts (CAFs) play a role in the progression of malignant tumors. They are formed by conversion of fibroblasts to smooth muscle α-actin-positive (SMA-positive) myofibroblasts. Polyamines are known to change the arrangement of the actin cytoskeleton by binding to the anionic actin. We tested the effect of the synthetic polyamine BPA-C8 on the transition of human dermal fibroblasts to myofibroblasts induced either by TGF-β1 alone or by TGF-β1 together with adhesion/growth-regulatory galectin-1. Pre-existing CAFs, myofibroblasts from pancreatitis, and rat smooth muscle cells were also exposed to BPA-C8. BPA-C8 impaired myofibroblast formation from activated fibroblasts, but it had no effect on cells already expressing SMA. BPA-C8 also reduced the occurrence of an extracellular matrix around the activated fibroblasts. The reported data thus extend current insights into polyamine activity, adding interference with tumor progression to the tumor-promoting processes warranting study.
- MeSH
- aktiny metabolismus MeSH
- fibroblasty účinky léků patologie MeSH
- galektin 1 metabolismus MeSH
- krysa rodu rattus MeSH
- kultivované buňky MeSH
- lidé MeSH
- myofibroblasty účinky léků patologie MeSH
- nádorové buňky kultivované MeSH
- nádory farmakoterapie metabolismus patologie MeSH
- polyaminy chemie farmakologie MeSH
- škára cytologie účinky léků MeSH
- transformující růstový faktor beta1 metabolismus MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
BACKGROUND/AIM: Stromal cells in the tumor microenvironment are primarily considered as sources of promalignant factors. The objective of our study was to define the effect of extracellular matrix (ECM) produced by normal dermal or cancer-associated fibroblasts exposed to adhesion/growth-regulatory lectin galectin-1 on human umbilical vein endothelial cells (HUVECs). MATERIALS AND METHODS: Fibroblasts were cultured for 10 days with lectin, followed by removing cellular constituents after an osmotic shock. Freshly-isolated HUVECs were placed on the ECM. In parallel, HUVECs were seeded on untreated and gelatin-coated surfaces as controls. A positive control for growth of HUVECs culture using medium supplemented with vascular endothelial growth factor completed the test panel. Cells were kept in contact to the substratum for two days and then processed for immunocytochemistry. RESULTS: HUVECs seeded on fibroblast-generated ECM presented a comparatively high degree of proliferation. Furthermore, contact to substratum produced by tumor-associated fibroblasts led to generation of a meshwork especially rich in fibronectin. CONCLUSION: Galectin-1 is apparently capable to trigger ECM production favorable for growth of HUVECs, prompting further work on characterizing structural features of the ECM and in situ correlation of lectin presence, ECM constitution and neoangiogenesis.
- MeSH
- endoteliální buňky pupečníkové žíly (lidské) fyziologie MeSH
- extracelulární matrix metabolismus MeSH
- fibroblasty fyziologie MeSH
- galektin 1 farmakologie MeSH
- kultivované buňky MeSH
- lidé MeSH
- nádorové mikroprostředí * MeSH
- proliferace buněk účinky léků MeSH
- vaskulární endoteliální růstový faktor A farmakologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
INTRODUCTION: One route of translating the information encoded in the glycan chains of cellular glycoconjugates into physiological effects is via receptor (lectin) binding. A family of endogenous lectins, sharing folding, a distinct sequence signature and affinity for β-galactosides (thus termed galectins), does so effectively in a context-dependent manner. AREAS COVERED: An overview is given on the multifunctional nature of galectins, with emphasis on galectin-1. The broad range of functions includes vital processes such as adhesion via glycan bridging, glycoconjugate transport or triggering signaling relevant, for example, for growth regulation. Besides distinct glycoconjugates, this lectin can also interact with certain proteins so that it can target counterreceptors at all sites of location, that is, in the cytoplasm and/or nucleus, at both sides of the membrane or extracellularly. Approaches to strategically exploit galectin activities with therapeutic intentions are outlined. EXPERT OPINION: The wide versatility of sugar coding and the multifunctionality of galectin-1 explain why considering to turn the protein into a therapeutic target is an ambitious aim. Natural pathways shaped by physiologic master regulators (e.g., the tumor suppressor p16(INK4a)) are suggested to teach inspiring lessons as to how the lectin might be recruited to clinical service.
- MeSH
- biologický transport účinky léků MeSH
- buněčná adheze účinky léků MeSH
- cílená molekulární terapie MeSH
- galektin 1 antagonisté a inhibitory metabolismus MeSH
- galektiny antagonisté a inhibitory metabolismus MeSH
- glykokonjugáty metabolismus MeSH
- lidé MeSH
- nádorové proteiny antagonisté a inhibitory metabolismus MeSH
- nádory farmakoterapie prevence a kontrola MeSH
- proliferace buněk účinky léků MeSH
- protinádorové látky farmakologie terapeutické užití MeSH
- racionální návrh léčiv * MeSH
- signální transdukce účinky léků MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
- Klíčová slova
- reparace, glykobiologie, mezibuněčné interakce,
- MeSH
- fibroblasty cytologie MeSH
- galektin 1 * aplikace a dávkování fyziologie sekrece MeSH
- galektin 3 * aplikace a dávkování fyziologie sekrece MeSH
- hojení ran * fyziologie MeSH
- kůže růst a vývoj zranění MeSH
- lidé MeSH
- mezibuněčné spoje MeSH
- myši zranění MeSH
- nádorová transformace buněk MeSH
- prospektivní studie MeSH
- regenerace MeSH
- techniky in vitro MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši zranění MeSH
- zvířata MeSH
- Publikační typ
- grafy a diagramy MeSH
Previously, we found that treatment of cutaneous wounds with Atropa belladonna L. (AB) revealed shortened process of acute inflammation as well as increased tensile strength and collagen deposition in healing skin wounds (Gál et al. 2009). To better understand AB effect on skin wound healing male Sprague-Dawley rats were submitted to one round full thickness skin wound on the back. In two experimental groups two different concentrations of AB extract were daily applied whereas the control group remained untreated. For histological evaluation samples were removed on day 21 after surgery and stained for wide spectrum cytokeratin, collagen III, fibronectin, galectin-1, and vimentin. In addition, in the in vitro study different concentration of AB extract were used to evaluate differences in HaCaT keratinocytes proliferation and differentiation by detection of Ki67 and keratin-19 expressions. Furthermore, to assess ECM formation of human dermal fibroblasts on the in vitro level fibronectin and galectin-1 were visualized. Our study showed that AB induces fibronectin and galectin-1 rich ECM formation in vitro and in vivo. In addition, the proliferation of keratinocytes was also increased. In conclusion, AB is an effective modulator of skin wound healing. Nevertheless, further research is needed to find optimal therapeutic concentration and exact underlying mechanism of action.
- MeSH
- Atropa belladonna chemie MeSH
- časové faktory MeSH
- extracelulární matrix metabolismus účinky léků MeSH
- fibroblasty metabolismus patologie účinky léků MeSH
- fibronektiny metabolismus MeSH
- galektin 1 metabolismus MeSH
- hojení ran účinky léků MeSH
- keratin-19 metabolismus MeSH
- keratinocyty metabolismus patologie účinky léků MeSH
- kolagen typ III metabolismus MeSH
- krysa rodu rattus MeSH
- kultivované buňky MeSH
- kůže metabolismus patologie účinky léků zranění MeSH
- lidé MeSH
- modely nemocí na zvířatech MeSH
- penetrující rány farmakoterapie metabolismus patologie MeSH
- potkani Sprague-Dawley MeSH
- rostlinné extrakty farmakologie chemie izolace a purifikace MeSH
- rozpouštědla chemie MeSH
- vimentin metabolismus MeSH
- voda chemie MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- lidé MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
