Necati Gunay

Biomarkers and cytokines of bone turnover: extensive evaluation in a cohort of patients with ankylosing spondylitis

BackgroundAnkylosing spondylitis (AS) is a chronic inflammatory disease of spine and sacroiliac joints; it is characterized by new bone formation, and the disease processes can be accompanied by osteoporosis. In the present study, we investigated changes in bone mineral density (BMD) and in the levels of various bone turnover-related biomarkers and cytokines in a cohort of AS patients, with regard to clinical parameters, disease activity, and treatment regimen.Methods55 AS patients and 33 healthy controls included in the study. Spinal mobility was assessed by the Bath Ankylosing Spondylitis Metrology Index (BASMI), and radiologic changes were scored by the Bath Ankylosing Spondylitis Radiologic Index (BASRI). Patients were also evaluated with the Bath Ankylosing Spondylitis Functional Index (BASFI) and the Bath Ankylosing Spondylitis Disease Activity Index (BASDAI). Bone mineral density (BMD) assessed by dual energy X-ray absorptiometry. Various biomarkers and cytokines of bone turnover including osteoprotegerin (OPG), serum band 5 tartrate-resistant acid phosphatase (TRAP-5), soluble receptor activator of nuclear factor kappa-B ligand (sRANKL), secreted frizzled-related protein 1 (sFRP-1), Dickkopf-related protein 1 (DKK-1), and sclerostin were studied.ResultsThe levels of TRAP-5, NTX, sRANKL, sclerostin, sFRP-1, DKK-1, and IFNγ, were similar between the patients and controls (p > 0.05), while BMD of femoral neck, and OPG levels were significantly lower in AS patients (p < 0.05). In a subgroup analysis, patients with active disease had significantly higher concentrations of OPG compared with the inactive group. Rest of the biomarkers and cytokines of bone turnover were similar between the active and inactive disease groups. Subgroup analysis of patients receiving anti-TNFα agents and conventional therapy revealed that OPG concentrations were significantly lower in the patients receiving biological drugs, while BAP and DKK-1 were significantly higher in the patients treated with conventional agents.ConclusionsIn this cross-sectional study we showed that OPG levels were significantly lower in AS patients compared to healthy subjects. On the other hand, the levels of wingless (Wnt) signal pathway inhibitors seem not altered. Ectopic bone formation in AS may be related to dysfunction of these molecules at the cellular level.

Saccharomyces boulardii expresses neuraminidase activity selective for α2,3‐linked sialic acid that decreases Helicobacter pylori adhesion to host cells

Helicobacter pylori is a major causative agent of gastritis and peptic ulcer disease and is an established risk factor for gastric malignancy. Antibiotic combination therapy can eradicate H. pylori. As these same regimens can evoke adverse effects and resistance, new alternative therapies or adjunctive treatments are needed. A probiotic approach may provide a novel strategy for H. pylori treatment. In the current study, two probiotic bacteria, Lactobacillus acidophilus and Lactobacillus reuteri, and a probiotic yeast, Saccharomyces boulardii, were evaluated for their ability to influence H. pylori viability, adherence to gastric and duodenal cells, as well as the effect of S. boulardii on cell surface expression of sialic acid. Our results indicate that S. boulardii contains neuraminidase activity selective for α(2‐3)‐linked sialic acid. This neuraminidase activity removes surface α(2‐3)‐linked sialic acid, the ligand for the sialic acid‐binding H. pylori adhesin, which in turn, inhibits H. pylori adherence to duodenal epithelial cells.

Fetuin-A and interleukin-18 levels in ankylosing spondylitis.

Aim:  Interleukin‐18 (IL‐18) and fetuin‐A have been implicated in atherosclerosis. Preliminary evidence suggests that ankylosing spondylitis (AS) is associated with an increased risk of atherosclerosis. The aim of the present study was to investigate possible abnormalities in IL‐18 and fetuin‐A levels in AS.

Fetuin-A is related to syndesmophytes in patients with ankylosing spondylitis: a case control study

OBJECTIVES: New bone formation is one of the hallmark characteristics of ankylosing spondylitis, which is thereby associated with syndesmophytes. Fetuin-A is a molecule that is abundantly found in calcified tissues and it shows high affinity for calcium phosphate minerals and related compounds. Considering the role of fetuin-A in the regulation of calcified matrix metabolism, we compared the fetuin-A levels in ankylosing spondylitis patients with syndesmophytes with those in patients without syndesmophytes and in healthy controls. We also studied other biomarkers that are thought to be related to syndesmophytes. METHODS: Ninety-four patients (49 patients without syndesmophytes, 67.3% male, 40.7±8.7 years; 45 patients with syndesmophytes, 71.1% M, 43.9±9.9 years) and 68 healthy controls (44.2±10.6 years and 70.6% male) were included in this study. Syndesmophytes were assessed on the lateral radiographs of the cervical and lumbar spine. The serum levels of fetuin-A, dickkopf-1, sclerostin, IL-6, high-sensitivity C-reactive protein and bone morphogenetic protein-7 were measured with an enzyme-linked immunosorbent assay. RESULTS: Patients with syndesmophytes had significantly higher levels of fetuin-A compared with patients without syndesmophytes and controls (1.16±0.13, 1.05±0.09 and 1.08±0.13 mg/ml, respectively). However, fetuin-A was not different between the patients without syndesmophytes and controls. Bone morphogenetic protein-7 was significantly lower; dickkopf-1 was significantly higher in patients with ankylosing spondylitis compared with controls. The sclerostin concentrations were not different between the groups. In regression analysis, fetuin-A was an independent, significant predictor of syndesmophytes. CONCLUSION: Our results suggest that fetuin-A may a role in the pathogenesis of bony proliferation in ankylosing spondylitis.

Evaluation of various endothelial biomarkers in ankylosing spondylitis

Atherosclerosis has been shown to be increased in chronic inflammatory diseases including ankylosing spondylitis (AS). Impaired endothelial function, the first step in atherosclerosis, may be reflected by changes in various endothelial biomarkers of hemostasis and the release of several cellular adhesion molecules or cytokines. In this study, we investigated changes in the levels of various possible markers with regard to disease activity and treatment regimen with/without anti-TNF-α drugs. Fifty-six AS patients (44 males) and 27 controls (19 males) with no known cardiovascular risk factors were included in the study. Spinal mobility was assessed by the Bath Ankylosing Spondylitis Metrology Index, and patients were evaluated with the Bath Ankylosing Spondylitis Functional Index and the Bath Ankylosing Spondylitis Disease Activity Index. Cytokines and various endothelial biomarkers were measured in serum samples using commercially available ELISA kits. Age, sex, BMI, waist circumference, fasting glucose, MAP, lipids are all similar between patients and controls. von Willebrand factor (vWF), soluble thrombomodulin (sTM), and urotensin (UT-II) were found to be significantly higher in the sera of the patients compared to the controls. Treatment with anti-TNF-α compared to conventional therapy and disease activity in AS patients seemed to have no effect on the blood levels of UT-II, sTM, CD146, vWF, plasminogen activator inhibitor-1, tissue plasminogen activator, or the thrombin–antithrombin complex. The increased UT-II, sTM, and vWF in AS patient sera regardless of treatment and disease activity suggest an increased tendency for atherosclerosis.

Anti toxic effect of broccoli extract on stannous dichloride toxicity

PURPOSE Since Technetium-99m ((99m)Tc) has favorable physical and chemical characteristics, it is widely used radioisotope in Nuclear Medicine. However, stannous dichloride (SnCl(2)) has been widely used as a reducing agent in labeling procedure of pharmaceutical with radionuclide, it has been realized that SnCl(2) have genotoxic and cytotoxic effects on biological systems. In previous studies, it has been shown that some herbal extract can reduce genotoxic and cytotoxic effects of SnCl(2). In the present study, it is aimed to evaluate the effect of the broccoli extract on the survival of E. coli ATCC 25922 strain against to toxic effects of SnCl(2). METHODS Broccoli was extracted with methanol extraction. HPLC and TLC analysis of broccoli extract were performed. Then antitoxicity and dose response assays were performed on bacterial strain. RESULTS The broccoli extract had dose dependent protective effect against SnCl(2) toxic effect on E. coli. CONCLUSIONS The consumption of broccoli may alter the stannous dichloride toxicity. Broccoli extract may use as a new protective strategies against the toxic effect of SnCl(2) on patients who were taken (99m)Tc radiopharmaceuticals.

Effects of vancomycin, daptomycin, and tigecycline on coagulase-negative staphylococcus biofilm and bacterial viability within biofilm: an in vitro biofilm model

Bacteria may hide in a hydrated polysaccharide matrix known as a biofilm. The structure of the bacterial biofilm renders phagocytosis difficult and increases antibiotic resistance. We hypothesized that repeated doses of antibiotics have an effect on bacteria within the biofilm and that it could inhibit or eradicate biofilm formation. Two clinical biofilm-positive coagulase-negative staphylococcus isolates were evaluated. The effects of antibiotics on preformed and nascent biofilm and on bacterial eradication within the biofilm were determined using different doses of vancomycin, daptomycin, and tigecycline for different durations in an in vitro biofilm model. Vancomycin neither penetrated the biofilm nor had any microbicidal effect on bacteria within the biofilm. Daptomycin had a microbicidal effect on bacteria within the biofilm but had no effect on biofilm inhibition and eradication (independent from dose and treatment time). Tigecycline inhibited and eradicated biofilm formation and had a microbicidal effect on bacteria within the biofilm. In conclusion, (i) biofilm formation appeared to be a major barrier to vancomycin activity, (ii) daptomycin had an antimicrobial effect on the bacteria within the biofilm but not on the biofilm burden, and (iii) tigecycline had effects both on bacteria within the biofilm and on biofilm burden. Thus, both tigecycline and daptomycin might be promising candidates for the treatment of biofilm infections.

AB0744 There is NO Relationship between Glucagon like Peptide-1 and Inflammation in Psoriasis and Psoriatic Arthritis: Table 1.

Background Several studies have found a higher prevalence of type 2 diabetes mellitus in patients with psoriasis and psoriatic arthritis (PsA). Interestingly, a complete remission of psoriasis has been observed following immediately after the gastric bypass surgery in obese diabetic patients before any weight loss could have occurred, most likely due to the increased levels of GLP-1. There have been also diabetic cases who have showed improvements in psoriasis under the treatment with GLP-1 receptor agonists and with dipeptidyl peptidase-IV (DPP-IV) inhibitors.GLP-1 was suggested to have anti-inflammatory effects in addition to its effects on glucose homeostasis. Objectives To investigate the GLP-1 level and its relationship with inflammation in patients with psoriasis and PsA. Methods This study included non-diabetic PsA patients and healthy controls. Disease activity was assessed in the patients by using “Composite Psoriatic Disease Activity Index (CPDAI)” which assessed five domains of disease: peripheral arthritis, dactylitis, enthesitis, axial involvement and skin findings. High-sensitive C-reactive protein (hs-CRP) levels were also investigated for the assessment of the disease activity. Fasting blood GLP-1 levels were measured in PsA patients by using ELISA method and compared with those measured in the controls. Results There were 97 PsA patients who fulfilled the CASPAR criteria. Fifty-seven healthy sex, age- and –body mass index (BMI) matched hospital workers were evaluated as controls (Table 1).14 patients had predominantly axial and 52 had predominantly peripheral disease. 22 (17.5%) were receiving corticosteroids. 12 patients (7.8%) were on anti-TNF treatment, 78 (%80.4) were on methotrexate.There was no statistically significant difference in the GLP-1 levels between PsA patients and healthy controls. GLP-1 levels in patients with active disease were also not different from inactive patients and controls. No difference was determined in GLP-1 levels between patients with predominantly axial and predominantly peripheral disease and healthy controls. GLP-1 levels in patients with psoriasis and PsA were not correlated with the other disease activity scores including BASDAI, DAS28, PASI and hsCRP levels. There was also no correlation between GLP-1 level and functional disease index (BASFI) and also health assessment parameters (HAQ, ASQoL). The subgroup analysis in patients who were not taking glucocorticoid treatment (n:27) revealed the similar results. Table 1. Demographic, clinical and laboratory features of the patients and controls Psoriatic arthritis, n=97 Healthy controls, n=57 P value Median age (yrs) 48 (25–65) 43 (31–57) 0.09 Sex, M/F 32/65 24/33 0.29 Median disease duration (yrs) 4 (0–43) Smoking status, % 28.9 40.4 0.15 GLP-1 (pmol/L) 15.2 (3.6–58.6) 15.3 (3–56.3) 0.77 Insulin (μIU/ml) 7.4 (2–24.7) 5.7 (2.5–14.2) 0.02 HOMA-IR 1.6 (0.4–5.6) 1.3 (0.4–3.3) 0.054 ESR (mm/h) 26 (2–100) 12 (3–35) <0.001 hs-CRP (mg/L) 4.9 (0.7–65.2) 1.5 (0.2–6.8) <0.001 * GLP-1: glucagon like polipeptid-1; HOMA-IR: insulin resistance; ESR: erythrocyte sedimentation rate; hs-CRP: high sensitive C-reactive protein. Conclusions The results of this study suggests that there is no significant relationship between GLP-1 and inflammatory process in patients with psoriasis and PsA. Disclosure of Interest None declared DOI 10.1136/annrheumdis-2014-eular.4514

P01-021 – Macrophage migration inhibitory factory in FMF

Familial Mediterranean Fever (FMF) is an autoinflammatory disorder characterized by recurrent, inflammatory, self-limited episodes of fever and serositis. Neutrophils are one of the key players in the pathophysiology of FMF. Macrophage migration inhibitory factor (MIF) is a pleiotropic cytokine involved in several inflammatory processes including innate and adaptive immune responses . In addition, MIF has been shown to regulate trafficking of inflammatory cells includingneutrophis to the sites of inflammation. Because its association with innate immunity, leukocyte trafficking, and inflammation MIF may be considered as an attractive cytokine in the pathogenesis of FMF.

THU0462 Increased Oxidative Stress and Macrophage Migration Inhibitory Factor in Patients with Familial Mediterranean Fever

Background Familial Mediterranean fever (FMF) is an autoinflammatory disorder characterized by recurrent, inflammatory, self-limited episodes of fever and serositis. Neutrophils are one of the key players in the pathophysiology of FMF. Macrophage migration inhibitory factor (MIF) is a pleiotropic cytokine involved in several inflammatory processes including innate and adaptive immune responses. In addition, MIF has been shown to regulate trafficking of inflammatory cells including neutrophils to the sites of inflammation. Because its association with innate immunity, leukocyte trafficking, and inflammation MIF may be considered as an attractive cytokine in the pathogenesis of FMF. In recent years, increasing attention has been focused on the role of oxidative stress in the pathogenesis of inflammatory rheumatic diseases. As neutrophils are one of the major sources for free radicals some researchers studied oxidative stress in FMF. Objectives In this study we aimed to investigate MIF levels and its relationship with oxidative stress and M694V mutations in patients with FMF. Methods Fifty one unrelated attack free FMF patients (14 M and 27 F, 32.8±8.7 years) and 30 healthy controls (16 M and 14 F, 32.7±7 years) were included in the study. Serum MIF were studied and allele frequency of M694V was calculated. Serum total oxidant status (TOS) and total anti-oxidant status (TAS) were also studied. Results Age, sex distribution, anthropometrical indices, smoking status, serum lipids and TAS were similar between patients and controls. However; CRP, ESR, MIF, and TOS were significantly higher in the patients’ group. Comparison of patients with and without M694V mutation revealed that MIF and TOS levels were not different between the groups. Regression analysis showed that none of the variables including disease duration, CRP, ESR, TOS, TAS and BMI were predicting MIF concentrations (P > 0.05) Conclusions We found increased concentrations of MIF and oxidative stress in patients with FMF. Increased MIF levels were significantly correlated with oxidative stress and its levels were independent from the inflammatory activity. M694V mutations seem no effect on MIF and oxidative stress. Disclosure of Interest None Declared