Tofacitinib is an oral Janus kinase (JAK) inhibitor for the treatment of rheumatoid arthritis (RA). The pathways affected by tofacitinib and the effects on gene expression in situ are unknown. ...Therefore, tofacitinib effects on synovial pathobiology were investigated.
A randomised, double-blind, phase II serial synovial biopsy study (A3921073; NCT00976599) in patients with RA with an inadequate methotrexate response. Patients on background methotrexate received tofacitinib 10 mg twice daily or placebo for 28 days. Synovial biopsies were performed on Days -7 and 28 and analysed by immunoassay or quantitative PCR. Clinical response was determined by disease activity score and European League Against Rheumatism (EULAR) response on Day 28 in A3921073, and at Month 3 in a long-term extension study (A3921024; NCT00413699).
Tofacitinib exposure led to EULAR moderate to good responses (11/14 patients), while placebo was ineffective (1/14 patients) on Day 28. Tofacitinib treatment significantly reduced synovial mRNA expression of matrix metalloproteinase (MMP)-1 and MMP-3 (p<0.05) and chemokines CCL2, CXCL10 and CXCL13 (p<0.05). No overall changes were observed in synovial inflammation score or the presence of T cells, B cells or macrophages. Changes in synovial phosphorylation of signal transducer and activator of transcription 1 (STAT1) and STAT3 strongly correlated with 4-month clinical responses (p<0.002). Tofacitinib significantly decreased plasma CXCL10 (p<0.005) at Day 28 compared with placebo.
Tofacitinib reduces metalloproteinase and interferon-regulated gene expression in rheumatoid synovium, and clinical improvement correlates with reductions in STAT1 and STAT3 phosphorylation. JAK1-mediated interferon and interleukin-6 signalling likely play a key role in the synovial response.
NCT00976599.
Malondialdehyde-acetaldehyde adducts (MAA) have been implicated in atherosclerosis. The purpose of this study was to investigate the role of MAA in atherosclerotic disease. Serum samples from ...controls (n = 82) and patients with; non-obstructive coronary artery disease (CAD), (n = 40), acute myocardial infarction (AMI) (n = 42), or coronary artery bypass graft (CABG) surgery due to obstructive multi-vessel CAD (n = 72), were collected and tested for antibody isotypes to MAA-modifed human serum albumin (MAA-HSA). CAD patients had elevated relative levels of IgG and IgA anti-MAA, compared to control patients (p<0.001). AMI patients had a significantly increased relative levels of circulating IgG anti-MAA-HSA antibodies as compared to stable angina (p<0.03) or CABG patients (p<0.003). CABG patients had significantly increased relative levels of circulating IgA anti-MAA-HSA antibodies as compared to non-obstructive CAD (p<0.001) and AMI patients (p<0.001). Additionally, MAA-modified proteins were detected in the tissue of human AMI lesions. In conclusion, the IgM, IgG and IgA anti-MAA-HSA antibody isotypes are differentially and significantly associated with non-obstructive CAD, AMI, or obstructive multi-vessel CAD and may serve as biomarkers of atherosclerotic disease.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
•IL-6, soluble IL-6 receptor, and glycoprotein 130 are associated with cardiac injury.•C-reactive protein is elevated 24h postmyocardial infarction.•Association of ventricle fibrillation with IL-6 ...and hsCRP may play a role in screening patients.•These biomarkers may help predict vulnerable plaque prior to acute myocardial infarction.
Biomarkers such as interleukin-6 (IL-6), soluble interleukin-6 receptor (sIL-6R), and high sensitive C-reactive protein (hsCRP) have been reported to be elevated in acute myocardial infarction (AMI). The aim of this study is to determine the relationship between these markers during AMI, as well as their relationship to clinical parameters in an effort to discern their predictive potential in cardiac events.
Serum was collected from 73 patients with; AMI, stable coronary artery disease (CAD), and controls during cardiac catheterization. Biomarker levels were determined and correlated with clinical data. IL-6 (11.75pg/ml, P<0.05) and sIL-6R (41,340pg/ml, P=0.05) were elevated in AMI compared with CAD and controls. At presentation, hsCRP was elevated in AMI patients (4.69mg/L) compared to controls (2.69mg/L, P<0.05); however, there was a significant decrease in hsCRP between AMI (4.69mg/L) and CAD patients (7.4mg/L, P<0.05). After 24h post-AMI hsCRP levels were increased compared to stable CAD (60.46mg/L, P<0.05) and were preceded by increased IL-6 at presentation. Soluble Gp130 (sGp130) showed no significant change between AMI, CAD, and control patients. However, sGp130 positively correlated with peak troponin in AMI (R=0.587, P<0.01), and negatively correlated with previous AMI (R=−0.382, P<0.05). Circulating monocyte mRNA expression isolated from selected AMI patients showed an increase in IL-6 mRNA (5.28-fold, P<0.01) and a decrease in both IL-6R (0.374-fold, P<0.01) and sGp130 mRNA (0.38-fold, P<0.01) as compared to CAD and controls.
Results demonstrate that IL-6 and sIL-6R are associated with AMI and cardiac injury. These data support the hypothesis that trans-IL-6 receptor binding may alter intracellular signaling, and blocking of IL-6 receptor binding may be pathogenic in AMI. These data may be predictive of mechanism(s) by which plaques become unstable and rupture.
Abstract only Introduction Malondialdehyde/Acetaldehyde (MAA) modified proteins have been suggested to play a role in the development/progression of atherosclerosis. Circulating antibodies directed ...against these proteins have recently been shown to be associated with the severity of the disease. More specifically, the isotype of the antibody to MAA correlated with either an acute MI (IgG) or stable plaque formation (IgA) formation. MAA is thought to form as a result of the oxidation of fat(s) and thus the concentration and antibody response should reflect the amount of fat in the diet. Objective The purpose of this study was to evaluate the antibody responses to MAA modified proteins following immunization and high fat western diet feeding in rats. Methods Male Sprague Dawley rats were immunized with MAA-modified protein weekly for 5 weeks and then assayed for antibodies to these proteins. Animals were then separated into the following groups: chow sham, chow MAA immunized, high fat sham, and high fat MAA immunized. The high fat animals were fed a Western diet with 2-thiouracil for 12 weeks, bled every 3 weeks, and serum assayed for the presence of circulating MAA antibodies. Results Prior to feeding with high fat diet, rats immunized with MAA-modified protein had a significant increase (P<0.001) in serum antibodies directed against these modified proteins compared to controls (N of 4 per group). Following feeding of high fat diet antibody concentrations increased 6 fold in the high fat MAA immunized group compared to the chow MAA immunized group (P<0.05). Antibodies in the high fat sham and chow sham had only minimal increases in antibodies to these proteins. Conclusions These data demonstrate that following immunization with MAA-modified proteins, circulating antibodies are produced that increase following consumption of a high fat Western diet. It suggests that MAA-modified proteins are produced at low levels following normal diet, producing antibodies which act as a normal clearance method for altered protein. When high fat consumption increases these antibody levels are increased in response to the oxidative stress. Implications Use of these antibodies as a biomarker in the future may help predict the onset or progression of atherosclerosis.
Abstract only Introduction: Oxidized proteins and the formation of foam cells are central aspects in the development and progression of atherosclerosis. Malondialdehyde acetaldehyde (MAA) modified ...low density lipoprotein (LDL) is generated when lipoproteins are exposed to the oxidative product malondialdehyde. These MAA-adducted proteins have been shown to bind scavenger receptors, up-regulate adhesion molecules, induce cytokine expression, and initiate antibody and T-cell responses in both human and animal models of atherosclerotic disease. However, the induction of foam cells using these antigens has been untested. Objective: The purpose of this study was to evaluate the ability of MAA-adducted proteins on LDL to induce foam cell formation and upregulate innate inflammatory mediators of atherosclerosis in human peripheral monocytes, mouse macrophage, and human aortic endothelial cell lines. Methods: Mouse aortic endothelial (2167), mouse macrophage (J774) and human peripheral monocytes were incubated with LDL, oxidized-LDL, MAA-LDL, or MAA-human albumin (25 and 50 ng/ml) for 24 or 48 hours and evaluated. Cells were accessed for uptake using Oil Red O, triglyceride production, and cholesterol esters using filipin. Real-time semi-quantitative RT-PCR and ELISA was performed to evaluate IL-6 and monocyte chemotactic protein-1 (MCP-1) mRNA expression and protein secretion. MAA binding was also assessed using an anti-MAA monoclonal antibody. Results: LDL uptake is increased in aortic endothelial cells, J774 and human monocytes as evidenced by filipin and Oil Red O staining for esterified cholesterol LDL as compared to sham controls. Triglyceride data show a 5.1 fold increase in monocytes and a 6 fold increase in aortic endothelial cells increase following MAA-LDL incubation over control p<0.001. Additionally, IHC for MAA-adducted proteins after incubation with MAA-modified demonstrate surface binding and intracellular uptake of MAA-adducted proteins. Incubation of MAA-albumin and MAA-LDL with aortic endothelial cells results in significant IL-6 and MCP-1 mRNA synthesis and protein expression 5 fold for IL-6 p<0.001 and 3 fold for MCP-1 p<0.01 Conclusions: These data show that MAA-modified proteins bind to and are processed by endothelial cells, macrophage, and human monocytes. MAA-modified proteins result in the production of innate immune mediators of tissue inflammation and inflammation recruitment. The direct effect of MAA in forming “foam cells” as evidenced by increased triglyceride and cholesterol is supportive for a role of MAA-modified proteins in the progression of atherosclerosis. The nature and impact of MAA-modification is hypothesized to be central to cellular apoptosis, autophagy and/or necrosis. Implications: MAA-modified proteins may be directly pathogenic in the development of plaque inflammation and progression of atherosclerosis.
Abstract only Introduction: Oxidized proteins have been implicated in the development and progression of atherosclerosis. Malondialdehyde (MDA)-acetaldehyde (AA) adduct (MAA), is produced and is the ...dominant epitope formed following incubation of proteins with the oxidative product MDA. Additionally, these MAA-modified proteins have been detected in JCR atherosclerotic rat aortic tissue and the human model of atherosclerosis. MAA-modified proteins have been implicated in the progression of atherosclerotic disease. Objective: The purpose of this study was to evaluate the association of MAA-adducted proteins and circulating IgM, IgG and IgA anti-MAA antibody isotypes to patients with normal coronary arteries and patients with stable and unstable atherosclerotic lesions. Methods: Over a six-month period, serum samples from normal controls (n=82), stable angina (n=42), acute myocardial infarction (AMI) (n=41), and coronary artery bypass graph surgery (CABG) (n=72) patients were collected and tested for the presence of anti-MAA antibody isotypes. All samples were collected prior to heparinization, intervention and/or bypass pump initiation. Aortic punch biopsies from CABG patients were subjected to immunohistochemical (IHC) staining using a monoclonal mouse anti-MAA antibody and detection by confocal microscopy. Results: Normal control patients had a significantly lower circulating anti-MAA IgG (97 ng/ml, SE=6.9) and IgA (82 ng/ml) as compared to patients with coronary artery disease (p<0.001). AMI patients had a significantly increased level of circulating anti-MAA IgG antibodies (242 ng/ml, SE=30.5) compared to stable angina (186 ng/ml, SE= 20.7) (p<0.04) or CABG patients (163 ng/ml, SE=14.6) (p<0.004). Serum samples from patients with CABG had significantly increased levels of circulating anti-MAA IgA antibodies (2495 ng/ml, SE=334) compared to stable angina (367 ng/ml, SE=64.4) (p<0.001) or AMI patients (361 ng/ml, SE=65.0) (p<0.001). Anti-MAA IgM antibodies were significantly different across the groups in similar fashion to IgG results. Confocal microscopy of aortic punch biopsies confirms an increased level of the MAA-adducts within the interstitial spaces of the aorta media. Conclusions: These data show that MAA-modified proteins are present in atherosclerotic tissues and there is a significant increase in the levels of circulating anti-MAA antibodies (IgM, IgG and IgA) in patients with coronary artery disease. Anti-MAA IgM and IgG phenotypes are significantly increased in patients who present with an AMI compared to normal coronary artery and stable CAD patients, whereas, the anti-MAA IgA phenotype is significantly increased in patients who present for CABG compared to all other groups. The immunoglobulin phenotype (IgM, IgG and/or IgA) is hypothesized secondary to differences in antigenic sensitization (Th1 vs. Th2) of MAA-modified proteins in diseased tissue. Implications: Anti-MAA IgM, IgG and IgA antibody isotypes and MAA-modified proteins may serve as biomarkers for subclinical atherosclerotic disease (IgM, IgG and IgA) as well as differentiate CAD patients who have stable (IgA) and unstable (IgG) atherosclerotic plaques.
Biomarkers such as interleukin-6 (IL-6), soluble interleukin-6 receptor (sIL-6R), and high sensitive C-reactive protein (hsCRP) have been reported to be elevated in acute myocardial infarction (AMI). ...The aim of this study is to determine the relationship between these markers during AMI, as well as their relationship to clinical parameters in an effort to discern their predictive potential in cardiac events. Serum was collected from 73 patients with; AMI, stable coronary artery disease (CAD), and controls during cardiac catheterization. Biomarker levels were determined and correlated with clinical data. IL-6 (11.75pg/ml, P<0.05) and sIL-6R (41,340pg/ml, P=0.05) were elevated in AMI compared with CAD and controls. At presentation, hsCRP was elevated in AMI patients (4.69mg/L) compared to controls (2.69mg/L, P<0.05); however, there was a significant decrease in hsCRP between AMI (4.69mg/L) and CAD patients (7.4mg/L, P<0.05). After 24h post-AMI hsCRP levels were increased compared to stable CAD (60.46mg/L, P<0.05) and were preceded by increased IL-6 at presentation. Soluble Gp130 (sGp130) showed no significant change between AMI, CAD, and control patients. However, sGp130 positively correlated with peak troponin in AMI (R=0.587, P<0.01), and negatively correlated with previous AMI (R=−0.382, P<0.05). Circulating monocyte mRNA expression isolated from selected AMI patients showed an increase in IL-6 mRNA (5.28-fold, P<0.01) and a decrease in both IL-6R (0.374-fold, P<0.01) and sGp130 mRNA (0.38-fold, P<0.01) as compared to CAD and controls. Results demonstrate that IL-6 and sIL-6R are associated with AMI and cardiac injury. These data support the hypothesis that trans-IL-6 receptor binding may alter intracellular signaling, and blocking of IL-6 receptor binding may be pathogenic in AMI. These data may be predictive of mechanism(s) by which plaques become unstable and rupture.
N-acetylcysteine and fenoldopam are commonly prescribed for prevention of contrast mediated nephropathy, however, comparative superiority of either agent is unknown.
In a prospective, randomized, ...parallel-group trial, adult cardiac catheterization patients at the university and veterans' hospitals with pre-existing stable renal insufficiency were randomized to
N-acetylcysteine 600 mg orally twice daily for 4 doses or fenoldopam 0.1 mcg/kg/min intravenously for a minimum of 8 h. All patients received intravenous hydration with normal saline (5% dextrose in normal saline for diabetics on insulin). Randomization was stratified for diabetes. The primary endpoint was mean change in Scr at 72 h. Secondary endpoint was the incidence of contrast-induced nephropathy (25% increase above baseline Scr or absolute increase of 0.5 mg/dL).
Study termination occurred after ninety-five patients (mean age 68
±
10 years, female 25%, diabetic 42%, mean baseline Scr 1.5
±
0.4 mg/dL) were randomized, with 84 completing follow-up (44
N-acetylcysteine, 40 fenoldopam). Overall, there were no significant differences in mean change in Scr at 72 h (
N-acetylcysteine 0.20
±
0.72 vs. fenoldopam 0.08
±
0.48 mg/dL,
p
=
0.4) or incidence of contrast-induced nephropathy (
N-acetylcysteine 5 vs fenoldopam 8,
p
=
0.4). No differences were detected in subgroup analyses for diabetes, baseline Scr >
1.7 or 2.0 mg/dL, gender, age >
70 years, or contrast volume >
150 mL. Results were similar after multivariate adjustment for diabetes, contrast volume, heart failure and gender.
Our randomized comparison failed to demonstrate a significant difference in the abilities of
N-acetylcysteine and fenoldopam to prevent the decline in renal function or the incidence of contrast-induced nephropathy during cardiac catheterization.
Objectives. We hypothesized that intravascular ultrasound may identify significant coronary artery narrowing in the mildly diseases artery of patients with or one- or two-vessel coronary artery ...disease.
Bacground. Necropsy studies have revealed that coronary angiography may underestimate stenosis severity in vessels that appear mildly diseased. Intravascular ultratound has been shown to detect atherosclerotic changs in a angiographically normal coronary arteries and to correlate better with histologic findings.
Methods. In 20 patients, we performed intravascular ultrasound imaging (3.5F catheter, 30-MHz transducer) in 37 coronary arteries that were considered mildly diseased (<50% diameter narrowing) by qualitative angiography. The angiographic diagnosis was no significant coronary artery in eight patients, one-vessel disease to seven and two-vessel disease in five. Each vessel, except for the left main coronary artery, was divided into proximal, mid and distal segments. Percent area narrowing and minimal lumen diameter were subsequently quantified by both ultrasound and quantitative angiography.
Results. Mean maximal arterial area narrowing by ultrasound in the 67 segments studied was 36 ± 20% (range 0% to 80.2%) and 19 ± 23% (range 0% to 82%) by quantitative angiography of these same (p < 0.001, paired ttest). Mean minimal lumen diameter of the segments was 3.3 ± 0.9 mm by ultrasound and 2.7 ± 0.8 mm by quantitative angiography. In 10 patients there were 19 angiographically mildly diseased segments where the percent arterial area narrowing by ultrasound was ≥50%. Intravascular ultrasound revealed that the more proximal (reference) segmnt had >25% intimal thickening in 12 of the 19 underestimated segments. In six stenosed segments (32%), total vessel area increased compared with that of the adjacent proximal vessel segment because of compensatory dilation.
Conclusions. Intravascular ultrasound identified potentially significant coronary artery disease in vessels that appear to be only mildly diseased by angiography.
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