Additional file 1: Figure S1. of Rheumatoid factor, not antibodies against citrullinated proteins, is associated with baseline disease activity in rheumatoid arthritis clinical trials

Disease activity at baseline in the rituximab database (IMAGE trial of early rheumatoid arthritis). Stratified analysis by the presence or absence of rheumatoid factor (RF) and of anti-citrullinated peptide antibodies (ACPA) (x-axis categories) as well as by the use, or not, of glucocorticoids. SDAI simplified disease activity index. Table S1. Disease activity indices at baseline in subgroups by RF and ACPA positivity, as well as by glucocorticoid use, or not. ACPA antibody against citrullinated peptides, CDAI clinical disease activity index, DAS Disease Activity Score, RF rheumatoid factor, SDAI simplified disease activity index. (DOCX 70 kb

Additional file 2: Figure S1. of Predictors of remission with etanercept-methotrexate induction therapy and loss of remission with etanercept maintenance, reduction, or withdrawal in moderately active rheumatoid arthritis: results of the PRESERVE trial

Summary of patient disposition in the open-label and double-blind periods. CDAI Clinical Disease Activity Index, DAS28 Disease Activity Score based on a 28-joint count, ETN etanercept, LDA low disease activity, MTX methotrexate, QW once weekly, SDAI Simplified Disease Activity Index. (EPS 1688 kb

Presentation_1_Determination of Autoantibody Isotypes Increases the Sensitivity of Serodiagnostics in Rheumatoid Arthritis.PDF

<p>Anti-citrullinated protein antibodies (ACPA) and rheumatoid factor (RF) are the most commonly used diagnostic markers of rheumatoid arthritis (RA). These antibodies are predominantly of the immunoglobulin (Ig) M (RF) or IgG (ACPA) isotype. Other subtypes of both antibodies—particularly IgA isotypes and other autoantibodies—such as RA33 antibodies—have been repeatedly reported but their diagnostic value has still not been fully elucidated. Here, we investigated the prevalence of IgA, IgG, and IgM subtypes of RF, ACPA, and RA33 antibodies in patients with RA. To determine the diagnostic specificity and sensitivity sera from 290 RA patients (165 early and 125 established disease), 261 disease controls and 100 healthy subjects were tested for the presence of IgA, IgG, and IgM isotypes of RF, ACPA, and RA33 by EliA™ platform (Phadia AB, Uppsala, Sweden). The most specific antibodies were IgG-ACPA, IgA-ACPA, and IgG-RF showing specificities >98%, closely followed by IgG- and IgA-RA33 while IgM subtypes were somewhat less specific, ranging from 95.8% (RA33) to 90% (RF). On the other hand, IgM-RF was the most sensitive subtype (65%) followed by IgG-ACPA (59.5%) and IgA-RF (50.7%). Other subtypes were less sensitive ranging from 35 (IgA-ACPA) to 6% (IgA-RA33). RA33 antibodies as well as IgA-RF and IgA-ACPA were found to increase the diagnostic sensitivity of serological testing since they were detected also in seronegative patients reducing their number from 109 to 85. Moreover, analyzing IgM-RF by EliA™ proved more sensitive than measuring RF by nephelometry and further reduced the number of seronegative patients to 76 individuals. Importantly, among antibody positive individuals, RA patients were found having significantly more antibodies (≥3) than disease controls which generally showed one or two antibody species. Thus, increasing the number of autoantibodies in serological routine testing provides valuable additional information allowing to better distinguish between RA and other rheumatic disorders, also in patients not showing antibodies in current routine diagnostics. In conclusion, testing for multiple autoantibody specificities increases the diagnostic power of autoimmune diagnostics and could further support physicians in clinical decision-making.</p

Study of pheonotype of the H9c2 cells derived from rat cardiac myoblasts

Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Martina Kutichová Supervisor: Doc. PharmDr. Tomáš Šimůnek, Ph.D. Supervisor - specialist: Doc. MUDr. Michaela Adamcová, Ph.D. Title of diploma thesis: Study of phenotype H9c2 cells derived from rat cardiac myoblasts The cell-based in vitro investigation represents an important and frequently employed platform in current biomedical research. In experimental cardiovascular research, enzymatically dispersed neonatal or adult cardiomyocytes from various animal species are considered as a "gold standard" for such experiments. However, besides isolated primary cultures, permanent cell lines are becoming extremely popular among cardiovascular scientists. H9c2(2-1) is the first permanent cardiomyocyte-derived cell line. Although these cells are known since the mid-seventies, there is not much information about their phenotype. This study followed the results of the shotgun proteomic analysis, which identified about 1500 proteins. The aim of this thesis was to continue with obtained proteomic results with focus on detection of specific proteins by Western blot method. H9c2 cell were studied for the presence of several muscle and/or cardiac specific proteins in both non-proliferating H9c2..

Presentation_3_Determination of Autoantibody Isotypes Increases the Sensitivity of Serodiagnostics in Rheumatoid Arthritis.PDF

<p>Anti-citrullinated protein antibodies (ACPA) and rheumatoid factor (RF) are the most commonly used diagnostic markers of rheumatoid arthritis (RA). These antibodies are predominantly of the immunoglobulin (Ig) M (RF) or IgG (ACPA) isotype. Other subtypes of both antibodies—particularly IgA isotypes and other autoantibodies—such as RA33 antibodies—have been repeatedly reported but their diagnostic value has still not been fully elucidated. Here, we investigated the prevalence of IgA, IgG, and IgM subtypes of RF, ACPA, and RA33 antibodies in patients with RA. To determine the diagnostic specificity and sensitivity sera from 290 RA patients (165 early and 125 established disease), 261 disease controls and 100 healthy subjects were tested for the presence of IgA, IgG, and IgM isotypes of RF, ACPA, and RA33 by EliA™ platform (Phadia AB, Uppsala, Sweden). The most specific antibodies were IgG-ACPA, IgA-ACPA, and IgG-RF showing specificities >98%, closely followed by IgG- and IgA-RA33 while IgM subtypes were somewhat less specific, ranging from 95.8% (RA33) to 90% (RF). On the other hand, IgM-RF was the most sensitive subtype (65%) followed by IgG-ACPA (59.5%) and IgA-RF (50.7%). Other subtypes were less sensitive ranging from 35 (IgA-ACPA) to 6% (IgA-RA33). RA33 antibodies as well as IgA-RF and IgA-ACPA were found to increase the diagnostic sensitivity of serological testing since they were detected also in seronegative patients reducing their number from 109 to 85. Moreover, analyzing IgM-RF by EliA™ proved more sensitive than measuring RF by nephelometry and further reduced the number of seronegative patients to 76 individuals. Importantly, among antibody positive individuals, RA patients were found having significantly more antibodies (≥3) than disease controls which generally showed one or two antibody species. Thus, increasing the number of autoantibodies in serological routine testing provides valuable additional information allowing to better distinguish between RA and other rheumatic disorders, also in patients not showing antibodies in current routine diagnostics. In conclusion, testing for multiple autoantibody specificities increases the diagnostic power of autoimmune diagnostics and could further support physicians in clinical decision-making.</p

Additional file 1: Table S1. of Nicotinic acetylcholine receptors modulate osteoclastogenesis

Primer sequences of genes investigated with quantitative real time PCR. (JPEG 27 kb

SNPs associated with seropositivity in RA.

<p>Redundancy discrimination analysis plot showing that IRF5, CD28 and CTLA4 are associated with seropositivity in RA patients. RF+–rheumatoid factor positive RA patients; RF-–rheumatoid factor negative RA patients; ACPA+–anti-citrullinated peptides antibodies positive RA patients; ACPA-–anti-citrullinated peptides antibodies negative RA patients; SE (0,1,2)—number of shared epitope coding alleles in HLA-DRB1 gene (✧); IRF5 (CC, CT, TT)—genotypes in IRF5 gene (C risk allele) (▷); CD28 (CC, CT, TT)–genotypes in CD28 gene (C risk allele) (◁); CTLA4 (AG, GG, AA)–genotypes in CTLA4 gene (G risk allele) (◊). Diagram reading clue: Symbols are genetic factors. Large bold symbols represent genotypes significantly influencing the presence of RF and ACPA. Small empty symbols represent other genotypes of selected genes. Direction of arrow indicates which serologic status is associated with the genetic parameters and the length of the arrow indicates the magnitude of the association.</p

The genetic discrimination of RA patients and controls.

<p>Linear discrimination analysis diagram shows that shared epitope and single nucleotide polymorphisms in PTPN22, STAT4, IRF5 and PADI4 genes significantly discriminated between RA patients and healthy controls. RA—RA patients; C—control group; SE (0,1,2)—number of SE coding allele in HLA-DRB1 gene (✧); IRF5 (CC, CT, TT)—genotypes in IRF5 gene (C risk allele) (◁); PADI4 (TT, CT, CC)–genotypes in PADI4 gene (T risk allele) (▽); PTPN22 (CC, CT, TT)–genotypes in PTPN22 gene (A risk allele) (△); STAT4 (GG, GT, TT)–genotypes in STAT4 gene (T risk allele) (☐). Diagram reading clue: Small circles represent individual cases. Large grey circles—centroids—represent subject groups (RA patients and controls). Symbols are genetic factors. Large bold symbols represent genotypes significantly influencing the distribution of subjects. Small empty symbols represent other genotypes of selected genes. The closer to the group centroid the gene symbol lies, the stronger is its impact on the classification of subjects to particular group.</p

A comment on 'Operationalizing Max Weber's probability concept of class situation: the concept of social class' by Ken Smith

Schematic representation of APC activation in wt and myeloid pten -/- animals. In wt, APCs are activated and initiate autoimmunity by activating and polarizing T cells toward the Th17 lineage. PTEN-deficient APCs fail to induce autoimmunity due to sustained PI3K signaling, leading to altered signaling and reduced polarizing cytokine expression. (JPEG 294 kb

Factors associated with clinical severity in RA.

<p>Redundancy analysis plot showing that risk alleles in AFF3 gene, together with ACPA positivity are associated with higher clinical severity of RA. ACPA—anti-citrullinated peptides antibodies (□); <i>AFF3</i> (TT, AT, AA)–genotypes in <i>AFF3</i> gene (T risk allele) (▽). Diagram reading clue: Symbols are genetic and serologic factors. Large bold symbols represent genotypes and antibody presence significantly influencing the clinical parameters of disease severity (DAS28, CRP, ESR, TJC, SJC, HAQ-DI). Small empty symbols represent other factors and genotypes of selected genes. Direction of arrow indicates which of the clinical factors are associated with the genetic and serologic parameters and the length of the arrow indicates the magnitude of the association.</p