Valvular heart disease leads to ventricular pressure and/or volume overload. Pressure overload leads to fibrosis, which might regress with its resolution, but the limits and details of this reverse remodeling are not known. To gain more insight into the extent and nature of cardiac fibrosis in valve disease, we analyzed needle biopsies taken from the interventricular septum of patients undergoing surgery for valve replacement focusing on the expression and distribution of major extracellular matrix protein involved in this process. Proteomic analysis performed using mass spectrometry revealed an excellent correlation between the expression of collagen type I and III, but there was little correlation with the immunohistochemical staining performed on sister sections, which included antibodies against collagen I, III, fibronectin, sarcomeric actin, and histochemistry for wheat germ agglutinin. Surprisingly, the immunofluorescence intensity did not correlate significantly with the gold standard for fibrosis quantification, which was performed using Picrosirius Red (PSR) staining, unless multiplexed on the same tissue section. There was also little correlation between the immunohistochemical markers and pressure gradient severity. It appears that at least in humans, the immunohistochemical pattern of fibrosis is not clearly correlated with standard Picrosirius Red staining on sister sections or quantitative proteomic data, possibly due to tissue heterogeneity at microscale, comorbidities, or other patient-specific factors. For precise correlation of different types of staining, multiplexing on the same section is the best approach.

• Klíčová slova
• Collagen, Fibronectin, Fibrosis, Pressure overload, Valvular heart disease,
• MeSH
• aortální insuficience metabolismus   patologie   chirurgie   MeSH
• aortální stenóza * metabolismus   patologie   chirurgie   MeSH
• extracelulární matrix - proteiny * metabolismus   analýza   MeSH
• fibróza * metabolismus   patologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mezikomorová přepážka patologie   metabolismus   MeSH
• senioři MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• extracelulární matrix - proteiny * MeSH

The aim of our work was to confirm an immunohistochemical profile of routine markers of epithelial and neuroendocrine differentiation in eleven cases of Merkel cell carcinoma, as well as to study the expression of two markers of early phases of neuronal differentiation, namely reelin and class III beta-tubulin, markers which have not yet been studied in Merkel cell carcinomas. In all the investigated tumours the characteristic "dot-like" pattern of cytokeratin 20 immunoexpression, as well as negative immunostaining for cytokeratin 7 and thyroid transcription factor 1 (TTF-1) were disclosed; all the tumours showed neuroendocrine differentiation, expressing either neuron specific enolase (NSE) or chromogranin A(CgA), or both. An interesting finding was observed when the anti-cytokeratin monoclonal antibody MNF 116 was used. The characteristic "dot-like" pattern was detected in high proportion of tumours, including two samples of local recurrence of one of the carcinomas, where neoplastic cells have lost the expression of cytokeratin 20. The majority (91%) of Merkel cell carcinomas included in our group showed positive immunodetection of class III beta-tubulin when TU-20 antibody was used, while TuJ-1 immunostaining was surprisingly negative in all the investigated tumours. Detection of reelin was negative in almost all the studied Merkel cell carcinomas except for cases, where neoplastic cells revealed weak focal immunostaining in a minor portion of neoplastic cells.

• MeSH
• extracelulární matrix - proteiny analýza   MeSH
• imunohistochemie MeSH
• keratiny analýza   MeSH
• lidé středního věku MeSH
• lidé MeSH
• Merkelův nádor chemie   patologie   MeSH
• molekuly buněčné adheze neuronové analýza   MeSH
• nádorové biomarkery analýza   MeSH
• nádory kůže chemie   patologie   MeSH
• protein reelin MeSH
• proteiny nervové tkáně analýza   MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• serinové endopeptidasy analýza   MeSH
• tubulin analýza   MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• extracelulární matrix - proteiny MeSH
• keratiny MeSH
• molekuly buněčné adheze neuronové MeSH
• nádorové biomarkery MeSH
• protein reelin MeSH
• proteiny nervové tkáně MeSH
• RELN protein, human MeSH Prohlížeč
• serinové endopeptidasy MeSH
• TUBB3 protein, human MeSH Prohlížeč
• tubulin MeSH

BACKGROUND: Pentosidine, an advanced glycation end product, increasingly accumulates in articular cartilage with age, and contributes to the pathogenesis of osteoarthritis (OA). Increased pentosidine concentrations are associated with inflammatory disorders-for example, rheumatoid arthritis. OBJECTIVE: To compare pentosidine serum concentrations in patients with knee OA and in healthy volunteers and to determine a relationship between pentosidine and cartilage oligomeric matrix protein (COMP)-a marker of articular cartilage destruction. METHODS: Paired serum and synovial fluid samples were obtained by arthrocentesis from 38 patients with knee OA and from 38 healthy volunteers. Pentosidine concentration was measured by reverse phase high performance liquid chromatography with fluorescent detection and COMP was determined by sandwich ELISA. RESULTS: Significantly increased serum pentosidine (p<0.01) and COMP (p<0.05) levels were detected in the patients with OA compared with the control group. Serum pentosidine correlated significantly with synovial fluid pentosidine (p<0.001). Pentosidine in synovial fluid (p<0.05) and in serum (p<0.05) correlated significantly with synovial fluid COMP. Pentosidine and COMP concentrations did not correlate significantly with the radiological stage of the disease. CONCLUSION: Increased pentosidine serum concentration in patients with OA and its correlation with the cartilage destruction marker COMP in synovial fluid suggests that pentosidine may be important in OA pathology and is a new potential OA marker.

• MeSH
• arginin analogy a deriváty   analýza   krev   MeSH
• artróza kolenních kloubů krev   diagnostické zobrazování   metabolismus   MeSH
• biologické markery analýza   krev   MeSH
• chrupavkový oligomerní matrixový protein MeSH
• extracelulární matrix - proteiny analýza   MeSH
• glykoproteiny analýza   MeSH
• kloubní chrupavka chemie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• lysin analogy a deriváty   analýza   krev   MeSH
• matriliny MeSH
• průřezové studie MeSH
• radiografie MeSH
• senioři MeSH
• stupeň závažnosti nemoci MeSH
• synoviální tekutina chemie   MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• arginin MeSH
• biologické markery MeSH
• chrupavkový oligomerní matrixový protein MeSH
• extracelulární matrix - proteiny MeSH
• glykoproteiny MeSH
• lysin MeSH
• matriliny MeSH
• pentosidine MeSH Prohlížeč
• TSP5 protein, human MeSH Prohlížeč

The extracellular matrix (ECM) plays a critical role in influencing the biological behavior of brain tumors and the diagnostic detection of ECM components in ependymomas might be of prognostic value. In the present study we evaluated immunohistochemically the expression of a spectrum of ECM glycoproteins (tenascin, vitronectin, fibronectin, laminin, collagen types II, IV and VI) in a series of 36 pediatric intracranial ependymomas. The distribution of the ECM glycoproteins was evaluated both within the tumor tissue and at the tumor invasion front, and the prognostic value of the results was tested in a survival analysis. The expression of most of the ECM glycoproteins was associated only with blood vessels. Tenascin and vitronectin were found in a more diffuse pattern around the tumor cells and at the tumor invasion fronts of several cases. The progression-free survival was significantly decreased for patients with tenascin positive tumors (in any of the studied compartments) and for the tumors with vitronectin accumulation at their invasion fronts. In one ependymoma containing foci of cartilage with metaplastic ossification we demonstrated that collagen types II and VI and tenascin were present in ECM of both the cartilage and the ependymoma, and were accompanied by areas of necrosis and dystrophic calcifications. We suggest, that the rare simultaneous production of the specific ECM components might lead to the formation of chondroid areas in ependymomas. An abundant production of some ECM glycoproteins (tenascin and vitronectin) is present in a proportion of ependymomas and its immunohistochemical detection is of prognostic relevance.

• MeSH
• dítě MeSH
• ependymom patologie   MeSH
• extracelulární matrix - proteiny analýza   biosyntéza   MeSH
• imunohistochemie MeSH
• kojenec MeSH
• lidé MeSH
• mladiství MeSH
• nádory mozku patologie   MeSH
• následné studie MeSH
• předškolní dítě MeSH
• přežití bez známek nemoci MeSH
• prognóza MeSH
• tenascin analýza   biosyntéza   MeSH
• Check Tag
• dítě MeSH
• kojenec MeSH
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• předškolní dítě MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• extracelulární matrix - proteiny MeSH
• tenascin MeSH

BACKGROUND: Cartilage oligomeric matrix protein/thrombospondin 5 (COMP/TSP 5) is one of the most promising serologic markers with regard to an ability to prognose development of osteoarthritis (OA). Our aim was to map the epitopes of three monoclonal antibodies (mAb) to COMP and to develop and characterize a sandwich enzyme-linked immunosorbent assay (ELISA) for measuring COMP levels in human body fluids. METHODS: COMP was digested with trypsin and the NH(2)-terminal sequence of the fragments recognized by each of the mAbs was determined. Steric competition among the mAbs was tested with an antibody capture assay. A sandwich ELISA was developed using unlabeled mAb 16-F12 as a capture antibody, and mAb 17-C10 labeled with biotin as the second antibody. RESULTS: Epitopes of the three mAbs were mapped to three different domains within the COMP subunit (16-F12, NH(2)-terminal domain; 17-C10, EGF-like domain; 12-C4, COOH-terminal domain). These epitopes did not overlap. mAbs 17-C10 and 12-C4 yielded similar serum COMP results when used as the secondary antibodies. Serum COMP levels measured with the new sandwich ELISA using mAbs 16-F12 and 17-C10 correlated strongly with results based on an inhibition ELISA with mAb 17-C10 alone (r(2) = 0.836; P < 0.0001). We characterized the new sandwich ELISA with regards to inter- and intra-assay variability, the range of COMP levels that can be expected in human synovial fluids (SF) and sera (controls and OA and rheumatoid arthritis (RA) patients), and the day-to-day and diurnal variability of COMP levels in sera. CONCLUSIONS: We have developed and characterized a sandwich ELISA for COMP that is sensitive and yields highly reproducible COMP results upon analysis of human sera and synovial fluids.

• MeSH
• chrupavkový oligomerní matrixový protein MeSH
• ELISA metody   MeSH
• extracelulární matrix - proteiny analýza   imunologie   MeSH
• glykoproteiny analýza   imunologie   MeSH
• lidé MeSH
• mapování epitopu MeSH
• matriliny MeSH
• monoklonální protilátky imunologie   MeSH
• synoviální tekutina chemie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, U.S. Gov't, P.H.S. MeSH
• Názvy látek
• chrupavkový oligomerní matrixový protein MeSH
• extracelulární matrix - proteiny MeSH
• glykoproteiny MeSH
• matriliny MeSH
• monoklonální protilátky MeSH
• TSP5 protein, human MeSH Prohlížeč

The endoneurial extracellular matrix (ECM) is produced by Schwann cells and fibroblasts under the control of axons. Dorsal and ventral spinal roots contain different types of axons, but information is not available on differences in the composition of their ECM. A comparison was made of the intensity of immunofluorescence staining of chondroitin sulfate proteoglycan, fibronectin, tenascin and thrombospondin in the endoneurial ECM of rat dorsal and ventral spinal roots. Sections of dorsal and ventral roots were incubated simultaneously for indirect immunofluorescence detection of the epitopes studied. Brightness of immunofluorescence staining was assessed by computer-assisted image analysis using interactive segmentation of digitized images to select areas to be analyzed. Our results revealed quantitative differences in the composition of endoneurial ECM of spinal dorsal and ventral roots, probably due to the presence of different types of axons. The ECM composition of the endoneurium in dorsal and ventral roots may be related with the creation of extrinsic conditions that support differential regeneration of afferent and motor axons after injury.

• MeSH
• chondroitinsulfát proteoglykany analýza   MeSH
• extracelulární matrix - proteiny * analýza   MeSH
• extracelulární matrix chemie   MeSH
• fibronektiny analýza   MeSH
• fluorescenční protilátková technika nepřímá metody   MeSH
• krysa rodu Rattus MeSH
• míšní kořeny chemie   cytologie   MeSH
• periferní nervy chemie   cytologie   MeSH
• počítačové zpracování obrazu metody   MeSH
• potkani Wistar MeSH
• tenascin analýza   MeSH
• thrombospondiny analýza   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• chondroitinsulfát proteoglykany MeSH
• extracelulární matrix - proteiny * MeSH
• fibronektiny MeSH
• tenascin MeSH
• thrombospondiny MeSH

The dorsal and ventral spinal roots contain different types of axons. The endoneurial extracellular matrix (ECM) among them is produced by Schwann cells and fibroblasts under the control of the axons. Chondroitin sulfate proteoglycan, fibronectin, tenascin-C, and thrombospondin are common components of the endoneurial ECM involved in the normal function as well as regeneration of the peripheral nerve. The present paper demonstrates a comparison of immunofluorescence staining for chondroitin sulfate proteoglycan, fibronectin, tenascin-C, and thrombospondin in the endoneurium of the rat dorsal and ventral spinal roots. Sections through the dorsal and ventral roots were cut simultaneously and adhered to the same microscopic slide. They were incubated simultaneously and the intensity of immunofluorescence staining was assessed by computer-assisted image analysis using interactive segmentation of digitized pictures to select the areas of measurement. The measurement of the immunofluorescence brightness revealed that the endoneurium of the dorsal roots was immunostained for the studied molecules at a higher intensity than in the ventral roots. The results suggest quantitative differences of the endoneurial content of the spinal dorsal and ventral roots probably corresponding to the presence of various types of axons. On the other hand, the different concentration of ECM molecules in the endoneurium of dorsal and ventral roots might be related to the formation of extrinsic conditions differently supporting regeneration of afferent and motor axons after their injury.

• MeSH
• axony chemie   MeSH
• chondroitinsulfát proteoglykany analýza   MeSH
• diagnostické zobrazování metody   MeSH
• extracelulární matrix - proteiny analýza   MeSH
• fibronektiny analýza   MeSH
• fluorescenční protilátková technika MeSH
• imunohistochemie MeSH
• krysa rodu Rattus MeSH
• míšní kořeny chemie   MeSH
• periferní nervy chemie   MeSH
• potkani Wistar MeSH
• tenascin analýza   MeSH
• thrombospondiny analýza   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH
• Názvy látek
• chondroitinsulfát proteoglykany MeSH
• extracelulární matrix - proteiny MeSH
• fibronektiny MeSH
• tenascin MeSH
• thrombospondiny MeSH