- MeSH
- Ameloblasts MeSH
- Extracellular Matrix Proteins MeSH
- Research Support as Topic MeSH
- Cloning, Molecular MeSH
- Humans MeSH
- Sequence Analysis MeSH
- Check Tag
- Humans MeSH
- Publication type
- Comparative Study MeSH
- Geographicals
- Czech Republic MeSH
Matrix Gla protein (MGP) je cirkulující protein s nízkou molekulární hmotností, který působí jako přirozený inhibitor kalcifikace. Řadíme ho do skupiny proteinů označených jako vitamin K dependentní proteiny. Jeho hlavní funkcí je prevence ukládání vápníku do měkkých tkání. Pro správné fungování musí být MGP dostatečně karboxylován a fosforylován, což je proces, který vyžaduje vitamin K2 jako kofaktor gamma-glutamylkarboxylasy. Nedostatek vitaminu K se projevuje kardiovaskulárním onemocněním, nedostatečnou mineralizací kostní tkáně a vznikem kalcifikujících deposit v měkkých tkáních. Cílem práce bylo navržení metod pro sledování hladiny MGP a defosforylovaného nedostatečně karboxylovaného MGP (dp-uc MGP) za využití imunochemické detekce a aplikovat je na analýzu reálných pacientských vzorků pro použití v klinických laboratořích, kde by tyto markery mohly sloužit jako potencionální markery závažnosti kloubních onemocnění, kardiovaskulárních onemocnění a také jako markery vypovídající o stavu vitaminu K v organismu.
Matrix Gla protein (MGP) is a circulating protein with a low molecular weight that acts as a natural inhibitor of calcification. It belongs to the group of vitamin K‐dependent proteins. For its proper function, MGP must undergo vitamin K-dependent carboxylation and phosphorylation. Its main function is the prevention of soft-tissue calcification. It requires vitamin K2 as a cofactor for gamma glutamyl carboxylase. Vitamin K insufficiency is manifested by cardiovascular diseases, insufficient mineralization of bone tissue and the formation of calcifying deposits in soft tissues. The aim of this study was to design methods for monitoring the levels of MGP and dp-uc MGP by using immunochemical detection and to apply them to the analysis of real samples for use in clinical laboratories. These markers could serve as potential markers of the severity of joint diseases, cardiovascular diseases and also as markers indicating the status of vitamin K in organism.
- MeSH
- Adult MeSH
- Endometrium physiology MeSH
- In Situ Hybridization methods MeSH
- Humans MeSH
- Matrix Metalloproteinases physiology MeSH
- Menstrual Cycle MeSH
- Specimen Handling MeSH
- Gene Expression Profiling MeSH
- Check Tag
- Adult MeSH
- Humans MeSH
- Female MeSH
- Publication type
- Review MeSH
- Comparative Study MeSH
Východisko. Matrix metaloproteázy (MMPs) patří mezi proteolytické enzymy. Jednou z jejich funkcí je i štěpení bazálních membrán buněk a extracelulární matrix. U maligních nádorů se tak mohou uplatnit v procesu invazivity a metastázování. Vznikají převážně ve stromálních buňkách (fibroblastech a endoteliích) reaktivně na přítomnost nádorových buněk. Ve vztahu k NSCLC jsou nejčastěji zmiňovány MMP-2 (gelatináza A), MMP-9 (gelatináza B) a především MMP-11 (stromelysin 3). Metody a výsledky. Zkoumali jsme vztah mezi expresí výše zmíněných matrix metaloproteáz pouze ve stromálních buňkách a 5letým přežitím u 80 nemocných po kurativní resekci pro NSCLC ve stadiu I dle TNM. Exprese MMP-2 byla asociována s 5letým přežitím, ale bez signifikantní korelace. Žádná korelace nebyla zjištěna u MMP-9. Statisticky téměř významný vztah byl zjištěn mezi expresí MMP-11 a 5letým přežitím. Závěry. Exprese MMP-11 ve stromálních buňkách kurativně zresekovaných pacientů s NSCLC v I. stadiu může být užitečná v predikci jejich prognózy.
Background. Matrix metalloproteinases (MMPs) belong to proteolytic enzymes. Degradation of the cell basement membrane and the extracellular matrix is one of their functions. In malignant tumors they can hypothetically contribute to the invasion and metastasis formation. They are mostly produced by stromal cells (fibroblasts and endothelial cells) as a response to the presence of tumor cells. MMP-2 (gelatinase A), MMP-9 (gelatinase B) and MMP-11 (stromelysin 3) are often mentioned in regard to Non-small Cell Lung Cancer (NSCLC). Methods and Results. The relation between the expression of the above-mentioned matrix metalloproteinases in stromal cells and the cancer-related survival in 80 patients after curative resection of NSCLC in stage I according to TNM was studied. The expression of MMP-2 was associated with cancer-related survival but without significant correlation. No correlation was found in MMP-9. There was a statistically near-significant relation between the expression of MMP-11 and cancer-related survival. Conclusions. The expression of MMP-11 in stromal cells in surgically treated NSCLC patients in stage I appears useful for evaluation of their prognosis.
Zymography is an electrophoretic method in which proteins are separated in a polyacrylamide gel in the presence of sodium dodecyl sulfate (SDS-PAGE). This method is used for the detection of enzymatic activity and molecular characterization of proteins. In contrast to the standard SDS-PAGE method, a substrate is incorporated into the gel during zymography, which is subsequently cleaved by target proteases. Many studies have focused on the development and progression of inflammatory diseases affecting the gastrointestinal tract, emphasizing the role of the largest group of proteases, matrix metalloproteinases (MMPs). The most used classification of this group of enzymes (by researchers in MMP biology) is based in part on the historical evaluation of the substrate specificity of MMPs and in part on the cellular localization of MMPs. MMPs are thus classified into the groups of collagenases, gelatinases, stromelysins, matrilysins, membrane-type MMPs (MT-MMPs), and others. An important group of MMPs are gelatinases which are involved in the breakdown of collagen type IV and gelatin of extracellular matrix and participate in the regulation of various physiological or pathological processes such as morphogenesis, angiogenesis, tissue repair, cirrhosis, arthritis, and metastasis. The present study's objective was to determine the amount of active MMP-9 and MMP-2 forms in tissue samples using zymography. The patient group was according to histology findings divided into the benign tumor (control) group (8 patients), and the malignant tumor group (24 patients). The respondents in the malignant tumor group were further divided according to the standard TNM classification. The results of this study confirmed that MMP-2, unlike MMP-9, can be used as a prognostic biomarker of CRC, because only the expression of active MMP-2 confirmed statistically significant differences between individual stages of CRC. Moreover, MMP-2 seems to play a more important role in higher stages of CRC. Substantial disparities in the determination of active MMPs between the observed groups support the assumption for the integration of zymography into clinical diagnostics of CRC together with molecular and other studies.
- MeSH
- Extracellular Matrix metabolism MeSH
- Colorectal Neoplasms * diagnosis metabolism MeSH
- Humans MeSH
- Matrix Metalloproteinase 2 * metabolism MeSH
- Matrix Metalloproteinase 9 MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
Závěrečná zpráva o řešení grantu Interní grantové agentury MZ ČR
51 l., [31] l. příl. : il., tab. ; 31 cm
Cílem práce je analýza aktivity MMP-2,MMP-9 a jejich inhibitorů a hodnotit vztah k aktivitě, formě choroby, tíži klinického postižení a ovlivnění léčbou.Určit asociační vztah genetických polymorfismů lokalizovaných v kandidátních genech s MS. Soubor budetvořen asi 80-ti pacienty s roztroušenou sklerózou dle Mc Donaldových kriterií jistou, kontrolní soubor bude složen z 50-ti zdravých dobrovolníků. Stanovení hladin MMP-2,MMP-9,TIMP-2 a TIMP-9 v periferní krvi bude provedeno metodou ELISA. Hladiny budouměřeny na počátku sledovaného období, dále vždy při atace a na konci sledovaného období. Genotypizace MMPs bude provedena metodami molekulární biologie.Bude odebráno 10 ml periferní krve s následnou izolací DNA a RNA Bude použita metoda PCR, restrikční analýza a heteroduplexová analýza.MMPs jsou slibným biologickým markerem, který by umožňoval laboratorní monitoraci onemocnění a hodnocení efektu léčby u pacientů s RS.; The aim of this study is to analyse the activity of MMP-2, MMP-9 and their inhibitors in MS and to assess their relationship to the disease activity, its form, and any correlation with the clinical disability and therapeutic influences and further to toexamine the possible association between genetic polymorphism in candidate genes with multiple sclerosis.The study is to include 80 MS patients and 50 healthy controls. MMP-2, MMP-9, TIMP-2 TIMP-1 serum levels will be quantified by ELISA. Serum levels will beassessed at the beginning of the study, during exacerbations and at the end of the study. Genetic analysis will be performed by standard molecular biology practice.DNA and RNA will be isolated from blood.The PCR method,restrict and heteroduplex analysis are to be employed. MMPs are a promising biological marker that enables us to assess the activity of the disease and to monitor the effects of treatment.
- MeSH
- Enzyme-Linked Immunosorbent Assay MeSH
- Genotype MeSH
- Immunogenetic Phenomena MeSH
- Matrix Metalloproteinase Inhibitors MeSH
- Matrix Metalloproteinase 2 analysis blood MeSH
- Matrix Metalloproteinase 9 analysis blood MeSH
- Polymerase Chain Reaction MeSH
- Polymorphism, Genetic MeSH
- Multiple Sclerosis diagnosis therapy MeSH
- Conspectus
- Biochemie. Molekulární biologie. Biofyzika
- NML Fields
- neurologie
- biologie
- NML Publication type
- závěrečné zprávy o řešení grantu IGA MZ ČR
Matrix-assisted laser desorption/ionisation (MALDI) of small molecules is challenging and in most cases impossible due to interferences from matrix ions precluding analysis of molecules <300-500 Da. A common matrix such as ferulic acid belongs to an important class of compounds associated with antioxidant activity. If the shared phenolic structure is related to the propensity as an active MALDI matrix then it follows that direct laser desorption/ionisation should be possible for polyphenols. Indeed matrix-less laser desorption/ionisation mass spectrometry is achieved whereby the analyte functions as a matrix and was used to monitor low molecular weight compounds in wine samples. Sensitivity ranging from 0.12-87 pmol/spot was achieved for eight phenolic acids (4-coumaric, 4-hydroxybenzoic, caffeic, ferulic, gallic, protocatechuic, syringic, vanillic) and 0.02 pmol/spot for trans-resveratrol. Additionally, 4-coumaric, 4-hydroxybenzoic, caffeic, ferulic, gallic, syringic, vanillic acids and trans-resveratrol were identified in wine samples using accurate mass measurements consistent with reported profiles based on liquid chromatography (LC)/MS. Minimal sample pre-treatment make the technique potentially appropriate for fingerprinting, screening and quality control of wine samples. Copyright (c) 2009 John Wiley & Sons, Ltd.
This study is focused on the analysis of extracellular DNA (eDNA) from a biofilm matrix formed by Staphylococcus aureus, Listeria monocytogenes, and Salmonella enterica. The presence of eDNA in the biofilm of all the studied strains was confirmed by confocal laser scanning microscopy using fluorescent dyes with high affinity to nucleic acid. The protocol for eDNA isolation from the biofilm matrix was established, and subsequent characterization of the eDNA was performed. The purified eDNA obtained from the biofilm matrix of all three microorganisms was compared to the genomic DNA (gDNA) isolated from relevant planktonic grown cells. The process of eDNA isolation consisted of biofilm cultivation, its collection, sonication, membrane filtration, dialysis, lyophilisation, and extraction of DNA separated from the biofilm matrix with cetyltrimethylammonium bromide. An amplified fragment length polymorphism (AFLP) was used for comparing eDNA and gDNA. AFLP profiles showed a significant similarity between eDNA and gDNA at the strain level. The highest similarity, with a profile concordance rate of 94.7% per strain, was observed for S. aureus, L. monocytogenes, and S. enterica exhibited lower profiles similarity (78% and 60%, respectively). The obtained results support the hypothesis that the eDNA of studied bacterial species has its origin in the gDNA.
