Myocarditis: A Rheumatologic Perspective

Myocarditis is an uncommon complication in patients with autoimmune rheumatic diseases. The majority of cases refer to postmortem findings. The mechanism of myocardial damage in Connective Tissue Diseases (CTDs) depends on the pathophysiology of underlying disease. Systemic inflammation, impaired microvascular circulation and vasculitis affect myocardial remodeling process, cause repeated focal ischemia resulting in hypertrophy, fibrosis of myocardium, conductive system and thus reduced contractility. Additionally, immunological abnormalities, coexisting myositis and the degree of disease activity are predictors of myocarditis progression. Clinical manifestations range from subclinical to severe forms. Early recognition is important for institution of appropriate immunomodulatory therapy

Experimental uncertainty budget for concrete compressive strength test based on a multifactorial analysis

The objective of the study is to introduce an experimental un­cer­tainty budget process for concrete compressive strength test, based on a pro­to­col that incorporates effects of multiple factors significant for the measure­ment result. The proposed procedure is rather useful for laboratories seeking accreditation according to ISO/IEC 17025, in order to emphasize the contri­bu­tion of type A uncertainty estimations, rather than relying on type B estimations that are unable to address the correlation between those factors. Two indepen­dent experiments were performed. Experiment I is proposed as a simple, suit­ably designed, reproducibility trial for laboratories performing EN 12390 test method, i.e. when a specified nominal curing age is targeted, following experi­mental design on multiple uncertainty parameters. A sensitivity analysis was introduced based on a semi-empirical multifactorial regression model (experi­ment II) for concrete compressive strength as a function of specimen’s curing age and W/C ratio. The present study is an effort towards an integrated and standardized method for experimental, semi-empirical multifactorial re­gression estimation of the uncertainty budget for the EN 12390 test method, being useful, also, as a baseline for internal quality control programs when ad­justed for the specific characteristics of concrete specimens tested by a laboratory

Intermembrane crosstalk drives inner membrane protein organization in Escherichia coli

Gram-negative bacteria depend on energised protein complexes that connect the two membranes of the cell envelope. However, β-barrel outer-membrane proteins (OMPs) and α-helical inner-membrane proteins (IMPs) display quite different organisation. OMPs cluster into islands that restrict their lateral mobility, while IMPs generally diffuse throughout the cell. Here, using live cell imaging of Escherichia coli, we demonstrate that when transient, energy-dependent transmembrane connections are formed, IMPs become subjugated by the inherent organisation of OMPs and that such connections impact IMP function. We show that while establishing a translocon for import, the colicin ColE9 sequesters the IMPs of the proton motive force (PMF)-linked Tol-Pal complex into islands mirroring those of colicin-bound OMPs. Through this imposed organisation, the bacteriocin subverts the outer-membrane stabilising role of Tol-Pal, blocking its recruitment to cell division sites and slowing membrane constriction. The ordering of IMPs by OMPs via an energised inter-membrane bridge represents an emerging functional paradigm in cell envelope biology

Strategy to characterize the number and type of repeating EPIYA phosphorylation motifs in the carboxyl terminus of CagA protein in Helicobacter pylori clinical isolates

Cytotoxin-associated gene A (CagA) diversity with regard to EPIYA-A, -B, -C, or -D phosphorylation motifs may play an important role in Helicobacter pylori pathogenesis, and therefore determination of these motifs in H. pylori clinical isolates can become a useful prognostic tool. We propose a strategy for the accurate determination of CagA EPIYA motifs in clinical strains, based upon one-step PCR amplification using primers that Hank the EPIYA coding region. We thus analyzed 135 H. pylori isolates derived from 75 adults and 60 children Greek patients. A total of 34 cases were found to be EPIYA PCR negative and were consequently verified as cagA negative by cagA-specific PCR, empty-site cagA PCR, and Western blotting. Sequencing of the remaining 101 PCR-positive amplicons confirmed that an accurate prediction of the number of EPIYA motifs on the basis of size distribution of the PCR products was feasible in all cases. Furthermore, our assay could identify closely related H. pylori subclones within the same patient, harboring different numbers of EPIYA repeats. The prevalence of CagA proteins with three EPIYA motifs (ABC) or four EPIYA motifs (ABCC) was the same within the adult and children groups. However, CagA species with more than four EPIYA motifs were observed exclusively within adults (8.6%), suggesting that CagA-positive strains may acquire additional EPIYA-C motifs throughout adulthood. Our strategy requires no initial cagA screening of the clinical isolates and can accurately predict the number of EPIYA repeats in single or multiple closely related subclones bearing different numbers of EPIYA motifs in their CagA, which may coexist within the same patient

Strategy To Characterize the Number and Type of Repeating EPIYA Phosphorylation Motifs in the Carboxyl Terminus of CagA Protein in Helicobacter pylori Clinical Isolates

Cytotoxin-associated gene A (CagA) diversity with regard to EPIYA-A, -B, -C, or -D phosphorylation motifs may play an important role in Helicobacter pylori pathogenesis, and therefore determination of these motifs in H. pylori clinical isolates can become a useful prognostic tool. We propose a strategy for the accurate determination of CagA EPIYA motifs in clinical strains, based upon one-step PCR amplification using primers that flank the EPIYA coding region. We thus analyzed 135 H. pylori isolates derived from 75 adults and 60 children Greek patients. A total of 34 cases were found to be EPIYA PCR negative and were consequently verified as cagA negative by cagA-specific PCR, empty-site cagA PCR, and Western blotting. Sequencing of the remaining 101 PCR-positive amplicons confirmed that an accurate prediction of the number of EPIYA motifs on the basis of size distribution of the PCR products was feasible in all cases. Furthermore, our assay could identify closely related H. pylori subclones within the same patient, harboring different numbers of EPIYA repeats. The prevalence of CagA proteins with three EPIYA motifs (ABC) or four EPIYA motifs (ABCC) was the same within the adult and children groups. However, CagA species with more than four EPIYA motifs were observed exclusively within adults (8.6%), suggesting that CagA-positive strains may acquire additional EPIYA-C motifs throughout adulthood. Our strategy requires no initial cagA screening of the clinical isolates and can accurately predict the number of EPIYA repeats in single or multiple closely related subclones bearing different numbers of EPIYA motifs in their CagA, which may coexist within the same patient