The identification and characterization of a novel protein, c19orf10, in the synovium

Joint inflammation and destruction have been linked to the deregulation of the highly synthetic fibroblast-like synoviocytes (FLSs), and much of our current understanding of the mechanisms that underlie synovitis has been collected from studies of FLSs. During a proteomic analysis of FLS cells, we identified a novel protein, c19orf10 (chromosome 19 open reading frame 10), that was produced in significant amounts by these cells. The present study provides a partial characterization of c19orf10 in FLSs, synovial fluid, and the synovium. Murine monoclonal and chicken polyclonal antibodies were produced against recombinant human c19orf10 protein and used to examine the distribution of c19orf10 in cultured FLSs and in synovial tissue sections from patients with rheumatoid arthritis or osteoarthritis. The intracellular staining pattern of c19orf10 is consistent with localization in the endoplasmic reticulum/Golgi distribution. Sections of rheumatoid arthritis and osteoarthritis synovia expressed similar patterns of c19orf10 distribution with perivascular and synovial lining staining. Double-staining in situ analysis suggests that fibroblast-like synovial cells produced c19orf10, whereas macrophages, B cells, or T cells produced little or none of this protein. There is evidence of secretion into the vascular space and the extracellular matrix surrounding the synovial lining. A competitive enzyme-linked immunosorbent assay confirmed the presence of microgram levels of c19orf10 in the synovial fluids of patients with one of various arthropathies. Collectively, these results suggest that c19orf10 is an FLS-derived protein that is secreted into the synovial fluid. However, the significance of this protein in synovial biology remains to be determined. The absence of known structural motifs or domains and its relatively late evolutionary appearance raise interesting questions about its function

The synovial proteome: analysis of fibroblast-like synoviocytes

The present studies were initiated to determine the protein expression patterns of fibroblast-like synovial (FLS) cells derived from the synovia of rheumatoid arthritis patients. The cellular proteins were separated by two-dimensional polyacrylamide gel electrophoresis and the in-gel digested proteins were analyzed by matrix-assisted laser desorption ionization mass spectrometry. A total of 368 spots were examined and 254 identifications were made. The studies identified a number of proteins that have been implicated in the normal or pathological FLS function (e.g. uridine diphosphoglucose dehydrogenase, galectin 1 and galectin 3) or that have been characterized as potential autoantigens in rheumatoid arthritis (e.g. BiP, colligin, HC gp-39). A novel uncharacterized protein product of chromosome 19 open reading frame 10 was also detected as an apparently major component of FLS cells. These results demonstrate the utility of high-content proteomic approaches in the analysis of FLS composition

Dihydrolipoamide dehydrogenase from porcine heart catalyzes NADH-dependent scavenging of nitric oxide

Abstract Dihydrolipoamide dehydrogenase (DLDH; EC 1.8.1.4) from porcine heart is capable of using nitric oxide (NO) as an electron acceptor, with NADH as the electron donor, forming nitrate in the reaction. NADPH was not effective as an electron donor. The reaction had a pH optimum near 6 and was not inhibited by cyanide or diphenyleneiodonium ions. The K m for NADH was 10 lM, while that for NO was 0.5 lM. The rate of NO conversion was comparable to the rate of lipoamide conversion (200 lmol min À1 mg À1 protein at pH 6). Cytochrome c or myoglobin were poor electron acceptors by themselves but, in the presence of methylene blue, DLDH had an activity of 5-7 lmol min À1 mg À1 protein with these substrates, indicating that DLDH can act also as a methemoglobin reductase. While the K m of DLDH for NO is relatively low, it is in the physiological range of NO levels encountered in the tissue. The enzyme may, therefore, have a significant role in modifying NO levels under specific cell conditions

Big Data Fusion Model for Heterogeneous Financial Market Data (FinDF)

The dawn of big data has seen the volume, variety, and velocity of data sources increase dramatically. Enormous amounts of structured, semi-structured and unstructured heterogeneous data can be garnered at a rapid rate, making analysis of such big data a herculean task. This has never been truer for data relating to financial stock markets, the biggest challenge being the 7 Vs of big data which relate to the collection, pre-processing, storage and real-time processing of such huge quantities of disparate data sources. Data fusion techniques have been adopted in a wide number of fields to cope with such vast amounts of heterogeneous data from multiple sources and fuse them together in order to produce a more comprehensive view of the data and its underlying relationships. Research into the fusing of heterogeneous financial data is scant within the literature, with existing work only taking into consideration the fusing of text-based financial documents. The lack of integration between financial stock market data, social media comments, financial discussion board posts and broker agencies means that the benefits of data fusion are not being realised to their full potential. This paper proposes a novel data fusion model, inspired by the data fusion model introduced by the Joint Directors of Laboratories, for the fusing of disparate data sources relating to financial stocks. Data with a diverse set of features from different data sources will supplement each other in order to obtain a Smart Data Layer, which will assist in scenarios such as irregularity detection and prediction of stock prices

NATO Advanced Research Workshop on Methods and Mechanisms for Producing Ions from Large Molecules

A NATO Advanced Research Workshop on Methods and Mechanisms for Producing Ions from Large Molecules was held at Minaki Lodge, Minaki, Ontario, Canada, from 24 to 28 June 1990. The workshop was hosted by the time-of-flight group of the Department of Physics at the University of Manitoba, and was attended by 64 invited participants from around the world. Twenty-nine invited talks were given and 19 papers were presented as posters. Of the 48 contributions, 38 are included in these proceedings. The conference was organized to study the rapidly changing field of mass spectrometry of biomolecules. Particle-induced desorption (especially with MeV particles) has been the most effective method of producing molecular ions from biomolecules. An important part of the workshop was devoted to recent developments in this field, particularly to progress in understanding the fundamentals of the desorption process. In this respect, the meeting was similar to previous conferences in Marburg, FRG (1978); Paris, F (1980); Uppsala, S (1981); College Station, USA (1983,1984); Wangerooge, FRG (1986); Orsay, F (1988); Spiekeroog, FRG (1989); and to the IFOS series of meetings at Munster, FRG (1981,1983,1985,1987) and L6vAnger, S (1989). As in the most recent of these meetings, there was some emphasis on new developments, particularly cluster bombardment. A departure from the concentration on particle bombardment processes at this conference was inspired by the dramatic results obtained with two new methods for producing molecular ions from large molecules: matrix-assisted laser desorption and electrospray

Identification of a novel bond between a histidine and the essential tyrosine in catalase HPII of Escherichia coli

8 pages, 7 figures, 1 table.-- PMID: 9144772 [PubMed].-- PMCID: PMC2143697.A bond between the N delta of the imidazole ring of His 392 and the C beta of the essential Tyr 415 has been found in the refined crystal structure at 1.9 A resolution of catalase HPII of Escherichia coli. This novel type of covalent linkage is clearly defined in the electron density map of HPII and is confirmed by matrix-assisted laser desorption/ionization mass spectrometry analysis of tryptic digest mixtures. The geometry of the bond is compatible with both the sp3 hybridization of the C beta atom and the planarity of the imidazole ring. Two mutated variants of HPII active site residues, H128N and N201H, do not contain the His 392-Tyr 415 bond, and their crystal structures show that the imidazole ring of His 392 was rotated, in both cases, by 80 degrees relative to its position in HPII. These mutant forms of HPII are catalytically inactive and do not convert heme b to heme d, suggesting a relationship between the self-catalyzed heme conversion reaction and the formation of the His-Tyr linkage. A model coupling the two processes and involving the reaction of one molecule of H2O2 on the proximal side of the heme with compound 1 is proposed.n. Thwiso rk was supported by an operating grant from the Natural Sciences and Engineering Research Council (NSERC (OGP9600) to P.C.L. and a postgraduate scholarship from NSERC to A.H. Work in Barcelona was supported by grants PB92-0707 and PB95-0218 to I.F. J.B. was the recipient of a doctoral fellowship from the Generalitat de Catalunya. Data collection in Hamburg was supporbteyd the Human Capital Mobility Project on contract CHGE-CT93-0040Peer reviewe