Bead arrays for antibody and complement profiling reveal joint contribution of antibody isotypes to C3 deposition

The development of antigen arrays has provided researchers with great tools to identify reactivities against self or foreign antigens from body fluids. Yet, these approaches mostly do not address antibody isotypes and their effector functions even though these are key points for a more detailed understanding of disease processes. Here, we present a bead array-based assay for a multiplexed determination of antigen-specific antibody levels in parallel with their properties for complement activation. We measured the deposition of C3 fragments from serum samples to reflect the degree of complement activation via all three complement activation pathways. We utilized the assay on a bead array containing native and citrullinated peptide antigens to investigate the levels of IgG, IgM and IgA autoantibodies along with their complement activating properties in serum samples of 41 rheumatoid arthritis patients and 40 controls. Our analysis revealed significantly higher IgG reactivity against the citrullinated fibrinogen β and filaggrin peptides as well as an IgA reactivity that was exclusive for citrullinated fibrinogen β peptide and C3 deposition in rheumatoid arthritis patients. In addition, we characterized the humoral immune response against the viral EBNA-1 antigen to demonstrate the applicability of this assay beyond autoimmune conditions. We observed that particular buffer compositions were demanded for separate measurement of antibody reactivity and complement activation, as detection of antigen-antibody complexes appeared to be masked due to C3 deposition. We also found that rheumatoid factors of IgM isotype altered C3 deposition and introduced false-positive reactivities against EBNA-1 antigen. In conclusion, the presented bead-based assay setup can be utilized to profile antibody reactivities and immune-complex induced complement activation in a high-throughput manner and could facilitate the understanding and diagnosis of several diseases where complement activation plays role in the pathomechanism

Anti-citrullinated protein antibodies cause arthritis by cross-reactivity to joint cartilage

Today, it is known that autoimmune diseases start a long time before clinical symptoms appear. Anti-citrullinated protein antibodies (ACPAs) appear many years before the clinical onset of rheumatoid arthritis (RA). However, it is still unclear if and how ACPAs are arthritogenic. To better understand the molecular basis of pathogenicity of ACPAs, we investigated autoantibodies reactive against the C1 epitope of collagen type II (CII) and its citrullinated variants. We found that these antibodies are commonly occurring in RA. A mAb (ACC1) against citrullinated C1 was found to cross-react with several noncitrullinated epitopes on native CII, causing proteoglycan depletion of cartilage and severe arthritis in mice. Structural studies by X-ray crystallography showed that such recognition is governed by a shared structural motif "RG-TG" within all the epitopes, including electrostatic potential-controlled citrulline specificity. Overall, we have demonstrated a molecular mechanism that explains how ACPAs trigger arthritis

The human secretome

The proteins secreted by human cells (collectively referred to as the secretome) are important not only for the basic understanding of human biology but also for the identification of potential targets for future diagnostics and therapies. Here, we present a comprehensive analysis of proteins predicted to be secreted in human cells, which provides information about their final localization in the human body, including the proteins actively secreted to peripheral blood. The analysis suggests that a large number of the proteins of the secretome are not secreted out of the cell, but instead are retained intracellularly, whereas another large group of proteins were identified that are predicted to be retained locally at the tissue of expression and not secreted into the blood. Proteins detected in the human blood by mass spectrometry-based proteomics and antibody-based immuno-assays are also presented with estimates of their concentrations in the blood. The results are presented in an updated version 19 of the Human Protein Atlas in which each gene encoding a secretome protein is annotated to provide an open-access knowledge resource of the human secretome, including body-wide expression data, spatial localization data down to the single-cell and subcellular levels, and data about the presence of proteins that are detectable in the blood

Evaluation of serum MMP-9 as predictive biomarker for antisense therapy in Duchenne

Duchenne Muscular Dystrophy (DMD) is a severe muscle disorder caused by lack of dystrophin. Predictive biomarkers able to anticipate response to the therapeutic treatments aiming at dystrophin re-expression are lacking. The objective of this study is to investigate Matrix Metalloproteinase-9 (MMP-9) as predictive biomarker for Duchenne. Two natural history cohorts were studied including 168 longitudinal samples belonging to 66 patients. We further studied 1536 samples obtained from 3 independent clinical trials with drisapersen, an antisense oligonucleotide targeting exon 51: an open label study including 12 patients; a phase 3 randomized, double blind, placebo controlled study involving 186 patients; an open label extension study performed after the phase 3. Analysis of natural history cohorts showed elevated MMP-9 levels in patients and a significant increase over time in longitudinal samples. MMP-9 decreased in parallel to clinical stabilization in the 12 patients involved in the open label study. The phase 3 study and subsequent extension study clarified that the decrease in MMP-9 levels was not predictive of treatment response. These data do not support the inclusion of serum MMP-9 as predictive biomarker for DMD patients

Physical, chemical and microbiological analyses of water fountains in Villages, Zonguldak Central District

This study was conducted in 23 villages of the central district of Zonguldak in 2016, aiming to evaluate the compatibility of the quality of water which is used in villages. Samples obtained from the water resources of the villages and consumed in the fountains were analysed according to the physical, chemical, and microbiological standards specified in the legal regulations. 7 fountains per village were selected randomly and 161 samples were collected. All of the samples were in compliance with the standards of taste and odour qualities, pH and electrical conductivity. None of the samples were suitable for the amount of free residual chlorine. 34 (21.1%) of 161 samples for turbidity, 20 (12.4%) samples for aluminum, 3 (1.9%) samples for colour parameter, 129 (80.1%) samples for Coliform bacteria, 74 (46.0%) samples for Escherichia coli, 14 (8.7%) for Clostridium perfringens (+ spores) were not suitable according to standards. When all the parameters were evaluated together, 131 (81.4%) samples were not suitable for drinking and using. Coliform bacteria and Escherichia coli levels were significantly higher in fountains on roads than fountains in settlements. It was determined that the presence of a protected area around the water resource was not a significant effect on bacteriological pollution. As a result, it can be connoted that fountains in rural areas in Zonguldak should be controlled, monitored and disinfected in order to protect public health.This descriptive study was performed in 23 villages located in the central district of Zonguldak, and the budget of the study was financially supported by the Bulent Ecevit University Department of Scientific Research Projects. In this study, 161 water samples were collected on March-April 2016, from 7 fountains per village. The number of water samples was determined according to the budget of the research and fountains were selected by using simple random sampling. Of the fountains, 129 (80.1%) are in the settlement area and 32 (19.9%) are on the roads. Physical and chemical analyses (smell, taste, ammonium, free residual chlorine, pH, conductivity, turbidity, aluminum, colour ) were performed in Bulent Ecevit University Science and Technology Application and Research Centre and the microbiological analyses (Coliform bacteria, Escherichia coli, Clostridium perfringens) were performed in Public Health Laboratory of the Ministry of Health. The physical, chemical and microbiological analyses of the samples were conducted to find out whether they were conformed with the standards specified in the RWHC. Data were evaluated with SPSS for Windows 19.0, the descriptive data were presented as arithmetical mean ± standard deviation and frequencies. Independent samples t-test was used in comparison of groups, and p < 0.05 was accepted as significant

Descriptive characteristics of coal workers' pneumoconiosis cases in Turkey

[No abstract available

Holmium Laser Thermal Keratoplasty for Hyperopia in Eyes Overcorrected With Laser in situ Keratomileusis for Myopia

PURPOSE: To assess the efficacy and safety of holmium laser thermal keratoplasty (Ho:LTK) for hyperopia in eyes overcorrected after laser in situ keratomileusis (LASIK) for myopia

Holmium laser thermal keratoplasty for hyperopia in eyes overcorrected with laser in situ keratomileusis for myopia

PURPOSE: To assess the efficacy and safety of holmium laser thermal keratoplasty (Ho:LTK) for hyperopia in eyes overcorrected after laser in situ keratomileusis (LASIK) for myopia

Non-invasive mechanical ventilation with spinal anesthesia for cesarean delivery

We present the successful use of perioperative non-invasive mechanical ventilation in a morbidly obese pregnant woman with bronchial asthma, severe preeclampsia and pulmonary edema undergoing an emergency cesarean delivery with spinal anesthesia. The combination of non-invasive mechanical ventilation with neuraxial anesthesia may be of value in selected parturients with acute or chronic respiratory insufficiency requiring surgery. © 2010 Elsevier Ltd. All rights reserved