WorldWideScience

Sample records for anti-cyclic citrullinated peptide-positive

  1. Strong combined gene-environment effects in anti-cyclic citrullinated peptide-positive rheumatoid arthritis

    DEFF Research Database (Denmark)

    Pedersen, Line Merete Blak; Jacobsen, Søren; Garred, Peter;

    2007-01-01

    To study the role of shared epitope (SE) susceptibility genes, alone and in combination with tobacco smoking and other environmental risk factors, for risk of subtypes of rheumatoid arthritis (RA) defined by the presence or absence of serum antibodies against cyclic citrullinated peptides (CCPs)....

  2. Profiling anti-cyclic citrullinated peptide antibodies in patients with juvenile idiopathic arthritis

    Directory of Open Access Journals (Sweden)

    Tebo Anne E

    2012-08-01

    Full Text Available Abstract Background Anti-citrullinated protein/peptide antibodies (ACPA, have high specificity for rheumatoid arthritis (RA. Some children with juvenile idiopathic arthritis (JIA, phenotypically resemble RA and test positive for rheumatoid factor (RF a characteristic biomarker of RA. We investigated the prevalence of ACPA and its relationship to other serologic markers associated with RA in a well-characterized JIA cohort. Methods Cases were 334 children with JIA, 30 of whom had RF + polyarticular JIA. Sera from all cases and 50 healthy pediatric controls were investigated by ELISA at a single time point for anti-cyclic citrullinated peptide (anti-CCP IgG, RF IgM, IgA and IgG, anti-RA33 IgG, and antinuclear antibodies (ANA. Comparisons between cases and controls were made using Chi-square or Fisher exact tests and T-tests. Results The prevalence of RF was 8% among controls, and 12% among cases (ns. The prevalence of ACPA was 2% in controls and 14.3% in cases (OR 8.2, p Conclusions ACPAs are detectable in 14% of children with JIA. Children with positive ACPA but negative RF are frequent, and may define a distinct subset of children with JIA. ACPA testing should be included in the classification of JIA.

  3. Anti-cyclic citrullinated peptide antibodies in children with Juvenile Idiopathic Arthritis

    Science.gov (United States)

    Hamooda, Mohamed; Fouad, Hala; Galal, Nermeen; Sewelam, Nadia; Megahed, Dina

    2016-01-01

    Aim The purpose of present study was to access the prevalence of anti-cyclic citrullinated peptide (anti-CCP) antibodies in children with Juvenile Idiopathic Arthritis (JIA), and to investigate the clinical significance and diagnostic value of the anti-CCP antibodies in correlation with age, sex & activity. Methods This case-control study was performed on 50 patients with JIA in addition to 40 sex and age-matched children as a control group. The participants were recruited from rheumatology Outpatient Clinic of Cairo University Specialized Pediatric Hospital. Patients were subjected to full history taking, clinical examination, routine laboratory investigations and x-rays on involved joints. Both patients and controls underwent assay of anti-CCP antibodies by AxSYM Anti-CCP IgG Microparticle Enzyme Immunoassay (MEIA) which is a semi-quantitative determination of the IgG class of autoantibodies specific to cyclic citrullinated peptide (CCP) in patients’ serum or plasma. Data were analyzed using Mann-Whitney U test, ANOVA, and independent-samples t-test by SPSS version 15. Results Anti-CCP positivity was identified amongst patients with JIA, particularly those JIA patients experiencing RF positive polyarticular disease onset. Above all, it is important that anti-CCP positivity and bone erosions, degree of joint damage, and ESR levels were significantly correlated. Conclusion Anti-CCP could be utilized as a valuable marker in the polyarticular form of JIA to direct early, and could be aggressive therapeutic intervention.

  4. Anti-cyclic Citrullinated Peptide Antibody (Anti-CCP and Diagnostic Value for Rheumatoid Arthritis

    Directory of Open Access Journals (Sweden)

    Mehmet Agilli

    2014-02-01

    Full Text Available Rheumatoid arthritis (RA is an inflammatory multisystem disease of unknown etiology characterized by chronic destructive synovitis. It and #8217;s prevalence is about 1% all over the world. Serologic markers are also important beside some clinical situations upon RA diagnosis. Today, the most commonly used laboratory test is rheumatoid factor (RF in patients with suspected RA. RF is sensitive but not a specific biomarker for diagnosing RA. Early diagnosis of RA is essential to prevent of progressive joint damage. In recent years, anticyclic citrullinated peptide/protein antibody (anti-CCP attracts the attention as a remarkable biomarker for early diagnosis. Anti-CCP which is a family of anti-citrullinated protein antibodies (ACPA family, showed quite satisfactory specificity in the diagnosis of RA. Due to the prescence of ACPA was included to 2010 RA diagnostic criteria, in a manner of speaking, importance of anti-CCP was registered. [TAF Prev Med Bull 2014; 13(1.000: 83-88

  5. Circulating anti-cyclic citrullinated peptide antibody in patients with rheumatoid arthritis and chronic obstructive pulmonary disease.

    Science.gov (United States)

    Yang, Deng-Ho; Tu, Chuan-Chou; Wang, Shou-Cheng; Wei, Cheng-Chung; Cheng, Ya-Wen

    2014-07-01

    Anti-cyclic citrullinated peptide (anti-CCP) antibody is highly specific for diagnosing rheumatoid arthritis (RA). Cigarette smoking is a lifestyle and environmental factor associated with anti-CCP production and is strongly associated with chronic obstructive pulmonary disease (COPD). This study assessed levels of anti-CCP antibodies and rheumatoid factor (RF) among patients with RA and COPD. The study sample comprised 63 patients with RA and 70 patients with COPD, all of whom underwent assessment of anti-CCP antibody and RF levels. Testing revealed that 54.2% of RA patients and 0% of COPD patients were positive for anti-CCP antibodies. Additionally, 82.5% of RA patients and 42% of COPD patients were positive for RF. Among RA patients, levels of anti-CCP antibodies were higher among smokers than among nonsmokers (242.7 ± 128.3 vs. 68.1 ± 112.1, P < 0.001). COPD patients had low titers of RF but were negative for anti-CCP antibodies. The presence of anti-CCP antibodies was a reliable serologic marker in RA diagnosis and was associated with cigarette smoking.

  6. Anti-Cyclic Citrullinated Peptide (Anti-CCP and Anti-Mutated Citrullinated Vimentin (Anti-MCV Relation with Extra-Articular Manifestations in Rheumatoid Arthritis

    Directory of Open Access Journals (Sweden)

    Laura Gonzalez-Lopez

    2014-01-01

    Full Text Available We evaluated the association between anti-cyclic citrullinated peptide antibodies (anti-CCP and anti-mutated citrullinated vimentin antibodies (anti-MCV with the presence of extra-articular (ExRA manifestations in 225 patients with rheumatoid arthritis (RA. Ninety-five patients had ExRA and 130 had no ExRA. There was no association of anti-CCP and anti-MCV levels with the presence of ExRA as total group (P=0.40 and P=0.91, resp.. Making an analysis of individual manifestations, rheumatoid nodules were associated with positivity for rheumatoid factor (RF; (P=0.01, anti-CCP (P=0.048, and anti-MCV (P=0.02. Instead, RF, anti-CCP, or anti-MCV were not associated with SS, chronic anemia, or peripheral neuropathy. Levels of anti-CCP correlated with the score of the Health Assessment Questionnaire-Disability Index (HAQ-Di (r=0.154, P=0.03, erythrocyte sedimentation rate (ESR; (r=0.155, P=0.03, and RF (P=0.254, P<0.001, whereas anti-MCV titres only correlated with RF (r=0.169, P=0.02. On adjusted analysis, ExRA was associated with longer age (P=0.015, longer disease duration (P=0.007, higher DAS-28 score (P=0.002, and higher HAQ-DI score (P=0.007, but serum levels of anti-CCP and anti-MCV were not associated. These findings show the need to strengthen the evaluation of the pathogenic mechanisms implied in each specific ExRA manifestation.

  7. Significance of Anti-cyclic Citrullinated Peptide Autoantibodies in Immune-mediated Inflammatory Skin Disorders with and without Arthritis

    Science.gov (United States)

    Grover, Chander; Kashyap, Bineeta; Daulatabad, Deepashree; Dhawan, Amit; Kaur, Iqbal R

    2016-01-01

    Background: Anti-cyclic citrullinated peptides (CCPs) are autoantibodies directed against citrullinated peptides. Rheumatoid factor (RF), an antibody against the Fc portion of IgG, is known to form immune complexes and contribute to the etiopathogenesis of various skin disorders. C-reactive protein (CRP), an acute-phase protein, increases following secretion of interleukin-6 from macrophages and T cells. Anti-CCP, RF, and CRP are well-established immune-markers, their diagnostic potential in immune-mediated skin disorders remains less widely studied. Aims and Objectives: To determine the correlation between anti-CCP, RF, and CRP in immune-mediated inflammatory skin diseases. Materials and Methods: About 61 clinically diagnosed cases of various immune-mediated skin diseases (psoriasis [n = 38], connective tissue diseases such as systemic lupus erythematosus and systemic sclerosis [n = 14], and immunobullous disorders including pemphigus vulgaris and pemphigus foliaceus [n = 9]) were included in the study. These patients were subclassified on the basis of presence or absence of arthritis. Arthritis was present in nine cases of psoriasis and seven connective tissue disorder patients. Detection of serum anti-CCP was done using enzyme-linked immunosorbent assay, whereas CRP and RF levels were detected using latex agglutination technique. Results: Of the 61 specimens, 14.75% had elevated serum anti-CCP levels. RF and CRP levels were elevated in 18.03% and 39.34% specimens, respectively. RF was elevated in 13.16% of inflammatory and 42.88% of connective tissue disorders, whereas anti-CCP was raised in 10.53% of inflammatory and 35.71% of connective tissue disorders. CRP positivity was highest in connective tissue disorders (50%), followed by 39.47% in inflammatory and 22.22% in immunobullous conditions. In none of the immunobullous patients, anti-CCP or RF levels were found to be elevated. Association of the presence of arthritis with elevated anti-CCP was found to be

  8. Usefulness of anti-cyclic citrullinate peptide antibody determination in synovial fluid analysis of patients with rheumatoid arthritis

    Directory of Open Access Journals (Sweden)

    G. Valesini

    2011-09-01

    Full Text Available Objective: To assess the role of anti-cyclic citrullinated peptide (CCP antibody detection in synovial fluid (SF of RA patients compared to OA patients. Methods: We evaluated in 25 RA subjects and 14 OA patients, presenting a knee-joint effusion, the main clinical and laboratory parameters including the number of painful and/or swollen joints, Ritchie index, morning stiffness, ESR, CRP and analysis of SF obtained by therapeutic arthrocentesis. IgG anti-CCP (ELISA, rheumatoid factor (RF and total IgG (nephelometry method were measured in SF and paired serum samples. Results: We found anti-CCP antibodies and RF in 64% (16/25 and 60% (15/25 of RA sera, respectively; 72% (18/25 of RA patients were positive for anti-CCP antibodies or RF. We found a higher SF/serum ratio for anti-CCP (p<0.004 compared to that for total IgG. The calculation of anti-CCP concentration as IgG anti-CCP (units/total IgG (g L-1 revealed higher values in SF than in serum (p<0.046 in RA patients. Among these, correlation analysis showed that anti-CCP/total IgG values in SF correlated with the relative concentration of serum anti-CCP/total IgG (rs=0.842; p<0.00001 and serum anti-CCP antibody levels (rs=0.799; p<0.0001. We did not find any correlation between SF anti-CCP levels and the main characteristics of SF as well as the clinical or laboratory parameters. Conclusion: Our study give evidence for a preferential production of anti-CCP antibodies at RA joint level, confirming the pathogenetic role of these autoantibodies. Moreover, SF determination of anti-CCP, corrected for the total amount of the corresponding immunoglobulin, may be helpful as diagnostic tool in selected cases.

  9. Frequency and diagnostic significance of anti-cyclic citrullinated peptide antibodies (ACCP and anti-modified citrullinated vimentin antibodies (AMCV in children with early juvenile arthritis

    Directory of Open Access Journals (Sweden)

    S O Salugina

    2008-01-01

    Full Text Available Objective. To determine frequency of anti-cyclic citrullinated peptide antibodies (ACCP and anti-modified citrullinated vimentin antibodies (AMCV elevation and their diagnostic significance in children with early juvenile arthritis (JA. Material and methods. ACCP were evaluated in serum of 80 pts with early JA (36 girls, 44 boys, mean age 8,5±5,03 years, AMCV — in 85 pts with early JA (49 girls and 36 boys aged from 1,5 to 16 years (mean age 8,7±4,9 years. Disease duration in all children was less than 6 months. Control group included 54 grown up pts with early rheumatoid arthritis (RA, 27 - with undifferentiated arthritis (UDA and 37 conditionally healthy children. АССР was assessed by immuno-enzyme assay (IEA with commercial kits “Axis Shield Diagnostics" (Great Britain, upper normal limit 5,0 U/ml. AMCV was examined by IEA with commercial kits “Orgentec Diagnostics” (Germany, upper normal limit — 25 U/ml. Results. ACCP was elevated in 7 children with early JA (8,8%. Frequency was higher than in healthy children but lower than in grown up pts with early RA and comparable with UDA. In juvenile rheumatoid arthritis (JRA ACCP were more frequent than in juvenile chronic arthritis (JCA. Concentration was higher in rheumatoid factor (RF positive pts with polyarticular JA. AMCV level was elevated in in 23 (27,1% pts with early JA (more frequent than in healthy donors but less frequent than in grown up pts with early RA and UDA. AMCV was significantly more frequent in JRA than in JCA and in RF positive than in RF negative pts. AMCV concentration in JA was higher than in healthy children but lower than in grown up pts with RA. It was also higher in RF+ than RF- JA. ACCP and AMCV correlated with swollen joint count, tender joint count and RF. AMCV also correlated with ESR and CRP. Conclusion. In pts with early JA ACCP and AMCV are equally or more frequent than RF. In spite of low sensitivity they have high specificity for JRA in contrast

  10. The Diagnostic Utility of Anti-cyclic Citrullinated Peptide Antibodies, Matrix Metalloproteinase-3, Rheumatoid Factor, Erythrocyte Sedimentation Rate, and C-reactive Protein in Patients with Erosive and Non-erosive Rheumatoid Arthritis

    Directory of Open Access Journals (Sweden)

    O. Shovman

    2005-01-01

    Full Text Available Objective: To compare the diagnostic utility of laboratory variables, including matrix metalloproteinase-3 (MMP-3, anti-cyclic citrullinated peptide (CCP antibodies, rheumatoid factor (RF, erythrocyte sedimentation rate (ESR, and C-reactive protein (CRP in patients with erosive and non-erosive rheumatoid arthritis (RA.

  11. Anti-cyclic citrullinated peptide (CCP) antibody in patients with wood-smoke-induced chronic obstructive pulmonary disease (COPD) without rheumatoid arthritis.

    Science.gov (United States)

    Sigari, Naseh; Moghimi, Nasrin; Shahraki, Farhad Saber; Mohammadi, Shilan; Roshani, Daem

    2015-01-01

    Citrullination, a post-translational modification of proteins, is increased in inflammatory processes and is known to occur in smokers. It can induce anti-cyclic citrullinated peptide (CCP) antibodies, the most specific serologic marker for rheumatoid arthritis. Thus far, the incidence of autoimmunity in patients with wood-smoke-induced chronic obstructive pulmonary disease (COPD) resulting in anti-CCP production has not been examined. We hypothesise that anti-CCP antibody level in these patients should be higher than that in healthy subjects. A total of 112 non-rheumatoid arthritis patients, including 56 patients with wood-smoke-induced COPD and 56 patients with tobacco-induced COPD, and 56 healthy non-smoker controls were included. The serum anti-CCP antibody levels were measured and compared between the groups and against smoke exposure and clinical characteristics. The mean anti-CCP antibody levels in wood-smoke-induced COPD group were significantly higher than those in tobacco-induced COPD group (p = 0.03) and controls (p = 0.004). Furthermore, 8 (14.2 %) patients with wood-smoke-induced COPD, 4 (7.14 %) with tobacco-induced COPD and 2 (3.57 %) controls exceeded the conventional cut-off of anti-CCP antibody positivity. No relationship was found between the anti-CCP antibody level and age, gender, duration of disease, Pack-years of smoking, and duration of exposure to wood smoke. Moreover, correlations between anti-CCP antibodies and severity of airflow limitation, CAT scores, mMRC scores of dyspnoea, and GOLD staging of COPD severity were not significant. Wood-smoke-induced COPD could significantly increase the anti-CCP antibody level in non-rheumatoid arthritis patients when compared with that in patients with tobacco-induced COPD and healthy controls.

  12. Anti-Cyclic Citrullinated Peptide Antibodies and Severity of Interstitial Lung Disease in Women with Rheumatoid Arthritis

    Directory of Open Access Journals (Sweden)

    Alberto Daniel Rocha-Muñoz

    2015-01-01

    Full Text Available Objective. To evaluate whether serum titers of second-generation anticyclic citrullinated peptide antibodies (anti-CCP2 are associated with the severity and extent of interstitial lung disease in rheumatoid arthritis (RA-ILD. Methods. In across-sectional study, 39 RA-ILD patients confirmed by high-resolution computed tomography (HRCT were compared with 42 RA without lung involvement (RA only. Characteristics related to RA-ILD were assessed in all of the patients and serum anti-CCP2 titers quantified. Results. Higher anti-CCP2 titers were found in RA-ILD compared with RA only (medians 77.9 versus 30.2 U/mL, P<0.001. In the logistic regression analysis after adjustment for age, disease duration (DD, smoke exposure, disease activity, functioning, erythrocyte sedimentation rate, and methotrexate (MTX treatment duration, the characteristics associated with RA-ILD were higher anti-CCP2 titers (P=0.003 and + RF (P=0.002. In multivariate linear regression, the variables associated with severity of ground-glass score were anti-CCP2 titers (P=0.02 and with fibrosis score DD (P=0.01, anti-CCP2 titers (P<0.001, and MTX treatment duration (P<0.001. Conclusions. Anti-CCP2 antibodies are markers of severity and extent of RA-ILD in HRCT. Further longitudinal studies are required to identify if higher anti-CCP2 titers are associated with worst prognosis in RA-ILD.

  13. Mannose-binding lectin gene polymorphisms are associated with disease activity and physical disability in untreated, anti-cyclic citrullinated peptide-positive patients with early rheumatoid arthritis

    DEFF Research Database (Denmark)

    Jacobsen, Søren; Garred, Peter; Madsen, Hans Ole;

    2009-01-01

    high producers (YA/YA). Anti-CCP was present in 93 patients (59%). High scores of disease activity, C-reactive protein-based DAS28 (p=0.02), and physical disability by HAQ (p=0.01) were associated with high MBL2 expression genotypes in a gene-dose dependent way, but only in anti-CCP-positive patients......OBJECTIVE: To study the association between polymorphisms in the mannose-binding lectin gene (MBL2) and disease activity, physical disability, and joint erosions in patients with newly diagnosed rheumatoid arthritis (RA). METHODS: Patients with early RA (n=158) not previously treated with disease...... activity by Disease Activity Score-28 (DAS28 score), physical disability by Health Assessment Questionnaire (HAQ) score, and erosive changes in hands and feet (Sharp-van der Heijde score). RESULTS: Eight patients were homozygous MBL2 defective (O/O), 101 belonged to an intermediate group, and 49 were MBL2...

  14. Prevalência de anticorpos contra peptídeos cíclicos citrulinados na artrite idiopática juvenil The prevalence of anti-cyclic citrullinated peptide antibodies in juvenile idiopathic arthritis

    Directory of Open Access Journals (Sweden)

    Sandra H. Machado

    2005-12-01

    Full Text Available OBJETIVOS: Avaliar a presença de anticorpos contra peptídeos cíclicos citrulinados em uma coorte de pacientes com artrite idiopática juvenil. MÉTODOS: A presença de anticorpos contra peptídeos cíclicos citrulinados foi avaliada por ensaio imunoenzimático (ELISA no soro de pacientes com artrite idiopática juvenil com idade inferior a 18 anos, acompanhados no ambulatório de reumatologia pediátrica do Hospital de Clínicas de Porto Alegre, com tempo de diagnóstico de doença de, no mínimo, 6 meses. Também foi estudada a presença do fator reumatóide IgM e do fator antinuclear em células Hep-2 RESULTADOS: Foram analisadas amostras séricas de 45 pacientes com artrite idiopática juvenil. A presença de títulos elevados de anticorpos contra peptídeos cíclicos citrulinados foi encontrada somente no soro de uma criança (2%, a qual apresentava quadro de poliartrite com fator reumatóide reagente. CONCLUSÕES: O anticorpo contra peptídeos cíclicos citrulinados pode ser detectado em crianças com artrite idiopática juvenil, mas em freqüência muito inferior aos adultos com artrite reumatóide. Torna-se importante avaliar se anticorpos contra peptídeos cíclicos citrulinados podem identificar os pacientes com artrite idiopática juvenil com potencial de evolução para artrite reumatóide do adulto.OBJECTIVES: To assess the presence of anti-cyclic citrullinated peptide antibodies in a cohort of patients with juvenile idiopathic arthritis. METHODS: Anti-cyclic citrullinated peptide antibodies was tested for with an enzyme linked immunoabsorbent assay (ELISA in serum samples of patients from the Hospital de Clínicas de Porto Alegre, all less than 18 years old and with previous diagnosis for at least 6 months. IgMRF (rheumatoid factor and antinuclear antibodies in Hep-2 cells were also assayed. RESULTS: Serum samples were analyzed from 45 patients. The presence of high levels of anti-cyclic citrullinated peptide antibodies was found

  15. Macrophage-inducible C-type lectin is associated with anti-cyclic citrullinated peptide antibodies-positive rheumatoid arthritis in men

    Institute of Scientific and Technical Information of China (English)

    WU Xin-yu; GUO Jian-ping; YIN Fang-rui; LU Xiao-lan; LI Ru; HE Jing; LIU Xu; LI Zhan-guo

    2012-01-01

    Background Macrophage-inducible C-type lectin (MINCLE) is an important member of C-type lectin superfamily,which has been shown evidence for susceptibility to arthritis in animal models.We aimed to investigate the possible association of MINCLE with rheumatoid arthritis (RA) susceptibility in Chinese Hart population.Methods Haplotypes from HapMap database (Chinese Hart Beijing,CHB) were used to select tag-single nucleotide polymorphism (SNP) (r2=0.8) residing in MINCLE gene.A total of 563 patients with RA and 404 healthy controls were TagMan genotyped for SNP rs10841845.Association analyses were performed on the whole data set and on RA subsets based on gender difference and the status of anti-cyclic citrullinated peptide (anti-CCP) antibody in RA patients.Association statistics were calculated by age and sex adjusted logistic regression.Results Overall,MINCLE SNP rs10841845 was not associated with susceptibility to RA.However,following anti-CCP stratification,rs10841845 GG genotypes conferred a significantly protective effects against anti-CCP-positive RA (OR 0.65,95% CI 0.430-0.995,P=0.048).Following gender stratification,SNP rs10841845 G allele appeared to insert its RA protective effect only in male patients,both at allele level (G vs.A OR 0.66,95% CI 0.46-0.93,P=0.018) and at genotype level (GG vs.AA+AG,OR 0.429,95% CI 0.20-0.95,P=0.036).Notably,the male RA protective effect of rs10841845 G allele was only seen in anti-CCP-positive RA (G vs.A:OR 0.64,95% CI 0.43-0.96,P=0.029; GG vs.AA+AG:OR 0.375,95% Cl 0.14-0.94,P=0.038).Furthermore,we observed a significant reduction of Disease Activity Score (DAS) 28 score (3.91±0.70 vs.5.66±0.31,P=0.022) and serum C-reactive protein levels (31.64±24.13 vs.91.80±12.02,P=0.012)in male anti-CCP-positive RA patients carrying rs10841845 GG genotype,compared with patients carrying AA+AG genotypes.Conclusions Our study provides the evidence for a gender specific association between MINCLE rs10841845 and RA

  16. 抗环瓜氨酸肽抗体在幼年特发性关节炎中的诊断意义%Significance of anti-cyclic citrullinated antibodies in Juvenile Idiopathic arthritis

    Institute of Scientific and Technical Information of China (English)

    刘小惠; 陈月; 刘建国; 蔡素芬; 邓宇虹

    2013-01-01

    Objective To investigate the anti-cyclic citrullinated peptide(anti-cyclic citrullinated petide) antibodies,anti-ker-atin antibodies(anti-keratin antibody,AKA) and anti-perinuclear factor antibody(antiperinuclear factor,APF) joint detection in the diagnosis of juvenile idiopathic arthritis in value. Methods 61 cases of children with JIA and 20 patients with other rheumatic diseases in children,40 cases of healthy children with three detection,and comparative analysis. Results Three testing positive indicators of children with JIA were significantly higher than normal,healthy children;the difference was statistically significant(P<0.01). In the group of children with JIA anti-CCP,AKA,APF a sensitivity of 55.73%,39.34%,26.23%,the sensitivity of anti-CCP was higher than antibodies AKA and APF;the difference was statistically significant (P<0.05). Conclusion Although the sensitivi-ty of anti-CCP antibodies in children with JIA is lower than the sensitivity of adults RA ,but the diagnosis of JIA has a higher sen-sitivity and specificity,especially for polyarticular and oligoarticular JIA diagnosis more valuable.%目的:探讨抗环瓜氨酸肽(anti-cyclic citrullinated petide)抗体、抗角蛋白抗体(anti-keratin antibody,AKA)和抗核周因子抗体(antiperinuclear factor,APF)联合检测在诊断幼年特发性关节炎中的价值。方法对61例JIA患儿与40例健康患儿进行3项检测,并进行比较分析。结果幼年特发性关节炎(juvenile idiopathic arthritis,JIA)患儿的3项检测指标阳性率均明显高于正常健康患儿,差异有统计学意义(P<0.01),在JIA组患儿中抗CCP、AKA、APF的敏感性分别为55.73%、39.34%、26.23%,抗CCP抗体的敏感性高于AKA和APF,差异有统计学意义(P<0.05)。结论抗CCP抗体在JIA患儿中的敏感性虽不及成人类风湿性关节炎(rheumatoid arthritis,RA),但对JIA的诊断仍具有较高的敏感性和特异性,特别是对多关

  17. The status of rheumatoid factor and anti-cyclic citrullinated peptide antibody are not associated with the effect of anti-TNFα agent treatment in patients with rheumatoid arthritis: a meta-analysis.

    Directory of Open Access Journals (Sweden)

    Qianwen Lv

    Full Text Available OBJECTIVES: This meta-analysis was conducted to investigate whether the status of rheumatoid factor (RF and anti-cyclic citrullinated peptide (anti-CCP antibody are associated with the clinical response to anti-tumor necrosis factor (TNF alpha treatment in rheumatoid arthritis (RA. METHODS: A systemic literature review was performed using the MEDLINE, SCOPUS, Cochrane Library, ISI Web of Knowledge, and Clinical Trials Register databases, and Hayden's criteria of quality assessment for prognostic studies were used to evaluate all of the studies. The correlation between the RF and anti-CCP antibody status with the treatment effect of anti-TNFα agents was analyzed separately using the Mantel Haenszel method. A fixed-effects model was used when there was no significant heterogeneity; otherwise, a random-effects model was applied. Publication bias was assessed using Egger's linear regression and a funnel plot. RESULTS: A total of 14 studies involving 5561 RA patients meeting the inclusion criteria were included. The overall analysis showed that the pooled relative risk for the predictive effects of the RF and anti-CCP antibody status on patient response to anti-TNFα agents was 0.98 (95% CI: 0.91-1.05, p=0.54 and 0.88 (95% CI: 0.76-1.03, p=0.11, respectively, with I(2 values of 43% (p=0.05 and 67% (p<0.01, respectively. Subgroup analyses of different anti-TNFα treatments (infliximab vs. etanercept vs. adalimumab vs. golimumab, response criteria (DAS28 vs. ACR20 vs. EULAR response, follow-up period (≥ 6 vs. <6 months, and ethnic group did not reveal a significant association for the status of RF and anti-CCP. CONCLUSIONS: Neither the RF nor anti-CCP antibody status in RA patients is associated with a clinical response to anti-TNFα treatment.

  18. Detecting Anti-cyclic Citrullinated Peptide Antibody in Patients with Connective Tissue Diseases%抗环瓜氨酸肽抗体在混合性结缔组织病中的检测及意义

    Institute of Scientific and Technical Information of China (English)

    尹耕; 岑筱敏; 杨闵; 谢其冰

    2011-01-01

    Objective To determine the clinical significance of anti-cyclic citrullinated peptide antibody (antiCCP) in mixed connective tissue diseases (MCTD).Methods Enzyme linked immunosorbent assay was performed to detect anti-CCP in 57 patients with MCTD, 78 with rheumatoid arthritis (RA), 64 with systemic lupus erythematosus (SLE), 56 with polymyositis/dermatomyositis (PM/DM), 53 with sjogren syndrome (SS) and with 33 systemic sclerosis (SSc).The association between anti-CCP and clinical features of MCTD was analysed.Results Anti-CCP was detected in 87.5% RA cases, 15.8% MCTD cases, 57.1% MCTD with RA cases, 14.1% SLE cases, 15.2% PM/DM cases, 18.9% SS cases and 9.1% SSc cases.Patients with RA (or MCTD with RA) were more likely to be anti-CCP positive than those without RA (P<0.05).The MCTD patients with positive anti-CCP had higher prevalence of RA and SS related manifestations than those MCTD patients with negative anti-CCP (P<0.05).The MCTD patients with RA had higher prevalence of RA-related symptoms, diffuse sclerosis and positive anti-CCP than those MCTD patients without RA (P<0.01).Significant deviation of disease spectrum was found in the MCTD patients with RA compared with the anti-CCP positive MCTD patients without RA.Conclusion High titer of anti-CCP in combination with RA, SLE and SSc manifestations in MCTD patients can be an indicator of erosive arthritis.%目的 探讨抗环瓜氨酸肽(CCP)抗体在混合性结缔组织病(MCTD)中的临床价值.方法 采用酶联免疫吸附法分别检测57例MCTD、78例类风湿关节炎(RA)、64例系统性红斑狼疮(SLE)、56例多发性肌炎/皮肌炎(PM/DM)、53例干燥综合征(SS)、33例系统性硬化病(SSc)患者血清的抗CCP抗体水平,分析比较抗CCP抗体与MCTD各种临床特征的关系.结果 抗CCP抗体在RA、MCTD、符合RA诊断标准的MCTD、SLE、PM/DM、SS、SSc患者中的阳性率分别为87.5%、15.8%、57.1%、14.1%、15.2%、18.9%、9.1%,其中RA组及符

  19. 抗CCP抗体和RF联检在RA诊疗中的临床价值%Clinical Value of Combined Detection of Serum Anti-Cyclic Citrullinated Peptide Antibody(Anti-CCP Ab)and Rheumatoid Factor(RF)in Diagnosis of Rheumatoid Arthritis

    Institute of Scientific and Technical Information of China (English)

    葛文亮

    2010-01-01

    目的:研究抗环瓜氨酸肽(anti-cyclic citrullinated peptide,Anti-CCP)(抗CCP抗体)和RF的检测在类风湿关节炎(RA)诊疗中的临床价值.方法:分别用酶联免疫吸附试验(ELISA)、BeckMan全自动蛋白分析仪同时检测早期RA组(病程<1年)42例,RA组(病程>1年)40例,非RA对照组40例患者血清抗CCP抗体和RF.结果:早期RA组、RA组的抗CCP抗体、RF阳性率显著高于非RA对照组(P<0.05)RA组抗CCP水平显著高于早期RA组(P<0.01),两者RF无显著差别(P>0.05).RA组与早期RA组CCP抗体与RF二者无相关性.结论:联检抗CCP抗体、RF有助于类风湿的早期诊断和预测病情的进展.

  20. The diagnostic value of magnetic resonance imaging combined with anti-cyclic citrullinated peptide antibody in early rheumatoid arthritis%磁共振成像联合抗环瓜氨酸肽抗体对早期类风湿关节炎的诊断价值

    Institute of Scientific and Technical Information of China (English)

    范智斌; 张建新; 王峻; 温鸿雁

    2012-01-01

    Objective To investigate the diagnostic value of magnetic resonance imaging (MRI) combined with anti-cyclic citrullinated peptide (CCP) antibody for early rheumatoid arthritis (RA). Methods Eighty-three patients with polyarthritis (male: n=12, female: n=71) underwent MR scanning of the hands and wrists. All MR imaging was performed using spin echo (SE) and short time inversion recovery (STIR) sequences. Anti-CCP serum levels of patients were determined by ELISA. Results During the first visit and the regular checkups, 69 patients met the diagnostic criteria of ACR and were confirmed with RA. Notably, 51 patients (74%) with positive anti-CCP antibody. MRI showed 43 RA patients with synovitis. Notably, there were 38 cases of bone marrow edema and 29 cases of bone destruction in the synovitis group. Conclusion MRI scanning combined with detection of anti-CCP antibody may be helpful in the early diagnosis of RA.%目的 结合血清抗环瓜氨酸肽(CCP)抗体的水平研究磁共振成像(MRI)对类风湿关节炎(RA)手关节病变的诊断价值.方法 收集符合多关节肿痛患者83例,男12例,女71例,行双手及双腕关节MRI扫描,均采用自旋回波(SE)、短时反转恢复(STIR)序列.采用酶联免疫吸附试验(ELISA)测定患者血清抗CCP抗体的水平.结果 初诊及随诊中病情进展达到美国风湿病学会(ACR)RA诊断标准可确诊的患者69例,其中抗CCP抗体阳性者51例,阳性率为74%.43例RA患者MRI显示滑膜炎,其中,滑膜炎组38例有骨髓水肿,29例有骨质破坏.结论 MRI扫描结合抗CCP抗体检测有助于RA的早期诊断.

  1. 手腕部MRI联合抗环瓜氨酸肽抗体检测在早期RA诊断中的应用价值%Application of wrist MRI combined anti-cyclic citrullinated peptide antibody detection in the diagnosis of early rheumatoid arthritis

    Institute of Scientific and Technical Information of China (English)

    王旭荣; 王军; 金聂; 吕金纯; 蒋晓彬; 陈晓军; 邱乾德

    2015-01-01

    目的 探讨手腕部MRI联合抗环瓜氨酸肽抗体(抗-CCP抗体)检测在早期类风湿性关节炎(RA)诊断中的应用价值.方法 选取本院风湿免疫科早期RA患者45例作为RA组,非RA患者45例作为非RA组,健康体检者43例作为对照组,全部行手腕部MRI检查并采用ELISA方法检测抗-CCP抗体,同时搜集患者临床症状、体征、实验室指标及MRI表现,对结果进行统计学分析.结果 RA组患者MRI阳性率和血清抗-CCP抗体阳性率明显高于非RA组和对照组,差异具有统计学意义(P<0.05);MRI与抗-CCP抗体对早期RA的灵敏度和特异度分别是88.88%和68.88%、82.22%和91.11%,两者联合检测灵敏度(64.44%)较单独检测有所降低,但特异度高达100%;MRI滑膜评分分值与抗-CCP抗体水平呈正相关(rs=0.612,P<0.05);MRI异常征象与关节疾病活动性评分(DAS28)呈正相关(rs=0.521,P<0.05),抗-CCP抗体阳性率与DAS28呈正相关(rs=0.541,P<0.05).结论 MRI和抗-CCP抗体联合检测可提高RA早期诊断的准确性,亦为动态评估RA病情变化提供了检测依据.%Objective To study the clinical value of wrist magnetic resonance imaging (MRI) combined anti-cyclic citrullinated peptide antibody detection in the diagnosis of early rheumatoid arthritis (RA).Methods Forty five patients with early RA were selected as RA group,45 cases of patients without rheumatoid arthritis as non-RA group,and 43 cases of people with normal examination as control group.All subjects were given wrist MRI and anti-cyclic citrullinated peptide (CCP) antibody with the enzyme linked immunosorbent assay (ELISA).At the same time,clinical symptoms,physical signs,MRI manifestations,and laboratory indicators were collected.All results were statistically analyzed.Results Positive rate of MRI lesions and serum anti-CCP antibody in RA group were significantly higher than non-RA group and control group (P <0.05).The sensitivity and specificity of MRI (or anti

  2. Comparative Analysis on the Diagnostic Value of Anti-Glucose-6-Phosphate Isomerase Antibodies and Anti-Cyclic Citrullinated Peptide Antibodies for Rheumatoid Arthrits%抗GPI抗体和抗CCP抗体对RA诊断价值的比较分析

    Institute of Scientific and Technical Information of China (English)

    胡忠圣; 赵枰; 张克霞; 郁超; 张秀琳

    2011-01-01

    Objective To assess the diagnostic value of anti-cyclic citrullinated peptide antibodies and anti-gIucose-6-phosphate isomerase antibodies for rheumatoid arthrits ( RA) . Methods The levels of seunn anti-CCP and anti-GPI in 42 patients with BA; 32patients with other rheumatic diseases and 30 normal controls were determined by ELISA. The diagnostic value of these two antibodies for RA were compared by receiver operating characteristic curve (ROC). Results The median levels of anti-CCP were significantly higher in rheumatoid arthrits group(283.0U/mI)than those in other rheumatic diseases group( 12. 4U/ml) and healthy controls group (11. 2U/ml) ( P <0.01) . The RA group serum anti GPI level( 1. 68 ± 1. 50mg/L) was slightly higher than that in other rheumatic dis-eases(0.71 ±0. 77mg/L) and healthy controls(0. 43 ±0. 24mg/L) (P <0. 01) . According to receiver operating characteristic curve a-nalysis; area under the curve of anti-GPIwas 0. 819 ; standard error was 0. 046; 95% CI(0. 729 -0. 909) garea under the curve of anti-CCP was 0. 829; standard error was 0.045; 95%CI(0. 741 -0.916);the diagnostic value of them are similar. In RA the sensitivity of anti-GPI super to that of anti-CCP; but show lower specific than anti-CCP. Conclusion The level of anti-GPI have high value as same as anti-CCP in diagnosing RA.%目的:评价抗环瓜氨酸肽(CCP)抗体和抗葡萄糖-6-磷酸异构酶(GPI)抗体对类风湿关节炎(RA)的诊断价值.方法:用酶联免疫吸附试验(ELISA)分别测定RA患者42例、其他风湿病患者32例以及健康对照者30例血清中的抗CCP抗体和抗GPI抗体,并应用R0C曲线比较两者对RA的诊断价值.结果:RA组血清抗CCP抗体水平(中位数)为283.0U/ml,与其他风湿病组(12.4U/ml)和健康对照组(11.2U/ml)比较,差异有统计学意义(P<0.01).RA组血清GPI水平(1.68±1.50)mg/L明显高于其他风湿性疾病组(0.71±0.77)mg/L和健康对照组(0.43±0.24)mg/L 差异也有统计学意义(P<0.01).

  3. Relationship Between Anti-cyclic Citrullinated Peptide Antibody and Interstitial Pulmonary Fibrosis Associated with Rheumatoid Arthritis%血清抗环瓜氨酸肽抗体与类风湿关节炎合并肺间质纤维化的相关性分析

    Institute of Scientific and Technical Information of China (English)

    陈燕; 张会英

    2011-01-01

    To investigate the relationship between the expression of anti-cyclic citrullinated peptide (anti-CCP) antibody in serum and interstitial pulmonary fibrosis (IPF) secondary from rheumatoid arthritis (RA). 87 RA patients were divided into two groups: 22 cases with IPF secondary from RA (RA-IPF) and 45 cases with sim ple RA. The anti-CCP antibody in patients sera were determined by Enzyme linked immunosorbent assay (ELISA) and rheumatoid factor (RF) were assessed by immunoturbidimetry method. The relationship between the two sera auto-antibodies and IPF associated with RA was analyzed. The results showed that the percentage of patients with high level anti-CCP antibody ( > 300U/mL) or high level RF( > 1000 U/mL) in RA-IPF group was significantly higher than that in simple RA group (P < 0.05 ). Higher positive rate of RF was also found in RA-IPF compared to patients without IPF (P < 0. 05 ). The level of anti-CCP antibody in serum in RA-IPF pa tients was negatively correlated with RF. RA-IPF patients were older than those patients without IPF, but there are no differences of ESR, CRP, IgA, IgG, IgM and DAS28 between the two groups of RA patients with or without IPF. The results indicate that IPF secondary from RA are more common in old patients. High level of anti-CCP antibody and RF are possibly related to the progression of IPF in RA patients.%分析类风湿关节炎(rheumatoid arthritis,RA)患者血清中抗环瓜氨酸肽(CCP)抗体水平与类风湿关节炎患者合并肺间质纤维化的关系.选取RA患者87例,其中合并肺间质纤维化(IPF)22例,单纯RA45例,采用酶联免疫吸附实验(ELISA)检测血清抗CCP抗体滴度,免疫比浊法检测类风湿因子(RF)滴度,并分析两者与RA-IPF关系.RA-IPF组抗CCP抗体高滴度(>300IU/mL)患者所占的百分比明显高于单纯RA组,P<0.05.RA-IPF组RF阳性率和高滴度(>1000IU/mL)患者的百分率明显高于单纯RA组,P<0.05,但抗CCP抗体与RF之间没有明显相关性.RA

  4. Significance of anti-cyclic citrullinated peptide antibody and magnetic resonance imaging of metacarpophalangeal joints and wrist in early rheumatoid arthritis%抗环瓜氨酸肽抗体和关节磁共振成像对早期类风湿关节炎的意义

    Institute of Scientific and Technical Information of China (English)

    岳涛; 程鹏; 范晓蕾; 孙红梅; 周嘉陵; 何东仪; 陈继红; 张湛明

    2011-01-01

    目的 研究抗环瓜氨酸肽抗体(抗CCP)和核磁共振成像(MRI)对早期类风湿关节炎(RA)的诊断价值.方法 2007年1月至2009年6月上海光华医院对早期RA组 94例、不典型单关节炎组24例及对照组35例行掌指关节及腕关节MR扫描.同时搜集患者的临床资料及抗CCP抗体、类风湿因子(RF-IgM)等实验室指标.统计并分析MRI征象及OMERACT评分同临床检查之间的关系.结果 抗CCP抗体在早期RA中敏感性55.3%,特异性88.6%.根据MRI所示,滑膜增生对早期RA的敏感性和特异性分别为100%、71.4%;骨髓水肿为25.5%、94.3%;骨侵蚀为88.3%、65.7%.其中骨髓水肿的特异性最高,表现出骨髓水肿与抗CCP抗体阳性有一定关联.关节MRI对于早期RA有很高的诊断价值,对于骨破坏及滑膜炎的的敏感性远高于传统X线检查.与掌指关节相比,腕关节的MRI骨破坏更明显.结论 早期RA的腕骨骨髓水肿与抗CCP抗体阳性相关,抗CCP抗体和MRI对于早期RA的特异性均较高,分别优于RF与X线平片,有利于RA的早期诊断.二者联合检查能减少早期RA的漏诊率.%Objective To study the diagnostic value of anti-cyclic citrullinated peptide antibody (anti-CCP) and magnetic resonance imaging (MRI) of metacarpophalangeal joints(MCP)and wrist in early rheumatoid arthritis(RA).Methods MRI of MCP and wrist joint, laboratory indices of anti-CCP and rheumatic factor (RF) were performed and recorded in the 94 early-stage RA patients, 24 non-typical monoarthritis and 35 other arthritis.The MRI findings and OMERACT (outcome measures in rheumatoid arthritis clinical trials) score were analyzed in comparison with their clinical and laboratory indices.Results The sensitivity of anti-CCP, synonitis, bone erosion and bone erosion was 55.3%, 100%, 25.5% and 88.3% respectively in early-stage RA patients.The specificity was 88.6%, 71.4%, 94.3% and 65.7% respectively.There was significant difference between early-stage RA group and other

  5. 以抗环瓜氨酸肽抗体改进对1987年美国风湿病学会关于类风湿关节炎分类标准的探讨%Evaluation of ACR 1987 criteria and the role of anti-cyclic citrullinated peptide antibodies for the diagnosis of rheumatoid arthritis

    Institute of Scientific and Technical Information of China (English)

    赵金霞; 王志敏; 栗占国

    2009-01-01

    Objective To revise the American College of Rheumatology classification criteria for rheumatoid arthritis(RA)with anti-cyclic citrullinated peptide(anti-CCP)antibodies and to evaluate its utility in the diagnosis of Chinese patients.Methods All patients from the Department of Rheumatology and Immunology of Peking University People's Hospital who had arthritis complaints in recent two years were enrolled.Patients were divided into RA group and non-RA group according to the clinical diagnosis by experienced rheumatologists.The diagnostic value of ACR criteria and the anti-CCP revised criteria(RA-6,RA-7 and RA-8)were evaluated by analyzing the clinical and laboratory parameters.Results A total of 604 patients were included in the study.312 patients were diagnosed as RA and 292 were diagnosed as other rheumatic diseases by rheumatologists.For those patients who had disease course for less than 2 years,the sensitivity of 1987 ACR criteria,RA-6,RA-7 and RA-8 criteria was 82.0%,91.0%.87.0%and 87.0%,respectively.The specificity of them was 95.6%,83.9%,95.6%and 95.6%.respectively.The sensitivity of 1987 ACR criteria,RA-6,RA-7 and RA-8 criteria for all the RA patients was 92.3%,96.8%.94.6%and 94.6%,respectively.The speciflcity of them was 92.8%,83.6%,92.8%and 92.8%.respectively.Conclusion The 1987 ACR criteria have high sensitivity and specificity in established RA.but its sensitivity in early RA is low.The RA-6criteria can improve the sensitivity dramatically but with reduced specificity.The RA-7 criteria can increase the sensitivity without sacrifice the specificity,especially in early RA patients.It may be used as a new set of classification criteria in clinical practice.%目的 改进1987年美国风湿病学会(ACR)修订的类风湿关节炎(RA)分类标准,增加抗环瓜氨酸肽(CCP)抗体和(或)保留类风湿结节或放射学改变等,探讨不同条件下的标准(分别称为RA-6、RA-7以及RA-8)对RA诊断的敏感性和特异性.方法 选取2006-2008年

  6. Meta-Analysis: Diagnostic Accuracy of Anti-Cyclic Citrullinated Peptide Antibody for Juvenile Idiopathic Arthritis

    Directory of Open Access Journals (Sweden)

    Yan Wang

    2015-01-01

    Full Text Available Objective. To estimate the diagnostic accuracy of the anti-CCP test in JIA and to evaluate factors associated with higher accuracy. Methods. Two investigators performed an extensive search of the literature published between January 2000 and January 2014. The included articles were assessed by the Quality Assessment of Diagnostic Accuracy Studies tool. The meta-analysis was performed using a summary ROC (SROC curve and a bivariate random-effect model to estimate sensitivity and specificity across studies. Results. The bivariate meta-analysis yielded a pooled sensitivity and specificity of 10% (95% confidence interval (CI: 6.0%–15.0% and 99.0% (95% CI: 98.0%–100.0%. The area under the SROC curve was 0.96. Sensitivity estimates were highly heterogeneous, which was partially explained by the higher sensitivity in the rheumatoid factor-positive polyarthritis (RF+ PA subtype (48.0%; 95% CI: 31.0%–65.0% than in the other subtypes (17.0%; 95% CI: 14.0%–20.0% and the higher sensitivity of the Inova assay (17.0%; 95% CI: 14.0%–20.%% than the other assays (0.05%; 95% CI: 2.0%–11.0%. Conclusions. Anti-CCP antibody test has a high specificity for the diagnosis of JIA. The sensitivity of this test is low and varies across populations but is higher in RF+ PA than in other JIA subtypes.

  7. THE CLINICAL AND DIAGNOSTIC VALUE OF ANTI-CYCLIC CITRULLINATED PEPTIDE ANTIBODIES IN EARLY JUVENILEARTHRITIS

    Directory of Open Access Journals (Sweden)

    S O Salugina

    2009-01-01

    Conclusion. In patients with early JA, the detection rate of CCPA is significantly higher than that in healthy children and comparable with that of RF. CCPAs have a high specificity for the diagnosis of JRA (an independent nosological entity within JA are a risk factor of polyarthritis. The early detection of CCPA alone or in combination with RF in JA patients may serve the basis for the early use of active, frequently aggressive therapy.

  8. Immune recognition of citrullinated epitopes.

    Science.gov (United States)

    Nguyen, Hai; James, Eddie A

    2016-10-01

    Conversion of arginine into citrulline is a post-translational modification that is observed in normal physiological processes. However, abnormal citrullination can provoke autoimmunity by generating altered self-epitopes that are specifically targeted by autoantibodies and T cells. In this review we discuss the recognition of citrullinated antigens in human autoimmune diseases and the role that this modification plays in increasing antigenic diversity and circumventing tolerance mechanisms. Early published work demonstrated that citrullinated proteins are specifically targeted by autoantibodies in rheumatoid arthritis and that citrullinated peptides are more readily presented to T cells by arthritis-susceptible HLA class II 'shared epitope' proteins. Emerging data support the relevance of citrullinated epitopes in other autoimmune diseases, including type 1 diabetes and multiple sclerosis, whose susceptible HLA haplotypes also preferentially present citrullinated peptides. In these settings, autoimmune patients have been shown to have elevated responses to citrullinated epitopes derived from tissue-specific antigens. Contrasting evidence implicates autophagy or perforin and complement-mediated membrane attack as inducers of ectopic citrullination. In either case, the peptidyl deiminases responsible for citrullination are activated in response to inflammation or insult, providing a mechanistic link between this post-translational modification and interactions with the environment and infection. As such, it is likely that immune recognition of citrullinated epitopes also plays a role in pathogen clearance. Indeed, our recent data suggest that responses to citrullinated peptides facilitate recognition of novel influenza strains. Therefore, increased understanding of responses to citrullinated epitopes may provide important insights about the initiation of autoimmunity and recognition of heterologous viruses. PMID:27531825

  9. Interrelationships between glutamine and citrulline metabolism

    Science.gov (United States)

    This article analyzes the contribution of glutamine to the synthesis of citrulline and reviews the evidence that glutamine supplementation increases citrulline production. Glutamine supplementation has been proposed in the treatment of critically ill patients; however, a recent large multicenter ran...

  10. Low-field magnetic resonance imaging or combined ultrasonography and anti-cyclic citrullinated peptide antibody improve correct classification of individuals as established rheumatoid arthritis

    DEFF Research Database (Denmark)

    Pedersen, Jens K; Lorenzen, Tove; Ejbjerg, Bo;

    2014-01-01

    (RA). METHODS: In 53 individuals from a population-based, cross-sectional study, historic fulfilment of the American College of Rheumatology (ACR) 1987 criteria ("classification") or RA diagnosed by a rheumatologist ("diagnosis") were used as standard references. The sensitivity, specificity and Area...... under Curve for Receiver Operating Characteristics curves (ROC-area: (sensitivity + specificity)/2) were calculated for "current fulfilment of the ACR 1987 criteria" (list format), "adapted ACR 1987 criteria" (list format, substituting IgM rheumatoid factor with ACPA and clinical joint swelling...... and erosions on radiography with synovitis and erosions detected by US on a semi-quantitative scale), and RA MRI scoring System (RAMRIS) scores on low-field MRI in the unilateral hand. RESULTS: For the ACR 1987 criteria the ROC-area was 75% (sensitivity/specificity = 50%/100%) (with "classification...

  11. Determination of citrulline in watermelon rind.

    Science.gov (United States)

    Rimando, Agnes M; Perkins-Veazie, Penelope M

    2005-06-17

    Watermelon (Citrullus vulgaris Schrad.) is a natural and rich source of the non-essential amino acid citrulline. Citrulline is used in the nitric oxide system in humans and has potential antioxidant and vasodilatation roles. A method using gas chromatography-mass spectrometry (GC-MS) was developed to separate citrulline from glutamic acid, which co-elute when analyzed by high performance liquid chromatography. Watermelons were analyzed by GC-MS to determine the citrulline content among varieties, types, flesh colors, and tissues. Citrulline content ranged from 3.9 to 28.5 mg/g dry weight (dwt) and was similar between seeded and seedless types (16.6 and 20.3 mg/g dwt, respectively). Red flesh watermelons had slightly less citrulline than the yellow or orange flesh watermelons (7.4, 28.5 and 14.2 mg/g dwt, respectively). Rind contained more citrulline than flesh on a dry weight basis (24.7 and 16.7 mg/g dwt, respectively) but a little less on a fresh weight (fwt) basis (1.3 and 1.9 mg/g fwt, respectively). These results indicate that watermelon rind, an underutilized agricultural waste, offers a source of natural citrulline. PMID:16007998

  12. Optimizing the identification of citrullinated peptides by mass spectrometry

    DEFF Research Database (Denmark)

    Bennike, Tue; Lauridsen, Kasper B.; Olesen, Michael Kruse;

    2013-01-01

    findings, we present a strategy for verifying citrullinated sites in complex samples post processing, in proteomics shotgun experiments. By requiring a missed cleavage for the identification of citrullinated peptides, we demonstrate that 64% of false-positively annotated citrullination sites could...... be removed. We furthermore demonstrated likely pitfalls of applying the strategy. In conclusion, manual annotation of citrullinated peptide spectra remains essential to ensure correct annotation. Implementing a missed cleavage requirement significantly reduces the number of spectra needing manual...

  13. Autoimmunity to citrullinated type II collagen in rheumatoid arthritis

    OpenAIRE

    Yoshida, Mamoru; TSUJI, Michiko; Kurosaka, Daitaro; Kurosaka, Daisaburo; Yasuda, Jun; Ito, Yoshitaka; Nishizawa, Tetsuro; Yamada, Akio

    2006-01-01

    The production of autoantibodies to citrullinated type II collagen and the citrullination of type II collagen were analyzed in rheumatoid arthritis. Autoantibodies to citrullinated type II collagen were detected in 78.5% of serum samples from 130 rheumatoid arthritis patients. Autoantibodies to native noncitrullinated type II collagen were detected in 14.6% of serum samples, all of which were positive for anti-citrullinated type II collagen antibodies. Serum samples were also positive for ant...

  14. Peptidylarginine Deiminases and Citrullination in the Central Nervous System

    OpenAIRE

    浅賀,宏昭

    2008-01-01

    Citrullination is one of the most recently described posttranslational modifications. It is the conversion of arginine residues in proteins to citrulline residues catalyzed by peptidylarginine deiminases(PADs;EC3.5.3.15)in a calcium-dependant fashion. Formation of citrulline residues drastically alters the structure and function of proteins and polypeptides.

  15. Intestinal and hepatic metabolism of glutamine and citrulline in humans.

    Science.gov (United States)

    van de Poll, Marcel C G; Ligthart-Melis, Gerdien C; Boelens, Petra G; Deutz, Nicolaas E P; van Leeuwen, Paul A M; Dejong, Cornelis H C

    2007-06-01

    Glutamine plays an important role in nitrogen homeostasis and intestinal substrate supply. It has been suggested that glutamine is a precursor for arginine through an intestinal-renal pathway involving inter-organ transport of citrulline. The importance of intestinal glutamine metabolism for endogenous arginine synthesis in humans, however, has remained unaddressed. The aim of this study was to investigate the intestinal conversion of glutamine to citrulline and the effect of the liver on splanchnic citrulline metabolism in humans. Eight patients undergoing upper gastrointestinal surgery received a primed continuous intravenous infusion of [2-(15)N]glutamine and [ureido-(13)C-(2)H(2)]citrulline. Arterial, portal venous and hepatic venous blood were sampled and portal and hepatic blood flows were measured. Organ specific amino acid uptake (disposal), production and net balance, as well as whole body rates of plasma appearance were calculated according to established methods. The intestines consumed glutamine at a rate that was dependent on glutamine supply. Approximately 13% of glutamine taken up by the intestines was converted to citrulline. Quantitatively glutamine was the only important precursor for intestinal citrulline release. Both glutamine and citrulline were consumed and produced by the liver, but net hepatic flux of both amino acids was not significantly different from zero. Plasma glutamine was the precursor of 80% of plasma citrulline and plasma citrulline in turn was the precursor of 10% of plasma arginine. In conclusion, glutamine is an important precursor for the synthesis of arginine after intestinal conversion to citrulline in humans.

  16. Glutamine: precursor or nitrogen donor for citrulline synthesis?

    Science.gov (United States)

    Marini, Juan C; Didelija, Inka Cajo; Castillo, Leticia; Lee, Brendan

    2010-07-01

    Although glutamine is considered the main precursor for citrulline synthesis, the current literature does not differentiate between the contribution of glutamine carbon skeleton vs. nonspecific nitrogen (i.e., ammonia) and carbon derived from glutamine oxidation. To elucidate the role of glutamine and nonspecific nitrogen in the synthesis of citrulline, l-[2-(15)N]- and l-[5-(15)N]glutamine and (15)N-ammonium acetate were infused intragastrically in mice. The amino group of glutamine labeled the three nitrogen groups of citrulline almost equally. The amido group and ammonium acetate labeled the ureido and amino groups of citrulline, but not the delta-nitrogen. D(5)-glutamine also infused in this arm of the study, which traces the carbon skeleton of glutamine, was utilized poorly, accounting for only 0.2-0.4% of the circulating citrulline. Dietary glutamine nitrogen (both N groups) incorporation was 25-fold higher than the incorporation of its carbon skeleton into citrulline. To investigate the relative contributions of the carbon skeleton and nonspecific carbon of glutamine, arginine, and proline to citrulline synthesis, U-(13)C(n) tracers of these amino acids were infused intragastrically. Dietary arginine was the main precursor for citrulline synthesis, accounting for approximately 40% of the circulating citrulline. Proline contribution was minor (3.4%), and glutamine was negligible (0.4%). However, the glutamine tracer resulted in a higher enrichment in the ureido group, indicating incorporation of nonspecific carbon from glutamine oxidation into carbamylphosphate used for citrulline synthesis. In conclusion, dietary glutamine is a poor carbon skeleton precursor for the synthesis of citrulline, although it contributes both nonspecific nitrogen and carbon to citrulline synthesis.

  17. N-Carbamoylputrescine, a citrulline-derived polyamine, is not a significant citrulline metabolite in rats.

    Science.gov (United States)

    Ramani, D; Nakib, S; Chen, H; Garbay, C; Loukaci, A; Cynober, L; De Bandt, J P

    2012-04-01

    Citrulline, a key amino acid of the urea cycle, has been shown to play a regulatory role in protein and energy metabolism in mammals. We questioned whether N-carbamoyl-putrescine (NCP), the decarboxylated derivative of citrulline, could play a role in the biological properties of this amino acid. To evidence the presence of NCP in mammalian tissues, we developed a sensitive reverse-phase high-performance liquid chromatography (HPLC) with fluorimetric detection method with precolumn dansyl derivatization and solid-phase extraction for the determination of NCP together with polyamines in biological samples. Dansyl NCP was identified with a 5.85-min retention time. Linearity was obtained in a concentration range of 0.125 to 12.5 μM. Intraday and day-to-day relative coefficients of variation ranged from 8.9% to 12.3% and from 14% to 14.3%, respectively. Recovery rates in serum ranged from 75% to 83%. Thereafter, we used this method to search for the presence of NCP in serum, muscle, liver, jejunum, and ileum in rats after both short-term intraperitoneal injection and long-term oral citrulline supplementation. We failed to detect NCP in these animals. These data suggest that NCP is not a significant citrulline metabolite in rats.

  18. Extrarenal citrulline disposal in mice with impaired renal function

    Science.gov (United States)

    The endogenous synthesis of arginine, a semiessential amino acid, relies on the production of citrulline by the gut and its conversion into arginine by the kidney in what has been called the "intestinal-renal axis" for arginine synthesis. Although the kidney is the main site for citrulline disposal,...

  19. Citrullination regulates pluripotency and histone H1 binding to chromatin

    DEFF Research Database (Denmark)

    Christophorou, Maria A; Castelo-Branco, Gonçalo; Halley-Stott, Richard P;

    2014-01-01

    Citrullination is the post-translational conversion of an arginine residue within a protein to the non-coded amino acid citrulline. This modification leads to the loss of a positive charge and reduction in hydrogen-bonding ability. It is carried out by a small family of tissue-specific vertebrate...

  20. Mitochondria: role of citrulline and arginine supplementation in MELAS syndrome.

    Science.gov (United States)

    El-Hattab, Ayman W; Emrick, Lisa T; Chanprasert, Sirisak; Craigen, William J; Scaglia, Fernando

    2014-03-01

    Mitochondria are found in all nucleated human cells and generate most of the cellular energy. Mitochondrial disorders result from dysfunctional mitochondria that are unable to generate sufficient ATP to meet the energy needs of various organs. Mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes (MELAS) syndrome is a frequent maternally inherited mitochondrial disorder. There is growing evidence that nitric oxide (NO) deficiency occurs in MELAS syndrome and results in impaired blood perfusion that contributes significantly to several complications including stroke-like episodes, myopathy, and lactic acidosis. Both arginine and citrulline act as NO precursors and their administration results in increased NO production and hence can potentially have therapeutic utility in MELAS syndrome. Citrulline raises NO production to a greater extent than arginine, therefore, citrulline may have a better therapeutic effect. Controlled studies assessing the effects of arginine or citrulline supplementation on different clinical aspects of MELAS syndrome are needed.

  1. Diagnostic Value of Anti-cyclic Citrullinated Peptide Antibody and RheumatoidFacter in Patients with Rheumatoid Arthritis%抗CCP抗体与RF联合检测在类风湿关节炎中的诊断价值

    Institute of Scientific and Technical Information of China (English)

    席作明; 孙涛

    2010-01-01

    目的 探讨抗环瓜氨酸肽(CCP)抗体和类风湿因子(RF)检测在类风湿关节炎(RA)诊断中的意义.方法 用ELISA方法检测108例(RA 62例,非RA 46例,及正常人30例)血清中的抗CCP抗体,免疫比浊法测RF,并比较抗CCP抗体与RF对于RA的敏感性和特异性.结果 抗CCP抗体在RA组患者血清中的阳性率51.8%,明显高于非RA组(3.21%)和正常对照组(0.0%),经t检验P<0.05.抗CCP抗体对诊断RA的敏感性和特异性分别为50.1%、46.2%,在62例确诊为RA患者的血清中抗CCP抗体与RF重叠阳性率为55.1%,两者含量呈正相关.结论 抗CCP抗体对RA具有很高的特异性,可视为新的RA血清学诊断指标,它与RA联合检测更能提高RA的早期诊断率.

  2. Citrullination regulates pluripotency and histone H1 binding to chromatin

    Science.gov (United States)

    Christophorou, Maria A.; Castelo-Branco, Gonçalo; Halley-Stott, Richard P.; Oliveira, Clara Slade; Loos, Remco; Radzisheuskaya, Aliaksandra; Mowen, Kerri A.; Bertone, Paul; Silva, José C. R.; Zernicka-Goetz, Magdalena; Nielsen, Michael L.; Gurdon, John B.; Kouzarides, Tony

    2014-03-01

    Citrullination is the post-translational conversion of an arginine residue within a protein to the non-coded amino acid citrulline. This modification leads to the loss of a positive charge and reduction in hydrogen-bonding ability. It is carried out by a small family of tissue-specific vertebrate enzymes called peptidylarginine deiminases (PADIs) and is associated with the development of diverse pathological states such as autoimmunity, cancer, neurodegenerative disorders, prion diseases and thrombosis. Nevertheless, the physiological functions of citrullination remain ill-defined, although citrullination of core histones has been linked to transcriptional regulation and the DNA damage response. PADI4 (also called PAD4 or PADV), the only PADI with a nuclear localization signal, was previously shown to act in myeloid cells where it mediates profound chromatin decondensation during the innate immune response to infection. Here we show that the expression and enzymatic activity of Padi4 are also induced under conditions of ground-state pluripotency and during reprogramming in mouse. Padi4 is part of the pluripotency transcriptional network, binding to regulatory elements of key stem-cell genes and activating their expression. Its inhibition lowers the percentage of pluripotent cells in the early mouse embryo and significantly reduces reprogramming efficiency. Using an unbiased proteomic approach we identify linker histone H1 variants, which are involved in the generation of compact chromatin, as novel PADI4 substrates. Citrullination of a single arginine residue within the DNA-binding site of H1 results in its displacement from chromatin and global chromatin decondensation. Together, these results uncover a role for citrullination in the regulation of pluripotency and provide new mechanistic insights into how citrullination regulates chromatin compaction.

  3. Glutamine: precursor or nitrogen donor for citrulline synthesis?

    Science.gov (United States)

    Glutamine (Gln) is considered the main precursor for citrulline (Cit) synthesis, but no attempts have been made to differentiate the contribution of Gln carbon (Gln-C) skeleton vs. the nonspecific contribution through NH3 and CO2. To study the contribution of dietary Gln-N to the synthesis of Cit, t...

  4. Organic anion transporter OAT1 is involved in renal handling of citrulline.

    Science.gov (United States)

    Nakakariya, Masanori; Shima, Yoichiro; Shirasaka, Yoshiyuki; Mitsuoka, Keisuke; Nakanishi, Takeo; Tamai, Ikumi

    2009-07-01

    Because citrulline plasma concentration is elevated in kidney failure, citrulline could be a biomarker of renal insufficiency, although the mechanism regulating its disposition in the kidney has not been clarified. In rat kidney slices, citrulline uptake was apparently Na(+) dependent, saturable with K(m) 556 microM, and significantly inhibited by anionic (PAH) and cationic (TEA) compounds, but not by probenecid at 1 mM. Preincubation of kidney slices with glutarate increased citrulline uptake, while such an increase was not observed after preincubation of the slices in Na(+)-free buffer. This result suggested that a sodium-dependent dicarboxylate cotransporter is involved in citrulline uptake by rat kidney slices. In studies using transporter-overexpressing cells, human organic anion transporter 1 (OAT1) and rat Oat1 exhibited citrulline transport activity with K(m) values of 238 and 373 microM, respectively, while other OATs and organic cation transporters (OCTs) did not transport citrulline. Based on the relative activity factor method, the contribution of rat Oat1 to the overall uptake of citrulline in rat kidney slices was approximately 70%. Moreover, the interaction among citrulline, PAH, and probenecid uptakes via rat Oat1 suggested that there are multiple functional sites on Oat1 and that the citrulline site may be distinct from the PAH and probenecid site. Thus OAT1/Oat1 appears to be one of the major contributors to renal basolateral uptake of citrulline, and impaired activities of these transporters may contribute substantially to the increase in plasma citrulline in renal failure. Accordingly, citrulline may be useful for diagnosis of kidney function as is creatinine. PMID:19403644

  5. Citrulline as a Biomarker in the Murine Total-Body Irradiation Model: Correlation of Circulating and Tissue Citrulline to Small Intestine Epithelial Histopathology.

    Science.gov (United States)

    Jones, Jace W; Tudor, Gregory; Li, Fei; Tong, Yan; Katz, Barry; Farese, Ann M; MacVittie, Thomas J; Booth, Catherine; Kane, Maureen A

    2015-11-01

    The use of plasma citrulline as a biomarker for gastrointestinal acute radiation syndrome via exposure to total-body irradiation in a murine model was investigated. The radiation exposure covered lethal, mid-lethal, and sub-lethal gastrointestinal acute radiation syndrome. Plasma citrulline profiles were generated over the first 6 d following total-body irradiation exposure of 6-15 Gy. In addition, plasma citrulline was comprehensively evaluated in the context of matching small intestine citrulline and histopathology. Higher plasma citrulline was significantly associated with lower irradiation doses over the first 6 d following the irradiation insult. Furthermore, higher plasma citrulline was significantly associated with higher crypt survival. The correlation of the plasma citrulline to crypt survival was more robust for higher irradiation doses and for later time points. The data suggested plasma citrulline was most informative for reflecting gastrointestinal injury resulting from exposure to 9-15 Gy total-body irradiation covering time-points 2-5 d post the irradiation insult.

  6. Plasma citrulline levels predict intestinal toxicity in patients treated with pelvic radiotherapy

    Energy Technology Data Exchange (ETDEWEB)

    Onal, Cem; Kotek, Ayse; Arslan, Gungor; Topkan, Erkan (Dept. of Radiation Oncology, Baskent Univ. Faculty of Medicine, Adana (Turkey)), E-mail: hcemonal@hotmail.com; Unal, Birsel (Dept. of Biochemistry, Baskent Univ. Faculty of Medicine, Ankara (Turkey)); Yavuz, Aydin; Yavuz, Melek (Dept. of Radiation Oncology, Akdeniz Univ. Faculty of Medicine, Antalya (Turkey))

    2011-11-15

    Background. Radiotherapy (RT) for abdominal and pelvic malignancies often causes severe small bowel toxicity. Citrulline concentrations are known to decrease with intestinal failure. We thus evaluated the feasibility of plasma citrulline levels in predicting radiation-induced intestinal toxicity. Material and methods. Fifty-three patients (36 prostate cancer, 17 endometrial cancer) who received 45 Gy pelvic RT using conventional fractionation were prospectively evaluated. Patients with prostate cancer received an additional 25-30.6 Gy conformal boost. Plasma citrulline levels were assessed on day 0, mid- (week 3) and post-RT (week 8), and four months post-RT. Dose-volume histogram, citrulline concentration changes, and weekly intestinal toxicity scores were analyzed. Results. Mean age was 63 years (range: 43-81 years) and mean baseline citrulline concentration was 38.0 +- 10.1 mumol/l. Citrulline concentrations were significantly reduced at week 3 (27.4 +- 5.9 mumol/l; p < 0.0001), treatment end (29.9 +- 8.8 mumol/l; p < 0.0001), and four months post-treatment (34.3 +- 12.1; p 0.01). The following factor pairs were significantly positively correlated: Citrulline concentration/mean bowel dose during, end of treatment, and four months post-RT; dose-volume parameters/citrulline change groups; cumulative mean radiation dose/intestinal toxicity at end and four months post-RT; citrulline changes/intestinal toxicity during and end of RT. Citrulline concentration changes significantly differed during treatment according to RTOG intestinal toxicity grades (p < 0.0001). Although the citrulline changes differed significantly within RTOG intestinal toxicity grades (p = 0.003), the difference between Grade 0 and Grade 1 did not differ significantly at the end of the treatment. At four months after RT, no significant differences were apparent. Conclusion. Citrulline-based assessment scores are objective and should be considered in measuring radiation-induced intestinal toxicity

  7. Restoration of impaired nitric oxide production in MELAS syndrome with citrulline and arginine supplementation.

    Science.gov (United States)

    El-Hattab, Ayman W; Hsu, Jean W; Emrick, Lisa T; Wong, Lee-Jun C; Craigen, William J; Jahoor, Farook; Scaglia, Fernando

    2012-04-01

    Mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes (MELAS) syndrome is one of the most common mitochondrial disorders. Although the pathogenesis of stroke-like episodes remains unclear, it has been suggested that mitochondrial proliferation may result in endothelial dysfunction and decreased nitric oxide (NO) availability leading to cerebral ischemic events. This study aimed to assess NO production in subjects with MELAS syndrome and the effect of the NO precursors arginine and citrulline. Using stable isotope infusion techniques, we assessed arginine, citrulline, and NO metabolism in control subjects and subjects with MELAS syndrome before and after arginine or citrulline supplementation. The results showed that subjects with MELAS had lower NO synthesis rate associated with reduced citrulline flux, de novo arginine synthesis rate, and plasma arginine and citrulline concentrations, and higher plasma asymmetric dimethylarginine (ADMA) concentration and arginine clearance. We conclude that the observed impaired NO production is due to multiple factors including elevated ADMA, higher arginine clearance, and, most importantly, decreased de novo arginine synthesis secondary to decreased citrulline availability. Arginine and, to a greater extent, citrulline supplementation increased the de novo arginine synthesis rate, the plasma concentrations and flux of arginine and citrulline, and NO production. De novo arginine synthesis increased markedly with citrulline supplementation, explaining the superior efficacy of citrulline in increasing NO production. The improvement in NO production with arginine or citrulline supplementation supports their use in MELAS and suggests that citrulline may have a better therapeutic effect than arginine. These findings can have a broader relevance for other disorders marked by perturbations in NO metabolism.

  8. L-citrulline immunostaining identifies nitric oxide production sites within neurons

    Science.gov (United States)

    Martinelli, G. P. T.; Friedrich, V. L. Jr; Holstein, G. R.

    2002-01-01

    The cellular and subcellular localization of L-citrulline was analyzed in the adult rat brain and compared with that of traditional markers for the presence of nitric oxide synthase. Light, transmission electron, and confocal laser scanning microscopy were used to study tissue sections processed for immunocytochemistry employing a monoclonal antibody against L-citrulline or polyclonal anti-neuronal nitric oxide synthase sera, and double immunofluorescence to detect neuronal nitric oxide synthase and L-citrulline co-localization. The results demonstrate that the same CNS regions and cell types are labeled by neuronal nitric oxide synthase polyclonal antisera and L-citrulline monoclonal antibodies, using both immunocytochemistry and immunofluorescence. Short-term pretreatment with a nitric oxide synthase inhibitor reduces L-citrulline immunostaining, but does not affect neuronal nitric oxide synthase immunoreactivity. In the vestibular brainstem, double immunofluorescence studies show that many, but not all, neuronal nitric oxide synthase-positive cells co-express L-citrulline, and that local intracellular patches of intense L-citrulline accumulation are present in some neurons. Conversely, all L-citrulline-labeled neurons co-express neuronal nitric oxide synthase. Cells expressing neuronal nitric oxide synthase alone are interpreted as neurons with the potential to produce nitric oxide under other stimulus conditions, and the subcellular foci of enhanced L-citrulline staining are viewed as intracellular sites of nitric oxide production. This interpretation is supported by ultrastructural observations of subcellular foci with enhanced L-citrulline and/or neuronal nitric oxide synthase staining that are located primarily at postsynaptic densities and portions of the endoplasmic reticulum. We conclude that nitric oxide is produced and released at focal sites within neurons that are identifiable using L-citrulline as a marker. Copyright 2002 IBRO.

  9. The effect of citrulline and arginine supplementation on lactic acidemia in MELAS syndrome.

    Science.gov (United States)

    El-Hattab, Ayman W; Emrick, Lisa T; Williamson, Kaitlin C; Craigen, William J; Scaglia, Fernando

    2013-12-01

    Mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes (MELAS) syndrome is a mitochondrial disorder in which nitric oxide (NO) deficiency may play a role in the pathogenesis of several complications including stroke-like episodes and lactic acidosis. Supplementing the NO precursors arginine and citrulline restores NO production in MELAS syndrome. In this study we evaluated the effect of arginine or citrulline on lactic acidemia in adults with MELAS syndrome. Plasma lactate decreased significantly after citrulline supplementation, whereas the effect of arginine supplementation did not reach statistical significance. These results support the potential therapeutic utility of arginine and citrulline in MELAS syndrome and suggest that citrulline supplementation may be more efficacious. However, therapeutic efficacy of these compounds should be further evaluated in clinical trials.

  10. Structure and pathogenicity of antibodies specific for citrullinated collagen type II in experimental arthritis

    DEFF Research Database (Denmark)

    Uysal, Hüseyin; Bockermann, Robert; Nandakumar, Kutty S;

    2009-01-01

    Antibodies to citrulline-modified proteins have a high diagnostic value in rheumatoid arthritis (RA). However, their biological role in disease development is still unclear. To obtain insight into this question, a panel of mouse monoclonal antibodies was generated against a major triple helical...... collagen type II (CII) epitope (position 359-369; ARGLTGRPGDA) with or without arginines modified by citrullination. These antibodies bind cartilage and synovial tissue, and mediate arthritis in mice. Detection of citrullinated CII from RA patients' synovial fluid demonstrates that cartilage-derived CII...

  11. Anticorpos antipeptídeos citrulinados e fator reumatoide em pacientes sudaneses com infecção por Leishmania donovani Anti-citrullinated peptide antibodies and rheumatoid factor in Sudanese patients with Leishmania donovani infection

    Directory of Open Access Journals (Sweden)

    Erik Ahlin

    2011-12-01

    Full Text Available OBJETIVO: Este estudo avaliou a presença de anticorpos antipeptídeos citrulinados cíclicos (anti-CCP, fator reumatoide (FR e imunocomplexos circulantes (ICC em pacientes sudaneses infectados por Leishmania donovani. PACIENTES E MÉTODOS: Os soros foram coletados de pacientes infectados por Leishmania (n = 116 e de sudaneses saudáveis (n = 93. Dezenove pacientes sudaneses com artrite reumatoide (AR e anti-CCP+ foram incluídos como controles positivos. Os níveis de ICC e anti-CCP foram medidos por ELISA. Para avaliar a reatividade citrulina-específica foi usada a placa-controle com peptídeos-controle cíclicos contendo arginina em vez de citrulina. RESULTADOS: Entre os pacientes infectados por Leishmania e os pacientes com AR e anti-CCP+, a maioria (86% era positiva para FR, enquanto a frequência de positividade para ICC foi maior entre pacientes com leishmaniose visceral (LV (LV 38%; AR e anti-CCP+ 24%. Quando foi analisada a reatividade anti-CCP, 12% dos pacientes com LV foram positivos. Os níveis de anti-CCP entre os pacientes com LV correlacionaram-se bem com os níveis de ICC encontrados (r = 0,65; P OBJECTIVE: The present study evaluated the presence of anti-cyclic citrullinated peptides antibodies (anti-CCP, rheumatoid factor (RF, and circulating immune complexes (CIC in Sudanese patients infected with the Leishmania donovani parasite. PATIENTS AND METHODS: Sera were collected from Leishmania infected patients (n = 116 and healthy Sudanese (n = 93. Nineteen Sudanese anti-CCP+ RA patients were included as positive controls. Levels of CIC and anti-CCP were measured by ELISA. Control plate with cyclic control peptides containing arginine instead of citrulline was used to evaluate citrulline specifi c reactivity. RESULTS: Among Leishmania-infected patients and anti-CCP+ RA patients, most were RF positive (86%, while the frequency of CIC positivity was higher among visceral leishmaniasis (VL patients (VL 38%; anti-CCP+ RA 24%. When

  12. Circulating levels of citrullinated and MMP-degraded vimentin (VICM) in liver fibrosis related pathology

    DEFF Research Database (Denmark)

    Vassiliadis, E.; Oliveira, C. P.; Alvares-da-Silva, M. R.;

    2012-01-01

    Aim: To investigate whether increased levels of vimentin citrullinated peptides identified by MS in articular cartilage can be measured in pathologies other than rheumatoid arthritis and be utilised for diagnostic purposes. Methods: A monoclonal antibody against the sequence RLRSSVPGV-citrulline ......Aim: To investigate whether increased levels of vimentin citrullinated peptides identified by MS in articular cartilage can be measured in pathologies other than rheumatoid arthritis and be utilised for diagnostic purposes. Methods: A monoclonal antibody against the sequence RLRSSVPGV......-citrulline (VICM) was developed and evaluated in a carbon tetrachloride (CCl4) (n=52 + 28 controls) rat model of liver fibrosis and two clinical cohorts of adult patients with hepatitis C (HCV) (n=92) and non-alcoholic fatty liver disease (NAFLD) (n=62), and compared to healthy controls. Results: In CCl4-treated...

  13. Circulating levels of citrullinated and MMP-degraded vimentin (VICM) in liver fibrosis related pathology

    DEFF Research Database (Denmark)

    Vassiliadis, Efstathios; Oliveira, Claudia P; Alvares-da-Silva, Mario R;

    2012-01-01

    AIM: To investigate whether increased levels of vimentin citrullinated peptides identified by MS in articular cartilage can be measured in pathologies other than rheumatoid arthritis and be utilised for diagnostic purposes. METHODS: A monoclonal antibody against the sequence RLRSSVPGV-citrulline ......AIM: To investigate whether increased levels of vimentin citrullinated peptides identified by MS in articular cartilage can be measured in pathologies other than rheumatoid arthritis and be utilised for diagnostic purposes. METHODS: A monoclonal antibody against the sequence RLRSSVPGV......-citrulline (VICM) was developed and evaluated in a carbon tetrachloride (CCl(4)) (n=52 + 28 controls) rat model of liver fibrosis and two clinical cohorts of adult patients with hepatitis C (HCV) (n=92) and non-alcoholic fatty liver disease (NAFLD) (n=62), and compared to healthy controls. RESULTS: In CCl(4...

  14. Characterization and Localization of Citrullinated Proteoglycan Aggrecan in Human Articular Cartilage.

    Directory of Open Access Journals (Sweden)

    Tibor T Glant

    Full Text Available Rheumatoid arthritis (RA is an autoimmune disease of the synovial joints. The autoimmune character of RA is underscored by prominent production of autoantibodies such as those against IgG (rheumatoid factor, and a broad array of joint tissue-specific and other endogenous citrullinated proteins. Anti-citrullinated protein antibodies (ACPA can be detected in the sera and synovial fluids of RA patients and ACPA seropositivity is one of the diagnostic criteria of RA. Studies have demonstrated that RA T cells respond to citrullinated peptides (epitopes of proteoglycan (PG aggrecan, which is one of the most abundant macromolecules of articular cartilage. However, it is not known if the PG molecule is citrullinated in vivo in human cartilage, and if so, whether citrulline-containing neoepitopes of PG (CitPG can contribute to autoimmunity in RA.CitPG was detected in human cartilage extracts using ACPA+ RA sera in dot blot and Western blot. Citrullination status of in vitro citrullinated recombinant G1 domain of human PG (rhG1 was confirmed by antibody-based and chemical methods, and potential sites of citrullination in rhG1 were explored by molecular modeling. CitPG-specific serum autoantibodies were quantified by enzyme-linked immunosorbent assays, and CitPG was localized in osteoarthritic (OA and RA cartilage using immunohistochemistry.Sera from ACPA+ RA patients reacted with PG purified from normal human cartilage specimens. PG fragments (mainly those containing the G1 domain from OA or RA cartilage extracts were recognized by ACPA+ sera but not by serum from ACPA- individuals. ACPA+ sera also reacted with in vitro citrullinated rhG1 and G3 domain-containing fragment(s of PG. Molecular modeling suggested multiple sites of potential citrullination within the G1 domain. The immunohistochemical localization of CitPG was different in OA and RA cartilage.CitPG is a new member of citrullinated proteins identified in human joints. CitPG could be found in

  15. Extensive Citrullination Promotes Immunogenicity of HSP90 through Protein Unfolding and Exposure of Cryptic Epitopes.

    Science.gov (United States)

    Travers, Timothy S; Harlow, Lisa; Rosas, Ivan O; Gochuico, Bernadette R; Mikuls, Ted R; Bhattacharya, Sanjoy K; Camacho, Carlos J; Ascherman, Dana P

    2016-09-01

    Post-translational protein modifications such as citrullination have been linked to the breach of immune tolerance and clinical autoimmunity. Previous studies from our laboratory support this concept, demonstrating that autoantibodies targeting citrullinated isoforms of heat shock protein 90 (HSP90) are associated with rheumatoid arthritis complicated by interstitial lung disease. To further explore the relationship between citrullination and structural determinants of HSP90 immunogenicity, we employed a combination of ELISA-based epitope profiling, computational modeling, and mass-spectrometric sequencing of peptidylarginine deiminase (PAD)-modified protein. Remarkably, ELISAs involving selected citrullinated HSP90β/α peptides identified a key epitope corresponding to an internal Arg residue (R502 [HSP90β]/R510 [HSP90α]) that is normally buried within the crystal structure of native/unmodified HSP90. In vitro time/dose-response experiments reveal an ordered pattern of PAD-mediated deimination events culminating in citrullination of R502/R510. Conventional as well as scaled molecular dynamics simulations further demonstrate that citrullination of selected Arg residues leads to progressive disruption of HSP90 tertiary structure, promoting exposure of R502/R510 to PAD modification and subsequent autoantibody binding. Consistent with this process, ELISAs incorporating variably deiminated HSP90 as substrate Ag indicate a direct relationship between the degree of citrullination and the level of ex vivo Ab recognition. Overall, these data support a novel structural paradigm whereby citrullination-induced shifts in protein structure generate cryptic epitopes capable of bypassing B cell tolerance in the appropriate genetic context.

  16. Citrulline as a Biomarker in the Non-human Primate Total- and Partial-body Irradiation Models: Correlation of Circulating Citrulline to Acute and Prolonged Gastrointestinal Injury.

    Science.gov (United States)

    Jones, Jace W; Bennett, Alexander; Carter, Claire L; Tudor, Gregory; Hankey, Kim G; Farese, Ann M; Booth, Catherine; MacVittie, Thomas J; Kane, Maureen A

    2015-11-01

    The use of plasma citrulline as a biomarker for acute and prolonged gastrointestinal injury via exposure to total- and partial-body irradiation (6 MV LINAC-derived photons; 0.80 Gy min) in nonhuman primate models was investigated. The irradiation exposure covered gastrointestinal injuries spanning lethal, mid-lethal, and sub-lethal doses. The acute gastrointestinal injury was assessed via measurement of plasma citrulline and small intestinal histopathology over the first 15 d following radiation exposure and included total-body irradiation at 13.0 Gy, 10.5 Gy, and 7.5 Gy and partial-body irradiation at 11.0 Gy with 5% bone marrow sparing. The dosing schemes of 7.5 Gy total-body irradiation and 11.0 Gy partial-body irradiation included time points out to day 60 and day 180, respectively, which allowed for correlation of plasma citrulline to prolonged gastrointestinal injury and survival. Plasma citrulline values were radiation-dependent for all radiation doses under consideration, with nadir values ranging from 63-80% lower than radiation-naïve NHP plasma. The nadir values were observed at day 5 to 7 post irradiation. Longitudinal plasma citrulline profiles demonstrated prolonged gastrointestinal injury resulting from acute high-dose irradiation had long lasting effects on enterocyte function. Moreover, plasma citrulline did not discriminate between total-body or partial-body irradiation over the first 15 d following irradiation and was not predictive of survival based on the radiation models considered herein.

  17. Plasma arginine and ornithine are the main citrulline precursors in mice infused with arginine-free diets.

    Science.gov (United States)

    Marini, Juan C; Didelija, Inka Cajo; Castillo, Leticia; Lee, Brendan

    2010-08-01

    Dietary arginine is the main dietary precursor for citrulline synthesis, but it is not known if other precursors can compensate when arginine is absent in the diet. To address this question, the contributions of plasma and dietary precursors were determined by using multitracer protocols in conscious mice infused i.g. either an arginine-sufficient diet [Arg(+)] or an arginine-free diet [Arg(-)]. The plasma entry rate of citrulline and arginine did not differ between the 2 diet groups (156 +/- 6 and 564 +/- 30 micromol kg(-1) h(-1), respectively); however, the entry rate of ornithine was greater in the mice fed the Arg(+) than the Arg(-) diet (332 +/- 33 vs. 180 +/- 16 micromol kg(-1) h(-1)). There was a greater utilization of plasma ornithine for the synthesis of citrulline (49 +/- 4 vs. 36 +/- 3 micromol kg(-1) h(-1), 30 +/- 3% vs. 24 +/- 2% of citrulline entry rate) in the mice fed the Arg(-) diet than the Arg(+) diet. The utilization of plasma arginine did not differ between the 2 diet groups for citrulline synthesis, either through plasma ornithine (approximately 29 +/- 3 micromol kg(-1) h(-1)) or at the site of citrulline synthesis (approximately 12 +/- 3 micromol kg(-1) h(-1)). The contribution of dietary proline to the synthesis of citrulline was mainly at the site of citrulline production (17 +/- 1 micromol kg(-1) h(-1)), rather than through plasma ornithine (5 +/- 0.4 micromol kg(-1) h(-1)). Dietary glutamine was utilized only at the site of citrulline synthesis (4 +/- 0.2 micromol kg(-1) h(-1)). Dietary glutamine and proline made a greater contribution to the synthesis of citrulline in mice fed the Arg(-) diet but remained minor sources for citrulline production. Plasma arginine and ornithine are able to support citrulline synthesis during arginine-free feeding.

  18. The periodontium of periodontitis patients contains citrullinated proteins which may play a role in ACPA (anti-citrullinated protein antibody) formation

    NARCIS (Netherlands)

    Nesse, Willem; Westra, Johanna; van der Wal, Jacqueline E.; Abbas, Frank; Nicholas, Anthony P.; Vissink, Arjan; Brouwer, Elisabeth; Westra J., [No Value

    2012-01-01

    Aim To determine the presence and location (stroma versus epithelium) of citrullinated proteins in periodontitis tissue as compared to non-periodontitis tissue and synovial tissue of RA patients. Materials & Methods Periodontitis, healthy periodontal and RA-affected synovial tissue samples were coll

  19. Citrullination-acetylation interplay guides E2F-1 activity during the inflammatory response.

    Science.gov (United States)

    Ghari, Fatemeh; Quirke, Anne-Marie; Munro, Shonagh; Kawalkowska, Joanna; Picaud, Sarah; McGouran, Joanna; Subramanian, Venkataraman; Muth, Aaron; Williams, Richard; Kessler, Benedikt; Thompson, Paul R; Fillipakopoulos, Panagis; Knapp, Stefan; Venables, Patrick J; La Thangue, Nicholas B

    2016-02-01

    Peptidyl arginine deiminase 4 (PAD4) is a nuclear enzyme that converts arginine residues to citrulline. Although increasingly implicated in inflammatory disease and cancer, the mechanism of action of PAD4 and its functionally relevant pathways remains unclear. E2F transcription factors are a family of master regulators that coordinate gene expression during cellular proliferation and diverse cell fates. We show that E2F-1 is citrullinated by PAD4 in inflammatory cells. Citrullination of E2F-1 assists its chromatin association, specifically to cytokine genes in granulocyte cells. Mechanistically, citrullination augments binding of the BET (bromodomain and extra-terminal domain) family bromodomain reader BRD4 (bromodomain-containing protein 4) to an acetylated domain in E2F-1, and PAD4 and BRD4 coexist with E2F-1 on cytokine gene promoters. Accordingly, the combined inhibition of PAD4 and BRD4 disrupts the chromatin-bound complex and suppresses cytokine gene expression. In the murine collagen-induced arthritis model, chromatin-bound E2F-1 in inflammatory cells and consequent cytokine expression are diminished upon small-molecule inhibition of PAD4 and BRD4, and the combined treatment is clinically efficacious in preventing disease progression. Our results shed light on a new transcription-based mechanism that mediates the inflammatory effect of PAD4 and establish the interplay between citrullination and acetylation in the control of E2F-1 as a regulatory interface for driving inflammatory gene expression. PMID:26989780

  20. Myelin Basic Protein Citrullination in Multiple Sclerosis: A Potential Therapeutic Target for the Pathology.

    Science.gov (United States)

    Yang, Lei; Tan, Dewei; Piao, Hua

    2016-08-01

    Multiple sclerosis (MS) is a multifactorial demyelinating disease characterized by neurodegenerative events and autoimmune response against myelin component. Citrullination or deimination, a post-translational modification of protein-bound arginine into citrulline, catalyzed by Ca(2+) dependent peptidylarginine deiminase enzyme (PAD), plays an essential role in physiological processes include gene expression regulation, apoptosis and the plasticity of the central nervous system, while aberrant citrullination can generate new epitopes, thus involving in the initiation and/or progression of autoimmune disorder like MS. Myelin basic protein (MBP) is the major myelin protein and is generally considered to maintain the stability of the myelin sheath. This review describes the MBP citrullination and its consequence, as well as offering further support for the "inside-out" hypothesis that MS is primarily a neurodegenerative disease with secondary inflammatory demyelination. In addition, it discusses the role of MBP citrullination in the immune inflammation and explores the potential of inhibition of PAD enzymes as a therapeutic strategy for the disease.

  1. Monoclonal L-citrulline immunostaining reveals nitric oxide-producing vestibular neurons

    Science.gov (United States)

    Holstein, G. R.; Friedrich, V. L. Jr; Martinelli, G. P.

    2001-01-01

    Nitric oxide is an unstable free radical that serves as a novel messenger molecule in the central nervous system (CNS). In order to understand the interplay between classic and novel chemical communication systems in vestibular pathways, the staining obtained using a monoclonal antibody directed against L-citrulline was compared with the labeling observed using more traditional markers for the presence of nitric oxide. Brainstem tissue from adult rats was processed for immunocytochemistry employing a monoclonal antibody directed against L-citrulline, a polyclonal antiserum against neuronal nitric oxide synthase, and/or NADPH-diaphorase histochemistry. Our findings demonstrate that L-citrulline can be fixed in situ by vascular perfusion, and can be visualized in fixed CNS tissue sections by immunocytochemistry. Further, the same vestibular regions and cell types are labeled by NADPH-diaphorase histochemistry, by the neuronal nitric oxide synthase antiserum, and by our anti-L-citrulline antibody. Clusters of L-citrulline-immunoreactive neurons are present in subregions of the vestibular nuclei, including the caudal portion of the inferior vestibular nucleus, the magnocellular portion of the medial vestibular nucleus, and the large cells in the ventral tier of the lateral vestibular nucleus. NADPH-diaphorase histochemical staining of these neurons clearly demonstrated their multipolar, fusiform and globular somata and long varicose dendritic processes. These results provide support for the suggestion that nitric oxide serves key roles in both vestibulo-autonomic and vestibulo-spinal pathways.

  2. L-citrulline supplementation reverses the impaired airway relaxation in neonatal rats exposed to hyperoxia

    Directory of Open Access Journals (Sweden)

    Sopi Ramadan B

    2012-08-01

    Full Text Available Abstract Background Hyperoxia is shown to impair airway relaxation via limiting L-arginine bioavailability to nitric oxide synthase (NOS and reducing NO production as a consequence. L-arginine can also be synthesized by L-citrulline recycling. The role of L-citrulline supplementation was investigated in the reversing of hyperoxia-induced impaired relaxation of rat tracheal smooth muscle (TSM. Methods Electrical field stimulation (EFS, 2–20 V-induced relaxation was measured under in vitro conditions in preconstricted tracheal preparations obtained from 12 day old rat pups exposed to room air or hyperoxia (>95% oxygen for 7 days supplemented with L-citrulline or saline (in vitro or in vivo. The role of the L-citrulline/L-arginine cycle under basal conditions was studied by incubation of preparations in the presence of argininosuccinate synthase (ASS inhibitor [α-methyl-D, L-aspartate, 1 mM] or argininosuccinate lyase inhibitor (ASL succinate (1 mM and/or NOS inhibitor [Nω-nitro-L-arginine methyl ester; 100 μM] with respect to the presence or absence of L-citrulline (2 mM. Results Hyperoxia impaired the EFS-induced relaxation of TSM as compared to room air control (p ; 0.5 ± 0.1% at 2 V to 50.6 ± 5.7% at 20 V in hyperoxic group: 0.7 ± 0.2 at 2 V to 80.0 ± 5.6% at 20 V in room air group. Inhibition of ASS or ASL, and L-citrulline supplementation did not affect relaxation responses under basal conditions. However, inhibition of NOS significantly reduced relaxation responses (p in vivo and in vitro also reversed the hyperoxia-impaired relaxation. The differences were significant (p ; 0.8 ± 0.3% at 2 V to 47.1 ± 4.1% at 20 V without L-citrulline; 0.9 ± 0.3% at 2 V to 68.2 ± 4.8% at 20 V with L-citrulline. Inhibition of ASS or ASL prevented this effect of L-citrulline. Conclusion The results indicate the presence of an L-citrulline/L-arginine cycle in the airways of rat pups

  3. Effects of L-citrulline diet on stress-induced cold hypersensitivity in mice

    Directory of Open Access Journals (Sweden)

    Yoshinori Kobayashi

    2014-01-01

    Full Text Available Background: L-citrulline is an amino acid discovered in watermelon (Citrullus lanatus, Cucurbitaceae and is a known component of the nitric oxide (NO cycle that plays an important role in adjusting blood circulation and supplying NO and a key component of the endothelium-derived relaxing factor. Objective: The objective of this study is to evaluate the effect of L-citrulline on a newly established stress-induced cold hypersensitivity mouse model. Materials and Methods: When normal mice were forced to swim in water at 25°C for 15 min, their core body temperature dropped to 28.9°C, and then quickly recovered to normal temperature after the mice were transferred to a dry cage at room temperature (25°C. A 1-h immobilization before swimming caused the core body temperature to drop to ca. 24.1°C (4.8°C lower than normal mice, and the speed of core body temperature recovery dropped to 57% of the normal control. We considered this delay in recovery from hypothermia to be a sign of stress-induced cold hypersensitivity. Similar cold hypersensitivity was induced by administration of 50 mM L-NG-nitroarginine methyl ester, a NO synthesis inhibitor. Results: In this study, we showed that recovery speed from the stress-induced hypothermia remarkably improved in mice fed a 1% L-citrulline-containing diet for 20 days. Furthermore, the nonfasting blood level of L-arginine and L-citrulline increased significantly in the L-citrulline diet group, and higher serum nitrogen oxide levels were observed during recovery from the cold. Conclusions: These results suggested that oral L-citrulline supplementation strengthens vascular endothelium function and attenuates stress-induced cold hypersensitivity by improving blood circulation.

  4. Contribution of peptide backbone to Anti-citrulline-dependent antibody reactivity

    DEFF Research Database (Denmark)

    Trier, Nicole Hartwig; Dam, Catharina; Olsen, Dorthe;

    2015-01-01

    Rheumatoid arthritis (RA) is one of the most common autoimmune diseases, affecting approximately 1–2% of the world population. One of the characteristic features of RA is the presence of autoantibodies. Especially the highly specific anti-citrullinated peptide antibodies (ACPAs), which have been ...... homology rather than sequence homology are favored between citrullinated epitopes. These findings are important in relation to clarifying the etiology of RA and to determine the nature of ACPAs, e.g. why some Cit-Gly-containing sequences are not targeted by ACPAs....

  5. Glutamine supplementation, citrulline production, and de novo arginine synthesis: Is there a relation?

    Science.gov (United States)

    We would like to comment on the recent publications by Buijs et al. The authors hypothesized that a parenteral supplement of glutamine stimulates citrulline formation and enhances de novo arginine synthesis. To test this hypothesis, they conducted an experiment with stable isotopes in patients under...

  6. Impaired nitric oxide production in children with MELAS syndrome and the effect of arginine and citrulline supplementation.

    Science.gov (United States)

    El-Hattab, Ayman W; Emrick, Lisa T; Hsu, Jean W; Chanprasert, Sirisak; Almannai, Mohammed; Craigen, William J; Jahoor, Farook; Scaglia, Fernando

    2016-04-01

    Mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes (MELAS) syndrome is one of the most frequent maternally inherited mitochondrial disorders. The pathogenesis of this syndrome is not fully understood and believed to result from several interacting mechanisms including impaired mitochondrial energy production, microvasculature angiopathy, and nitric oxide (NO) deficiency. NO deficiency in MELAS syndrome is likely to be multifactorial in origin with the decreased availability of the NO precursors, arginine and citrulline, playing a major role. In this study we used stable isotope infusion techniques to assess NO production in children with MELAS syndrome and healthy pediatric controls. We also assessed the effect of oral arginine and citrulline supplementations on NO production in children with MELAS syndrome. When compared to control subjects, children with MELAS syndrome were found to have lower NO production, arginine flux, plasma arginine, and citrulline flux. In children with MELAS syndrome, arginine supplementation resulted in increased NO production, arginine flux, and arginine concentration. Citrulline supplementation resulted in a greater increase of these parameters. Additionally, citrulline supplementation was associated with a robust increase in citrulline concentration and flux and de novo arginine synthesis rate. The greater effect of citrulline in increasing NO production is due to its greater ability to increase arginine availability particularly in the intracellular compartment in which NO synthesis takes place. This study, which is the first one to assess NO metabolism in children with mitochondrial diseases, adds more evidence to the notion that NO deficiency occurs in MELAS syndrome, suggests a better effect for citrulline because of its greater role as NO precursor, and indicates that impaired NO production occurs in children as well as adults with MELAS syndrome. Thus, the initiation of treatment with NO precursors may be

  7. Increased plasma citrulline in mice marks diet-induced obesity and may predict the development of the metabolic syndrome.

    Directory of Open Access Journals (Sweden)

    Manuela Sailer

    Full Text Available In humans, plasma amino acid concentrations of branched-chain amino acids (BCAA and aromatic amino acids (AAA increase in states of obesity, insulin resistance and diabetes. We here assessed whether these putative biomarkers can also be identified in two different obesity and diabetic mouse models. C57BL/6 mice with diet-induced obesity (DIO mimic the metabolic impairments of obesity in humans characterized by hyperglycemia, hyperinsulinemia and hepatic triglyceride accumulation. Mice treated with streptozotocin (STZ to induce insulin deficiency were used as a type 1 diabetes model. Plasma amino acid profiling of two high fat (HF feeding trials revealed that citrulline and ornithine concentrations are elevated in obese mice, while systemic arginine bioavailability (ratio of plasma arginine to ornithine + citrulline is reduced. In skeletal muscle, HF feeding induced a reduction of arginine levels while citrulline levels were elevated. However, arginine or citrulline remained unchanged in their key metabolic organs, intestine and kidney. Moreover, the intestinal conversion of labeled arginine to ornithine and citrulline in vitro remained unaffected by HF feeding excluding the intestine as prime site of these alterations. In liver, citrulline is mainly derived from ornithine in the urea cycle and DIO mice displayed reduced hepatic ornithine levels. Since both amino acids share an antiport mechanism for mitochondrial import and export, elevated plasma citrulline may indicate impaired hepatic amino acid handling in DIO mice. In the insulin deficient mice, plasma citrulline and ornithine levels also increased and additionally these animals displayed elevated BCAA and AAA levels like insulin resistant and diabetic patients. Therefore, type 1 diabetic mice but not DIO mice show the "diabetic fingerprint" of plasma amino acid changes observed in humans. Additionally, citrulline may serve as an early indicator of the obesity-dependent metabolic

  8. An in silico model of enterocytic glutamine to citrulline conversion pathway.

    Science.gov (United States)

    Bensaci, J; Curis, E; Nicolis, I; de Bandt, J-P; Bénazeth, S

    2012-10-01

    Enterocyte is one of the main sites of amino acids metabolism and particularly of the citrulline biosynthesis. Working at the cellular scale and applying ordinary differential equations (ODEs) formalism, we have built a mathematical model of the enterocytic glutamine to citrulline conversion in the fasting state. This model enables us to test different physiopathological scenarios of enzyme activity loss. Results from two different approaches were compared: a standard approach (KA) based on the Michaelis-Menten assumptions and an association-dissociation approach (VH) based on the kinetic mass action law. For both approaches, ODEs system was numerically solved using Mathematica™. In both cases, the model correctly predicts the physiological plasma citrulline steady-state, but the two approaches present clear differences for metabolites of enzymes having a complex mechanism, challenging the validity of the KA approach in such cases. When physiopathological scenarios of enzyme activity loss are simulated, both approaches predict a very sharp transition from the physiological citrulline plasma level to the lack of its production: the concentration profiles of these simulations show a clear threshold of which characteristics vary with the involved enzyme. Moreover, amongst all enzymes included in the model, the ornithine aminotransferase (OAT) shows the highest sensitivity in the system whatever the approach used. This model points out the limits of the Michaelis-Menten approach to model complex enzyme mechanisms. It highlights the key role of OAT in the studied citrulline synthesis pathway and also suggests an order of magnitude about the optimal ratio of enzyme concentrations in this pathway. PMID:22399052

  9. Evaluation of Clinical Usefulness of Antikeratin Antibody, Antiperinuclear Factor and Anti-Cyclic Citrullinated Peptide Antibody in Juvenile Rheumatoid Arthritis%血清抗角蛋白抗体、抗核周因子和抗环瓜氨酸肽抗体在幼年类风湿关节炎应用中的评价

    Institute of Scientific and Technical Information of China (English)

    韩彤昕; 李彩凤; 何晓琥; 王江; 邝伟英; 周怡芳

    2007-01-01

    目的 评价抗角蛋白抗体(AKA)、抗核周因子(APF)和抗环瓜氨酸肽抗体(anti-CCP)在幼年类风湿关节炎(JRA)应用中的意义.方法 收集76例JRA及54例非JRA患儿,健康对照30例血清,测定其AKA、APF和抗CCP水平,对JRA诊断敏感性、特异性进行评价,并对JRA患儿中3种抗体阳性和阴性组的临床症状、体征及实验室检查指标进行比较,并作统计学分析.结果 AKA、APF和抗CCP及联合检测ROC曲线下面积略高于0.5, 但Pa>0.05.单一抗体检测灵敏度为30%~50%,联合检测灵敏度无增加.单一抗体检测的特异度为50%~80%,联合检测特异度增加至80%~85%.JRA组AKA(+)病例较多出现关节强直,心包积液,骨质稀疏,肺炎支原体抗体阳性率相对高,且ASO、ESR、CRP水平较高,Pa<0.05;APF(+)病例较多出现关节强直,骨质稀疏,肺炎支原体抗体阳性率相对高,而且ASO、ESR、CRP水平较高,有显著差异(P<0.05),抗CCP(+)病例与抗CCP(-)病例比较,在临床表现、实验室检查方面无显著差异.结论 AKA、APF、抗CCP抗体对JRA缺乏早期诊断意义及特异性,AKA、APF对判断疾病的活动性、病理损害程度和预后有临床意义.

  10. The Comparision of Anti-cyclic Citrullinated Peptide Antibody and Rheumatoid Factor in Rheumatoid Arthritis Diagnosis%抗环瓜氨酸多肽抗体和类风湿因子在类风湿性关节炎诊断中的比较

    Institute of Scientific and Technical Information of China (English)

    周厚清; 许赤多; 张辉; 董敏

    2009-01-01

    目的:检测抗环瓜氨酸肽(CCP)抗体在类风湿性关节炎(RA)中的阳性率,探讨抗CCP抗体检测在RA中的意义.方法:用ELISA方法检测30例RA,35例其他风湿性疾病,20例非风湿性疾病中的抗CCP抗体的分布.用免疫比浊法测定类风湿因子(RF),比较抗CCP抗体与RF的相关性.结果:30例RA病人中,抗CCP抗体的阳性率为73.33%.抗CCP抗体对RA的敏感性和特异性分别为73.33%和94.54%.RF的阳性率为76.67%.RF抗体对RA的敏感性和特异性分别为76.67%和80.0%.抗CCP抗体与RF之间呈正相关性.结论:抗CCP抗体对RA具有良好的敏感性和特异性,可视为RA新的血清学诊断指标,能用于RA的诊断.同时联合检测抗CCP抗体和RF有助于提高RA检测的敏感性和特异性.

  11. Value of anti-cyclic peptide containing citrulline antibody for the early diagnosis of rheumatoid arthritis%类风湿关节炎早期诊断中抗环瓜氨酸肽抗体检测的意义

    Institute of Scientific and Technical Information of China (English)

    葛艳玲; 宋慧; 刘武征; 杨凯楠

    2015-01-01

    目的 探讨单独或联合检测抗环瓜氨酸肽抗体(anti-CCP)、类风湿因子(RF)对类风湿关节炎(RA)早期诊断的价值. 方法 选取2012-2014年门诊及住院患者1 961例,已确诊为RA患者共509例(RA组),非RA患者共1 028例(非RA组),初步诊断非RA,随诊确诊为RA患者424例(初诊非RA随诊为RA组),分别采用速率散射比浊法检测RF;电化学发光法检测anti-CCP.结果 RA组与非RA组比较,单独检测anti-CCP与联合检测anti-CCP/RF的敏感度比较差异无统计学意义(P>0.05),而特异度(88.6%比60.4%)比较差异有统计学意义(P0.05), but the specificity between two groups (88.6%vs. 60.4%) was significant difference ( P<0.05). There were significant differences in the sensitivity (81.7% vs. 74.3%) and specificity (88.6% vs. 66.0%) between by using anti-CCP antibody alone and RF alone.In firstly was not diagnosed RA but later was diagnosed RA group, there were significantly difference in sensitivity (98.3%vs. 82.1%) and specificity (91.6%vs. 81.5%) by using anti-CCP antibody alone and RF alone. Conclusion There is important clinical value by using anti-CCP antibody alone for the early diagnosis of RA.

  12. The 2007 ESPEN Sir David Cuthbertson Lecture: amino acids between and within organs. The glutamate-glutamine-citrulline-arginine pathway.

    Science.gov (United States)

    Deutz, Nicolaas E P

    2008-06-01

    In daily practice, the plasma concentration of amino acids is usually viewed as a parameter of production. However, both a high production and/or a reduced disposal capacity can result in an increased plasma concentration. In this presentation, I will discuss my research on interorgan relationships of the amino acids glutamate, glutamine, citrulline and arginine to explain the regulation of the plasma arginine level. The reduced glutamine disposal during liver failure is related to enhanced plasma glutamine level without any change in muscle and gut production or consumption rate. In contrast during sepsis, a small reduction in plasma glutamine is related to a substantially enhanced organ glutamate and glutamine production or consumption rate. These observations are a good example that plasma levels are directly related to production or consumption rates. Because glutamine breakdown in the gut produces citrulline, there is a good relation between the amount of metabolically active gut tissue and gut and whole body citrulline production. Arginine is produces from citrulline in the kidney and a reduced gut glutamine to citrulline conversion during sepsis explains the reduced de novo arginine production that is related to the reduced plasma arginine level. The interorgan route between muscle, gut, liver and kidney of the amino acids glutamate, glutamine, citrulline and arginine is a very good example of how complicated the regulation of plasma amino acid levels can be. However, in-depth research is necessary and will give us important clues to new nutritional strategies.

  13. Serum metabolomics identifies citrulline as a predictor of adverse outcomes in an equine model of gut-derived sepsis.

    Science.gov (United States)

    Steelman, Samantha M; Johnson, Philip; Jackson, Amy; Schulze, James; Chowdhary, Bhanu P

    2014-05-15

    Acute laminitis is an inflammatory disease of the equine foot that often occurs secondarily to sepsis or systemic inflammation associated with gastrointestinal disease. It has been suggested that laminitis is similar to multiple organ dysfunction syndrome in humans, although in horses the weight-bearing laminar epithelium of the foot appears to be the tissue most sensitive to insult and the first "organ" to fail. Metabolomics performed on serum samples collected before (Con) and after (Lmn) experimental induction of gastrointestinal-associated sepsis in six horses detected 1,177 metabolites of both mammalian and bacterial origin in equine serum. Network and correlation analyses suggested a dysregulation of fatty acid metabolism in the Lmn group, as well as an accumulation of organic acids such as lactate. Furthermore, concentrations of the amino acid citrulline were decreased in Lmn samples from all study animals, suggesting that citrulline might be useful as a biomarker to identify critically ill animals that are at risk of developing laminitis. We therefore established normal ranges of plasma citrulline concentrations in a separate group of horses (n = 36) and tested the ability of citrulline to predict adverse outcomes (laminitis or death) in critically ill horses (n = 23). Plasma citrulline was significantly lower in critically ill horses that went on to experience adverse outcomes (n = 6). Further study is required to accurately determine a diagnostic cutoff, but the present data are suggestive of the predictive value of citrulline as a biomarker for laminar failure in equine sepsis. PMID:24619519

  14. Intestinal absorption in lysinuric protein intolerance: impaired for diamino acids, normal for citrulline.

    OpenAIRE

    Rajantie, J.; Simell, O.; Perheentupa, J

    1980-01-01

    Lysinuric protein intolerance (LPI) is an autosomal recessive defect of diamino acid transport characterised by massive diaminoaciduria, especially lysinuria, with hyperammonaemia after heavy nitrogen intake. The defect has previously been demonstrated in the kidney, and is probably present in the liver cells. To evaluate the effect of the LPI gene on the net intestinal absorption of the diamino acids and citrulline, separate oral loads of each were given to controls, and to subjects heterozy...

  15. Does Citrulline Have Protective Effects on Liver Injury in Septic Rats?

    Directory of Open Access Journals (Sweden)

    Bin Cai

    2016-01-01

    Full Text Available Citrulline (Cit supplementation was proposed to serve as a therapeutic intervention to restore arginine (Arg concentrations and improve related functions in sepsis. This study explored whether citrulline had positive effects on liver injury and cytokine release in the early stages of sepsis. The cecal ligation and puncture (CLP model was utilized in our study. Rats were divided into four groups: normal, Cit, CLP, and CLP+Cit. The CLP group and CLP+Cit group were separated into 6-, 12-, and 24-hour groups, according to the time points of sacrifice after surgery. Intragastric administration of L-citrulline was applied to rats in Cit and CLP+Cit groups before surgery. Serum AST and ALT levels and levels of MDA, SOD, NO, and iNOS in the liver tissues were evaluated. Plasma concentrations of Cit and Arg were assessed using HPLC-MS/MS. Serum concentrations of cytokines and chemokines were calculated by Luminex. Results showed SOD activities of CLP+Cit groups were significantly higher than that of CLP groups, contrasting with the MDA and NO levels which were significantly lower in CLP+Cit groups than in CLP groups. In addition, plasma concentrations of TNF-α, IL-6, and IL-1β were significantly lower in the CLP+Cit 6-hour group than in the CLP 6-hour group.

  16. Quantitative analysis of 15N labeled positional isomers of glutamine and citrulline via electrospray ionization tandem mass spectrometry of their dansyl derivatives

    Science.gov (United States)

    The enteral metabolism of glutamine and citrulline are intertwined because, while glutamine is one of the main fuel sources for the enterocyte, citrulline is one of its products. It has been shown that the administration of 15N labeled glutamine results in the incorporation of the 15N label into cit...

  17. Acute Citrulline-Malate Supplementation and High-Intensity Cycling Performance.

    Science.gov (United States)

    Cunniffe, Brian; Papageorgiou, Maria; OʼBrien, Barbara; Davies, Nathan A; Grimble, George K; Cardinale, Marco

    2016-09-01

    Cunniffe, B, Papageorgiou, M, O'Brien, B, Davies, NA, Grimble, GK, and Cardinale, M. Acute citrulline-malate supplementation and high-intensity cycling performance. J Strength Cond Res 30(9): 2638-2647, 2016-Dietary L-citrulline-malate (CM) consumption has been suggested to improve skeletal muscle metabolism and contractile efficiency, which would be expected to predispose exercising individuals to greater fatigue resistance. The purpose of this study was to examine the effects of CM supplementation on acid-base balance and high-intensity exercise performance. In a double-blind, placebo-controlled, crossover study, 10 well-trained males consumed either 12 g of CM (in 400 ml) or lemon sugar-free cordial (placebo [PL]) 60 minutes before completion of 2 exercise trials. Each trial consisted of subjects performing 10 (×15 seconds) maximal cycle sprints (with 30-second rest intervals) followed by 5 minutes recovery before completing a cycle time-to-exhaustion test (TTE) at 100% of individual peak power (PP). Significant increases in plasma concentrations of citrulline (8.8-fold), ornithine (3.9-fold), and glutamine (1.3-fold) were observed 60 minutes after supplementation in the CM trial only (p ≤ 0.05) and none of the subjects experienced gastrointestinal side-effects during testing. Significantly higher exercise heart rates were observed in CM condition (vs. PL) although no between trial differences in performance related variables (TTE: [120 ± 61 seconds CM vs. 113 ± 50 seconds PL]), PP or mean power, ([power fatigue index: 36 ± 16% CM vs. 28 ± 18% PL]), subjective rating of perceived exertion or measures of acid-base balance (pH, lactate, bicarbonate, base-excess) were observed (p > 0.05). This study demonstrated that acute supplementation of 12 g CM does not provide acute ergogenic benefits using the protocol implemented in this study in well-trained males. PMID:26808848

  18. L-citrulline protects from kidney damage in type 1 diabetic mice.

    OpenAIRE

    Romero, Maritza J.; Lin eYao; Supriya eSridhar; Anil eBhatta; Huijuan eDou; Ganesan eRamesh; Michael W Brands; Pollock, David M.; Caldwell, Ruth B.; Cederbaum, Stephen D.; C Alvin eHead; Zsolt eBagi; Rudolf eLucas; Robert William Caldwell

    2013-01-01

    Rationale. Diabetic nephropathy is a major cause of end-stage renal disease, associated with endothelial dysfunction. Chronic supplementation of L-arginine (L-arg), the substrate for endothelial nitric oxide synthase (eNOS), failed to improve vascular function. L-citrulline (L-cit) supplementation not only increases L-arg synthesis, but also inhibits cytosolic arginase I (Arg I), a competitor of eNOS for the use of L-arg, in the vasculature. Aims. To investigate whether L-cit treatment reduc...

  19. l-Citrulline Protects from Kidney Damage in Type 1 Diabetic Mice

    OpenAIRE

    Romero, Maritza J.; Yao, Lin; Sridhar, Supriya; Bhatta, Anil; Dou, Huijuan; Ramesh, Ganesan; Michael W Brands; Pollock, David M.; Caldwell, Ruth B.; Cederbaum, Stephen D.; Head, C. Alvin; Bagi, Zsolt; Lucas, Rudolf; Caldwell, Robert W.

    2013-01-01

    Rationale: Diabetic nephropathy (DN) is a major cause of end-stage renal disease, associated with endothelial dysfunction. Chronic supplementation of l-arginine (l-arg), the substrate for endothelial nitric oxide synthase (eNOS), failed to improve vascular function. l-Citrulline (l-cit) supplementation not only increases l-arg synthesis, but also inhibits cytosolic arginase I, a competitor of eNOS for the use of l-arg, in the vasculature. Aims: To investigate whether l-cit treatment reduce...

  20. Newborn screening for dihydrolipoamide dehydrogenase deficiency: Citrulline as a useful analyte

    Directory of Open Access Journals (Sweden)

    Shane C. Quinonez

    2014-01-01

    Full Text Available Dihydrolipoamide dehydrogenase deficiency, also known as maple syrup urine disease (MSUD type III, is caused by the deficiency of the E3 subunit of branched chain alpha-ketoacid dehydrogenase (BCKDH, α-ketoglutarate dehydrogenase (αKGDH, and pyruvate dehydrogenase (PDH. DLD deficiency variably presents with either a severe neonatal encephalopathic phenotype or a primarily hepatic phenotype. As a variant form of MSUD, it is considered a core condition recommended for newborn screening. The detection of variant MSUD forms has proven difficult in the past with no asymptomatic DLD deficiency patients identified by current newborn screening strategies. Citrulline has recently been identified as an elevated dried blood spot (DBS metabolite in symptomatic patients affected with DLD deficiency. Here we report the retrospective DBS analysis and second-tier allo-isoleucine testing of 2 DLD deficiency patients. We show that an elevated citrulline and an elevated allo-isoleucine on second-tier testing can be used to successfully detect DLD deficiency. We additionally recommend that DLD deficiency be included in the “citrullinemia/elevated citrulline” ACMG Act Sheet and Algorithm.

  1. Citrullination as early-stage indicator of cell response to single-walled carbon nanotubes.

    Science.gov (United States)

    Mohamed, Bashir Mustafa; Movia, Dania; Knyazev, Anton; Langevin, Dominique; Davies, Anthony Mitchell; Prina-Mello, Adriele; Volkov, Yuri

    2013-01-01

    Single-walled carbon nanotubes (SWCNTs) have been widely explored as potential technologies for information systems and medical applications. The impact of SWCNTs on human health is of prime concern, if SWCNTs have a future in the manufacturing industry. This study proposes a novel, inflammation-independent paradigm of toxicity for SWCNTs, identifying the protein citrullination process as early-stage indicator of inflammatory responses of macrophages (THP-1) and of subtle phenotypic damages of lung epithelial (A549) cells following exposure to chemically-treated SWCNTs. Our results showed that, while most of the cellular responses of A549 cells exposed to SWCNTs are different to those of similarly treated THP-1 cells, the protein citrullination process is triggered in a dose- and time-dependent manner in both cell lines, with thresholds comparable between inflammatory (THP-1) and non-inflammatory (A549) cell types. The cellular mechanism proposed herein could have a high impact in predicting the current risk associated with environmental exposure to SWCNTs. PMID:23350031

  2. Differentiation of Neisseria gonorrhoeae isolates requiring proline, citrulline, and uracil by plasmid content, serotyping, and pulsed-field gel electrophoresis.

    OpenAIRE

    Ng, L K; Carballo, M.; Dillon, J A

    1995-01-01

    A combination of DNA macrorestriction analysis using pulsed-field gel electrophoresis and a serotyping method using three panels of monoclonal antibody was used to discriminate 43 epidemiologically unrelated Neisseria gonorrhoeae isolates requiring proline, citrulline, and uracil (PCU-) into 35 groups. This indicates that PCU- isolates of N. gonorrhoeae are not clonal.

  3. Apo AIV and Citrulline Plasma Concentrations in Short Bowel Syndrome Patients: The Influence of Short Bowel Anatomy

    Science.gov (United States)

    Targarona, Jordi; Ruiz, Jorge; García, Natalia; Oró, Denise; García-Villoria, Judit; Creus, Gloria; Pita, Ana M.

    2016-01-01

    Introduction Parenteral nutrition (PN) dependence in short bowel syndrome (SBS) patients is linked to the functionality of the remnant small bowel (RSB). Patients may wean off PN following a period of intestinal adaptation that restores this functionality. Currently, plasma citrulline is the standard biomarker for monitoring intestinal functionality and adaptation. However, available studies reveal that the relationship the biomarker with the length and function of the RSB is arguable. Thus, having additional biomarkers would improve pointing out PN weaning. Aim By measuring concomitant changes in citrulline and the novel biomarker apolipoprotein AIV (Apo AIV), as well as taking into account the anatomy of the RSB, this exploratory study aims to a better understanding of the intestinal adaptation process and characterization of the SBS patients under PN. Methods Thirty four adult SBS patients were selected and assigned to adapted (aSBS) and non-adapted (nSBS) groups after reconstructive surgeries. Remaining jejunum and ileum lengths were recorded. The aSBS patients were either on an oral diet (ORAL group), those with intestinal insufficiency, or on oral and home parenteral nutrition (HPN group), those with chronic intestinal failure. Apo AIV and citrulline were analyzed in plasma samples after overnight fasting. An exploratory ROC analysis using citrulline as gold standard was performed. Results Biomarkers, Apo AIV and citrulline showed a significant correlation with RSBL in aSBS patients. In jejuno-ileocolic patients, only Apo AIV correlated with RSBL (rb = 0.54) and with ileum length (rb = 0.84). In patients without ileum neither biomarker showed any correlation with RSBL. ROC analysis indicated the Apo AIV cut-off value to be 4.6 mg /100 mL for differentiating between the aSBS HPN and ORAL groups. Conclusions Therefore, in addition to citrulline, Apo AIV can be set as a biomarker to monitor intestinal adaptation in SBS patients. As short bowel anatomy is shown

  4. Accuracy and standardization of diagnostic methods for the detection of antibodies to citrullinated peptides

    Directory of Open Access Journals (Sweden)

    M. Tampoia

    2011-06-01

    Full Text Available Anti-citrullinated peptide antibodies (ACPA have a very high specificity for rheumatoid arthritis, much more than that of the rheumatoid factor. In addition, ACPA can be found in sera in the pre-clinical phase, are associated with more severe joint destruction and with higher disease activity. In recent years, keeping pace with new knowledge and with progress made in the antigenic composition of tests and in the characterization of immunogenic epitopes, many immunoenzymatic (ELISA methods of second and third generation have been produced and marketed commercially, and their use has spread among clinical laboratories. Today, completely automated methods are also available, which are easy to use and with a higher throughput, rendering the diagnostic utility of testing ever faster and more effective. This review takes into consideration the more important characteristics of the new ACPA-ELISA tests now commercially available, and also considers recent progress in standardizing test results.

  5. Branched-chain amino acids, arginine, citrulline alleviate central fatigue after 3 simulated matches in taekwondo athletes: a randomized controlled trial

    OpenAIRE

    Chen, I-Fan; Wu, Huey-June; Chen, Chung-Yu; Chou, Kuei-Ming; Chang, Chen-Kang

    2016-01-01

    Background The decline in cognitive performance has been shown after fatiguing exercise. Branched-chain amino acids (BCAA) have been suggested to alleviate exercise-induced central fatigue. Arginine and citrulline could remove the excess NH3 accumulation accompanied with BCAA supplementation by increasing nitric oxide biosynthesis and/or urea cycle. The purpose of this study is to investigate the effect of the combined supplementation of BCAA, arginine, and citrulline on central fatigue after...

  6. The Supplementation of Branched-Chain Amino Acids, Arginine, and Citrulline Improves Endurance Exercise Performance in Two Consecutive Days

    Directory of Open Access Journals (Sweden)

    I-Shiung Cheng, Yi-Wen Wang, I-Fan Chen, Gi-Sheng Hsu, Chun-Fang Hsueh, Chen-Kang Chang

    2016-09-01

    Full Text Available The central nervous system plays a crucial role in fatigue during endurance exercise. Branched-chain amino acids (BCAA could reduce cerebral serotonin synthesis by competing with its precursor tryptophan for crossing the blood brain barrier. Arginine and citrulline could prevent excess hyperammonemia accompanied by BCAA supplementation. This study investigated the combination of BCAA, arginine, and citrulline on endurance performance in two consecutive days. Seven male and three female endurance runners ingested 0.17 g·kg-1 BCAA, 0.05 g·kg-1 arginine and 0.05 g·kg-1 citrulline (AA trial or placebo (PL trial in a randomized cross-over design. Each trial contained a 5000 m time trial on the first day, and a 10000 m time trial on the second day. The AA trial had significantly better performance in 5000 m (AA: 1065.7 ± 33.9 s; PL: 1100.5 ± 40.4 s and 10000 m (AA: 2292.0 ± 211.3 s; PL: 2375.6 ± 244.2 s. The two trials reported similar ratings of perceived exertion. After exercise, the AA trial had significantly lower tryptophan/BCAA ratio, similar NH3, and significantly higher urea concentrations. In conclusion, the supplementation could enhance time-trial performance in two consecutive days in endurance runners, possibly through the inhibition of cerebral serotonin synthesis by BCAA and the prevention of excess hyperammonemia by increased urea genesis.

  7. Comparison of Two Assays to Determine Anti-Citrullinated Peptide Antibodies in Rheumatoid Arthritis in relation to Other Chronic Inflammatory Rheumatic Diseases: Assaying Anti-Modified Citrullinated Vimentin Antibodies Adds Value to Second-Generation Anti-Citrullinated Cyclic Peptides Testing

    Directory of Open Access Journals (Sweden)

    Miriam Lizette Díaz-Toscano

    2014-01-01

    Full Text Available Determination of anti-citrullinated peptide antibodies (ACPA plays a relevant role in the diagnosis of rheumatoid arthritis (RA. To date, it is still unclear if the use of several tests for these autoantibodies in the same patient offers additional value as compared to performing only one test. Therefore, we evaluated the performance of using two assays for ACPA: second-generation anti-citrullinated cyclic peptides antibodies (anti-CCP2 and anti-mutated citrullinated vimentin (anti-MCV antibodies for the diagnosis of RA. We compared three groups: RA (n=142, chronic inflammatory disease (CIRD, n=86, and clinically healthy subjects (CHS, n=56 to evaluate sensitivity, specificity, predictive values, and likelihood ratios (LR of these two assays for the presence of RA. A lower frequency of positivity for anti-CCP2 was found in RA (66.2% as compared with anti-MCV (81.0%. When comparing RA versus other CIRD, sensitivity increased when both assays were performed. This strategy of testing both assays had high specificity and LR+. We conclude that adding the assay of anti-MCV antibodies to the determination of anti-CCP2 increases the sensitivity for detecting seropositive RA. Therefore, we propose the use of both assays in the initial screening of RA in longitudinal studies, including early onset of undifferentiated arthritis.

  8. Diagnostic significance of combined testing of anti-cyclic citrullinated peptide antibody and hidden rheumatoid factor immunoglobulin M in patients with juvenile rheumatoid arthritis%抗环瓜氨酸肽抗体和隐匿性类风湿因子免疫球蛋白M型在诊断早期幼年类风湿关节炎中的意义

    Institute of Scientific and Technical Information of China (English)

    滕庆; 刘永革; 何晓琥

    2004-01-01

    目的检测抗环瓜氨酸肽(CCP)抗体及隐匿性类风湿因子IgM型(HRF-IgM),并探讨其在幼年类风湿关节炎(JRA)早期诊断中的临床意义.方法用人工合成CCP链为抗原检测抗CCP抗体;对27例早期诊断的JRA做动态检测,通过阳性预测值(PPV)和阴性预测值(NPV)确定抗CCP抗体和HRF-IgM对早期诊断的JRA的特异性和敏感性.结果抗CCP抗体和HRF-IgM总阳性率分别为58.5%、65.0%.后者敏感性要高于前者,病情越重或受累的关节越多,抗体检出率越高.对早期JRA的PPV、抗CCP抗体特异性要高于HRF-IgM.当两种实验联合应用时,对具有早期关节炎表现发展成JRA的PPV为93.7%.结论抗C℃P抗体和HRF-IgM在JRA患儿均有较高的检出率,并与疾病严重程度有关.抗CCP抗体与HRF-IgM联合应用时,可使JRA的PPV进一步提高.

  9. Asymmetric Dimethylarginine Is a Well Established Mediating Risk Factor for Cardiovascular Morbidity and Mortality-Should Patients with Elevated Levels Be Supplemented with Citrulline?

    Science.gov (United States)

    McCarty, Mark F

    2016-07-08

    The arginine metabolite asymmetric dimethylarginine (ADMA) is a competitive inhibitor and uncoupler of endothelial nitric oxide synthase (eNOS), an enzyme that acts in multifarious ways to promote cardiovascular health. This phenomenon likely explains, at least in part, why elevated ADMA has been established as an independent risk factor for cardiovascular events, ventricular hypertrophy, and cardiovascular mortality. Fortunately, the suppressive impact of ADMA on eNOS activity can be offset by increasing intracellular arginine levels with supplemental citrulline. Although the long-term impact of supplemental citrulline on cardiovascular health in patients with elevated ADMA has not yet been studied, shorter-term clinical studies of citrulline administration demonstrate effects suggestive of increased NO synthesis, such as reductions in blood pressure and arterial stiffness, improved endothelium-dependent vasodilation, increased erection hardness, and increased ejection fractions in patients with heart failure. Supplemental citrulline could be a practical option for primary or secondary prevention of cardiovascular events and mortality, as it is inexpensive, has a mild flavor, and is well tolerated in doses (3-6 g daily) that can influence eNOS activity. Large and long-term clinical trials, targeting patients at high risk for cardiovascular events in whom ADMA is elevated, are needed to evaluate citrulline's potential for aiding cardiovascular health.

  10. Genetic markers of rheumatoid arthritis susceptibility in anti-citrullinated peptide antibody negative patients

    Science.gov (United States)

    Viatte, Sebastien; Plant, Darren; Bowes, John; Lunt, Mark; Eyre, Stephen; Barton, Anne; Worthington, Jane

    2012-01-01

    Introduction There are now over 30 confirmed loci predisposing to rheumatoid arthritis (RA). Studies have been largely undertaken in patients with anticyclic citrullinated peptide (anti-CCP) positive RA, and some genetic associations appear stronger in this subgroup than in anti-CCP negative disease, although few studies have had adequate power to address the question. The authors therefore investigated confirmed RA susceptibility loci in a large cohort of anti-CCP negative RA subjects. Methods RA patients and controls, with serological and genetic data, were available from UK Caucasian patients (n=4068 anti-CCP positive, 2040 anti-CCP negative RA) and 13,009 healthy controls. HLA-DRB1 genotypes and 36 single nucleotide polymorphisms were tested for association between controls and anti-CCP positive or negative RA. Results The shared epitope (SE) showed a strong association with anti-CCP positive and negative RA, although the effect size was significantly lower in the latter (effect size ratio=3.18, p<1.0E-96). A non-intronic marker at TNFAIP3, GIN1/C5orf30, STAT4, ANKRD55/IL6ST, BLK and PTPN22 showed association with RA susceptibility, irrespective of the serological status, the latter three markers remaining significantly associated with anti-CCP negative RA, after correction for multiple testing. No significant association with anti-CCP negative RA was detected for other markers (eg, AFF3, CD28, intronic marker at TNFAIP3), though the study power for those markers was over 80%. Discussion In the largest sample size studied to date, the authors have shown that the strength of association, the effect size and the number of known RA susceptibility loci associated with disease is different in the two disease serotypes, confirming the hypothesis that they might be two genetically different subsets. PMID:22661644

  11. The clinical significance of antibody determination to cyclic citrullinated peptides in systemic sclerosis

    Directory of Open Access Journals (Sweden)

    Stamenković Bojana

    2012-01-01

    Full Text Available Introduction. Anti-citrullinated peptides antibodies (ACPA are present in 80% of sera of rheumatoid arthritis (RA patients with high specificity for diagnosis and prediction for the development of early erosive arthritis. A few studies have reported a low frequency ACPA in systemic sclerosis (SSc patients with the presence of arthritis. Objective. The aim of our study was to determine the frequency of ACPA in systemic sclerosis (SSc patients, their correlation with clinical manifestations and radiographic features. Methods. The study included 82 patients with SSc, mean age 54.4 years, 59 with the limited (lSSc and 23 with the diffuse (dSSc form of the disease. The control group included 28 healthy age and sex matched subjects. ACPA and rheumatoid factor (RF were determined in all SSc patients and healthy subjects in whom standard radiography of hands and wrists was also done. Results. The presence of ACPA was detected in 11 (13.4% of SSc patients. Their level was not increased in any of the controls. Positive RF was found in 15.9% of SSc patients. Arthritis was present in 17.1%, as well as marginal bone erosions. There was a statistically significant association between positive ACPA and arthritis (p<0.0001 and positive ACPA and marginal bone erosions (p=0.0002. Conclusion. The research confirmed the correlation between ACPA with clinical signs of arthritis and radiographic damage of hand joints. ACPA is a useful diagnostic marker in the identification of SSc patients with arthritis and anatomic bone damage enabling the use of adequate therapy in order to prevent joint damage and poor quality of life.

  12. Anti-citrullinated peptide antibodies in lupus patients with or without deforming arthropathy.

    Science.gov (United States)

    Damián-Abrego, G N; Cabiedes, J; Cabral, A R

    2008-04-01

    The objective was to study the association of antibodies against cyclic citrullinated peptides (anti-CCP) in patients with lupus articular damage. We studied 34 systemic lupus erythematosus patients (30 women) with (n = 14) or without (n = 20) deforming arthropathy. Anti-DNA and arthritis were mandatory inclusion criteria for both groups. As controls, 34 patients with rheumatoid arthritis and nine patients with rheumatoid arthritis and systemic lupus erythematosus (rhupus) were included. Anti-CCP and rheumatoid factor were determined by ELISA and nephelometry respectively. All patients had recent x-ray films of the hands that were evaluated according to Sharp's method. Systemic lupus erythematosus patients had a mean 6.50 +/- 0.86 (SD, range 5-8) American College of Rheumatology (ACR) criteria, rheumatoid arthritis patients met 5.38 +/- 0.60 (range 4-6) ACR criteria for rheumatoid arthritis and rhupus patients had 5.78 +/- 0.44 (range 5-6) criteria for rheumatoid arthritis and 5.11 +/- 0.78 (range 4-6) for systemic lupus erythematosus. Systemic lupus erythematosus patients, with or without deforming arthropathy, had normal serum anti-CCP concentrations. In contrast, rheumatoid arthritis and rhupus patients had 30- and 23-fold higher than normal amounts of anti-CCP (p lupus deforming arthropathy were more frequently positive for rheumatoid factor (65%) than patients with non-deforming arthritis (15%) (p = 0.005). Patients with lupus deforming arthropathy had similar frequency of erosions and mean Sharp's score than rhupus patients. Anti-CCP antibodies do not associate with lupus arthropathy, whether deforming, non-deforming or erosive.

  13. L-citrulline protects from kidney damage in type 1 diabetic mice.

    Directory of Open Access Journals (Sweden)

    Maritza J Romero

    2013-12-01

    Full Text Available Rationale. Diabetic nephropathy is a major cause of end-stage renal disease, associated with endothelial dysfunction. Chronic supplementation of L-arginine (L-arg, the substrate for endothelial nitric oxide synthase (eNOS, failed to improve vascular function. L-citrulline (L-cit supplementation not only increases L-arg synthesis, but also inhibits cytosolic arginase I (Arg I, a competitor of eNOS for the use of L-arg, in the vasculature. Aims. To investigate whether L-cit treatment reduces diabetic nephropathy in streptozotocin (STZ-induced type 1 diabetes in mice and rats and to study its effects on arginase II (ArgII function, the main renal isoform. Methods. STZ-C57BL6 mice received L-cit or vehicle supplemented in the drinking water. For comparative analysis, diabetic ArgII knock out mice and L-cit-treated STZ-rats were evaluated. Results. L-cit exerted protective effects in kidneys of STZ-rats, and markedly reduced urinary albumin excretion, tubulo-interstitial fibrosis and kidney hypertrophy, observed in untreated diabetic mice. Intriguingly, L-cit treatment was accompanied by a sustained elevation of tubular ArgII at 16 wks and significantly enhanced plasma levels of the anti-inflammatory cytokine IL-10. Diabetic ArgII knock out mice showed greater BUN levels, hypertrophy, and dilated tubules than diabetic wild type mice. Despite a marked reduction in collagen deposition in ArgII knock out mice, their albuminuria was not significantly different from diabetic wild type animals. L-cit also restored NO/ROS balance and barrier function in high glucose-treated monolayers of human glomerular endothelial cells. Moreover, L-cit also has the ability to establish an anti-inflammatory profile, characterized by increased IL-10 and reduced IL-1beta and IL-12(p70 generation in the human proximal tubular cells. Conclusions. L-cit supplementation established an anti-inflammatory profile and significantly preserved the nephron function during type 1

  14. Asymmetric Dimethylarginine Is a Well Established Mediating Risk Factor for Cardiovascular Morbidity and Mortality—Should Patients with Elevated Levels Be Supplemented with Citrulline?

    Science.gov (United States)

    McCarty, Mark F.

    2016-01-01

    The arginine metabolite asymmetric dimethylarginine (ADMA) is a competitive inhibitor and uncoupler of endothelial nitric oxide synthase (eNOS), an enzyme that acts in multifarious ways to promote cardiovascular health. This phenomenon likely explains, at least in part, why elevated ADMA has been established as an independent risk factor for cardiovascular events, ventricular hypertrophy, and cardiovascular mortality. Fortunately, the suppressive impact of ADMA on eNOS activity can be offset by increasing intracellular arginine levels with supplemental citrulline. Although the long-term impact of supplemental citrulline on cardiovascular health in patients with elevated ADMA has not yet been studied, shorter-term clinical studies of citrulline administration demonstrate effects suggestive of increased NO synthesis, such as reductions in blood pressure and arterial stiffness, improved endothelium-dependent vasodilation, increased erection hardness, and increased ejection fractions in patients with heart failure. Supplemental citrulline could be a practical option for primary or secondary prevention of cardiovascular events and mortality, as it is inexpensive, has a mild flavor, and is well tolerated in doses (3–6 g daily) that can influence eNOS activity. Large and long-term clinical trials, targeting patients at high risk for cardiovascular events in whom ADMA is elevated, are needed to evaluate citrulline’s potential for aiding cardiovascular health. PMID:27417628

  15. Asymmetric Dimethylarginine Is a Well Established Mediating Risk Factor for Cardiovascular Morbidity and Mortality—Should Patients with Elevated Levels Be Supplemented with Citrulline?

    Directory of Open Access Journals (Sweden)

    Mark F. McCarty

    2016-07-01

    Full Text Available The arginine metabolite asymmetric dimethylarginine (ADMA is a competitive inhibitor and uncoupler of endothelial nitric oxide synthase (eNOS, an enzyme that acts in multifarious ways to promote cardiovascular health. This phenomenon likely explains, at least in part, why elevated ADMA has been established as an independent risk factor for cardiovascular events, ventricular hypertrophy, and cardiovascular mortality. Fortunately, the suppressive impact of ADMA on eNOS activity can be offset by increasing intracellular arginine levels with supplemental citrulline. Although the long-term impact of supplemental citrulline on cardiovascular health in patients with elevated ADMA has not yet been studied, shorter-term clinical studies of citrulline administration demonstrate effects suggestive of increased NO synthesis, such as reductions in blood pressure and arterial stiffness, improved endothelium-dependent vasodilation, increased erection hardness, and increased ejection fractions in patients with heart failure. Supplemental citrulline could be a practical option for primary or secondary prevention of cardiovascular events and mortality, as it is inexpensive, has a mild flavor, and is well tolerated in doses (3–6 g daily that can influence eNOS activity. Large and long-term clinical trials, targeting patients at high risk for cardiovascular events in whom ADMA is elevated, are needed to evaluate citrulline’s potential for aiding cardiovascular health.

  16. Preventing and curing citrulline-induced autoimmune arthritis in a humanized mouse model using a Th2-polarizing iNKT cell agonist.

    Science.gov (United States)

    Walker, Kyle M; Rytelewski, Mateusz; Mazzuca, Delfina M; Meilleur, Shannon A; Mannik, Lisa A; Yue, David; Brintnell, William C; Welch, Ian; Cairns, Ewa; Haeryfar, S M Mansour

    2012-07-01

    Invariant natural killer T (iNKT) cells are innate lymphocytes with unique reactivity to glycolipid antigens bound to non-polymorphic CD1d molecules. They are capable of rapidly releasing pro- and/or anti-inflammatory cytokines and constitute attractive targets for immunotherapy of a wide range of diseases including autoimmune disorders. In this study, we have explored the beneficial effects of OCH, a Th2-polarizing glycolipid agonist of iNKT cells, in a humanized mouse model of rheumatoid arthritis (RA) in which citrullinated human proteins are targeted by autoaggressive immune responses in mice expressing an RA susceptibility human leukocyte antigen (HLA) DR4 molecule. We found for the first time that treatment with OCH both prevents and cures citrulline-induced autoimmune arthritis as evidenced by resolved ankle swelling and reversed histopathological changes associated with arthritis. Also importantly, OCH treatment blocked the arthritogenic capacity of citrullinated antigen-experienced splenocytes without compromising their global responsiveness or altering the proportion of splenic naturally occurring CD4(+)CD25(+)FoxP3(+) regulatory T cells. Interestingly, administering the Th1-promoting iNKT cell glycolipid ligand α-C-galactosylceramide into HLA-DR4 transgenic mice increased the incidence of arthritis in these animals and exacerbated their clinical symptoms, strongly suggesting a role for Th1 responses in the pathogenesis of citrulline-induced arthritis. Therefore, our findings indicate a role for Th1-mediated immunopathology in citrulline-induced arthritis and provide the first evidence that iNKT cell manipulation by Th2-skewing glycolipids may be of therapeutic value in this clinically relevant model, a finding that is potentially translatable to human RA. PMID:21912419

  17. Typing by serovar, antibiogram, plasmid content, riboprobing, and isoenzyme typing to determine whether Neisseria gonorrhoeae isolates requiring proline, citrulline, and uracil for growth are clonal.

    OpenAIRE

    Ng, L K; Dillon, J R

    1993-01-01

    Neisseria gonorrhoeae isolates requiring proline, citrulline, and uracil for growth (PCU-) have homogeneous phenotypes; most are plasmid-free, belong to few serovars, and are significantly associated with intermediate levels of susceptibility to penicillin, tetracycline, erythromycin, and cefoxitin. Because of their lack of variation by these criteria, molecular typing methods, ribotyping (restriction fragment length polymorphism [RFLP] of rRNA genes), and multilocus enzyme electrophoresis we...

  18. Shared Epitope Alleles Remain A Risk Factor for Anti-Citrullinated Proteins Antibody (ACPA) – Positive Rheumatoid Arthritis in Three Asian Ethnic Groups

    OpenAIRE

    Too Chun-Lai; Leonid Padyukov; Jasbir Singh Dhaliwal; Emeli Lundström; Abqariyah Yahya; Nor Asiah Muhamad; Lars Klareskog; Lars Alfredsson; Per Tobias Larsson; Shahnaz Murad

    2011-01-01

    BACKGROUND: To investigate the associations between HLA-DRB1 shared epitope (SE) alleles and rheumatoid arthritis in subsets of rheumatoid arthritis defined by autoantibodies in three Asian populations from Malaysia. METHODS: 1,079 rheumatoid arthritis patients and 1,470 healthy controls were included in the study. Levels of antibodies to citrullinated proteins (ACPA) and rheumatoid factors were assessed and the PCR-SSO method was used for HLA-DRB1 genotyping. RESULTS: The proportion of ACPA ...

  19. Roles of export genes cgmA and lysE for the production of L-arginine and L-citrulline by Corynebacterium glutamicum.

    Science.gov (United States)

    Lubitz, Dorit; Jorge, João M P; Pérez-García, Fernando; Taniguchi, Hironori; Wendisch, Volker F

    2016-10-01

    L-arginine is a semi-essential amino acid with application in cosmetic, pharmaceutical, and food industries. Metabolic engineering strategies have been applied for overproduction of L-arginine by Corynebacterium glutamicum. LysE was the only known L-arginine exporter of this bacterium. However, an L-arginine-producing strain carrying a deletion of lysE still accumulated about 10 mM L-arginine in the growth medium. Overexpression of the putative putrescine and cadaverine export permease gene cgmA was shown to compensate for the lack of lysE with regard to L-arginine export. Moreover, plasmid-borne overexpression of cgmA rescued the toxic effect caused by feeding of the dipeptide Arg-Ala to lysE-deficient C. glutamicum and argO-deficient Escherichia coli strains. Deletion of the repressor gene cgmR improved L-arginine titers by 5 %. Production of L-lysine and L-citrulline was not affected by cgmA overexpression. Taken together, CgmA may function as an export system not only for the diamine putrescine and cadaverine but also for L-arginine. The major export system for L-lysine and L-arginine LysE may also play a role in L-citrulline export since production of L-citrulline was reduced when lysE was deleted and improved by 45 % when lysE was overproduced.

  20. Single cell cloning and recombinant monoclonal antibodies generation from RA synovial B cells reveal frequent targeting of citrullinated histones of NETs

    Science.gov (United States)

    Corsiero, Elisa; Bombardieri, Michele; Carlotti, Emanuela; Pratesi, Federico; Robinson, William; Migliorini, Paola; Pitzalis, Costantino

    2016-01-01

    Objectives Rheumatoid arthritis (RA) is characterised by breach of self-tolerance towards citrullinated antigens with generation of anti-citrullinated peptide/proteins antibodies (ACPA). Currently, the nature and source of citrullinated antigens driving the humoral autoimmune response within synovial ectopic lymphoid structures (ELS) is a crucial unknown aspect of RA pathogenesis. Here we characterised the autoreactive B-cell response of lesional B cells isolated from ELS+RA synovium. Methods Single synovial tissue CD19+cells were Fluorescence Activated Cell Sorting (FACS)-sorted and VH/VL Ig genes cloned to generate recombinant monoclonal antibodies (rmAbs) from patients with ELS+/ACPA+RA. Results RA-rmAbs immunoreactivity analysis provided the following key findings: (1) in a chIP-based array containing 300 autoantigens and in a ‘citrullinome’ multiplex assay, a strong reactivity against citrullinated histones H2A/H2B (citH2A/H2B) was observed in ∼40% of RA-rmAbs, followed by cit-fibrinogen and cit-vimentin; (2) anti-citH2A/H2B-reactive RA-rmAbs (but not anti-citH2A/H2B negative) selectively recognised neutrophil extracellular traps (NETs) from peripheral blood and/or RA joint neutrophils; (3) anti-citH2A/citH2B and anti-NET immunobinding was dependent on affinity maturation and was completely abrogated following reversion of hypermutated IgVH/VL genes to germline sequences; (4) ELS+ (not ELS−) RA synovial tissues engrafted into Severe Combined ImmunoDeficiency (SCID) mice released human anti-citH2A/citH2B and anti-NET antibodies in association with the intra-graft expression of CXCL13 and lymphotoxin (LT)-β, two master regulators of ELS. Conclusion We provided novel evidence that B cells differentiated within synovial ELS in the RA joints frequent target deiminated proteins which could be generated during NETosis of RA synovial neutrophils including histones. Thus, NETs could represent a source of citrullinated antigens fuelling the ACPA autoimmune

  1. Environmental risk factors differ between rheumatoid arthritis with and without auto-antibodies against cyclic citrullinated peptides

    DEFF Research Database (Denmark)

    Pedersen, Line Merete Blak; Jacobsen, Søren; Klarlund, Mette;

    2006-01-01

    -CCP-antibodies. Associations between exposure variables and risk of anti-CCP-positive and anti-CCP-negative RA were evaluated using logistic regression. A series of RA subtype-specific risk factors were identified. Tobacco smoking (odds ratio [OR] = 1.65; 95% confidence interval: 1.03-2.64, for > 20 versus 0 pack......The aim of this study was to evaluate new and previously hypothesised non-genetic risk factors for serologic subtypes of rheumatoid arthritis (RA) defined by the presence or absence of auto-antibodies to cyclic citrullinated peptides (CCP). In a national case-control study, we included 515 patients......-fold increased risk of this subtype compared with normal-weight individuals (OR = 3.45; 1.73-6.87). Age at menarche was the only examined factor that was significantly associated with both serologic subtypes of RA (p-trends = 0.01); women with menarche at age > or = 15 years had about twice the risk...

  2. HLA-DRB1 genotypes and the risk of developing anti citrullinated protein antibody (ACPA positive rheumatoid arthritis.

    Directory of Open Access Journals (Sweden)

    Nathalie Balandraud

    Full Text Available OBJECTIVE: To provide a table indicating the risk for developing anti citrullinated protein antibody (ACPA positive rheumatoid arthritis (RA according to one's HLA-DRB1 genotype. METHODS: We HLA-DRB1 genotyped 857 patients with ACPA positive RA and 2178 controls from South Eastern and Eastern France and calculated Odds Ratios (OR for developing RA for 106 of 132 possible genotypes accounting for 97% of subjects. RESULTS: HLA-DRB1 genotypic ORs for developing ACPA positive RA range from 28 to 0.19. HLA-DRB1 genotypes with HLA-DRB1*04SE (HLA-DRB1*0404, HLA-DRB1*0405, HLA-DRB1*0408, HLA-DRB1*04∶01, HLA-DRB1*01 are usually associated with high risk for developing RA. The second HLA-DRB1 allele in genotype somewhat modulates shared epitope associated risk. We did not identify any absolutely protective allele. Neither the Reviron, nor the du Montcel models accurately explains our data which are compatible with the shared epitope hypothesis and suggest a dosage effect among shared epitope positive HLA-DRB1 alleles, double dose genotypes carrying higher ORs than single dose genotypes. CONCLUSION: HLA-DRB1 genotypic risk for developing ACPA positive RA is influenced by both HLA-DRB1 alleles in genotype. We provide an HLA-DRB1 genotypic risk table for ACPA positive RA.

  3. The feeding route (enteral or parenteral) affects the plasma response of the dipetide Ala-Gln and the amino acids glutamine, citrulline and arginine, with the administration of Ala-Gln in preoperative patients.

    Science.gov (United States)

    Melis, Gerdien C; Boelens, Petra G; van der Sijp, Joost R M; Popovici, Theodora; De Bandt, Jean-Pascal; Cynober, Luc; van Leeuwen, Paul A M

    2005-07-01

    Enhancement of depressed plasma concentrations of glutamine and arginine is associated with better clinical outcome. Supplementation of glutamine might be a way to provide the patient with glutamine, and also arginine, because glutamine provides the kidney with citrulline, from which the kidney produces arginine when plasma levels of arginine are low. The aim of the present study was to investigate the parenteral and enteral response of the administered dipeptide Ala-Gln, glutamine, citrulline and arginine. Therefore, seven patients received 20 g Ala-Gln, administered over 4 h, parenterally or enterally, on two separate occasions. Arterial blood samples were taken before and during the administration of Ala-Gln. ANOVA and a paired t test were used to test differences (Pglutamine was observed with parenteral infusion of the dipeptide, although enteral infusion also significantly increased plasma levels of glutamine. The highest plasma response of citrulline was observed with the enteral administration of the dipeptide, although parenteral administration also increased plasma levels of citrulline. Plasma arginine increased significantly with parenteral infusion, but not with enteral administration of Ala-Gln. In conclusion, administrations of Ala-Gln, parenteral or enteral, resulted in an increased plasma glutamine response, as compared with baseline. Interestingly, in spite of the high availability of citrulline with enteral administration of the dipeptide, only parenteral infusion of Ala-Gln increased plasma arginine concentration.

  4. Anticorpos antiproteínas citrulinadas e a artrite reumatóide Auto-antibodies to citrullinated proteins and rheumatoid arthritis

    Directory of Open Access Journals (Sweden)

    Renata Trigueirinho Alarcon

    2007-06-01

    Full Text Available Pacientes com artrite reumatóide (AR possuem uma variedade de auto-anticorpos no soro e no líquido sinovial. Entre esses auto-anticorpos, destacam-se aqueles direcionados a proteínas citrulinadas, que são específicos para AR, aparecem precocemente durante a evolução da enfermidade e são bastante úteis para auxiliar no diag-nóstico da doença. Entre os antígenos citrulinados reconhecidos por auto-anticorpos na AR, encontram-se a profilagrina, a filagrina e a vimentina. Células e tecidos ricos nessas proteínas serviram de substrato para os primeiros ensaios laboratoriais para detecção dessa classe de auto-anticorpos. A descoberta de que os epitopos reconhecidos por esses auto-anticorpos eram peptídeos contendo citrulina permitiu o desenvolvimento de uma plataforma baseada em ELISA. O formato de ELISA possibilitou maior padronização e reprodutibilidade dos ensaios, resultando em ampla aceitação mundial como os auto-anticorpos mais específicos e precoces para o diagnóstico da AR. Há controvérsia quanto à capacidade dos anticorpos contra proteínas citrulinadas predizerem a gravidade da doença. O papel dos antígenos citrulinados na fisiopatologia da artrite reumatóide é sugerido pela forte especificidade desses auto-anticorpos para a doença, pelo achado de proteínas citrulinadas na sinóvia inflamada, pela produção intra-articular desses auto-anticorpos e pela extrema afinidade de peptídeos citrulinados por moléculas de HLA-DRB1 que contêm o epitopo compartilhado. Esses achados acenam com a possibilidade de novas e fascinantes descobertas rumo à melhor compreensão da fisiopatologia da AR.Rheumatoid arthritis (RA patients have a variety of auto-antibodies in the serum and synovial fluid. Among these auto-antibodies, those directed against citrullinated proteins are distinguished because that are specific for RA, appear early during the evolution of the disease and they are important to assist in the diagnosis of

  5. Quantitative Metabolomic Analysis of Urinary Citrulline and Calcitroic Acid in Mice after Exposure to Various Types of Ionizing Radiation.

    Science.gov (United States)

    Goudarzi, Maryam; Chauthe, Siddheshwar; Strawn, Steven J; Weber, Waylon M; Brenner, David J; Fornace, Albert J

    2016-01-01

    With the safety of existing nuclear power plants being brought into question after the Fukushima disaster and the increased level of concern over terrorism-sponsored use of improvised nuclear devices, it is more crucial to develop well-defined radiation injury markers in easily accessible biofluids to help emergency-responders with injury assessment during patient triage. Here, we focused on utilizing ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) to identify and quantitate the unique changes in the urinary excretion of two metabolite markers, calcitroic acid and citrulline, in mice induced by different forms of irradiation; external γ irradiation at a low dose rate (LDR) of 3.0 mGy/min and a high dose rate (HDR) of 1.1 Gy/min, and internal exposure to Cesium-137 ((137)Cs) and Strontium-90 ((90)Sr). The multiple reaction monitoring analysis showed that, while exposure to (137)Cs and (90)Sr induced a statistically significant and persistent decrease, similar doses of external γ beam at the HDR had the opposite effect, and the LDR had no effect on the urinary levels of these two metabolites. This suggests that the source of exposure and the dose rate strongly modulate the in vivo metabolomic injury responses, which may have utility in clinical biodosimetry assays for the assessment of exposure in an affected population. This study complements our previous investigations into the metabolomic profile of urine from mice internally exposed to (90)Sr and (137)Cs and to external γ beam radiation. PMID:27213362

  6. Preventive effects of citrulline on Western diet-induced non-alcoholic fatty liver disease in rats.

    Science.gov (United States)

    Jegatheesan, Prasanthi; Beutheu, Stéphanie; Freese, Kim; Waligora-Dupriet, Anne-Judith; Nubret, Esther; Butel, Marie-Jo; Bergheim, Ina; De Bandt, Jean-Pascal

    2016-07-01

    A Western diet induces insulin resistance, liver steatosis (non-alcoholic fatty liver disease (NAFLD)) and intestinal dysbiosis, leading to increased gut permeability and bacterial translocation, thus contributing to the progression of NAFLD to non-alcoholic steatohepatitis. In the present study, we sought, in a model of Western diet-induced NAFLD, to determine whether citrulline (Cit), an amino acid that regulates protein and energy metabolism, could decrease Western diet-induced liver injuries, as well as the mechanisms involved. Sprague-Dawley rats were fed a high-fat diet (45 %) and fructose (30 %) in drinking water or a control diet associated with water (group C) for 8 weeks. The high-fat, high-fructose diet (Western diet) was fed either alone (group WD) or with Cit (1 g/kg per d) (group WDC) or an isonitrogenous amount of non-essential amino acids (group WDA). We evaluated nutritional and metabolic status, liver function, intestinal barrier function, gut microbiota and splanchnic inflammatory status. Cit led to a lower level of hepatic TAG restricted to microvesicular lipid droplets and to a lower mRNA expression of CCAAT-enhancer-binding protein homologous protein, a marker of endoplasmic reticulum stress, of pro-inflammatory cytokines Il6 (Plevels. In the colon, it decreased inflammation (Tnfα and Tlr4 expressions) and increased claudin-1 protein expression. This was associated with higher levels of Bacteroides/Prevotella compared with rats fed the Western diet alone. Cit improves Western diet-induced liver injuries via decreased lipid deposition, increased insulin sensitivity, lower inflammatory process and preserved antioxidant status. This may be related in part to its protective effects at the gut level. PMID:27197843

  7. Quantitative Metabolomic Analysis of Urinary Citrulline and Calcitroic Acid in Mice after Exposure to Various Types of Ionizing Radiation

    Directory of Open Access Journals (Sweden)

    Maryam Goudarzi

    2016-05-01

    Full Text Available With the safety of existing nuclear power plants being brought into question after the Fukushima disaster and the increased level of concern over terrorism-sponsored use of improvised nuclear devices, it is more crucial to develop well-defined radiation injury markers in easily accessible biofluids to help emergency-responders with injury assessment during patient triage. Here, we focused on utilizing ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS to identify and quantitate the unique changes in the urinary excretion of two metabolite markers, calcitroic acid and citrulline, in mice induced by different forms of irradiation; X-ray irradiation at a low dose rate (LDR of 3.0 mGy/min and a high dose rate (HDR of 1.1 Gy/min, and internal exposure to Cesium-137 (137Cs and Strontium-90 (90Sr. The multiple reaction monitoring analysis showed that, while exposure to 137Cs and 90Sr induced a statistically significant and persistent decrease, similar doses of X-ray beam at the HDR had the opposite effect, and the LDR had no effect on the urinary levels of these two metabolites. This suggests that the source of exposure and the dose rate strongly modulate the in vivo metabolomic injury responses, which may have utility in clinical biodosimetry assays for the assessment of exposure in an affected population. This study complements our previous investigations into the metabolomic profile of urine from mice internally exposed to 90Sr and 137Cs and to X-ray beam radiation.

  8. Acute ingestion of citrulline stimulates nitric oxide synthesis but does not increase blood flow in healthy young and older adults with heart failure.

    Science.gov (United States)

    Kim, Il-Young; Schutzler, Scott E; Schrader, Amy; Spencer, Horace J; Azhar, Gohar; Deutz, Nicolaas E P; Wolfe, Robert R

    2015-12-01

    To determine if age-associated vascular dysfunction in older adults with heart failure (HF) is due to insufficient synthesis of nitric oxide (NO), we performed two separate studies: 1) a kinetic study with a stable isotope tracer method to determine in vivo kinetics of NO metabolism, and 2) a vascular function study using a plethysmography method to determine reactive hyperemic forearm blood flow (RH-FBF) in older and young adults in the fasted state and in response to citrulline ingestion. In the fasted state, NO synthesis (per kg body wt) was ∼ 50% lower in older vs. young adults and was related to a decreased rate of appearance of the NO precursor arginine. Citrulline ingestion (3 g) stimulated de novo arginine synthesis in both older [6.88 ± 0.83 to 35.40 ± 4.90 μmol · kg body wt(-1) · h(-1)] and to a greater extent in young adults (12.02 ± 1.01 to 66.26 ± 4.79 μmol · kg body wt(-1) · h(-1)). NO synthesis rate increased correspondingly in older (0.17 ± 0.01 to 2.12 ± 0.36 μmol · kg body wt(-1) · h(-1)) and to a greater extent in young adults (0.36 ± 0.04 to 3.57 ± 0.47 μmol · kg body wt(-1) · h(-1)). Consistent with the kinetic data, RH-FBF in the fasted state was ∼ 40% reduced in older vs. young adults. However, citrulline ingestion (10 g) failed to increase RH-FBF in either older or young adults. In conclusion, citrulline ingestion improved impaired NO synthesis in older HF adults but not RH-FBF, suggesting that factors other than NO synthesis play a role in the impaired RH-FBF in older HF adults, and/or it may require a longer duration of supplementation to be effective in improving RH-FBF.

  9. Effects of type II collagen epitope carbamylation and citrullination in human leucocyte antigen (HLA)-DR4(+) monozygotic twins discordant for rheumatoid arthritis.

    Science.gov (United States)

    De Santis, M; Ceribelli, A; Cavaciocchi, F; Generali, E; Massarotti, M; Isailovic, N; Crotti, C; Scherer, H U; Montecucco, C; Selmi, C

    2016-09-01

    The aim of this study is to investigate the effect of the native, citrullinated or carbamylated type II human collagen T cell- and B cell-epitopes on the adaptive immune response in rheumatoid arthritis (RA). Peripheral blood T and B cells obtained from a human leucocyte D4-related (antigen DR4(-) HLA-DR4)(+) woman with early RA, her healthy monozygotic twin and an unrelated HLA-DR3(+) woman with early RA were analysed for activation (CD154/CD69), apoptosis (annexin/7-aminoactinomycin), cytokine production [interferon (IFN)γ/interleukin (IL)-17/IL-4/IL-10/IL-6] and functional phenotype (CD45Ra/CCR7) after stimulation with the collagen native T cell epitope (T261-273), the K264 carbamylated T cell epitope (carT261-273), the native B cell epitope (B359-369) or the R360 citrullinated B cell epitope (citB359-369), and the combinations of these. The T cell memory compartment was activated by T cell epitopes in both discordant DR4(+) twins, but not in the DR3(+) RA. The collagen-specific activation of CD4(+) T cells was induced with both the native and carbamylated T cell epitopes only in the RA twin. Both T cell epitopes also induced IL-17 production in the RA twin, but a greater IL-4 and IL-10 response in the healthy twin. The citrullinated B cell epitope, particularly when combined with the carbamylated T cell epitope, induced B cell activation and an increased IL-6/IL-10 ratio in the RA twin compared to a greater IL-10 production in the healthy twin. Our data suggest that circulating collagen-specific T and B cells are found in HLA-DR4(+) subjects, but only RA activated cells express co-stimulatory molecules and produce proinflammatory cytokines. Carbamylation and citrullination further modulate the activation and cytokine polarization of T and B cells. PMID:27314557

  10. The role of citrullinated proteins in the pathogenesis of rheumatoid arthritis%蛋白质瓜氨酸化及其在类风湿关节炎中的意义

    Institute of Scientific and Technical Information of China (English)

    钟兵; 方勇飞

    2015-01-01

    类风湿关节炎(rheumatoid arthritis,RA)的发病机制至今尚未阐明。 RA 患者体内发现越来越多的抗瓜氨酸化蛋白抗体(ACPA)提示瓜氨酸化可能参与其发病过程。本文从蛋白质的瓜氨酸化过程、生理功能,ACPA 在 RA 的诊断、预后及发病机制中的作用进行综述,试图更全面地了解蛋白质的瓜氨酸化在 RA 发病各环节中的作用。%The mechanism of rheumatoid arthritis(RA)has not been clarified.More and more anti-citrullinated protein antibod-ies(ACPA)have been found in RA patients ,suggesting that citrullinated proteins may be involved in the pathogenesis of RA .This paper reviews the process of protein citrullination ,physiological function and the role of ACPA in the diagnosis ,prognosis and pathogenesis of RA in order to get a more comprehensive understanding of the role of citrullinated proteins in the pathogenesis of RA .

  11. Arginine methylation and citrullination of splicing factor proline- and glutamine-rich (SFPQ/PSF) regulates its association with mRNA.

    Science.gov (United States)

    Snijders, Ambrosius P; Hautbergue, Guillaume M; Bloom, Alex; Williamson, James C; Minshull, Thomas C; Phillips, Helen L; Mihaylov, Simeon R; Gjerde, Douglas T; Hornby, David P; Wilson, Stuart A; Hurd, Paul J; Dickman, Mark J

    2015-03-01

    Splicing factor proline- and glutamine-rich (SFPQ) also commonly known as polypyrimidine tract-binding protein-associated-splicing factor (PSF) and its binding partner non-POU domain-containing octamer-binding protein (NONO/p54nrb), are highly abundant, multifunctional nuclear proteins. However, the exact role of this complex is yet to be determined. Following purification of the endogeneous SFPQ/NONO complex, mass spectrometry analysis identified a wide range of interacting proteins, including those involved in RNA processing, RNA splicing, and transcriptional regulation, consistent with a multifunctional role for SFPQ/NONO. In addition, we have identified several sites of arginine methylation in SFPQ/PSF using mass spectrometry and found that several arginines in the N-terminal domain of SFPQ/PSF are asymmetrically dimethylated. Furthermore, we find that the protein arginine N-methyltransferase, PRMT1, catalyzes this methylation in vitro and that this is antagonized by citrullination of SFPQ. Arginine methylation and citrullination of SFPQ/PSF does not affect complex formation with NONO. However, arginine methylation was shown to increase the association with mRNA in mRNP complexes in mammalian cells. Finally we show that the biochemical properties of the endogenous complex from cell lysates are significantly influenced by the ionic strength during purification. At low ionic strength, the SFPQ/NONO complex forms large heterogeneous protein assemblies or aggregates, preventing the purification of the SFPQ/NONO complex. The ability of the SFPQ/NONO complex to form varying protein assemblies, in conjunction with the effect of post-translational modifications of SFPQ modulating mRNA binding, suggests key roles affecting mRNP dynamics within the cell.

  12. 抗瓜氨酸化蛋白抗体联合检测在类风湿关节炎中诊断价值的分析%Combined detection of anti-citrullinated protein antibodies in the diagnosis of rheumatoid arthritis

    Institute of Scientific and Technical Information of China (English)

    赵金霞; 刘湘源; 王志敏; 栗占国

    2010-01-01

    Objective Antibodies against citrulline-containing peptides such as anti-perinuclear factor (APF), anti-keratin antibodies (AKA), anti-filaggrin antibodies (AFA) and anti-cyclic citruilinated peptide (CCP) antibodies are very specific in RA. In recent years, detection of APF, AKA and anti-CCP antibodies have been widely used in clinical practice. Studies on the combined detection of these ACPA in diagnosing RA are limited in number. The aim of this study is to detect combined examination of APF, AKA, and anti-CCP antibodies and compare their values in the diagnosis of rheumatoid arthritis. The significance of combined detection of these ACPAs in rheumatoid arthritis is also investigated. Methods Five hunndred and fifly-one patients who suffered from arthritic problems during the recent two years were selected from the Department of Rheumatology and Immunology of Peking University People's Hospital. 304 of the patients were RA and 247 were diagnosed to have other rheumatic diseases based on the corresponding classification criteria. AKA and APF were tested by indirect immunofluorescence assay. Anti-CCP antibodies were tested antibodies, AKA and APF tests for RA were 76.2%, 43.6%, and 34.5%, respectively, and the specificities were highest specificity (100%), but it had a rather low sensitivity (28.3%). When two of the three ACPA were positive, the sensitivity and specificity for the diagnosis of RA was 48.4% and 99.2%, respectively. When either anti-CCP antibodies, AKA, or APF was treated as an individual parameter, the sensitivity was slightly increased (77.3%). However, the specificity decreased to 94.7%. Conclusion Anti-CCP antibodies are the most helpful makers for the diagnosis of rheumatoid arthritis among the three ACPAs which are used in clinical practice. The combined detection of anti-CCP antibodies, AKA, and APF cannot increase the diagnostic sensitivity and specificity of rheumaotid arthritis.%目的 分别检测抗核周因子(APF)、抗角蛋白抗

  13. Citrulina plasmática como marcador de pérdida de masa enterocitaria en la enfermedad celíaca en la infancia Plasma citrulline as a marker of loss of enterocitary mass in coeliac disease in childhood

    Directory of Open Access Journals (Sweden)

    J. Blasco Alonso

    2011-08-01

    Full Text Available Introducción: La citrulina plasmática no está incorporada a las proteínas endógenas ni exógenas y constituye un teórico marcador de la atrofia vellositaria. El objetivo del estudio es relacionar los niveles plasmáticos de citrulina y arginina con la severidad de la afectación de la mucosa intestinal en pacientes celiacos. Material y métodos: Estudio transversal de cohortes en niños entre 16 meses y 14 años: 46 con enfermedad celíaca al diagnóstico; 9 celíacos siguiendo dieta sin gluten y 42 controles. Se determina concentración plasmática de aminoácidos, en mmol/L, y variables clínicas y analíticas asociadas. Resultados: No diferencias estadísticamente significativas en IMC, edad o función renal, con ligero incremento de esteatorrea en celíacos. Citrulina, arginina y glutamina plasmáticas significativamente más bajas en los casos (17,7 μmol/l, 38,7 μmol/l, 479,6 μmol/l respectivamente que en controles (28,9 μmol/l, 56,2 μmol/l, 563,7 μmol/l. Citrulina plasmática significativamente más baja en grados avanzados de atrofia (13,8 μmol/l vs 19,7 μmol/l, p Introduction: Plasma citrulline is not incorporated in endogenous or exogenous proteins so it is a theoretical marker of villous atrophy. Our aim was to correlate plasma citrulline levels with severity of villous atrophy in celiac patients. Methods: Observational case-control study longitudinal in children 16 month-old to 14 year-old: 48 with untreated celiac disease, 9 celiac children under gluten free diet and 35 non-celiac healthy children. Plasma amino acids concentration is determined, expressed in μmol/L, and so are other clinical and analytical data. Results: No statistically significative difference found in the referring to BMI, age or renal function. Small increase in fecal fat in celiac children. Citrulline, arginine and glutamine are significantly lower in cases (17.7 μmol/l, 38.7 μmol/l, 479.6 μmol/l respectively than in controls (28.9 μmol/l, 56

  14. NITRIC OXIDE (NO, CITRULLINE - NO CYCLE ENZYMES, GLUTAMINE SYNTHETASE AND OXIDATIVE STRESS IN ANOXIA (HYPOBARIC HYPOXIA AND REPERFUSION IN RAT BRAIN

    Directory of Open Access Journals (Sweden)

    M. Swamy, Mohd Jamsani Mat Salleh, K. N .S. Sirajudeen, Wan Roslina Wan Yusof, G. Chandran

    2010-01-01

    Full Text Available Nitric oxide is postulated to be involved in the pathophysiology of neurological disorders due to hypoxia/ anoxia in brain due to increased release of glutamate and activation of N-methyl-D-aspartate receptors. Reactive oxygen species have been implicated in pathophysiology of many neurological disorders and in brain function. To understand their role in anoxia (hypobaric hypoxia and reperfusion (reoxygenation, the nitric oxide synthase, argininosuccinate synthetase, argininosuccinate lyase, glutamine synthetase and arginase activities along with the concentration of nitrate /nitrite, thiobarbituric acid reactive substances and total antioxidant status were estimated in cerebral cortex, cerebellum and brain stem of rats subjected to anoxia and reperfusion. The results of this study clearly demonstrated the increased production of nitric oxide by increased activity of nitric oxide synthase. The increased activities of argininosuccinate synthetase and argininosuccinate lyase suggest the increased and effective recycling of citrulline to arginine in anoxia, making nitric oxide production more effective and contributing to its toxic effects. The decreased activity of glutamine synthetase may favor the prolonged availability of glutamic acid causing excitotoxicity leading to neuronal damage in anoxia. The increased formation of thiobarbituric acid reactive substances and decreased total antioxidant status indicate the presence of oxidative stress in anoxia and reperfusion. The increased arginase and sustained decrease of GS activity in reperfusion group likely to be protective.

  15. Association of anticyclic citrullinated peptide antibodies with extra-articular manifestations, gender, and tabagism in rheumatoid arthritis patients from southern Brazil.

    Science.gov (United States)

    Goeldner, Isabela; Skare, Thelma L; de Messias Reason, Iara T; Nisihara, Renato M; Silva, Marília B; da Rosa Utiyama, Shirley R

    2011-07-01

    Gender and environmental factors are known to influence the clinical heterogeneity and outcome of rheumatoid arthritis (RA). Some variables have been suggested to be associated with the severity of the disease, which can be of great value in the correct management of RA patients. The purpose of this study was to investigate the associations among anticyclic citrullinated antibody (anti-CCP2) positivity, extra-articular manifestations (EAM), gender, and tobacco exposure in a Brazilian RA population. We performed a transversal study comprising 156 RA patients which were investigated for EAM, functional class, presence of anti-CCP2, and IgM rheumatoid factor (IgM-RF). The determination of anti-CCP2 was performed using enzyme immunoassay (ELISA) kits and IgM-RF by latex agglutination test. Clinical and demographical data were obtained through review of charts. Anti-CCP positivity intensity was directly correlated with tobacco smoking, sex, and the development of rheumatoid nodules. Intense anti-CCP2 reaction was 19.8-fold higher in females vs. males, 2.7-fold higher in tobacco vs. non-tobacco users, 7.7-fold higher in female vs. male tobacco users, and 5.15-fold higher in patients with rheumatoid nodules. Tobacco smoking, gender, and rheumatoid nodules are significantly correlated with anti-CCP2 positivity in Brazilian RA patients. PMID:21340496

  16. Shared epitope alleles remain a risk factor for anti-citrullinated proteins antibody (ACPA--positive rheumatoid arthritis in three Asian ethnic groups.

    Directory of Open Access Journals (Sweden)

    Too Chun-Lai

    Full Text Available BACKGROUND: To investigate the associations between HLA-DRB1 shared epitope (SE alleles and rheumatoid arthritis in subsets of rheumatoid arthritis defined by autoantibodies in three Asian populations from Malaysia. METHODS: 1,079 rheumatoid arthritis patients and 1,470 healthy controls were included in the study. Levels of antibodies to citrullinated proteins (ACPA and rheumatoid factors were assessed and the PCR-SSO method was used for HLA-DRB1 genotyping. RESULTS: The proportion of ACPA positivity among Malay, Chinese and Indian rheumatoid arthritis patients were 62.9%, 65.2% and 68.6%, respectively. An increased frequency of SE alleles was observed in ACPA-positive rheumatoid arthritis among the three Asian ethnic groups. HLA-DRB1*10 was highly associated with rheumatoid arthritis susceptibility in these Asian populations. HLA-DRB1*0405 was significantly associated with susceptibility to rheumatoid arthritis in Malays and Chinese, but not in Indians. HLA-DRB1*01 did not show any independent effect as a risk factor for rheumatoid arthritis in this study and HLA-DRB1*1202 was protective in Malays and Chinese. There was no association between SE alleles and ACPA- negative rheumatoid arthritis in any of the three Asian ethnic groups. CONCLUSION: The HLA-DRB1 SE alleles increase the risk of ACPA-positive rheumatoid arthritis in all three Asian populations from Malaysia.

  17. EFSA NDA Panel (EFSA Panel on Dietetic Products, Nutrition and Allergies), 2014. Scientific Opinion on the substantiation of a health claim related to citrulline-malate and faster recovery from muscle fatigue after exercise pursuant to Article 13(5) of Regulation (EC) No 1924/2006

    DEFF Research Database (Denmark)

    Tetens, Inge

    the human study for the scientific substantiation of the claim. A health claim on citrulline-malate and faster recovery from muscle fatigue after exercise has already been assessed by the Panel with an unfavourable outcome. The additional information submitted by the applicant did not provide evidence...... on the scientific substantiation of a health claim related to citrulline-malate and faster recovery from muscle fatigue after exercise. The Panel considers that citrulline-malate is sufficiently characterised. The claimed effect proposed by the applicant is “improved recovery from muscle fatigue”. Faster recovery...... function. The evidence provided by the applicant did not establish that a faster reduction of blood lactate concentrations through a dietary intervention leads to faster recovery from muscle fatigue by contributing to the restoration of muscle function after exercise. No conclusions could be drawn from...

  18. Anti-citrullinated protein antibodies suppress let-7a expression in monocytes from patients with rheumatoid arthritis and facilitate the inflammatory responses in rheumatoid arthritis.

    Science.gov (United States)

    Lai, Ning-Sheng; Yu, Hui-Chun; Yu, Chia-Li; Koo, Malcolm; Huang, Hsien-Bin; Lu, Ming-Chi

    2015-12-01

    We hypothesized that anti-citrullinated protein antibodies (ACPAs) could affect the expression of miRNAs in monocytes and contribute to the inflammatory responses in rheumatoid arthritis (RA). The expression profiles of 270 human miRNAs, co-cultured with ACPAs or human immunoglobulin G (IgG), were analyzed using real-time polymerase chain reaction. Ten miRNAs exhibited differential expression in U937 cells after co-cultured with ACPAs compared with human IgG. The expression levels of these miRNAs were investigated in monocytes from 21 ACPA-positive RA patients and 13 controls. Among these miRNAs, the expression levels of let-7a was decreased in monocytes from ACPA-positive RA patients. The expression levels of let-7a showed a negative correlation with positivity of rheumatoid factor in patients sampled. We found that transfection of U937 cells with let-7a mimic suppressed K-Ras protein expression. In the ACPA-mediated signaling pathway, transfection of U937 cells with let-7a mimic suppressed the ACPA-enhanced phosphorylation of c-Jun N-terminal kinase (JNK), extracellular signal-regulated kinase 1/2 (ERK1/2), and the expression and secretion of interleukin (IL)-1β. In conclusion, ACPA-mediated decreased let-7a expression in monocytes from ACPA-positive RA patients. Decreased let-7a expression was associated with the positivity of RF in ACPA-positive RA patients. The decreased expression of let-7a could facilitate the inflammatory pathway via enhanced ACPA-mediated phosphorylation of ERK1/2 and JNK and increased expression of IL-1β through an increase in the expression of Ras proteins.

  19. l-Citrulline supplementation attenuates blood pressure, wave reflection and arterial stiffness responses to metaboreflex and cold stress in overweight men.

    Science.gov (United States)

    Figueroa, Arturo; Alvarez-Alvarado, Stacey; Jaime, Salvador J; Kalfon, Roy

    2016-07-01

    Combined isometric exercise or metaboreflex activation (post-exercise muscle ischaemia (PEMI)) and cold pressor test (CPT) increase cardiac afterload, which may lead to adverse cardiovascular events. l-Citrulline supplementation (l-CIT) reduces systemic arterial stiffness (brachial-ankle pulse wave velocity (baPWV)) at rest and aortic haemodynamic responses to CPT. The aim of this study was to determine the effect of l-CIT on aortic haemodynamic and baPWV responses to PEMI+CPT. In all, sixteen healthy, overweight/obese males (age 24 (sem 6) years; BMI 29·3 (sem 4·0) kg/m2) were randomly assigned to placebo or l-CIT (6 g/d) for 14 d in a cross-over design. Brachial and aortic systolic blood pressure (SBP), diastolic blood pressure (DBP) and mean arterial pressure (MAP), aortic augmented pressure (AP), augmentation index (AIx), baPWV, reflection timing (Tr) and heart rate (HR) were evaluated at rest and during isometric handgrip exercise (IHG), PEMI and PEMI+CPT at baseline and after 14 d. No significant effects were evident after l-CIT at rest. l-CIT attenuated the increases in aortic SBP and wave reflection (AP and AIx) during IHG, aortic DBP, MAP and AIx during PEMI, and aortic SBP, DBP, MAP, AP, AIx and baPWV during PEMI+CPT compared with placebo. HR and Tr were unaffected by l-CIT in all conditions. Our findings demonstrate that l-CIT attenuates aortic blood pressure and wave reflection responses to exercise-related metabolites. Moreover, l-CIT attenuates the exaggerated arterial stiffness response to combined metaboreflex activation and cold exposure, suggesting a protective effect against increased cardiac afterload during physical stress. PMID:27160957

  20. Use of anti-citrullinated peptide (Anti –CCP antibodies in distinguishing patients with systemic lupus erythematosus and rheumatoid arthritis

    Directory of Open Access Journals (Sweden)

    Harry Isbagio

    2004-12-01

    Full Text Available Diagnosis of Rheumatoid arthritis (RA and systemic lupus erythematosus (SLE can be confused in their initial stages. The joints, especially the hands, are commonly affected in both disorders, many patients with SLE are initially misdiagnosed as having RA Given that the outcome for the two diseases is diverse, it would be helpful to have serological marker to distinguish between them at onset. Anti-citrullinated peptide antibodies (anti-CCP have recently been described as highly specific for RA. The objective of this study is to confirm the specificity of anti-CCP antibodies and to determine whether they might distinguish patients with RA from those with SLE. This study is a cross sectional study on a group of patients with RA (n=27, SLE with arthritis (n=20, other autoimmune diseases (non-rheumatic diseases, n = 8, and healthy adults (n=20. Anti-CCP was determined by a commercial Elisa test and Rheumatoid factor (RF was determined by the standard slide latex test. The sensitivity and specificity of anti-CCP for the diagnosis of RA was 63.0% and 97.9% respectively, comparing with RF for RA that was 40.7 % and 85.4 %. Only 1 healthy adult was anti-CCP+, no anti-CCP was detected from SLE and other autoimmune disease. The mean of titer anti CCP in normal healthy adult, other autoimmune diseases, SLE and RA was 1.35 Normal 0 false false false EN-US X-NONE X-NONE MicrosoftInternetExplorer4 ± 2.04, 0.63 Normal 0 false false false EN-US X-NONE X-NONE MicrosoftInternetExplorer4 ± 0.59, 0.75 Normal 0 false false false EN-US X-NONE X-NONE MicrosoftInternetExplorer4 ± 0.59, and 38.17 Normal 0 false false false EN-US X-NONE X-NONE MicrosoftInternetExplorer4 ± 44.22 RU/ml, respectively. There was a highly significant difference between the mean of titer anti CCP for RA with others diseases (p <0.001. We conclude that detection of anti-CCP is very useful for the diagnosis of RA and distinguishing RA from SLE. (Med J Indones 2004; 13: 227-31Keywords

  1. Cartilage oligomeric matrix protein associates differentially with erosions and synovitis and has a different temporal course in cyclic citrullinated peptide antibody (anti-CCP)-positive versus anti-CCP-negative early rheumatoid arthritis

    DEFF Research Database (Denmark)

    Christensen, Anne F; Lindegaard, Hanne; Hørslev-Petersen, Kim;

    2011-01-01

    -suppressive effect. We aimed to compare circulating cartilage oligomeric matrix protein (COMP), a marker of cartilage turnover, in untreated anti-CCP-positive and anti-CCP-negative RA, and to study the temporal pattern of COMP through 4 years of treatment, including the relationship to imaging and clinical findings......OBJECTIVE: Cyclic citrullinated peptide antibody (anti-CCP)-positive and anti-CCP-negative rheumatoid arthritis (RA) have been suggested as 2 distinctive disease subsets with respect to disease activity and prognosis. Previously, we proposed that anti-CCP antibodies might have a chondrocyte......-CCP, Health Assessment Questionnaire, visual analog scale scores for pain and global and physician assessment, and magnetic resonance imaging (MRI) of the nondominant hand were recorded at baseline. COMP in serum was measured by ELISA at inclusion and serially through 4 years. RESULTS: Median baseline COMP...

  2. A weighted genetic risk score using all known susceptibility variants to estimate rheumatoid arthritis risk

    Science.gov (United States)

    Yarwood, Annie; Han, Buhm; Raychaudhuri, Soumya; Bowes, John; Lunt, Mark; Pappas, Dimitrios A; Kremer, Joel; Greenberg, Jeffrey D; Plenge, Robert; Worthington, Jane; Barton, Anne; Eyre, Steve

    2015-01-01

    Background There is currently great interest in the incorporation of genetic susceptibility loci into screening models to identify individuals at high risk of disease. Here, we present the first risk prediction model including all 46 known genetic loci associated with rheumatoid arthritis (RA). Methods A weighted genetic risk score (wGRS) was created using 45 RA non-human leucocyte antigen (HLA) susceptibility loci, imputed amino acids at HLA-DRB1 (11, 71 and 74), HLA-DPB1 (position 9) HLA-B (position 9) and gender. The wGRS was tested in 11 366 RA cases and 15 489 healthy controls. The risk of developing RA was estimated using logistic regression by dividing the wGRS into quintiles. The ability of the wGRS to discriminate between cases and controls was assessed by receiver operator characteristic analysis and discrimination improvement tests. Results Individuals in the highest risk group showed significantly increased odds of developing anti-cyclic citrullinated peptide-positive RA compared to the lowest risk group (OR 27.13, 95% CI 23.70 to 31.05). The wGRS was validated in an independent cohort that showed similar results (area under the curve 0.78, OR 18.00, 95% CI 13.67 to 23.71). Comparison of the full wGRS with a wGRS in which HLA amino acids were replaced by a HLA tag single-nucleotide polymorphism showed a significant loss of sensitivity and specificity. Conclusions Our study suggests that in RA, even when using all known genetic susceptibility variants, prediction performance remains modest; while this is insufficiently accurate for general population screening, it may prove of more use in targeted studies. Our study has also highlighted the importance of including HLA variation in risk prediction models. PMID:24092415

  3. Intestinal renal metabolism of L-citrulline and L-arginine following enteral or parenteral infusion of L-alanyl-L-[2,15N]glutamine or L-[2,15N]glutamine in mice.

    Science.gov (United States)

    Boelens, Petra G; van Leeuwen, Paul A M; Dejong, Cornelis H C; Deutz, Nicolaas E P

    2005-10-01

    Previously, we observed increased plasma arginine (ARG) concentrations after glutamine (GLN)-enriched diets, in combination with clinical benefits. GLN delivers nitrogen for ARG synthesis, and the present study was designed to quantify the interorgan relationship of exogenous L-GLN or GLN dipeptide, by enteral or parenteral route, contributing to intestinal citrulline (CIT) and renal de novo ARG synthesis in mice. To study this, we used a multicatheterized mouse model with Swiss mice (n = 43) in the postabsorptive state. Stable isotopes were infused into the jugular vein or into the duodenum {per group either free L-[2,(15)N]GLN or dipeptide L-ALA-L-[2,(15)N]GLN, all with L-[ureido-(13)C-(2)H(2)]CIT and L-[guanidino-(15)N(2)-(2)H(2)]ARG} to establish renal and intestinal ARG and CIT metabolism. Blood flow was measured using (14)C-para-aminohippuric acid. Net intestinal CIT release, renal uptake of CIT, and net renal ARG efflux was found, as assessed by arteriovenous flux measurements. Quantitatively, more de novo L-[2,(15)N]CIT was produced when free L-[2,(15)N]GLN was given than when L-ALA-L-[2,(15)N]GLN was given, whereas renal de novo L-[2,(15)N]ARG was similar in all groups. In conclusion, the intestinal-renal axis is hereby proven in mice in that L-[2,(15)N]GLN or dipeptide were both converted into de novo renal L-[2,(15)N]ARG; however, not all was derived from intestinal L-[2,(15)N]CIT production. In this model, the feeding route and form of GLN did not influence de novo renal ARG production derived from GLN.

  4. EFSA Panel on Dietetic Products, Nutrition and Allergies (NDA); Scientific Opinion on the substantiation of a health claim related to citrulline-malate and faster recovery from muscle fatigue after exercise pursuant to Article 13(5) of Regulation (EC) No 1924/2006

    DEFF Research Database (Denmark)

    Tetens, Inge

    Following an application from Biocodex, submitted pursuant to Article 13(5) of Regulation (EC) No 1924/2006 via the Competent Authority of Belgium, the Panel on Dietetic Products, Nutrition and Allergies was asked to deliver an opinion on the scientific substantiation of a health claim related...... to the methodological limitations of the study. A number of mechanistic, animal and in vitro studies were submitted. In the absence of evidence for an effect of consumption of citrulline-malate on a faster recovery from muscle fatigue after exercise in humans, these studies cannot be used as a source of data...

  5. Preparation of particulate cyclic citrullinated peptide antigens and optimization of experimental conditions%颗粒型环瓜氨酸肽抗原的制备及条件优化

    Institute of Scientific and Technical Information of China (English)

    付琳; 裘宇容; 姜云飞; 夏佳音; 王海芳

    2013-01-01

    目的 探讨颗粒型环瓜氨酸肽抗原的制备方法,并对实验条件进行优化.方法 通过碳二亚胺交联法将人工合成的CCP与BSA共价偶联,使偶联产物蛋白大小在70-170 KD之间,经紫外吸收光谱鉴定偶联成功.采用碳二亚胺两步法将偶联成功的CCP与BSA复合物又与聚苯乙烯微球偶联,优化偶联条件,根据蛋白偶联量和偶联效率判断偶联效果.结果 (1)BSA与CCP偶联质量比为1∶1,EDC与NHS用量比为2∶3,偶联pH为6.0,BSA与CCP偶联可以达到较好的效果.BSA与CCP偶联产物在70 KD-170 KD之间,紫外吸收峰有明显左移.(2)微球与偶联蛋白的吸附效率随着EDC用量的增大而增大,当EDC用量达到7.5 mg/ml时,偶联蛋白最大效率达到63.66%.(3)随着偶联时间的延长,蛋白偶联微球的效率不断增加,在偶联时间为0.5-2 h内,偶联效率增加最快,当反应时间2 h后,反应趋于平衡.(4)对不同偶联反应pH值条件进行筛选,pH为7.5时偶联效果最好,pH7.0-8.0为适宜偶联的pH范围.结论 CCP-BSA偶联产物蛋白与胶乳颗粒偶联,当EDC浓度为7.5 mg/ml,偶联时间为2 h,反应PH值为7.5,蛋白偶联效率可达到63.66%.%Objective To investigate the preparation method of particulate cyclic citrullinated peptide antigens,and optimize the experimental conditions. Methods The artificially synthesized CCP was covalent coupled with BSA by the method of carbodiimide cross linking,and the protein sizes of1 the coupled products were between 70 to 170 KD. The covalent coupling was tested by ultraviolet absorption spectrum. Then the successful coupling complex of BSA and CCP were covalently coupled to the microspheres,and the coupling conditions were optimized. The covalent coupling effect was determined according to the protein coupling quantity and the coupling efficiency. Results (1) When the mass ra tio of BSA and CCP was 1 : 1,the ratio of EDC and NHS was 2 : 3,and the pH of coupling was 6.0,the coupling result of BSA

  6. Presence of antibodies to cyclic citrullinated peptides in juvenile-onset systemic lupus erythematosus%儿童系统性红斑狼疮抗环瓜氨酸肽抗体检测及其临床意义

    Institute of Scientific and Technical Information of China (English)

    刘海英; 刘云锋; 关启鸿; 钟艳玲; 皮蕾; 张白杜; 郭彩娇; 曾华松

    2010-01-01

    Objective To determine the prevalence of antibodies to cyclic citrullinated peptides (antiCCP) in patients with juvenile-onset systemic lupus erythematosus (JSLE) and its potential clinical significance. Methods Anti-CCP was measured in sera from patients with JSLE (n=47), juvenile idiopathic arthritis (JIA, n=54) and the sera from age-matched healthy children (n=40) using the third generation of anti-CCP ELISA commercial kit. The association of anti-CCP with other laboratory parameters and clinical features, especially arthritic symptoms in JSLE was also analyzed. T-test, Mann-Whitney U test, Chi-square and Fisher's exact test were used for statistical analysis. Results Out of the 47 JSLE patients, 6 (13%) were anti-CCP positive, which was significantly higher than that of the healthy controls( 13% vs 0, P<0.05 ), but not different from that of the JIA group (26%, P=0.098). RF was more prevalent in JSLE patients with anti-CCP than patients without (83% vs 15%, P<0.01 ), but there was no difference in other laboratory parameters and the clinical features ineluding the occurrence of arthritis (67% vs 51%, P>0.05). As one of the initial symptoms, arthritis was observed in 25 of 47 JSLE patients and no one had developed deforming arthropathy.There was no statistical difference in anti-CCP positivity between JSLE patients with and without articular involvement ( 16% vs 9%, P>0.05 ). Anti-CCP was not detected in any of the 3 patients with JSLE who had experienced joint pain and limited activity during 3 years follow-up. Conclusion Anti-CCP could be detected in patients with JSLE. It is noteworthy when differentiate from juvenile idiopathic arthritis, but the presence of anti-CCP does not relate with the occurrence of arthritis at presentation and persistence of arthritis in JSLE.%目的 检测儿童系统性红斑狼疮(JSLE)患者血清抗环瓜氨酸肽(CCP)抗体水平,了解抗CCP抗体在该病中的阳性检出率以及探讨其与

  7. Left ventricular function in treatment-naive early rheumatoid arthritis

    DEFF Research Database (Denmark)

    Løgstrup, Brian B; Deibjerg, Lone K; Nielsen, Agnete Desirèe;

    2014-01-01

    BACKGROUND: The role of inflammation and anti-cyclic citrullinated peptide antibodies (anti-CCP) in the pathogenesis of cardiovascular disease in early rheumatoid arthritis (RA) remains unclear. Previous studies have suggested that both disease activity and disease duration are associated.......3±1.3 years and steroid- and disease-modifying antirheumatic drug (DMARD)-naive early RA were included. Disease activity was scored by the use of the Danish national DANBIO registry (number of swollen joints (NSJ (28)), number of tender joints (NTJ (28)), C-reactive protein (CRP) and Health Assessment...

  8. Increased soluble programmed death-1 (sPD-1) is associated with disease activity and radiographic progression in early rheumatoid arthritis

    DEFF Research Database (Denmark)

    Greisen, S R; Rasmussen, T K; Stengaard-Pedersen, K;

    2014-01-01

    to be implicated in disease activity in chronic rheumatoid arthritis (RA). METHOD: We measured sPD-1 at baseline and 9 months after treatment initiation in plasma from early RA patients (n = 34). We tested for correlations with the Disease Activity Score using 28 joint counts (DAS28), the Health Assessment...... Questionnaire (HAQ) score, immunoglobulin M rheumatoid factor (IgM-RF), anti-cyclic citrullinated peptide (anti-CCP) antibodies, C-reactive protein (CRP), interleukin-21 (IL-21), and total Sharp score (TSS). We also measured sPD-1 in plasma from healthy volunteers (HV) (n = 20) and in plasma and synovial fluid...

  9. 抗突变型瓜氨酸波形蛋白检测在类风湿关节炎诊断中的意义%Detection of the Antibodies of Anti-mutated Citrullinated Vimentin in Diagnosis of Rheumatoid Arthritis

    Institute of Scientific and Technical Information of China (English)

    陈霞

    2011-01-01

    Objective To analyze diagnostic value of anti-mutated citrullinated vimentin(anti-MCV),antibody for rheumatoid arthritis (RA). Methods Enzyme-linked immunosorbent assay(ELISA)was used to detect the level of serum anti-MCV antibody and anti-cyclin citrullinated peptides(CCP) antibody in 41 patients with RA,39 patients with other autoimmune diseases and 30 healthy controls.Mann-Whitney U test and Spearman correlation were used for statistical analysis, the diagnostic value of these two antibodies for RA were compared by receiver operating characteristic curve (ROC). Results The median level of anti-MCV was significantly higher in RA group (410. 00 U/ml) than that of other autoimmune diseases group (10. 60 U/ml) and healthy control group (8. 30 U/ml) (U=242. 0,P<0. 01,and (7 = 318. 0,P<0. 01,re-spectively). According to receiver operating characteristic curve analysis,area under the curve of anti-MCV was 0.901, standard error was 0. 032,95%CI (0. 838~0. 963) ;area under the curve of anti-CCP was 0. 918,standard error was 0. 027, 95%CI (0. 865~0. 970). The diagnostic value of them are similar. In RA the sensitivity of Anti-MCV super to that of Anti-CCP,but show lower specific than Anti-CCP. Anti-MCV has correlated to Anti-CCP(r=0. 615,P<0. 01). Conclusion Anti-MCV is a valuable index for the diagnosis of RA. And it could be a good new serum marker.%目的 探讨血清抗突变型瓜氨酸波形蛋白(MCV)抗体水平在诊断类风湿性关节炎(RA)中的意义.方法 用酶联免疫吸附试验(ELISA)测定41例RA患者、39例非RA患者和30例健康体检者血清抗MCV抗体、抗CCP抗体水平,采用Mann-Whitney U检验、Spearman相关分析,并用ROC曲线比较对RA的诊断价值.结果 RA组抗MCV抗体水平(中位数410.00 U/ml)明显高于非RA组(中位数10.60 U/ml)和健康对照组(中位数8.30 U/ml)(U值分别为242.0和318.0,P值均<0.01).抗MCV抗体ROC曲线下面积为0.901,标准误0.032,其95%的近似可信区间为0.838~0.963.

  10. Serum Levels of Anticyclic Citrullinated Peptide Antibodies, Interleukin-6, Tumor Necrosis Factor-α, and C-Reactive Protein Are Associated with Increased Carotid Intima-Media Thickness: A Cross-Sectional Analysis of a Cohort of Rheumatoid Arthritis Patients without Cardiovascular Risk Factors

    Science.gov (United States)

    Vázquez-Del Mercado, Mónica; Nuñez-Atahualpa, Lourdes; Figueroa-Sánchez, Mauricio; Gómez-Bañuelos, Eduardo; Rocha-Muñoz, Alberto Daniel; Martín-Márquez, Beatriz Teresita; Martínez-García, Erika Aurora; Macias-Reyes, Héctor; Gamez-Nava, Jorge Ivan; Navarro-Hernandez, Rosa Elena; Nuñez-Atahualpa, María Alejandra; Andrade-Garduño, Javier

    2015-01-01

    The main cause of death in rheumatoid arthritis (RA) is cardiovascular events. We evaluated the relationship of anticyclic citrullinated peptide (anti-CCP) antibody levels with increased carotid intima-media thickness (cIMT) in RA patients. Methods. Forty-five anti-CCP positive and 37 anti-CCP negative RA patients, and 62 healthy controls (HC) were studied. All groups were assessed for atherogenic index of plasma (AIP) and cIMT. Anti-CCP, C-reactive protein (CRP), and levels of tumor necrosis factor alpha (TNFα) and interleukin-6 (IL-6) were measured by enzyme-linked immunosorbent assay (ELISA). Results. The anti-CCP positive RA patients showed increased cIMT compared to HC and anti-CCP negative (P < 0.001). Anti-CCP positive versus anti-CCP negative RA patients, had increased AIP, TNFα and IL-6 (P < 0.01), and lower levels of high density lipoprotein cholesterol (HDL-c) (P = 0.02). The cIMT correlated with levels of anti-CCP (r = 0.513, P = 0.001), CRP (r = 0.799, P < 0.001), TNFα (r = 0.642, P = 0.001), and IL-6 (r = 0.751, P < 0.001). In multiple regression analysis, cIMT was associated with CRP (P < 0.001) and anti-CCP levels (P = 0.03). Conclusions. Levels of anti-CCP and CRP are associated with increased cIMT and cardiovascular risk supporting a clinical role of the measurement of cIMT in RA in predicting and preventing cardiovascular events. PMID:25821796

  11. Serum Levels of Anticyclic Citrullinated Peptide Antibodies, Interleukin-6, Tumor Necrosis Factor-α, and C-Reactive Protein Are Associated with Increased Carotid Intima-Media Thickness: A Cross-Sectional Analysis of a Cohort of Rheumatoid Arthritis Patients without Cardiovascular Risk Factors

    Directory of Open Access Journals (Sweden)

    Mónica Vázquez-Del Mercado

    2015-01-01

    Full Text Available The main cause of death in rheumatoid arthritis (RA is cardiovascular events. We evaluated the relationship of anticyclic citrullinated peptide (anti-CCP antibody levels with increased carotid intima-media thickness (cIMT in RA patients. Methods. Forty-five anti-CCP positive and 37 anti-CCP negative RA patients, and 62 healthy controls (HC were studied. All groups were assessed for atherogenic index of plasma (AIP and cIMT. Anti-CCP, C-reactive protein (CRP, and levels of tumor necrosis factor alpha (TNFα and interleukin-6 (IL-6 were measured by enzyme-linked immunosorbent assay (ELISA. Results. The anti-CCP positive RA patients showed increased cIMT compared to HC and anti-CCP negative (P<0.001. Anti-CCP positive versus anti-CCP negative RA patients, had increased AIP, TNFα and IL-6 (P<0.01, and lower levels of high density lipoprotein cholesterol (HDL-c (P=0.02. The cIMT correlated with levels of anti-CCP (r=0.513, P=0.001, CRP (r=0.799, P<0.001, TNFα (r=0.642, P=0.001, and IL-6 (r=0.751, P<0.001. In multiple regression analysis, cIMT was associated with CRP (P<0.001 and anti-CCP levels (P=0.03. Conclusions. Levels of anti-CCP and CRP are associated with increased cIMT and cardiovascular risk supporting a clinical role of the measurement of cIMT in RA in predicting and preventing cardiovascular events.

  12. Arginine, citrulline and nitric oxide metabolism in sepsis

    Science.gov (United States)

    Arginine has vasodilatory effects, via its conversion by nitric oxide (NO) synthase into NO, and immunomodulatory actions that play important roles in sepsis. Protein breakdown affects arginine availability, and the release of asymmetric dimethylarginine, an inhibitor of NO synthase, may therefore a...

  13. Rheumatoid arthritis and periodontitis; a possible link via citrullination

    NARCIS (Netherlands)

    de Smit, M J; Tonckens-Brouwer, Elisabeth; Vissink, A; van Winkelhoff, A J

    2011-01-01

    Rheumatoid Arthritis (RA) and chronic and aggressive periodontitis are chronic inflammatory disorders characterized by deregulation of the host inflammatory response. Increased secretion of pro-inflammatory mediators results in soft and hard tissue destruction of the synovium and periodontium respec

  14. Interactions between smoking, increased serum levels of anti-CCP antibodies, rheumatoid factors, and erosive joint disease in patients with early, untreated rheumatoid arthritis

    DEFF Research Database (Denmark)

    Krol, A.; Garred, P; Heegaard, N H H;

    2015-01-01

    OBJECTIVES: To determine to what extent shared epitopes, smoking, and anti-cyclic citrullinated peptide (anti-CCP) antibodies are associated with disease activity and erosive disease in patients with rheumatoid arthritis (RA) at disease onset. METHOD: RA patients not previously treated with disease...... or antibodies. All antibody levels measured were associated with smoking and shared epitopes. CONCLUSIONS: Shared epitopes and smoking were associated with the production of anti-CCP antibodies and rheumatoid factors of IgM and IgA isotypes, which again were associated with erosive disease at presentation only...... in smokers. As shared epitopes and smoking were not directly associated with erosive disease, smoking may enhance the development of erosive disease in RA at different levels or through separate pathways....

  15. Case of interstitial lung disease with anti-EJ and anti-CCP antibodies preceding rheumatoid arthritis.

    Science.gov (United States)

    Tomioka, Hiromi; Kaneko, Masahiro; Kogata, Yoshinori; Katsuyama, Eiji; Ishikawa, Seiko; Fujii, Takao

    2012-06-01

    Autoantibodies against aminoacyl-tRNA synthetases (ARSs) are highly specific for myositis and/or interstitial lung disease. We report a rare case of double positive antibodies (anti-EJ antibody, the least common among anti-aminoacyl-tRNA synthetase antibodies, and anti-cyclic citrullinated peptide antibody, reported to be specific for rheumatoid arthritis) in a patient who presented with interstitial lung disease and later developed rheumatoid arthritis. The patient did not have clinically apparent myositis over a period of careful follow-up of several years. The initial pulmonary pathologic findings showed a nonspecific interstitial pneumonia pattern, with the formation of lymphoid follicles, which should be recognized as the first manifestation of rheumatoid arthritis.

  16. Left ventricular deformation at 2-year follow-up in treatment-naive rheumatoid arthritis patients is associated with high inflammation parameters

    DEFF Research Database (Denmark)

    Logstrup, B. B.; Masic, D.; Laurbjerg, T. B.;

    2015-01-01

    : To assess LV function by speckle-tracking echocardiography and the amount of coronary calcium by coronary computer tomography (CCT) in relation to anti-cyclic citrullinated peptide antibody (anti-CCP) status during a 2-year follow-up period in a open cohort of treatment-naive RA patients. Methods: Thirty......-nine treatment-naive early RA patients (21 women) were included at time of diagnosis and followed for 2 years. Disease activity, IgM rheumatoid factor (IgM-RF) and anti-CCP was assessed at time of diagnosis and at 2-year follow-up. We assessed both the LV deformation by speckle-tracking echocardiography whereby...

  17. Differentiation of Boc-protected alpha,delta-/delta,alpha- and beta,delta-/delta,beta-hybrid peptide positional isomers by electrospray ionization tandem mass spectrometry.

    Science.gov (United States)

    Raju, G; Ramesh, V; Srinivas, R; Sharma, G V M; Shoban Babu, B

    2010-06-01

    Two new series of Boc-N-alpha,delta-/delta,alpha- and beta,delta-/delta,beta-hybrid peptides containing repeats of L-Ala-delta(5)-Caa/delta(5)-Caa-L-Ala and beta(3)-Caa-delta(5)-Caa/delta(5)-Caa-beta(3)-Caa (L-Ala = L-alanine, Caa = C-linked carbo amino acid derived from D-xylose) have been differentiated by both positive and negative ion electrospray ionization (ESI) ion trap tandem mass spectrometry (MS/MS). MS(n) spectra of protonated isomeric peptides produce characteristic fragmentation involving the peptide backbone, the Boc-group, and the side chain. The dipeptide positional isomers are differentiated by the collision-induced dissociation (CID) of the protonated peptides. The loss of 2-methylprop-1-ene is more pronounced for Boc-NH-L-Ala-delta-Caa-OCH(3) (1), whereas it is totally absent for its positional isomer Boc-NH-delta-Caa-L-Ala-OCH(3) (7), instead it shows significant loss of t-butanol. On the other hand, second isomeric pair shows significant loss of t-butanol and loss of acetone for Boc-NH-delta-Caa-beta-Caa-OCH(3) (18), whereas these are insignificant for its positional isomer Boc-NH-beta-Caa-delta-Caa-OCH(3) (13). The tetra- and hexapeptide positional isomers also show significant differences in MS(2) and MS(3) CID spectra. It is observed that 'b' ions are abundant when oxazolone structures are formed through five-membered cyclic transition state and cyclization process for larger 'b' ions led to its insignificant abundance. However, b(1)(+) ion is formed in case of delta,alpha-dipeptide that may have a six-membered substituted piperidone ion structure. Furthermore, ESI negative ion MS/MS has also been found to be useful for differentiating these isomeric peptide acids. Thus, the results of MS/MS of pairs of di-, tetra-, and hexapeptide positional isomers provide peptide sequencing information and distinguish the positional isomers.

  18. Somatostatin, substance P and calcitonin gene-related peptide-positive intramural nerve structures of the human large intestine affected by carcinoma.

    Directory of Open Access Journals (Sweden)

    Jerzy Kaleczyc

    2010-11-01

    Full Text Available The aim of this study was to investigate the arrangement and chemical coding of enteric nerve structures in the human large intestine affected by cancer. Tissue samples comprising all layers of the intestinal wall were collected during surgery form both morphologically unchanged and pathologically altered segments of the intestine (n=15, and fixed by immersion in buffered paraformaldehyde solution. The cryostat sections were processed for double-labelling immunofluorescence to study the distribution of the intramural nerve structures (visualized with antibodies against protein gene-product 9.5 and their chemical coding using antibodies against somatostatin (SOM, substance P (SP and calcitonin gene-related peptide (CGRP. The microscopic observations revealed distinct morphological differences in the enteric nerve system structure between the region adjacent to the cancer invaded area and the intact part of the intestine. In general, infiltration of the cancer tissue resulted in the gradual (depending on the grade of invasion first decomposition and reduction to final partial or complete destruction and absence of the neuronal elements. A comparative analysis of immunohistochemically labeled sections (from the unchanged and pathologically altered areas revealed a statistically significant decrease in the number of CGRP-positive neurons and nerve fibres in both submucous and myenteric plexuses in the transitional zone between morphologically unchanged and cancer-invaded areas. In this zone, a decrease was also observed in the density of SP-positive nerve fibres in all intramural plexuses. Conversely, the investigations demonstrated statistically insignificant differences in number of SP- and SOM-positive neurons and a similar density of SOM-positive nerve fibres in the plexuses of the intact and pathologically changed areas. The differentiation between the potential adaptive changes in ENS or destruction of its elements by cancer invasion should be a subject of further investigations.

  19. PADI4 Haplotypes in Association with RA Mexican Patients, a New Prospect for Antigen Modulation

    Directory of Open Access Journals (Sweden)

    Maria Guadalupe Zavala-Cerna

    2013-01-01

    Full Text Available Peptidyl arginine deiminase IV (PAD 4 is the responsible enzyme for a posttranslational modification called citrullination, originating the antigenic determinant recognized by anti-cyclic citrullinated peptide antibodies (ACPA. Four SNPs (single nucleotide polymorphisms have been described in PADI4 gene to form a susceptibility haplotype for rheumatoid arthritis (RA; nevertheless, results in association studies appear contradictory in different populations. The aim of the study was to analyze if the presence of three SNPs in PADI4 gene susceptibility haplotype (GTG is associated with ACPA positivity in patients with RA. This was a cross-sectional study that included 86 RA patients and 98 healthy controls. Polymorphisms PADI4_89, PADI4_90, and PADI4_92 in the PADI4 gene were genotyped. The susceptibility haplotype (GTG was more frequent in RA patients; interestingly, we found a new haplotype associated with RA with a higher frequency (GTC. There were no associations between polymorphisms and high scores in Spanish HAQ-DI and DAS-28, but we did find an association between RARBIS index and PADI4_89, PADI4_90 polymorphisms. We could not confirm an association between susceptibility haplotype presence and ACPA positivity. Further evidence about proteomic expression of this gene will determine its participation in antigenic generation and autoimmunity.

  20. The role of citrulline in patients following hematopoietic stem cell transplantation

    NARCIS (Netherlands)

    Herbers, A.H.E.

    2015-01-01

    Hematopoietic stem cell transplantation (HSCT) provides effective treatment of hematological malignancies and other disorders. However, the procedure temporarily compromises the immune system resulting in damage to the gastrointestinal (GI) tract, called mucosal barrier injury (MBI), and neutropenia

  1. Anti-citrullinated protein antibodies (ACPA) in rheumatoid arthritis : linking genetic predisposition to clinical outcome

    NARCIS (Netherlands)

    Woude, Diane van der

    2012-01-01

    Rheumatoid arthritis (RA) is a disease characterized by arthritis of mainly the small joints of the hands and feet, which is thought to be the result of an autoimmune response. It is the most common inflammatory arthritis with a prevalence of 0.5-1.0% in European and North-American populations 1. Th

  2. Citrulline as a Marker for Chemotherapy Induced Mucosal Barrier Injury in Pediatric Patients

    NARCIS (Netherlands)

    van Vilet, Michel J.; Tissing, Wim J. E.; Rings, Edmond H. H. M.; Koetse, Harma A.; Stellaard, Frans; Kamps, Willem A.; de Bont, Eveline S. J. M.

    2009-01-01

    Background. The Currently used National Cancer Institute (NCI) adverse events criteria for mucosal barrier injury (MBI) are insufficient for use in children. We searched for objective, easily measurable indicators for MBI in children with cancer. Purpose. In children with acute myeloid leukemia, var

  3. Impact de la citrulline sur le métabolisme du tissu adipeux

    OpenAIRE

    Joffin, Nolwenn

    2015-01-01

    Obesity is frequently associated with type 2 diabetes and cardiovascular diseases, related to metabolic and endocrine dysregulation of white adipose tissue (WAT). During aging, the loss of muscle mass may be associated with obesity and defines the concept of sarcopenic obesity. Treatments implemented to counteract these conditions showed a very partial success. It is therefore appropriate to develop original alternative strategies that could lead to targeted therapies. Our team studies the me...

  4. Citrullination of central nervous system proteins during the development of experimental autoimmune encephalomyelitis.

    NARCIS (Netherlands)

    Raijmakers, R.; Vogelzangs, J.H.P.; Croxford, J.L.; Wesseling, P.; Venrooij, W.J.W. van; Pruijn, G.J.M.

    2005-01-01

    Immunization of mammals with central nervous system (CNS)-derived proteins or peptides induces experimental autoimmune encephalomyelitis (EAE), a disease resembling the human autoimmune disease multiple sclerosis (MS). Both diseases are accompanied by destruction of a part of the of the myelin sheat

  5. The Multifaceted Aspects of Interstitial Lung Disease in Rheumatoid Arthritis

    Directory of Open Access Journals (Sweden)

    Lorenzo Cavagna

    2013-01-01

    Full Text Available Interstitial lung disease (ILD is a relevant extra-articular manifestation of rheumatoid arthritis (RA that may occur either in early stages or as a complication of long-standing disease. RA related ILD (RA-ILD significantly influences the quoad vitam prognosis of these patients. Several histopathological patterns of RA-ILD have been described: usual interstitial pneumonia (UIP is the most frequent one, followed by nonspecific interstitial pneumonia (NSIP; other patterns are less commonly observed. Several factors have been associated with an increased risk of developing RA-ILD. The genetic background plays a fundamental but not sufficient role; smoking is an independent predictor of ILD, and a correlation with the presence of rheumatoid factor and anti-cyclic citrullinated peptide antibodies has also been reported. Moreover, both exnovo occurrence and progression of ILD have been related to drug therapies that are commonly prescribed in RA, such as methotrexate, leflunomide, anti-TNF alpha agents, and rituximab. A greater understanding of the disease process is necessary in order to improve the therapeutic approach to ILD and RA itself and to reduce the burden of this severe extra-articular manifestation.

  6. Association of CD247 polymorphisms with rheumatoid arthritis: a replication study and a meta-analysis.

    Directory of Open Access Journals (Sweden)

    María Teruel

    Full Text Available Given the role of CD247 in the response of the T cells, its entailment in autoimmune diseases and in order to better clarify the role of this gene in RA susceptibility, we aimed to analyze CD247 gene variants previously associated with other autoimmune diseases (rs1052237, rs2056626 and rs864537 in a large independent European Caucasian population. However, no evidence of association was found for the analyzed CD247 single-nucleotide polymorphisms (SNPs with RA and with the presence/absence of anti-cyclic citrullinated polypeptide. We performed a meta-analysis including previously published GWAS data from the rs864537 variant, revealing an overall genome-wide significant association between this CD247 SNP and RA with anti-CCP (OR = 0.90, CI 95% = 0.87-0.93, Poverall = 2.1×10(-10. Our results show for first time a GWAS-level association between this CD247 polymorphism and RA risk.

  7. Vitamin D receptor gene polymorphism in rheumatoid arthritis and its association with atherosclerosis

    Directory of Open Access Journals (Sweden)

    Amal M El-Barbary

    2015-01-01

    Patients and methods This study included forty early rheumatoid arthritis patients and forty healthy controls. Disease activity score 28 (DAS-28, Modified Health Assessment Questionnaire (MHAQ, Carotid intima-media thickness (cIMT were assessed using B-mode ultrasound, Erythrocyte sedimentation rate (ESR, C reactive protein (CRP, Lipid profile, anti cyclic citrullinated PolyPeptid (anti-CCP, serum interleukin-6, Total serum vitamin D and genotype determination of BsmI, FokI polymorphism and allel frequency were measured. Results Vitamin D deficiency was observed in 25% of patients. There was no significant difference between RA patients and controls regarding the distribution of BsmI genotype frequencies and allele. However, a significant difference between rheumatoid arthritis patients and controls regarding the distribution of FokI genotype and allele frequencies was found. In addition, FokI polymorphism and the F allele was significantly associated with RA. anti-CCP, interleukin-6 levels, (cIMT and vitamin D deficiency were significantly higher in the presence of bb homozygote of BsmI genotypes and FF homozygote of FokI genotypes. A significant negative correlation between 25 hydroxy vitamin D levels with (DAS-28, ESR, (CRP, and IL-6 (P < 0.001. However, there was positive correlation between 25 hydroxyvitamin D levels and HDL-C (P < 0.001.

  8. リウマチ膠原病診療における抗環状シトルリン化ペプチド抗体(抗CCP抗体)の臨床的有用性の検討

    OpenAIRE

    下山, 久美子; 小川, 法良; 坂井, 知之; 澤木, 俊興; 河南, 崇典; 唐澤, 博美; 正木, 康史; 田中, 真生; 福島, 俊洋; 廣瀬, 優子; 梅原, 久範

    2007-01-01

    目的:近年,関節リウマチ(rheumatoid arthritis ; RA)の診療において,抗環状シトルリン化ペプチド抗体(anti-cyclic citrullinated peptide antibody:抗CCP抗体)は感度,特異度が高く注目されている.我々は抗CCP抗体のRA診療における有用性について検討した.対象と方法:多発関節痛症例115例(女性89例,男性26例)に関して抗CCP抗体,ESR, CRP, IgM-RF, IgG-RF, RAPA, CARF, MMP-3, C1q-IC, Stage, Class, Joint score, Sharp score, KL-6, SP-D,胸部CTを検討した.結果:抗CCP抗体は,特異度(93.5%)に優れていた.またRA群(45例)において,抗CCP抗体陽性群のSharp score (10.9±22.4)は,抗CCP抗体陰性群(1.7±1.8)と比べて有意に高値であり(P...

  9. Seasonal onychomadesis in an elderly gentleman

    Directory of Open Access Journals (Sweden)

    Supriya Venugopal

    2010-01-01

    Full Text Available A 79-year-old man with a history of dementia and hypertension initially presented with a ten year history of Beau’s lines and seasonal nail shedding of his fingernails only. He denied any exposure to heavy metals, unusual activities or food. He stated that the seasonal nail shedding had been occurring for the last 5-10 years. On examination, six out of ten fingernails had been affected. He had significant toenail dystrophy. Fungal cultures and PAS staining of the toenails were negative. Routine serum biochemistry and haematology results were normal. Serum arsenic, cadmium and lead levels were also normal. Vitamin B12, zinc, folate, iron studies, thyroid function studies and homocysteine levels were also normal. Rheumatoid factor and anti-cyclic citrullinated peptide antibody antibodies were negative. Bilateral hand X-ray showed osteoarthritic change and did not show any features of psoriatic arthropathy. We discuss the case of a 79-year-old man with seasonal nail shedding, curiously affecting his fingernails only.

  10. Joint Involvement in Primary Sjögren’s Syndrome: An Ultrasound “Target Area Approach to Arthritis”

    Directory of Open Access Journals (Sweden)

    Luis M. Amezcua-Guerra

    2013-01-01

    Full Text Available Objective. To characterize the ultrasound (US pattern of joint involvement in primary Sjögren’s syndrome (pSS. Methods. Seventeen patients with pSS, 18 with secondary Sjögren’s syndrome (sSS, and 17 healthy controls underwent US examinations of various articular regions. Synovitis (synovial hypertrophy/joint effusion, power Doppler (PD signals, and erosions were assessed. Results. In patients with pSS, synovitis was found in the metacarpophalangeal joints (MCP, 76%, wrists (76%, and knees (76%, while the proximal interphalangeal joints, elbows, and ankles were mostly unscathed. Intra-articular PD signals were occasionally detected in wrists (12%, elbows (6%, and knees (6%. Erosions were evident in the wrists of three (18% patients with pSS, one of these also having anti-cyclic citrullinated peptide (anti-CCP antibodies. While US synovitis does not discriminate between sSS and pSS, demonstration of bone erosions in the 2nd MCP joints showed 28.8% sensitivity and 100% specificity for diagnosing sSS; in comparison, these figures were 72.2 and 94.1% for circulating anti-CCP antibodies. Conclusions. In pSS, the pattern of joint involvement by US is polyarticular, bilateral, and symmetrical. Synovitis is the US sign most commonly found in patients with pSS, especially in MCP joints, wrists, and knees, and bone erosions also may occur.

  11. Analysis of foot structural damage in rheumatoid arthritis: clinical evaluation by validated measures and serological correlations

    Directory of Open Access Journals (Sweden)

    E. Bartoloni Bocci

    2011-06-01

    Full Text Available Objective: To examine foot involvement in rheumatoid arthritis (RA and to characterize structural alterations in patients with anti-cyclic citrullinated peptide (CCP antibody-positive and -negative disease. Methods: Seventy-eight patients with RA with foot pain were consecutively enrolled. The Manchester Hallux Valgus (MHV rating scale was used to evaluate the hallux valgus deformity degree. The Foot Posture Index (FPI6, a novel, foot-specific outcome measure, was adopted in order to quantify variation in the position of the foot. The findings were correlated with disease duration and presence or absence of anti-CCP antibodies. Results: About 84.6% patients had different degrees of hallux valgus and 65.4% subjects had a pronated foot. These two foot alterations were prevalently found in patients with long-standing disease and circulating anti-CCP antibodies. On the contrary, RA patients without anti-CCP and early disease essentially displayed a supinated foot without relevant hallux valgus deformity. Conclusion: Our findings allowed to identify different anatomic foot alterations in RA patients according to disease duration and negative prognostic factors such as anti-CCP antibodies. Our findings support the role of an accurate analysis of foot structural damage and may suggest the usefulness of a correct plantar orthosis prescription also in early phases of the disease.

  12. Monitoring myeloablative therapy-induced small bowel toxicity by serum citrulline concentration: a comparison with sugar permeability tests.

    NARCIS (Netherlands)

    Lutgens, L.C.; Blijlevens, N.M.A.; Deutz, N.E.; Donnelly, J.P.; Lambin, P.; Pauw, B.E. de

    2005-01-01

    BACKGROUND: Intestinal mucositis is an important cause of cancer treatment-related morbidity and mortality, carrying a serious economic burden. Currently, objective parameters are lacking that would enable the monitoring of gut damage in routine clinical practice, thus hindering the development of c

  13. Immunogenetic and immunological aspects of rheumatoid arthritis : DERAA and anti-citrulline reactivity can make the difference

    NARCIS (Netherlands)

    Feitsma, Anouk Leonie

    2010-01-01

    Rheumatoid arthritis (RA) is a chronic inflammation of several joints caused by autoimmunity. HLA molecules are most important risk factor involved in RA development. Regarding the risk of RA development, three variants can be discriminated; the shared epitope increases the risk, DERAA-containing HL

  14. Autoantibodies to cyclic citrullinated peptides predict progression to rheumatoid arthritis in patients with undifferentiated arthritis - A prospective cohort study

    NARCIS (Netherlands)

    Gaalen, F.A. van; Linn-Rasker, S.P.; Venrooij, W.J.W. van; Jong, B.A. de; Breedveld, F.C.; Verweij, C.L.; Toes, R.E.M.; Huizinga, T.W.J.

    2004-01-01

    Objective. Rheumatoid arthritis (RA) is a common, severe, chronic inflammatory joint disease. Since the disease may initially be indistinguishable from other forms of arthritis, early diagnosis can be difficult. Autoantibodies seen in RA can be detected years before clinical symptoms develop. In an

  15. Autoantibodies to cyclic citrullinated peptides predict progression to rheumatoid arthritis in patients with undifferentiated arthritis: a prospective cohort study.

    NARCIS (Netherlands)

    Gaalen, van FA; Linn-Rasker, SP; Venrooij, W.J.; Jong, B.A.; Breedveld, F.C.; Verweij, C.L.; Toes, RE; Huizinga, T.W.

    2004-01-01

    OBJECTIVE: Rheumatoid arthritis (RA) is a common, severe, chronic inflammatory joint disease. Since the disease may initially be indistinguishable from other forms of arthritis, early diagnosis can be difficult. Autoantibodies seen in RA can be detected years before clinical symptoms develop. In an

  16. Increased plasma citrulline in mice marks diet-induced obesity and may predict the development of the metabolic syndrome

    NARCIS (Netherlands)

    Sailer, M.; Dahlhoff, C.; Giesbertz, P.; Eidens, M.K.; Wit, de N.J.W.; Rubio-Aliaga, I.; Boekschoten, M.V.; Müller, M.R.; Daniel, H.

    2013-01-01

    Article About the Authors Metrics Comments Related Content Abstract Introduction Results Discussion Materials and Methods Supporting Information Acknowledgments Author Contributions References Reader Comments (0) Figures Abstract In humans, plasma amino acid concentrations of branched-chain amino ac

  17. Progress of anti-CCP antibody in the application of diagnosis of rheumatoid arthritis%抗CCP抗体在类风湿性关节炎诊断中的应用进展

    Institute of Scientific and Technical Information of China (English)

    梁海; 吴久鸿; 向卓; 张学辉

    2014-01-01

    目前类风湿关节炎的诊断检查有炎性标志物(包括血沉和C反应蛋白)检测,免疫复合物和补体检测,关节滑液检测以及自身抗体检测。这些指标经常伴随类风湿关节炎的活动而改变,早期诊断主要依靠的是自身抗体检测。抗环瓜氨酸肽( cyclic citrullinated peptide,CCP)抗体是一种抗角蛋白抗体,在类风湿性关节炎的敏感度和特异度较类风湿因子高。发病一年内,类风湿性关节炎患者大部分血清中可检测到抗CCP抗体的存在,同时抗CCP抗体阳性也可以用来预测严重破坏的类风湿性关节炎。因此,抗CCP抗体对于类风湿性关节炎的早期诊断及预后评估是极有帮助的。本文综述了近年来抗CCP抗体对于诊断类风湿关节炎的应用进展。%Currently the markers for the diagnosis of rheumatoid arthritis include inflammatory makers ( including blood sedimentation and C-reactive protein) , immune complexe and complement detection, detection of joint synovial fluid, and detection of autoantibodies.These indicators change with the activity of arthritis.Early diagnosis relies mainly on detection of autoantibodies. Anti-cyclic citrullinated peptide antibody( anti-CCP) is a kind of anti-keratin antibody, having higher sensitivity and specificity than the rheumatoid factor in diagnosis of rheumatoid arthritis.During the first year of rheumatoid arthritis, the existence of anti-CCP can be detected in the serum of most of these people.Positive anti-CCP can also be used to predict serious damage.Therefore, anti-CCP is extremely helpful in early diagnosis and prognosis assessment of rheumatoid arthritis.In this paper, the recent application of anti-CCP in the diagnosis of rheumatoid arthritis is reviewed.

  18. Efficacy of golimumab plus methotrexate in methotrexate-naïve patients with severe active rheumatoid arthritis.

    Science.gov (United States)

    Emery, Paul; Fleischmann, Roy M; Hsia, Elizabeth C; Xu, Stephen; Zhou, Yiying; Baker, Daniel

    2014-09-01

    The purpose of this study was to assess the treatment benefit of golimumab + methotrexate (MTX) vs. MTX monotherapy in MTX-naïve patients with severe active rheumatoid arthritis (RA). This was a post hoc analysis of MTX-naïve RA patients in the GO-BEFORE trial who were randomized to receive placebo + MTX (n = 160), golimumab 50 mg + MTX (n = 159), or golimumab 100 mg + MTX (n = 159). Subsets of patients with severe disease were identified using these baseline criteria: C-reactive protein (CRP) ≥1.5 mg/dL, CRP ≥3.0 mg/dL, swollen joint count (SJC) ≥10 and tender joint count (TJC) ≥12, SJC ≥ 20/TJC ≥ 12, 28-joint count Disease Activity Score using CRP (DAS28-CRP) >5.1, and anti-cyclic citrullinated peptide antibody-positive status. The treatment effect of golimumab + MTX vs. MTX alone was evaluated for these outcomes: the proportions of patients achieving ≥20, 50, and 70 % improvement in the American College of Rheumatology criteria; DAS28-CRP European League Against Rheumatism response; DAS28-CRP golimumab + MTX groups vs. placebo + MTX for all of the outcomes evaluated. Furthermore, the treatment effect of golimumab + MTX was consistently greater among patients in the severe disease subsets when compared with the overall GO-BEFORE trial population. The treatment benefit of golimumab + MTX vs. MTX monotherapy was most pronounced within the subsets of patients with CRP ≥3.0 mg/dL and SJC ≥ 20/TJC ≥ 12. Following treatment with golimumab + MTX, improvements in RA signs/symptoms and in progression of structural damage were evident for the overall GO-BEFORE population, with the treatment effect more pronounced among patients with severe active disease.

  19. Silibinin Improves the Effects of Methotrexate in Patients with Active Rheumatoid Arthritis: Pilot Clinical Study

    Science.gov (United States)

    Hussain, Saad Abdulrahman; Mortada, Ahmed Hashem; Jasim, Nazar Abdulateef; Gorial, Faiq Isho

    2016-01-01

    Objectives Our study sought to evaluate the effects of silibinin in patients with active rheumatoid arthritis (RA) treated with methotrexate (MTX). Methods We conducted a randomized multi-center, double-blind, placebo-controlled clinical trial over a 16-week treatment period at the Al-Sader and Baghdad Teaching Hospitals in Najaf and Baghdad, respectively. A total of 60 patients (30 of each sex) with active RA, already maintained on 12 mg MTX weekly for at least three consecutive months, were included in the study. Patients were randomly allocated to receive either 120 mg silibinin twice daily or a placebo, combined with their regular MTX regimen. The patients were evaluated by measuring disease activity score using the 28-joint Disease Activity Score, Simple Disease Activity Index, and Health Assessment Questionnaire–Disability Index scores at the start and end of the study. Blood samples were evaluated for the erythrocyte sedimentation rate (ESR), hemoglobin (Hb), high-sensitivity C-reactive protein (hs-CRP), creatine kinase (CK), anti-cyclic citrullinated peptide (CCP), and the serum cytokine levels of tumor necrosis factor (TNF)-α, interleukin (IL)-6, IL-8, IL-10, and IL-2. Results Silibinin significantly decreases the already elevated clinical scores compared to placebo treatment. ESR, IL-8, IL-6, TNF-α, anti-CCP, hs-CRP levels were significantly reduced. Additionally, the use of silibinin significantly increases Hb, IL-10, and IL-2 levels. Conclusion Silibinin may improve the effects of MTX on certain biochemical and clinical markers of patients with active RA. PMID:27403238

  20. Concomitância de artrite reumatoide e espondilite anquilosante em um único paciente: importância dos novos critérios de classificação Concurrent rheumatoid arthritis and ankylosing spondylitis in one patient: the importance of new classification criteria

    Directory of Open Access Journals (Sweden)

    Valderilio Feijó Azevedo

    2013-02-01

    Full Text Available Relatamos um caso de concomitância de espondilite anquilosante e artrite reumatoide em um paciente caucasiano de 65 anos, com achados clínicos de poliartrite simétrica com erosão de metacarpofalangeana ao raio X convencional e dor lombar infl amatória, HLA-B27+, associada à sacroiliíte. O paciente apresentou valores elevados de fator reumatoide e antipeptídeo citrulinado cíclico (anti-CCP. Realizamos uma revisão da literatura na qual as principais características de casos previamente reportados foram comparadas às deste caso. Este é o primeiro relato de caso de concomitância das duas doenças em que se utilizou teste laboratorial para dosagem do anti-CCP associado ao preenchimento das últimas versões dos critérios ASAS axial e ACR/EULAR para a classificação de espondilite anquilosante e artrite reumatoide, respectivamente.We report the case of concomitant ankylosing spondylitis and rheumatoid arthritis in a 65-year-old Caucasian male, who had symmetric polyarthritis with erosion of the metacarpophalangeal joint on conventional X-ray, infl ammatory low back pain with HLA-B27 positivity, and sacroiliitis. Laboratory analysis showed high levels of rheumatoid factor and anti-cyclic citrullinated peptide antibody (anti-CCP. Clinical features of previously reported cases were compared with those of our case. This is the first case report on the coexistence of both diseases in the same patient, for whom anti- CCP testing and the latest versions of axial ASAS criteria and ACR/EULAR criteria for the classification of ankylosing spondylitis and rheumatoid arthritis, respectively, were used.

  1. Serum levels of IL-17, IL-4, and INFγ in Serbian patients with early rheumatoid arthritis

    Science.gov (United States)

    Pavlovic, Voja; Dimic, Aleksandar; Milenkovic, Sasa; Krtinic, Dane

    2014-01-01

    Background: Rheumatoid arthritis (RA) is a chronic, systemic, inflammatory disease with autoimmune etiology, characterized by synovial inflammation and destruction of joint cartilage and bone. There are controversial data about the profile of interleukin-17 (IL-17A), interleukin-4 (IL-4), and interferon-gamma (INFγ), indicating in some studies the key role of IL-17, while in others the Th1 cytokines. Materials and Methods: Serum samples of 31 early RA patients were evaluated for erythrocytes sedimentation rate (ESR), rheumatoid factor (RF), C-reactive protein (CRP), anti-cyclic citrullinated peptide antibodies (anti-CCP), and for the tested cytokines (IL-17A, IL-4, and INFγ). Disease activity score (DAS28) calculation was done for all patients. Control serum samples were obtained from 29 healthy volunteers. Results: The levels of tested cytokines were significantly higher (IL-17A, p < 0.001; INFγ, p < 0.001; IL-4, p < 0.01) in patients with early RA, compared to the healthy controls. In early RA patients, a strong correlation of serum IL-17A was found with DAS28, ESR, and CRP. Also, significant negative correlation was found between serum INFγ levels and the DAS28 score, indicating that INFγ may play a key role in maintaining immune homeostasis in patients with RA. Conclusion: The mean serum IL-17A levels in patients with early RA, corresponded with the disease activity and severity. This might highlight the usefulness of the serum IL-17A level in defining the activity and predictive patterns, for aggressive disease therapy, and it might express specific therapeutically targets. PMID:24672560

  2. Serum levels of IL-17, IL-4, and INFγ in Serbian patients with early rheumatoid arthritis

    Directory of Open Access Journals (Sweden)

    Voja Pavlovic

    2014-01-01

    Full Text Available Background: Rheumatoid arthritis (RA is a chronic, systemic, inflammatory disease with autoimmune etiology, characterized by synovial inflammation and destruction of joint cartilage and bone. There are controversial data about the profile of interleukin-17 (IL-17A, interleukin-4 (IL-4, and interferon-gamma (INFγ, indicating in some studies the key role of IL-17, while in others the Th1 cytokines. Materials and Methods: Serum samples of 31 early RA patients were evaluated for erythrocytes sedimentation rate (ESR, rheumatoid factor (RF, C-reactive protein (CRP, anti-cyclic citrullinated peptide antibodies (anti-CCP, and for the tested cytokines (IL-17A, IL-4, and INFγ. Disease activity score (DAS28 calculation was done for all patients. Control serum samples were obtained from 29 healthy volunteers. Results: The levels of tested cytokines were significantly higher (IL-17A, p < 0.001; INFγ, p < 0.001; IL-4, p < 0.01 in patients with early RA, compared to the healthy controls. In early RA patients, a strong correlation of serum IL-17A was found with DAS28, ESR, and CRP. Also, significant negative correlation was found between serum INFγ levels and the DAS28 score, indicating that INFγ may play a key role in maintaining immune homeostasis in patients with RA. Conclusion: The mean serum IL-17A levels in patients with early RA, corresponded with the disease activity and severity. This might highlight the usefulness of the serum IL-17A level in defining the activity and predictive patterns, for aggressive disease therapy, and it might express specific therapeutically targets.

  3. E26 Transformation-Specific-1 (ETS1 and WDFY Family Member 4 (WDFY4 Polymorphisms in Chinese Patients with Rheumatoid Arthritis

    Directory of Open Access Journals (Sweden)

    Yiqun Zhang

    2014-02-01

    Full Text Available E26 transformation-specific-1 (ETS1 and WDFY family member 4 (WDFY4 are closely related with systemic lupus erythematosus. We hypothesized that ETS1 and WDFY4 polymorphisms may contribute to rheumatoid arthritis (RA susceptibility. We studied ETS1 rs1128334 G/A and WDFY4 rs7097397 A/G gene polymorphisms in 329 patients with RA and 697 controls in a Chinese population. Genotyping was done using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. When the WDFY4 rs7097397 AA homozygote genotype was used as the reference group, the AG genotype was associated with a significantly increased risk for RA. In the dominant model, when the WDFY4 rs7097397 AA homozygote genotype was used as the reference group, the AG/GG genotypes were associated with a significant increased susceptibility to RA. In stratification analyses, a significantly increased risk for RA associated with the WDFY4 rs7097397 AG genotype was evident among female patients, younger patients, C-reactive protein (CRP negative patients and both anti-cyclic citrullinated peptide antibody (ACPA positive patients and negative patients compared with the WDFY4 rs7097397 AA genotype. These findings suggested that WDFY4 rs7097397 A/G may be associated with the risk of RA, especially among younger, female patients, CRP-negative patients and both ACPA positive and negative patients. However, our results were obtained from a moderate-sized sample, and therefore this is a preliminary conclusion. To confirm these findings, validation by a larger study from a more diverse ethnic population is needed.

  4. Wrist ultrasound analysis of patients with early rheumatoid arthritis

    Directory of Open Access Journals (Sweden)

    J.A. Mendonça

    2011-01-01

    Full Text Available In the present study, we evaluated 42 wrists using the semi-quantitative scales power Doppler ultrasound (PDUS and gray scale ultrasound (GSUS with scores ranging from 0 to 3 and correlated the results with clinical, laboratory and radiographic data. Twenty-one patients (17 women and 4 men with rheumatoid arthritis according to criteria of the American College of Rheumatology were enrolled in the study from September 2008 to July 2009 at Universidade Estadual de Campinas (UNICAMP. The average disease duration was 14 months. The patients were 66.6% Caucasians and 33.3% non-Caucasians, with a mean age of 42 and 41 years, respectively. A dorsal longitudinal scan was performed by ultrasound on the radiocarpal and midcarpal joints using GE LOGIQ XP-linear ultrasound and a high frequency (8-10 MHz transducer. All patients were X-rayed, and the Larsen score was determined for the joints, with grades ranging from 0 to V. This study showed significant correlations between clinical, sonographic and laboratory data: GSUS and swollen right wrist (r = 0.546, GSUS of right wrist and swelling of left wrist (r = 0.511, PDUS of right wrist and pain in left wrist (r = 0.436, PDUS of right wrist and C-reactive protein (r = 0.466. Ultrasound can be considered a useful tool in the diagnosis of synovitis in early rheumatoid arthritis mainly when the anti-cyclic citrullinated peptide and rheumatoid factor are negative, and can lead to an early change in the therapeutic decision.

  5. Arginine methylation and citrullination of splicing factor proline- and glutamine-rich (SFPQ/PSF) regulates its association with mRNA

    OpenAIRE

    Snijders, Ambrosius P; Hautbergue, Guillaume M.; Bloom, Alex; Williamson, James C.; Minshull, Thomas C.; Phillips, Helen L.; Mihaylov, Simeon R.; Gjerde, Douglas T.; Hornby, David P; Stuart A Wilson; Hurd, Paul J.; Dickman, Mark J.

    2015-01-01

    Splicing factor proline- and glutamine-rich (SFPQ) also commonly known as polypyrimidine tract-binding protein-associated-splicing factor (PSF) and its binding partner non-POU domain-containing octamer-binding protein (NONO/p54nrb), are highly abundant, multifunctional nuclear proteins. However, the exact role of this complex is yet to be determined. Following purification of the endogeneous SFPQ/NONO complex, mass spectrometry analysis identified a wide range of interacting proteins, includi...

  6. Fragments of Citrullinated and MMP-degraded Vimentin and MMP-degraded Type III Collagen Are Novel Serological Biomarkers to Differentiate Crohn's Disease from Ulcerative Colitis

    DEFF Research Database (Denmark)

    Mortensen, Joachim Høg; Godskesen, Line Elbjerg; Jensen, Michael Dam;

    2015-01-01

    BACKGROUND AND AIMS: A hallmark of inflammatory bowel disease [IBD] is chronic inflammation, which leads to excessive extracellular matrix [ECM] remodelling and release of specific protein fragments, called neoepitopes. We speculated that the biomarker profile panel for ulcerative colitis [UC......-degraded vimentin [VICM] were studied with a competitive ELISA assay system in a cohort including 164 subjects [CD n = 72, UC n = 60, and non-IBD controls n = 32] and a validation cohort of 61 subjects [CD n = 46, and UC n = 15]. Receiver operating characteristic curve analysis and logistic regression modelling....... Furthermore, the biomarkers C1M [AUC = 0.81], C3M [AUC = 0.83], VICM [AUC = 0.83], and P1NP [AUC = 0.77] were best to differentiate UC from non-IBD. The best combinations of biomarkers to differentiate CD from UC and UC from non-IBD were VICM, C3M, C4M [AUC = 0.90] and VICM, C3M [AUC = 0.98] respectively...

  7. COMPARATIVE CLINICAL, LABORATORY, AND INSTRUMENTAL EVALUATION OF INTERSTITIAL LUNG CHANGES IN RHEUMATOID ARTHRITIS

    Directory of Open Access Journals (Sweden)

    D. V. Bestaev

    2014-01-01

    Full Text Available Interstitial lung disease (ILD is a common extra-articular manifestation of rheumatoid arthritis (RA. Objective: to study the associations of the data of high-resolution computed tomography (HRCT and the esti- mate of diffusing lung capacity (DLC with clinical and laboratory parameters in RA patients with and without ILD. Subjects and methods. 79 RA patients fulfilling the 1987 American College of Rheumatology criteria (61 women and 18 men admitted to the Nasonova Research Institute of Rheumatology were included. Results. HRCT revealed signs of ILD in 58 (73% cases. The patients with ILD were divided into three groups: 1 18 (31% patients with ground glass opacities; 2 34 (58.6% patients with fibrosis; 3 6 (10.4% patients with the honeycomb lung. Twenty-one (27% patients with ILD were included in Group 4. In the ILD patients with ground glass opacities, the levels of anti-cyclic citrullinated peptide (ACCP antibodies and rheumatoid factor (RF were much above those in the patients without ILD (240 [166; 410.5], 480 [140; 850.5] and 73 [31; 101], 330,5 [118.5; 604.8], respectively. In the patients with ILD, the concentration of C-reactive protein (CRP (46 [35; 91] was higher than that in those without ILD (24 [18; 31]; p < 0.05. In the ILD patients with ground glass opacities, DLC was considerably below that in those with ILD – 59.2±11.2 and 79.8±12.1% of the normal value, respectively (p < 0.001. Conclusion. The associations found between ACCP antibodies and DLC, DAS28 and DLC may suggest that ACCP antibodies are implicated in the pathogenesis of ILD and the lung is involved in the immunoinflammatory process. The high percent of smokers detected in our investigation confirms the considerable role of smoking in the pathogene- sis of RA-associated ILD. In the RA patients with ILD, ground glass opacities must be an indicator of the activity of an immunopathological process in the lung.

  8. Immunological effects of adalimumab in rheumatoid arthritis: Results of a Russian national study

    Directory of Open Access Journals (Sweden)

    Dmitri Evgenyevich Karateev

    2013-01-01

    Full Text Available A reduction in immunological parameters during therapy for rheumatoid arthritis (RA can yield new data on the mechanisms of anti-inflammatory action and be of great practical importance since this allows judgment of the depth of impact on the immunological process and therefore may be regarded as one of the components of improvement and remission. Objective: to study the effect of the tumor necrosis factor-a (TNF-а inhibitor adalimumab (ADA on a number of key immunological parameters in RA. Subjects and methods. The study included 100 patients (11 men and 89 women diagnosed with RA from 5 research centers. The patients were observed to have high RA activity: at baseline, DAS28 6.22+0.84 scores; C-reactive protein (CRP 37.1+34.7 mg/l. The mean number of disease-modifying antirheumatic drugs (DMARDs used by a patient in the history was 2.1. During 24-week treatment, the patients took ADA in a subcutaneous dose of 40 mg every 2 weeks both alone and in combination with DMARDs. The time course of changes in the serum levels of CRP, IGM rheumatoid factor (RF and in the concentration of anti-cyclic citrullinated peptide 2 (anti-CCP2 antibodies was determined by enzyme immunoassay (EIA using the Axis-Shield Diagnostics commercial kits (United Kingdom. The levels of TNF-a, interleukin (IL 6, 12, matrix metalloproteinase 3 (MMP3 were measured in pg/ml by EIA using the Bender MedSystems commercial kits (USA according to the manufacturer's directions. Results. During ADA therapy, there was a rapid reduction in the level of CRP from 34.3 to 11.317.2 mg/l following 2 weeks (p<0.001, which thereafter remained low (11.9 mg/l, with some fluctuations, until week 24 of the study. There was a significant reduction in blood RF levels from 169.24 to 111.97 at 24 weeks (p<0.001. After 12 weeks of ADA treatment, there was virtually a two-fold decrease in IL-6 content from 8.87 to 4.87 pg/ml and later on to 4.03 pg/ml at week 24 (p>0,05. The mean levels of anti-CCP2

  9. COMORBIDITY IN RHEUMATOID ARTHRITIS

    Directory of Open Access Journals (Sweden)

    T. A. Panafidina

    2014-01-01

    Full Text Available The peak onset of rheumatoid arthritis (RA is at 30-55 years of age. At this age, the patients have also other concomi- tant diseases (comorbidities that affect the course and prognosis of RA, the choice of its treatment policy, quality of life of the patients. Objective: to identify the most important and common comorbidities in patients with RA. Subjects and methods. Two hundred patients (median age 55 [46; 61] years were enrolled; there was a preponderance of women (82.5% with median disease duration 5 [1; 10] years, seropositive for IgM rheumatoid factor (83.0% and anti-cyclic citrullinated peptide antibodies (81.6% with moderate and high disease activity (median DAS28 value 3.9 [3.1; 4.9]. Varying degrees of destructive changes in hand and foot joints were radiologically detected in 71.2% of the patients; 64.5% of the patients had Functional Class II. Methotrexate was given to 69.5% of the patients; therapy with biological agents was used in 21.0% of the cases. 15.5% of the patients did not receive DMARD or biologics. 43.0% of the patients with RA received glucocorticoids. Results. Comorbidities were present in 72.0% of the patients with RA. The most common diseases were hypertension (60.0%, dyslipidemia (45.0%, fractures at various sites (29.5%, and coronary heart disease (21.0%. Myocardial infarction and stroke were observed in 1.5 and 1.0% of cases, respectively. There was diabetes mellitus (DM in 7.5% of the cases and osteoporosis in 15.5% of the patients. 81.7% of the patients with RA and hypertension and 80.0% of those with RA and DM received antihypertensive and sugar-lowering therapy, respectively. At the same time the RA patients with dyslipidemia and osteoporosis received specific drugs far less frequently (30.0 and 29.0%, respectively. Conclusion. Comorbidities are frequently encountered in RA. By taking into account the fact that cardiovascular dis- eases are a main cause of death in RA; it is necessary to adequately and timely

  10. Study of association of CD40-CD154 gene polymorphisms with disease susceptibility and cardiovascular risk in Spanish rheumatoid arthritis patients.

    Directory of Open Access Journals (Sweden)

    Mercedes García-Bermúdez

    Full Text Available OBJECTIVE: Rheumatoid arthritis (RA is a chronic inflammatory disease associated with increased cardiovascular (CV mortality. Since CD40-CD154 binding has direct consequences on inflammation process initiation, we aimed to replicate previous findings related to disease susceptibility in Spanish RA population. Furthermore, as the major complication in RA disease patients is the development of CV events due to accelerated atherosclerosis, and elevated levels of CD40L/CD154 are present in patients with acute myocardial infarction, we assessed the potential association of CD40 and CD154/CD40L gene variants with CV risk in Spanish RA patients. METHODS: One thousand five hundred and seventy-five patients fulfilling the 1987 ACR classification criteria for RA and 1600 matched controls were genotyped for the CD40 rs1883832, rs4810485 and rs1535045 and CD154 rs3092952 and rs3092920 gene polymorphisms, using predesigned TaqMan single nucleotide polymorphism genotyping assays. Afterwards, we investigated the influence of CD40-CD154 gene variants in the development of CV events. Also, in a subgroup of 273 patients without history of CV events, we assessed the influence of these polymorphisms in the risk of subclinical atherosclerosis determined by carotid ultrasonography. RESULTS: Nominally significant differences in the allele frequencies for the rs1883832 CD40 gene polymorphism between RA patients and controls were found (p=0.038. Although we did not observe a significant association of CD40-CD154 gene variants with the development of CV events, an ANCOVA model adjusted for sex, age at the time of the ultrasonography assessment, follow-up time, traditional CV risk factors and anti-cyclic citrullinated peptide antibodies disclosed a significant association (p=0.0047 between CD40 rs1535045 polymorphism and carotid intima media thickness, a surrogate marker of atherosclerosis. CONCLUSION: Data from our pilot study indicate a potential association of rs

  11. SMAD3 rs17228212 gene polymorphism is associated with reduced risk to cerebrovascular accidents and subclinical atherosclerosis in anti-CCP negative Spanish rheumatoid arthritis patients.

    Directory of Open Access Journals (Sweden)

    Mercedes García-Bermúdez

    Full Text Available Rheumatoid arthritis (RA is a complex polygenic inflammatory disease associated with accelerated atherosclerosis and increased risk of cardiovascular (CV disease. Previous genome-wide association studies have described SMAD3 rs17228212 polymorphism as an important signal associated with CV events. The aim of the present study was to evaluate for the first time the relationship between this gene polymorphism and the susceptibility to CV manifestations and its potential association with the presence of subclinical atherosclerosis assessed by the evaluation of carotid intima-media thickness (cIMT in patients with RA.One thousand eight hundred and ninety-seven patients fulfilling classification criteria for RA were genotyped for SMAD3 rs17228212 gene polymorphism through TaqMan genotyping assay. Also, subclinical atherosclerosis determined by the assessment of cIMT was analyzed in a subgroup of these patients by carotid ultrasonography.No statistically significant differences were observed when allele frequencies of RA patients with or without CV events were compared. Nevertheless, when RA patients were stratified according to anti-cyclic citrullinated peptide (anti-CCP status, we found that in RA patients who were negative for anti-CCP antibodies, the presence of C allele of SMAD3 rs17228212 polymorphism conferred a protective effect against the risk of cerebrovascular accident (CVA after adjustment for demographic and classic CV risk factors (HR [95%CI]=0.36 [0.14-0.94], p=0.038 in a Cox regression model. Additionally, correlation between the presence of C allele of SMAD3 rs17228212 polymorphism and lower values of cIMT was found after adjustment for demographic and classic CV risk factors (p-value=0.0094 in the anti-CCP negative RA patients.Our results revealed that SMAD3 rs17228212 gene variant is associated with lower risk of CVA and less severe subclinical atherosclerosis in RA patients negative for anti-CCP antibodies. These findings may have

  12. Usefulness of anti -CCP antibodies in rheumatic diseases in Indian patients

    Directory of Open Access Journals (Sweden)

    Gupta Rajiva

    2009-03-01

    Full Text Available Background: The usefulness of anti-cyclic citrullinated peptide antibodies (anti-CCP antibodies to identify rheumatic arthritis (RA from other rheumatic diseases presenting with joint pain is not well studied. Aims: We aimed to determine the sensitivity and specificity of anti-CCP antibodies in Indian RA patients with respect to non-RA rheumatic diseases and to study the relationship of anti-CCP antibodies and IgG, IgM and IgA rheumatoid factor in RA. Settings and Design: Case-control cross-sectional study carried out in the rheumatology division of All India Institute of Medical Sciences.Materials and Methods: Sixty-three patients with rheumatoid arthritis (RA and 51 patients with non-RA rheumatic diseases having joint pain were included in the study. Sera were tested for anti-CCP antibodies (IgG and IgA, IgM, IgG rheumatoid factor, using a commercially available enzyme-linked immunosorbent assay. Statistical Analysis: Statistical analysis was performed using SPSS statistical software version 11.5. Results: Fifty-four of 63 RA patients (85.71% were positive for anti-CCP antibodies. In the non-RA group, anti-CCP antibody was positive in only 5 of 51 patients (9.8%. Our study found a sensitivity of 85% and a specificity of 90.19% with regard to the use of anti-CCP antibodies assay in patients with joint pain to correctly identify RA. Anti-CCP antibodies positive patients did not have more erosive disease. IgM-RF-positive patients had more erosion when compared to the IgM-RF-negative group. Thirty-two of 57 (56.1% IgM-RF-positive patients had erosions, while no patient (0/6 patients had erosions in the IgM-RF-negative group (P = 0.01 Conclusion: Anti-CCP antibodies have high sensitivity and specificity for diagnosis of RA, in Indian patients. Anti-CCP antibodies positive patients did not have more erosive disease in our study.

  13. The value evaluation of two diagnosis indicators of rheumatoid arthritis%类风湿性关节炎两项诊断指标应用价值评价

    Institute of Scientific and Technical Information of China (English)

    栗晓芳; 侯小平; 张永良; 王俊明; 李萍

    2013-01-01

    目的 类风湿性关节炎(Rheumatoid Arthritis,RA)是一种最常见的自身免疫病,且致畸性较高,本实验探讨抗环瓜氨酸肽抗体(Anti-Cyclic Citrullinated Peptide antibodies)和类风湿因子(rheumatoid factor,RF)联合检测对RA的诊断价值及应用.方法 选取某院门诊及住院RA患者90例,非RA患者70例,健康体检者50例,分别用胶体金法检测血清中抗-CCP抗体、用速率散射比浊法检测血清RF,对二者进行比较.结果 抗-CCP抗体在RA组阳性率(81.1%),高于非RA患者组(4.2%)和健康体检组(0.0%) (P<0.05);RF在RA组的阳性率(90%),高于非RA患者组(17.1%)和健康体检组(0.0%) (P<0.05);RF诊断RA的敏感性(90%)高于抗-CCP抗体(81.1%)(P< 0.05);抗-CCP抗体诊断RA的特异性(97.5%)高于RF (89.2%),假阳性率(2.5%)低于RF(10.8%) (P<0.05);抗-CCP抗体和RF联合诊断RA的特异性98.3%,敏感性80%.结论 抗-CCP抗体对RA诊断具有高特异性,RF对RA诊断具有高敏感性,两者结合可以提高对RA诊断的特异性,从而有助于RA与其他结缔组织病相鉴别.

  14. 一种基于量子点检测抗CCP抗体的免疫荧光层析法%Development of an Immunochromatographic Test Strip for the Detection of Anti-CCP Antibody Based on Quantum Dots

    Institute of Scientific and Technical Information of China (English)

    郭利宁; 何红秋

    2013-01-01

    抗环瓜氨酸多肽(cyclic citrullinated peptide,CCP)抗体是类风湿关节炎(rheumatoid arthritis,RA)早期诊断的重要生物标志物.为了实现对RA的早期诊断,本研究建立了一种基于CdTe量子点标记技术检测抗CCP抗体的免疫荧光层析法.将CCP多肽与小牛血清白蛋白(bovine serum albumin,BSA)连接,再将CCP-BSA和羊抗鼠IgG分别在硝酸纤维素膜(nitrocellulose membrane,NC膜)上划线,作为检测线(test line,T线)和质控线(control line,C线).制备量子点并在量子点上标记鼠抗人IgG,喷在玻璃纤维上并烘干,最后组装大卡、切割并封装制成检测试纸条.应用该试纸条检测了RA患者及健康人血清临床样本200份,以酶联免疫吸附测定法(enzyme-linked immunosorbent assay,ELISA)为对照,计算免疫荧光层析法的检测灵敏度和特异性.结果显示,建立的量子点免疫荧光层析试纸条检测抗CCP抗体的灵敏度为97.5%,特异性为95.8%.该方法操作简单、快速,可实现床旁检测(point-of-care testing,POCT),能应用于RA的早期诊断.%Anti-cyclic citrullinated peptide antibody ( anti-CCP antibody) is an important biomarker for early detection of rheumatoid arthritis ( RA). For clinical diagnosis of RA in the early stage, an immunochromatographic test strip for anti-CCP antibody detection was developed based on quantum dots. First, CCP was linked to bovine serum albumin (BSA) ; and then, the CCP-BSA and goat anti-mouse IgG were dotted on the nitrocellulose membrane as test line ( T line) and control line ( C line) , respectively. Subsequently, quantum dots solution was prepared and conjugated to the mouse anti-human IgG antibody; further more, the marked quantum dots were sprayed to the glass fibers and dried. Last, all components of immunochromatographic test strip were assembled, cut and packaged. The test strip has been applied to the detection of 200 clinic serum samples from RA patients and healthy people, and the sensitivity

  15. Diagnostic value of combined detection of anti-CCP, anti-MCV antibodies and RF in rheumatoid arthritis%血清抗CCP抗体、抗MCV抗体与RF联合检测诊断类风湿关节炎的临床价值

    Institute of Scientific and Technical Information of China (English)

    许军华; 刘树业

    2013-01-01

    目的 探讨抗环瓜氨酸肽(CCP)抗体、抗突变型瓜氨酸波形蛋白(MCV)抗体和类风湿因子(RF)联合检测诊断类风湿关节炎(RA)的价值.方法 采用ELISA法检测101例RA患者(RA组)、55例非RA自身免疫性疾病患者(非RA组)和55例健康查体者(对照组)血清抗CCP抗体、抗MCV抗体和RF诊断RA水平,并进行比较.结果 RA组抗CCP抗体、抗MCV抗体和RF阳性率均显著高于非RA组和对照组(P均<0.01).RA组抗CCP抗体、抗MCV抗体和RF的敏感性分别为83.2%、70.3%和79.2%,特异性分别为93.6%、94.5%和75.4%.三项指标联合检测诊断RA的特异性达99.1%,敏感性降至67.3%.结论 抗CCP抗体、抗MCV抗体和RF联合检测可提高诊断RA的准确性.%Objective To investigate the clinical value of combined detection of anti-cyclic citrullinated peptide (antiCCP) antibodies,anti-mutated citrullinated vimentin (anti-MCV) antibodies and rheumatoid factor (RF) in rheumatoid arthritis (RA) diagnosis.Methods Anti-CCP,anti-MCV antibodies and RF were detected by ELISA in 101 RA patients(RA group),55 patients with other rheumatic diseases (non-RA group),and 55 healthy controls (control group).The differences of anti-CCP,anti-MCV antibodies and RF in the three groups were compared.Results The positive ratios of anti-CCP,anti-MCV antibodies and RF were significantly higher in the RA group than those in the non-RA group and control group (P < 0.01).In the RA group,the sensitivity of anti-CCP antibodies was 83.2% with 93.6% specificity,the sensitivity of anti-MCV antibodies was 70.3% and the specificity was 94.5%,the sensitivity of RF was 79.2% with 75.4% specificity.The specificity of combined detection of anti-CCP,anti-MCV antibodies and RF was increased to 99.1%,with a lower 67.3% sensitivity.Conclusions The combined detection of anti-CCP,anti-MCV antibodies and RF could improve the diagnostic accuracy of rheumatoid arthritis (RA).

  16. Relationship between Esophageal Hypersensitivity and Calcitonin Gene-related Peptide Positive Fibers in Lower Esophageal Sphincter Mucosa of Patients with Non-erosive Reflux Disease%非糜烂性反流病患者食管内脏高敏感性与食管下括约肌局部降钙素基因相关肽阳性神经纤维的关系

    Institute of Scientific and Technical Information of China (English)

    杨敏; 房殿春; 李兆申; 徐晓蓉; 邹多武; 孙振兴; 屠振兴; 许国铭; 龚燕芳

    2006-01-01

    背景:食管内脏高敏感性是非糜烂性反流病(NERD)最重要的病理生理特征之一,但引起食管内脏感觉过敏的确切机制仍不甚清楚.目的:通过测定NERD患者食管对机械扩张刺激和对酸刺激的敏感性变化和降钙素基因相关肽(CGRP)在食管下括约肌(LES)局部组织中的表达,探讨食管感知阈值与LES局部黏膜CGRP表达之间的关系.方法:采用Synectics内脏刺激器/电子气压泵行食管气囊扩张术以检测食管对机械扩张刺激的敏感性;采用食管酸灌注试验(bemstein test)检测食管对酸的敏感性;采用免疫组化法和图像分析技术观察LES局部组织中CGRP的表达.结果:根据对酸刺激和(或)机械扩张刺激的感知过敏,NERD患者可分为感知过敏组(21例)和感知正常组(10例).感知过敏组患者食管对气囊扩张刺激的初始感知阈值和最大耐受疼痛阈值较感知正常组和正常对照组显著降低(P<0.05).感觉过敏组LES黏膜中CGRP阳性纤维数和平均吸光度(A)值较感知正常组和正常对照组显著增高(P<0.05).LES局部组织中CGRP阳性产物A值与食管初始感知阈值和最大耐受疼痛阈值呈直线负相关(r分别为-0.68和-0.79,P<0.03).结论:多数NERD患者存在对食管机械扩张刺激和(或)对食管酸刺激感知过敏;感知过敏的NERD患者其LES局部黏膜中CGRP表达增加,提示LES肽能神经的改变可能与食管内脏高敏感性有关.

  17. Diagnostic significance of detection of anti-citrullinated peptide antibodies in juvenile rheumatoid arthritis%抗瓜氨酸肽抗体谱在幼年类风湿关节炎诊断中的意义

    Institute of Scientific and Technical Information of China (English)

    茹晋丽; 李小峰; 张莉芸; 魏华; 胡学芳; 牛红青

    2007-01-01

    目的 探讨抗环瓜氨酸肽抗体(抗CCP抗体)、类风湿因子(RF)、抗核周因子(APF)、抗角蛋白抗体(AKA)对幼年类风湿关节炎(JRA)诊断的意义,并与类风湿关节炎(RA)进行比较.方法 分别采用酶联免疫吸附试验(ELISA)、胶乳凝集法、间接免疫荧光法检测54例JRA患者、31例非RA对照组、116例成人RA患者血清中的抗CCP抗体、RF、APF、AKA.结果 54例JRA患者抗CCP抗体、RF、APF、AKA的敏感性分别为61.1%、57.4%、37.0%、18.5%,特异性分别为96.8%、93.6%、96.8%、100%.抗CCP抗体与RF在JRA的敏感性差异无统计学意义,但明显高于APF、AKA;4种抗体在JRA的特异性比较差异无统计学意义.RA患者中4种抗体敏感性分别为82.3%、78.3%、48.7%、25.4%,特异性分别为95.7%、73.7%、91.6%、94.0%.4种抗体在JRA诊断中的敏感性均明显低于RA组,RF在JRA患者中特异性明显高于RA患者,而其他3种抗体的特异性在两组患者比较差异无统计学意义.结论 虽然抗CCP抗体谱在JRA诊断中的敏感性不及RA组,但对JRA的诊断仍具有较好的敏感性和特异性,可用于JRA的诊断.

  18. Constitutive expression of citrulline biosynthetic genes in Corynebacterium glutamicum%瓜氨酸生产相关基因簇在谷氨酸棒杆菌中的组成型表达

    Institute of Scientific and Technical Information of China (English)

    谢红翠; 郝宁; 韦萍; 严明; 许琳

    2011-01-01

    构建1种组成型载体并将载体应用在表达瓜氨酸相关基因簇argCJBDF上.通过去除pXMJ19诱导型启动子上游阻遏蛋白lacⅠ基因的方法,构建组成型质粒pXMJ19 -lacⅠ,并将谷氨酸棒杆菌中合成瓜氨酸途径的基因簇argCJBDF克隆到改造过的组成型载体中,实现瓜氨酸合成相关基因簇argCJBDF在谷氨酸棒杆菌的组成型表达.结果表明:重组菌在30℃摇瓶发酵72 h后,N-乙酰谷氨酸激酶的酶活达到(0.323±0.015) U/mg,瓜氨酸的产量达到4.33 g/L.成功构建的组成型表达载体,实现了外源基因簇argCJBDF在谷氨酸棒杆菌中的组成型表达.

  19. Intérêt de la citrulline dans la prise en charge du sujet âgé dénutri : Etudes expérimentale et clinique

    OpenAIRE

    Faure, Cécile

    2010-01-01

    La dénutrition protéino-énergétique, qui touche une partie de la population âgée, accentue leur déclin musculaire (sarcopénie et dynapénie) et participe donc à l'augmentation de la morbi-mortalité. La prévention et le traitement de la dénutrition s'avèrent difficiles, lié au défaut de réponse à la renutrition observé chez les personnes âgées ; celui-ci est expliqué par une forte extraction splanchnique des acides aminés en période post-prandiale, responsable d'une moindre stimulation de la pr...

  20. Identification of a linear epitope recognized by a monoclonal antibody directed to the heterogeneous nucleoriboprotein A2

    DEFF Research Database (Denmark)

    Tronstrøm, Julie; Dragborg, Anette H.; Hansen, Paul Robert;

    2014-01-01

    Rheumatoid arthritis (RA) is a chronic autoimmune disorder, characterized by progressive joint destruction and disability. Classical autoantibodies of RA are rheumatoid factors and citrulline antibodies. Patients positive for these autoantibodies are usually associated with a progressive disease...... course. A subgroup of RA patients does not express citrulline antibodies, instead are approximately 35% of these anti-citrulline-negative patients reported to express autoantibodies to the heterogeneous nucleoriboprotein A2, a ribonucleoprotein involved in RNA transport and processing also referred...

  1. X-RAY PROGRESSION AND ITS IMPACT ON THE FUNCTIONAL STATUS OF PATIENTS WITH EARLY RHEUMATOID ARTHRITIS DURING A 5-YEAR FOLLOW-UP

    Directory of Open Access Journals (Sweden)

    Yu. A. Ermakova

    2015-01-01

    Full Text Available Objective: to assess progression of X-ray joint changes and their impact on functional status of patients with early rheumatoid arthritis (RA.Subjects and methods. The investigation enrolled 204 patients with early RA included in the RADICAL (Early Arthritis: Diagnosis, Outcomes, Criteria, Active Treatment program in the period 2003 to 2007. The duration of the disease at the inclusion was less than 2 years. RA was diagnosed according to 1987 American College of Rheumatology (ACR criteria. Rheumatoid factor was presentin 67.5% of the patients and anti-cyclic citrullinated peptide antibodies – in 57%. Women were 86.5%; median age was 49 [40; 58] years. The patients received traditional diseasemodifying antirheumatic drugs and biological agents (24%. Therapy response was monitored applying the tight control strategy. Main clinical and laboratory parameters, DAS28, and HAQ scores, and functional class and hand and foot X-ray were recorded every year.Results and discussion. After 5 years of follow-up, new erosions appeared in 57% of the patients. Joint erosions were present in 16.5% of the patients at the inclusion and in 73% following 5 years. At baseline, after 1, 2, 3, 4, and 5 years, the total erosion scores (modified Sharp score was equal to 0 [0; 0], 0 [0; 1], 0 [0; 4], 1 [0; 8], 3 [0; 12], 6 [0; 14], respectively. A significant functional improvement could be achieved just during the first year of follow-up. Thus, the median HAQ value decreased from 1.125 [0.625; 1.75] to 0.5 [0.125; 1.0], reaching the population level. Following 2, 3, 4, and 5 years, the median HAQ was 0.5 [0; 1.0], 0.5 [0; 1.0], 0.5 [0; 1.0], and 0.75 [0.125; 1.125], respectively. There was a weak positive correlation between total erosion scores and HAQ in the eаrly stage of the disease. However, it became more marked after 3, 4, and 5 years (correlation coefficient, 0.38, 0.39, and 0.40,respectively; p < 0.01. Conclusion.In spite of the stable pattern

  2. SUBCLINICAL INTERSTITIAL PULMONARY INJURY IN RHEUMATOID ARTHRITIS

    Directory of Open Access Journals (Sweden)

    D. V. Bestaev

    2015-01-01

    Full Text Available Subjects and methods. The study enrolled 61 inpatients diagnosed with RA (according to the 1987 American College of Rheumatology criteria who were treated at the V.A. Nasonova Research Institute of Rheumatology; in so doing, high-resolution computed tomography revealed lung changes as a ground glass pattern in 15 patients, reticular striation, traction bronchoectases, and lung tissue changes as honeycomb ones in 25 patients; no lung abnormalities were found in 21 patients. DAS28 was applied to determine the inflammatory activity of RA. The RA patients underwent X-ray studies of the hand, foot, and chest, by using accordingly X-Ray unit and spiral computed tomography scanner (section thickness, 0.65 mm. External respiration function (ERF indicators were studied with plethysmograph. IgM rheumatoid factor was measured using an immune nephelometer. Serum anti-cyclic citrullinated peptide antibodies were assayed by immunochemiluminescence technique on a Cobas e411 analyzer. The xMAP technology using a BioPlex200 analyzer was employed to determine the serum concentrations of 27 cytokines in 15 patients with subclinical IPI and in 25 with clinical IPI. Results and discussion. The major respiratory signs in patients with IPI proved to be cough (24 %, expectoration (20 %, dyspnea (16 %, and crepitation (64 % on auscultation. Three patients with subclinical IPI were found to have crepitation on auscultation. Respiratory symptoms were absent in the RA patients without IPI. It should be noted that there are a larger number of RA patients with a high smoking index among the RA patients with IPI than among those without IPI (p < 0.05. Investigation of ERF indicators revealed a statistically significantly lower lung diffusing capacity (LDC in the RA patients with subclinical IPI than in those without IPI (p < 0.05. Other ERF indicators showed no significant deviations of the reference values. LDC and total lung capacity appeared to be statistically

  3. Prevalence of Undiagnosed Diabetes in Rheumatoid Arthritis: an OGTT Study

    Science.gov (United States)

    Ursini, Francesco; Russo, Emilio; D’Angelo, Salvatore; Arturi, Franco; Hribal, Marta Letizia; D’Antona, Lucia; Bruno, Caterina; Tripepi, Giovanni; Naty, Saverio; De Sarro, Giovambattista; Olivieri, Ignazio; Grembiale, Rosa Daniela

    2016-01-01

    Abstract Rheumatoid arthritis (RA) is a chronic inflammatory disease characterized by an excess of cardiovascular disease (CVD) risk, estimated to be at least 50% greater when compared to the general population. Although the widespread diffusion of type 2 diabetes mellitus (T2DM) awareness, there is still a significant proportion of patients with T2DM that remain undiagnosed. Aim of this cross-sectional study was to evaluate the prevalence of undiagnosed diabetes and prediabetes in RA patients. For the present study, 100 consecutive nondiabetic RA patients were recruited. Age- and sex-matched subjects with noninflammatory diseases (osteoarthritis or fibromyalgia) were used as controls. After overnight fasting, blood samples were obtained for laboratory evaluation including serum glucose, total cholesterol, high-density lipoprotein (HDL)-cholesterol, low-density lipoprotein (LDL)-cholesterol, triglycerides, uric acid, erythrocyte sedimentation rate (ESR), high sensitivity C-reactive protein (hs-CRP), rheumatoid factor (RF), and anti-Cyclic Citrullinated Peptide Antibodies (ACPA). A standard Oral Glucose Tolerance Test (OGTT) with 75 g of glucose was performed and blood samples were collected at time 0, 30, 60, 90, and 120 minutes, for measurement of plasma glucose concentrations. The prevalence of impaired fasting glucose (IFG) (9/100 vs 12/100, P = 0.49), impaired glucose tolerance (IGT) (19/100 vs 12/100, P = 0.17), and concomitant IFG/IGT (5/100 vs 9/100, P = 0.27) was similar between groups, whereas the prevalence of diabetes was significantly higher in RA patients (10/100 vs 2/100, P = 0.02). In a logistic regression analysis, increasing age (OR = 1.13, 95% CI 1.028–1.245, P = 0.01) and disease duration (OR = 1.90, 95% CI 1.210–2.995, P = 0.005) were both associated with an increased likelihood of being classified as prediabetes (i.e. IFG and/or IGT) or T2DM. A ROC curve was built to evaluate the predictivity of disease

  4. 类风湿关节炎患者血纤维蛋白原免疫复合物的检测和意义%Significance of Detecting the Immune Complexes Contain Fibrinogen in Patients with Rheumatoid Arthritis

    Institute of Scientific and Technical Information of China (English)

    王艾丽; 吉滢; 郑田; 严孝岭; 刘国瑞; 贾煊

    2012-01-01

    Objective To explore the diagnostic value and significance of immune complexes contain fibrinogen(Fb-IC) in patients with rheumatoid arthritis (RA). Methods Fb-ICs were determined by ELISA using anti-human Fb antibody,the serum sample of 126 patients with RA,75 patients with non-RA diseases (lung cancer,systemic lupus erythematosus,undiffer-entiated connective tissue disease and kidney disease) and 34 healthy controls were involved in this study. The consistency of results was compared with that of Clq binding ELISA assay. The correlation of Fb-ICs to anti-cyclic citrullinated pepdide (CCP) and rheumatoid factor (RF) were analysed. Results The Fb-ICs positive rates of RA group,non-RA disease group and healthy control group were 53. 17% (67/126) ,10. 67% (8/75) and 5. 88% (2/34) respectively. The positive rate of RA group was significantly higher than other two groups (X2 = 36. 31 and 22. 16, P<0. 001). The sensitivity of Fb-ICs was 53. 17% with high specificity ( 90. 00%) in RA. The coincidence for both ELISA methods of anti- Fb and Clq was 75%,the results were related (Kappa values>0. 4). There was 55. 3% and 62. 2% Fb-ICs-positive patients with anti-CCP and RF-positive respectively. Conclusion Fb-ICs presents a reasonable sensitivity with high specificity to RA and it is valuable to RA diagnosis. More than 50% of anti-CCP antibodies and RF-positive RA patients existed Fb-ICs and it may be the pathogenic mechanism of RA and conduce to diagnostic value for RA.%目的 检测类风湿关节炎(RA)患者血清中含纤维蛋白原免疫复合物(Fb-IC),探讨Fb-IC在RA中的意义.方法 用抗Fb抗体建立ELISA法检测126例RA,75例其他疾病包括肺癌、系统性红斑狼疮(SLE)、未分化结缔组织病(UCTD)、肾脏疾病患者血清中Fb-ICs,另设健康对照34例.将该法结果与C1q结合法检测ICs的结果进行比较.分析RA患者Fb-IC与抗环瓜氨酸肽(CCP)抗体、类风湿因子( RF)的关系.结果 RA组、非RA疾病组和对

  5. THE FIRST RUSSIAN STRATEGIC STUDY OF PHARMACOTHERAPY FOR RHEUMATOID ARTHRITIS (REMARCA TRIAL: RESULTS OF 12-MONTH TREATMENT IN 130 PATIENTS

    Directory of Open Access Journals (Sweden)

    D. E. Karateev

    2014-01-01

    Full Text Available To introduce treat-to-target recommendations is an important task of modern rheumatology; however, there is still a diversity of serious problems relating to a scientific rationale and a clinical one for this strategy and to the possibilities of its implementation in real clinical practice, in the rheumatology service of the Russian Federation in particular, by taking into account the specific features of funding for high-tech medical care.Objective: to determine the efficiency and safety of combined therapy with subcutaneous methotrexate (MT and biological agents (BA when using the treat-to-target strategy in patients with active early and extended-stage rheumatoid arthritis (RA who have risk factors for a poor prognosis.Subjects and methods.The results of the REMARCA (Russian InvEstigation of MethotrexAte and biologicals in eaRly aCtive inflammatory Arthritis trial of 130 patients followed up for 12 months or more were given. There was a female preponderance; mean age 48.9±13.9 years, rheumatoid factor positivity (86.9%; anti-cyclic citrullinated peptide antibody positivity (89.2%. Seventy patients formed a subgroup of early RA (disease duration ≤6 months (mean 4.17±1.39 months; 60 patients were a subgroup of advanced-stage RA (disease duration >6 months (mean 30.8±32.7 months. In all the patients, therapy was initiated by using subcutaneous MT with its rapid dose escalation up to 20–30 mg/week and the achievement of the treatment goal (low disease activity or remission was checked every 3 months and depending on the result a decision had been taken to add or not to add a biological agent (BA (a tumor necrosis factor inhibitor or abatacept. If the former was insufficiently effective, it was substituted for a BA from another class.Results. Subcutaneous MT monotherapy provided remission or low disease activity in 49 (37.7% patients; a BA was given to 81 (62.3% patients. Following 6 and 12 months, low activity or remission

  6. Clinical Features and Risk Factors in Patients with Rheumatoid Arthritis Combined with Interstitial Lung Disease%类风湿关节炎合并肺间质病变的临床特点及相关因素

    Institute of Scientific and Technical Information of China (English)

    姜帆; 刘荣清

    2012-01-01

    目的 分析类风湿关节炎合并肺间质病变(RA-ILD)患者的临床特点及相关因素.方法 回顾性分析2006年9月-2011年5月我院住院及门诊病人93例RA-ILD及93例未合并ILD的类风湿关节炎(RA)患者的一般情况、临床特点及类风湿因子(RF)、血沉(ESR)、C反应蛋白(CRP)、抗核抗体(ANA)及抗角蛋白抗体(AKA)、抗环瓜氨酸肽(CCP)抗体、抗SSA/SSB抗体、红细胞计数(RBC)、血红蛋白(HGB)、血小板计数(PLT)进行比较.结果 RA-ILD组患者呼吸系统症状多数以咳嗽为首发症状.RA-ILD组患者年龄、RF滴度、吸烟构成比均显著高于无ILD的RA组,有统计学意义(P均<0.05).RA-ILD组病程、性别、ESR、CRP、ANA、抗SSA/SSB抗体,以及服用甲氨蝶呤与无ILD的RA组比较差异无统计学意义(P均>0.05).经过后退法建立Logistic回归分析模型,多因素分析显示年龄(OR=1.093,95%CI 1.053~1.136,P<0.05),年龄每增大1岁,危险性增加0.093倍.结论 年龄增大是RA发生ILD的相关因素,高龄的RA患者应警惕ILD的发生.%Objective To analyze the clinical features and risk factors of rheumatoid arthritis combined with interstitial lung disease (RA - ILD). Methods 93 cases with RA - ILDand 93 cases with RA without ILD from September,2006 to May,2011 were retrospectively analyzed. General,Clinical,rheumatoid factor (RF) , erythrocyte sedimentation rate (ESR) ,C -reactive protein (CRP) ,antinuclear antibody (ANA) ,anti -keratin antibody (AKA) ,anti -cyclic citrullinated peptide antibody (CCP) ,anti - SSA/anti -SSB,red blood cell (ESR) ,hemoglobin (HGB) and platelets (PLT) were compared between two groups. Results First symptoms of RA - ILD patients was cough. The age, RF level, prevalences of smoking in RA - ILD group were significantly higher than those in control group (P 0.05). The Logistic regression model showed that onset age were significantly associated with pulmonary disease ( OR = 1. 093 ,95% CI 1. 053 ~ 1. 136 , P < 0. 05

  7. Perfil dos pacientes com manifestações extra-articulares de artrite reumatoide de um serviço ambulatorial em Curitiba, Sul do Brasil Epidemiological profile of patients with extra-articular manifestations of rheumatoid arthritis from the city of Curitiba, South of Brazil

    Directory of Open Access Journals (Sweden)

    Mariana Costa Moura

    2012-10-01

    t and chi-square tests, and a significance level of 5% (α= 0.05. RESULTS: During the course of the disease, 120 patients (45.8% had ExRA. Pulmonary manifestation, rheumatoid nodules and Sjögren's syndrome were the most common manifestations found. Rheumatoid factor and anti-cyclic citrullinated peptide antibody were positive in most patients tested. Most patients were classified as Steinbrocker functional classes 1 and 2. The mean DAS-28 was 3.629, and the mean HAQ score, 1.12. Patients with ExRA had longer disease duration (P < 0.05, and current smoking habit associated with the presence of ExRA (P < 0.05. CONCLUSIONS: The prevalence of ExRA during disease course was 45.8%, and current smoking habit correlated with the presence of ExRA.

  8. 肿瘤标志物在类风湿性关节炎伴间质性肺病中的临床意义%Clinical value of tumor markers in rheumatoid arthritis with interstitial lung diseases

    Institute of Scientific and Technical Information of China (English)

    夏俊波; 严秀娟; 任振义; 陈灏; 王娇莉

    2012-01-01

    目的 探讨肿瘤标志物在类风湿性关节炎伴间质性肺病(RA-ILD)中的临床意义.方法 收集2008年1月至2011年1月住院的82例类风湿关节炎患者.根据是否伴有间质性肺病,将其分为RA-ILD组(38例)和RA组(44例),比较两组血清肿瘤标志物癌胚抗原(CEA)、糖蛋白抗原15-3(CA15-3)、糖蛋白抗原19-9(CA19-9)水平、抗环瓜氨酸肽(CCP)抗体水平和肺功能包括用力肺活量(FVC)、第1秒用力呼气量(FEV1)、一氧化碳弥散量(DL)占预计值百分比,分析两组患者肿瘤标志物表达差异以及RA-ILD组肿瘤标志物与肺功能、抗CCP抗体相关性.结果 RA-ILD组外周血CEA、CA15-3、CA19-9水平均明显高于RA组(均P<0.01);RA-ILD组FVC、FEV1和DLco占预计值百分比均明显低于RA组(均P<0.01),但RA-ILD组抗CCP抗体水平与RA组比较差异无统计学意义(P >0.05).RA-ILD组CEA水平与FVC、FEV1和DLco呈负相关(r=-0.499、-0.576、-0.512,均P<0.01),但CEA、CA15-3和CA19-9均与抗CCP抗体无相关性(r=-0.192、-0.264、-0.268,均P >0.05).结论 肿瘤标志物CEA、CA15-3和CA19-9在RA-ILD中表达升高,CEA可能反应RA-ILD肺间质受损的严重程度,而不是关节损害的严重程度.%Objective To assess the clinical significance of tumor markers (TMs) in rheumatoid arthritis (RA) with interstitial lung diseases (ILD). Methods Eighty-two RA inpatients were admitted in Hangzhou First People's Hospital from January 2008 to January 2011; among whom 38 were complicated by interstitial lung diseases (RA-ILD). Tumor markers (CEA, CA15-3, CA19-9), pulmonary function indexes (FVC, FEVi, DLco) and anti-cyclic citrullinated peptide antibody (anti-CCP) levels were measured in all patients. Correlation between TMs and pulmonary function indexes, level of anti-CCP antibody was analyzed by Spearman rank correlation. Results Serum levels of CEA, CA15-3 and CA19-9 in RA-ILD group were significantly higher than those in RA group (P0.05). Serum CEA level was

  9. Anticuerpo anticitrulina y manifestaciones extra articulares en artritis reumatoidea Anticitrulin antibody and the extra-articular manifestations in rheumatoid arthritis

    Directory of Open Access Journals (Sweden)

    María Jezabel Haye Salinas

    2013-02-01

    Full Text Available Los pacientes con artritis reumatidea (AR pueden desarrollar manifestaciones extra articulares (MExA, relacionadas a su morbi-mortalidad. Los anticuerpos anti-péptidos citrulinados cíclicos (ACCP son específicos para la AR y estan relacionados con el daño articular; y podrían tener rol patogénico en las MExA. Nuestro objetivo fue determinar la relación entre los anticuerpos ACCP y MExA en pacientes con AR. Se incluyeron 74 pacientes con diagnóstico de AR (ACR 1987 mayores de 18 años, de más de 6 meses de evolución, con MExA, y un control apareado por sexo y edad sin MExA por cada paciente. Las variables demográficas, clínicas y de laboratorio se compararon con test t, chi cuadrado o Mann-Whitney. Se realizó análisis multivariado; p ≤ 0.05. Los pacientes con MExA presentaron mayor título de anticuerpo ACCP (116 vs. 34, p A large proportion of rheumatoid arthritis (RA patients develop extra-articular manifestations (EAM, which are associated with morbidity and early mortality. Anti cyclic citrullinated peptide (ACCP antibody has proven to be highly specific for the diagnosis of RA, associated with severe joint damage and may have some role in the pathogenesis of EAM. The aim of this study was to determine the relationship between ACCP antibody and the presence of EAM in RA patients. Seventy four RA patients (ACR 1987 with EAM, > 18 years, more than 6 months duration were included, and an EAM free control, matched by sex and age, for each patient. Demographic, clinical and laboratory variables were compared using t-test, chi-square or Mann-Whitney test. Multivariate analysis was performed: p ≤ 0.05. Patients with EAM presented a greater value of ACCP antibody (116 vs. 34, p < 0.01 and rheumatoid factor (108 vs. 34.5, p < 0.01. Independent association with current smoking habit (p = 0.02, OR = 3.78, 95%: 1.17-12.2, RF positive (p = 0.04, OR 3.23, CI 95%: 1.04 to 11.8 and ACCP antibody positive (p = 0.04, OR 3.23, 95% CI: 1

  10. The application of quantitative MRI analysis in wrist bone erosion of rheumatoid arthritis%磁共振定量分析在类风湿关节炎手腕部骨侵蚀中的应用

    Institute of Scientific and Technical Information of China (English)

    王颖芳; 朱小春; 杜红卫; 吴红华; 蔡晓笑

    2012-01-01

    OBJECTIVE To analyse and evaluate the application of magnetic resonance quantitative analysis in wrist bone erosion of rheumatoid arthritis, to provide reference for future clinical applications. METHODS Randomly sampled 108 cases with wrist scans by MRI from January 2007 to October 2011, after 1 week, patients diagnosed with rheumatoid arthritis were as the research objects. All the research objects were grouped according to sex, age, course of disease, rheumatoid factor, anti-cyclic citrullinated peptide, C reactive protein, erythrocyte sedimentation rate and other indices, the used Mann-Whitney method to analyze the difference of bone erosion. RESULTS The results in this study showed 73 patients met the criteria as RA, accounted for about 67.59%; bone erosion scores in CRP positive/negative group, early/late period in RA group and ESR positive/negative groups were significantly different (P< 0.05); the bone erosion scores in CCP positive/negative groups were significantly different (P< 0.05). CONCLUSION MRI quantitative analysis provides an effective basis to diagnosis and prognosis of wrist bone erosion of rheumatoid arthritis, and could be widely used in clinical work in the future.%目的 对磁共振定量分析在类风湿关节炎手腕部骨侵蚀中的应用进行分析与评价,为今后的临床应用提供参考.方法 随机抽取在2007年1月~2011年10月这段时间里采用MRI对手腕部进行扫描的临床患者病例108例,在1周之后均进行了实验室检查,将确诊为类风湿关节炎(RA)的患者作为本次研究的研究对象.将所有研究对象按照性别、年龄、病程、类风湿因子、抗环瓜氨酸肽、C反应蛋白、血沉等观察指标进行分组,采用Mann-Whimey方法对各组间骨侵蚀的差异进行分析.结果 在本次研究中所接受检查的108名患者中有73例符合RA标准,约占67.59%;对于CRp阳/阴性组、早期/中晚期RA组以及ESR阳/阴性组间的骨侵蚀评分之间存

  11. THE FIRST RUSSIAN STRATEGIC STUDY OF PHARMACOTHERAPY FOR RHEUMATOID ARTHRITIS (REMARCA TRIAL: RESULTS OF 12-MONTH TREATMENT IN 130 PATIENTS

    Directory of Open Access Journals (Sweden)

    D. E. Karateev

    2015-01-01

    Full Text Available To introduce treat-to-target recommendations is an important task of modern rheumatology; however, there is still a diversity of serious problems relating to a scientific rationale and a clinical one for this strategy and to the possibilities of its implementation in real clinical practice, in the rheumatology service of the Russian Federation in particular, by taking into account the specific features of funding for high-tech medical care. Objective: to determine the efficiency and safety of combined therapy with subcutaneous methotrexate (MT and biological agents (BA when using the treat-to-target strategy in patients with active early and extended-stage rheumatoid arthritis (RA who have risk factors for a poor prognosis. Subjects and methods. The results of the REMARCA (Russian InvEstigation of MethotrexAte and biologicals in eaRly aCtive inflammatory Arthritis trial of 130 patients followed up for 12 months or more were given. There was a female preponderance; mean age 48.9±13.9 years, rheumatoid factor positivity (86.9%; anti-cyclic citrullinated peptide antibody positivity (89.2%. Seventy patients formed a subgroup of early RA (disease duration ≤6 months (mean 4.17±1.39 months; 60 patients were a subgroup of advanced-stage RA (disease duration >6 months (mean 30.8±32.7 months. In all the patients, therapy was initiated by using subcutaneous MT with its rapid dose escalation up to 20–30 mg/week and the achievement of the treatment goal (low disease activity or remission was checked every 3 months and depending on the result a decision had been taken to add or not to add a biological agent (BA (a tumor necrosis factor inhibitor or abatacept. If the former was insufficiently effective, it was substituted for a BA from another class. Results. Subcutaneous MT monotherapy provided remission or low disease activity in 49 (37.7% patients; a BA was given to 81 (62.3% patients. Following 6 and 12 months, low activity or remission according

  12. 异基因脐带间充质干细胞移植治疗类风湿关节炎疗效及安全性%Allogenic Derived Mesenchymal Stem Cells Transplantation for 4 Patients with Rheumatoid Arthritis

    Institute of Scientific and Technical Information of China (English)

    王秦; 李小峰; 马丽辉; 李军霞; 陈俊伟; 李芳; 薛丽巾; 白洁

    2011-01-01

    目的:探讨异基因脐带间充质干细胞(uhcMSC)移植治疗难治性类风湿关节炎(RA)的疗效及安全性.方法:从足月顺产供者脐带中分离培养MSC,给予3例RA患者行MSCs移植治疗,移植细胞数(1 x10(6)/kg体重).评价患者移植前后的临床表现和实验室检查指标的改变.结果:异基因MSC移植后,3例RA患者随访9-11个月,所有患者均无移植相关并发症.移植后1月疾病活动评分(DAS-28)由5.13降低为2.16(n=3),1例血液系统受累患者血细胞(中性粒细胞、红细胞、血小板)均上升.移植前后比较抗环瓜氨酸(CCP)抗体滴度降低.1例患者移植前后比较Th17细胞明显升高,Treg细胞降低.结论:异基因脐带MSC移植治疗难治性RA有效、安全.uhcMSC取材方便,扩增迅速,输注安全经济.%Objective To explore the clinical efficacy and safety of allogenic derived mesenchymal stem cells transplantation (MSCT) in patients with refractory rheumatoid arthritis (RA). Methods Three patients with refractory RA ( all were female patients) aged 43 - 47 were enrolled in the study and the Ethics Committee of the Affiliated The Shanxi Medical University approved informed consent was obtained for each patient The umbilical cord of healthy donors were aspirated and the mesenchymal stem cells ( MSC) were expanded in vitro. MSC 1 X 106/kg body weight was infused intra-venously for each patient Before MSCT, all patients were administrated with cyclophosphamide ( CTX) 400 - 600 mg. The clinical manifestations and laboratory tests were compared before and after MSCT. Results The three patients were followed up for nine to eleven months after MSCT. No patient had developed transplantation related complications. The rheumatoid arthritis disease activity scord ( DAS-28 ) score decreased from 5. 13 to 2. 16 one month after MSCT (n =3). Erythrocyte sedimentation rate (ESR) decreased significantly one month after MSCT (n =3). Three patients were followed up for nine months, anti-cyclic

  13. Analysis of change in serum anti-CCP antibody level and its diagnostic value in patients of early rheumatoid arthritis%早期类风湿关节炎患者血清抗CCP抗体水平的变化及其诊断价值

    Institute of Scientific and Technical Information of China (English)

    王蓉辉; 马丽辉

    2015-01-01

    Objective To investigate the change in serum anti-cyclic citrullinated peptide (anti-CCP) anti-body level and its diagnostic value in patients of early rheumatoid arthritis (RA). Methods One hundred and nine patients with early rheumatoid arthritis (RA group) in our hospital from June 2012 to May 2014, 92 patients without rheumatoid arthritis (non-RA group) and 80 healthy subjects (healthy group) were enrolled in the study. Enzyme-linked immunosorbent assay (ELISA) was used to test the anti-CCP antibody level in the three groups. Results Serum anti-CCP antibody level of RA group was (21.27 ± 10.32) IU/ml, significantly higher than that of non-RA group and the healthy group, and the differences were statistically significant (P0.05). The sensitivity of anti-CCP antibody diagnosis was 66.97%, and specificity was 84.30%, with the positive predic-tive value (+PV) of 73%, the negative predictive value (-PV) of 80.11%, and Youden index of 0.513. The receiver operating characteristic (ROC) curve for anti-CCP antibody in the diagnosis of RA was produced, with AUC=0.886, 95%confidence interval of (0.837~0.934), and the best bound value of 17.5 IU/ml. Conclusion Anti-CCP antibody levels are higher in RA patients, and anti-CCP antibody detection has a higher value in the early diagnosis of RA.%目的:探讨抗环瓜氨酸抗体(抗CCP抗体)在早期类风湿关节炎(RA)患者中的水平变化及其在诊断中的应用价值。方法选取2012年6月至2014年5月在我院就诊的早期RA患者109例(RA组),同时选取其他风湿病患者92例(非RA组)和健康体检者80例(健康组),采用酶联免疫吸附试验(ELISA)法检测三组受试者的抗CCP抗体水平。结果 RA组血清抗CCP抗体水平为(21.27±10.32) IU/ml,明显高于非RA组和健康组,差异均有统计学意义(P0.05);抗CCP抗体诊断的灵敏度为66.97%,特异度为84.30%,阳性预测值为73.00%,阴性预测值为80.11%,约登指数0.513;抗CCP

  14. 抗CCP、RF、CRP、ASO对类风湿关节炎的诊断意义%The significance of serum anti - CCP antibody, RF, CRP, ASO in the diagnosis of rheumatoid arthritis.

    Institute of Scientific and Technical Information of China (English)

    刘发河; 曾海莲; 郑小江; 杨春媚

    2011-01-01

    Objective To investigate the significance of serum anti - cyclic citrullinated peptide ( CCP ) antibody, C - reactive protein ( CRP ), antistreptolysin O ( ASO ) and rheumatoid factor ( RF ) in the diagnosis of rheumatoid arthritis ( RA ).Methods CCP was detected in serum samples from 115 patients with RA by ELISA and compared with serum samples from 131 cases of undifferentiated inflammatory arthritis ( UIA ) patients.CRP, ASO and RF were detected by immunoturbidimetry method.Results In RA , the positive rates of anti - CCP, CRP and RF were higher than UIA.The sensitivities of anti - CCP, CRP and RF for RA were 65.2% , 70.0% and 60.9% , respectively.The specificities of anti - CCP, RF and CRP for RA were 93.9%, 80.3% and 58.0%, respectively.Statistical test indicated that there were no significant differences in sensitivities of anti - CCP, CRP and RF, but the specificity of CCP was higher than those of RF and CRP for the diagnosis of RA.The positive rate of ASO in RA was only 12.5% and no significant difference in the positive rate of ASO between RA and UIA was found.Conclusion The anti - CCP assay has high sensitivity and specificity for the diagnosis of RA and the combined detection of anti - CCP, CRP and RF may further improve the sensitivity.ASO has no significance for the diagnosis of RA, but it can be an indication of streptococcal infection in RA patients.%目的 探讨抗环瓜氨酸肽(CCP)抗体、类风湿因子(RF)、C反应蛋白(CRP)、抗链球菌溶血素"O"(ASO)对类风湿关节炎(RA)的诊断意义.方法 收集115例RA患者和131例非RA的自身免疫病患者的血清,用酶联免疫吸附试验检测抗-CCP抗体,免疫比浊法检测RF、CRP、ASO.结果 抗CCP、RF、CRP的阳性率在RA组显著高于非RA组,抗-CCP对RA的敏感性为65.2%,CRP对RA的敏感性70.0%,RF的敏感性较抗-CCP和CRP稍低,为60.9%.特异性以抗CCP为最高93.9%,RF对RA的特异性为80.3%,CRP较低为58.0%,经统计学检验,抗CCP的灵

  15. CARDIOVASCULAR RISK IN PATIENTS WITH EARLY RHEUMATOID ARTHRITIS BEFORE DISEASE-MODIFYING ANTIRHEUMATIC THERAPY (PRELIMINARY DATA OF THE REMARCА STUDY

    Directory of Open Access Journals (Sweden)

    Yu. N. Gorbunova

    2014-01-01

    peptide-positive patients with early RA and high disease activity have high and very high cardiovascular risks.

  16. Grr1p is required for transcriptional induction of amino acid permease genes and proper transcriptional regulation of genes in carbon metabolism of Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    Eckert-Boulet, Nadine; Regenberg, Birgitte; Nielsen, Jens

    2005-01-01

    and a grr1 Delta strain and adding citrulline in the exponential phase. Whole-genome transcription analyses were performed on samples from each cultivation, both immediately before and 30 min after citrulline addition. Transcriptional induction of the AAP genes AGP1, BAP2, BAP3, DIP5, GNP1 and TAT1 is fully...

  17. Watermelon, phytochemicals and health

    Science.gov (United States)

    Watermelon fruit contains lycopene, a carotenoid pigment, and citrulline, an amino acid. These plant compounds may be helpful in preventing some chronic diseases. The amount of lycopene in watermelon ranges from 35 to 125 mg per kg of edible portion, and there is 2 to 4 mg per kg citrulline presen...

  18. Peptidylarginine deiminase activity in postmortem white matter of patients with multiple sclerosis

    NARCIS (Netherlands)

    De Keyser, J; Schaaf, M; Teelken, A

    1999-01-01

    The myelin sheath in multiple sclerosis (MS) appears to contain a higher proportion of the citrullinated isoform of myelin basic protein MBP-C8. In vitro, MBP-associated arginine is deiminated to citrulline by the enzyme peptidylarginine deiminase (PAD). We investigated PAD activity in white matter

  19. Autoanticorpos na artrite reumatoide inicial: coorte Brasília - resultados de uma análise seriada de três anos Autoantibodies in early rheumatoid arthritis: Brasília cohort - results of a three-year serial analysis

    Directory of Open Access Journals (Sweden)

    Licia Maria Henrique da Mota

    2011-12-01

    Full Text Available O valor diagnóstico e prognóstico da análise seriada dos anticorpos como fator reumatoide (FR, anticorpos antipeptídeos citrulinados cíclicos (anti-CCP e antivimentina citrulinada (anti-Sa não está definido nos pacientes com artrite reumatoide inicial (ERA. OBJETIVOS: Avaliar de forma prospectiva a presença de FR, anti-CCP e anti-Sa em pacientes com ERA. PACIENTES E MÉTODOS: Quarenta pacientes da coorte Brasília de ERA (menos de 12 meses foram avaliados e monitorados durante três anos. Os dados clínicos e demográficos foram registrados, além dos resultados (ELISA para FR (IgM, IgG e IgA, anti-CCP (CCP2, CCP3 e CCP3.1 e anti-Sa na avaliação inicial e aos 3, 6, 12, 18, 24 e 36 meses de acompanhamento. Comparações pelos testes t de Student e t pareado. RESULTADOS: A idade média foi de 45 anos, 90% dos pacientes do gênero feminino. No momento do diagnóstico, FR foi observado em 50% dos casos (FR IgA 42%, FR IgG 30% e FR IgM 50%, anti-CCP em 52,5% (não houve diferença entre CCP2, CCP3 e CCP3.1 e anti-Sa em 10%. Após três anos, não houve diferença na prevalência de FR e anti-CCP, mas a de anti-Sa aumentou para 17,5% (P = 0,001. CONCLUSÃO: A análise repetida do FR e anti-CCP, incluindo aqui diferentes isotipos, durante três anos de acompanhamento, não mostrou mudanças significativas. A terceira geração do anti-CCP não aumentou o valor diagnóstico dos testes de segunda geração.The diagnostic and prognostic value of the serial measurement of antibodies, such as rheumatoid factor (RF, anti-cyclic citrullinated peptide (anti-CCP, and anti-citrullinated vimentin (anti-Sa antibodies, has not been defined in early rheumatoid arthritis (ERA. OBJECTIVES: To prospectively assess the presence of RF, anti-CCP, and anti-Sa in ERA patients. PATIENTS AND METHODS: Forty ERA (less than 12 months patients of the Brasília cohort were evaluated and followed up for three years. Both clinical and demographic data were recorded, in

  20. Savings in Public Services after the Crisis: A Multilevel Analysis of Public Preferences in the EU27 (Working Paper 8)

    NARCIS (Netherlands)

    S.G.J. Van de Walle (Steven); S.R. Jilke (Sebastian)

    2012-01-01

    textabstractPolicy responses to the financial crisis can be divided into cyclical and anti-cyclical approaches. The former advocates reducing public spending in times of financial constraints. The latter approach advocates public spending to boost the economy. Using multinomial multilevel analysis o

  1. Raw bovine milk improves gut responses to feeding relative to infant formula in preterm piglets

    DEFF Research Database (Denmark)

    Li, Yang; Lykke, Mikkel; Chatterton, D E W;

    2014-01-01

    height) and function (increased nutrient absorption and enzyme activities, decreased gut permeability, nutrient fermentation, and NEC severity). BC further improved these effects relative to BM (lactase activity, lactose absorption, plasma citrulline, and tissue interleukin-8). WMP induced similar...

  2. Inhibition of insulin fibrillation by osmolytes: Mechanistic Insights

    OpenAIRE

    Sinjan Choudhary; Nand Kishore; Hosur, Ramakrishna V.

    2015-01-01

    We have studied here using a number of biophysical tools the effects of osmolytes, betaine, citrulline, proline and sorbitol which differ significantly in terms of their physical characteristics such as, charge distribution, polarity, H-bonding abilities etc, on the fibrillation of insulin. Among these, betaine, citrulline, and proline are very effective in decreasing the extent of fibrillation. Proline also causes a substantial delay in the onset of fibrillation in the concentration range (5...

  3. Enteral Glutamine Administration in Critically Ill Nonseptic Patients Does Not Trigger Arginine Synthesis

    Directory of Open Access Journals (Sweden)

    Mechteld A. R. Vermeulen

    2016-01-01

    Full Text Available Glutamine supplementation in specific groups of critically ill patients results in favourable clinical outcome. Enhancement of citrulline and arginine synthesis by glutamine could serve as a potential mechanism. However, while receiving optimal enteral nutrition, uptake and enteral metabolism of glutamine in critically ill patients remain unknown. Therefore we investigated the effect of a therapeutically relevant dose of L-glutamine on synthesis of L-citrulline and subsequent L-arginine in this group. Ten versus ten critically ill patients receiving full enteral nutrition, or isocaloric isonitrogenous enteral nutrition including 0.5 g/kg L-alanyl-L-glutamine, were studied using stable isotopes. A cross-over design using intravenous and enteral tracers enabled splanchnic extraction (SE calculations. Endogenous rate of appearance and SE of glutamine citrulline and arginine was not different (SE controls versus alanyl-glutamine: glutamine 48 and 48%, citrulline 33 versus 45%, and arginine 45 versus 42%. Turnover from glutamine to citrulline and arginine was not higher in glutamine-administered patients. In critically ill nonseptic patients receiving adequate nutrition and a relevant dose of glutamine there was no extra citrulline or arginine synthesis and glutamine SE was not increased. This suggests that for arginine synthesis enhancement there is no need for an additional dose of glutamine when this population is adequately fed. This trial is registered with NTR2285.

  4. Enteral Glutamine Administration in Critically Ill Nonseptic Patients Does Not Trigger Arginine Synthesis.

    Science.gov (United States)

    Vermeulen, Mechteld A R; Brinkmann, Saskia J H; Buijs, Nikki; Beishuizen, Albertus; Bet, Pierre M; Houdijk, Alexander P J; van Goudoever, Johannes B; van Leeuwen, Paul A M

    2016-01-01

    Glutamine supplementation in specific groups of critically ill patients results in favourable clinical outcome. Enhancement of citrulline and arginine synthesis by glutamine could serve as a potential mechanism. However, while receiving optimal enteral nutrition, uptake and enteral metabolism of glutamine in critically ill patients remain unknown. Therefore we investigated the effect of a therapeutically relevant dose of L-glutamine on synthesis of L-citrulline and subsequent L-arginine in this group. Ten versus ten critically ill patients receiving full enteral nutrition, or isocaloric isonitrogenous enteral nutrition including 0.5 g/kg L-alanyl-L-glutamine, were studied using stable isotopes. A cross-over design using intravenous and enteral tracers enabled splanchnic extraction (SE) calculations. Endogenous rate of appearance and SE of glutamine citrulline and arginine was not different (SE controls versus alanyl-glutamine: glutamine 48 and 48%, citrulline 33 versus 45%, and arginine 45 versus 42%). Turnover from glutamine to citrulline and arginine was not higher in glutamine-administered patients. In critically ill nonseptic patients receiving adequate nutrition and a relevant dose of glutamine there was no extra citrulline or arginine synthesis and glutamine SE was not increased. This suggests that for arginine synthesis enhancement there is no need for an additional dose of glutamine when this population is adequately fed. This trial is registered with NTR2285.

  5. Enteral Glutamine Administration in Critically Ill Nonseptic Patients Does Not Trigger Arginine Synthesis

    Science.gov (United States)

    Vermeulen, Mechteld A. R.; Brinkmann, Saskia J. H.; Buijs, Nikki; Beishuizen, Albertus; Bet, Pierre M.; Houdijk, Alexander P. J.; van Goudoever, Johannes B.; van Leeuwen, Paul A. M.

    2016-01-01

    Glutamine supplementation in specific groups of critically ill patients results in favourable clinical outcome. Enhancement of citrulline and arginine synthesis by glutamine could serve as a potential mechanism. However, while receiving optimal enteral nutrition, uptake and enteral metabolism of glutamine in critically ill patients remain unknown. Therefore we investigated the effect of a therapeutically relevant dose of L-glutamine on synthesis of L-citrulline and subsequent L-arginine in this group. Ten versus ten critically ill patients receiving full enteral nutrition, or isocaloric isonitrogenous enteral nutrition including 0.5 g/kg L-alanyl-L-glutamine, were studied using stable isotopes. A cross-over design using intravenous and enteral tracers enabled splanchnic extraction (SE) calculations. Endogenous rate of appearance and SE of glutamine citrulline and arginine was not different (SE controls versus alanyl-glutamine: glutamine 48 and 48%, citrulline 33 versus 45%, and arginine 45 versus 42%). Turnover from glutamine to citrulline and arginine was not higher in glutamine-administered patients. In critically ill nonseptic patients receiving adequate nutrition and a relevant dose of glutamine there was no extra citrulline or arginine synthesis and glutamine SE was not increased. This suggests that for arginine synthesis enhancement there is no need for an additional dose of glutamine when this population is adequately fed. This trial is registered with NTR2285. PMID:27200186

  6. INVESTIGATION OF CANDIDATE GENE POLYMORPHISMS IN AN IMMUNE RESPONSE AS MARKERS FOR THE RISK OF DEVELOPING RHEUMATOID ARTHRITIS AND PRODUCING AUTOANTIBODIES

    Directory of Open Access Journals (Sweden)

    I. A. Guseva

    2016-01-01

    Full Text Available Objective: to investigate the distribution of the genotypes and alleles of the PTPN22, TNFAIP3, CTLA4, TNFA, IL6, IL6R, IL10, MCP1, and ICAM1 genes in patients with rheumatoid arthritis (RA and in the control group of healthy individuals, to estimate their significance as molecular genetic markers for predisposition to RA; and to analyze the correlation between the gene polymorphisms included in the study and the production of anti-cyclic citrullinated peptide antibodies (ACCPA and IgM rheumatoid factor (RF.Subjects and methods. The investigation was conducted within the framework of the «Early arthritis: Diagnosis, outcome, criteria, active treatment program». The prospective follow-up study included 122 patients with RA fulfilling the 1987 American College of Rheumatology (ACR criteria; with disease duration of ≤ 2 years. 73 (59.8% patients were included during the first 6 months after the onset of the disease. 74 (60.7% and 81 (66.5% patients were found to be positive for ACCPA and IgM RF, respectively. 314 healthy blood donors served as a control group. A real-time polymerase chain reaction was used in the patients and control individuals to study the distribution of the polymorphic variants of PTPN22 (+1858 C >T, rs2476601, TNFAIP3 (rs675520, rs6920220, rs10499194, CTLA4 (+49A>G, rs231775 , TNFА (-308A>G, rs1800629, IL6 (-174G>C, rs1800795, IL6R (+358A>C, rs8192284, IL10 (-592A>C, rs1800872, -1082 A>G, rs1800896, MCP1/CCL2 (+2518A>G, rs1024611, and ICAM1 (721G>A, rs1799969 genes. Results and discussion. This analysis revealed an association of PTPN22 (+1858 C >T, rs2476601 and TNFAIP3 (rs675520, rs10499194 polymorphisms with the risk of RA (odds ratio (OR, 1.5; 95% confidence interval (CI, 1.0–2.3; p = 0.05; OR, 1.5; 95% CI, 1.1–2.0; p = 0.02; OR, 0.5; 95% CI, 0.4–0.8; p = 0.01, respectively. Further, there was a tendency towards a positive association of TNFAIP3 (rs6920220 and IL6R (rs8192284 polymorphisms with a predisposition

  7. 类风湿关节炎患者抑郁症发生的相关因素分析%Analysis on depression and the related factors in patients with rheumatoid arthritis

    Institute of Scientific and Technical Information of China (English)

    陈晓; 杨敏; 王天; 付睿; 曾小峰

    2013-01-01

    as having depression in this study.There were no significant differences in gender,age,marital status,rheumatoid factor (RF),anti-cyclic citrullinated peptide (Anti-CCP)positive,using of DMARDs and Prednisone dose between the depression and the non-depression group (P > 0.05).There were statistically significant differences in working conditions,cultural degree,annual household income,DAS28,HAQ score between the two groups (P < 0.05).Logistic regression analysis showed that annual household income,disease duration,DAS28,HAQ score were risk factors of depression.Conclusions Depression were frequently seen in RA patients,and were related to high disease activity,more disability and low income.Identifying associated predictors of depression in patients with RA can help diagnosis of depressive symptoms earlier as well as guide targeting treatment.

  8. 抗鼠科肉瘤病毒癌基因同源物B1抗体的检测及其在类风湿关节炎诊断中的意义%Significance of the detection of anti-v-raf murine sarcoma viral oncogene homologue B1 antibodies in the diagnostic practice of rheumatoid arthritis

    Institute of Scientific and Technical Information of China (English)

    张霞; 赵金霞; 孙琳; 李茹; 刘湘源; 栗占国

    2012-01-01

    Objective: To clarify the clinical significance of the antibody against v-raf murine sarcoma viral oncogene homologue Bl ( BRAF) in the diagnostic practice of rheumatoid arthritis (RA). Methods: In the study, 112 patients with RA, 112 patients with other rheumatic diseases,and 73 healthy individuals were recruited . With recombinant human BRAF protein as antigen, we examined the level of anti-BRAF antibody in all the patients by enzyme-linked immunosorbent assay ( ELISA) , The clinical data of the RA patients were collected simultaneously, and analysed statistically by using SPSS 13. 0. Results: The positive rate of anti-BRAF antibody was 53. 6% in the RA patients, which was significantly higher than that of the normal control group(4. 1% ,P <0. 01 ) and other rheumatic diseases groups (P all <0. 01) except osteoarthritis group. The titer of anti-BRAF antibody was also notably higher in the patients with RA than in other rheumatic diseases and normal control groups(P all <0. 01). The diagnostic sensitivity and specificity of anti-BRAF antibody for RA were 53. 6% and 84. 3% respectively. The positive rate of anti-BRAF antibody in rheumatoid factor, anti-cyclic citrullinated peptide antibody, antikera-tin antibody,antiperinuclear factor negative groups were 52. 6% ,38. 2% , 30. 3% and 31.0% respectively. It showed significant negative correlation between the titer of anti-BRAF antibody and patient' s age, disease duration and the level of CRP. Conclusion: The anti-BRAF antibody contributes to the diagnosis of RA, and may act as a supplement of other autoantibodies.%目的:了解抗鼠科肉瘤病毒癌基因同源物B1( v-raf murine sarcoma viral oncogene homologue B1,BRAF)抗体在类风湿关节炎( rheumatoid arthritis,RA)诊断中的临床意义.方法:纳入RA患者112例、健康对照73例、其他风湿病患者112例,以重组人BRAF蛋白为抗原,采用酶联免疫吸附法(enzyme-linked immunosorbent assays,ELISA)检测患者血清中的抗BRAF抗体水

  9. Consenso da Sociedade Brasileira de Reumatologia 2011 para o diagnóstico e avaliação inicial da artrite reumatoide 2011 Consensus of the Brazilian Society of Rheumatology for diagnosis and early assessment of rheumatoid arthritis

    Directory of Open Access Journals (Sweden)

    Licia Maria Henrique da Mota

    2011-06-01

    of RA Committee members of the Brazilian Society of Rheumatology. RESULTS AND CONCLUSIONS: The following ten reccommendations were established: 1 RA diagnosis should be established considering clinical findings and complementary test results; 2 Special attention should be given to the differential diagnosis of arthritis; 3 Rheumatoid factor (RF is an important diagnostic test, but has limited sensitivity and specificity, mainly in early RA; 4 Anti-CCP (anti-cyclic citrullinated peptide antibody is a marker with sensitivity similar to that of the RF, but with higher specificity, mainly in the initial phase of disease; 5 Although unspecific, acute-phase reactants should be measured in patients with clinical suspicion of RA; 6 Conventional radiography should be performed for diagnostic and prognostic assessment of the disease. When necessary and available, ultrasound and magnetic resonance may be used; 7 Rheumatoid arthritis classification criteria (ACR/EULAR 2010, although not yet validated, may be used as a guide to aid in diagnosing patients with early RA; 8 One of the combined disease activity indices should be used to assess disease activity; 9 At least one of the functional capacity assessment instruments, such as mHAQ or HAQ-DI, should be regularly used; 10 At the early assessment of the disease, the presence of worse prognostic factors, such as polyarticular involvement, high titers of RF and/or anti-CCP, and early joint erosion, should be investigated.

  10. 5例抗丙氨酰 tRNA 合成酶抗体阳性患者临床特征%Clinical Characters of Anti-alanyl-tRNA Synthetase Antibody

    Institute of Scientific and Technical Information of China (English)

    吴庆军; 张文; 李永哲; 田新平; 张; 赵岩; 曾小峰; 张奉春; 唐福林

    2014-01-01

    Objective The purpose of this study was to examine the clinical profile of patients with anti-alanyl-tRNA synthetase (anti-PL-1 2)antibody.Methods Five patients with anti-PL-1 2 antibody admitted into Peking Union Medical College Hospital were presented.The major clinical,serological,and radiographic data were analyzed.Results All 5 patients with anti-PL-1 2 antibody had evidence of interstitial lung disease (ILD)by clinical examination,imaging and pulmonary function tests.Clinical features presented were dry cough,gradual dyspnea,and crackles at lung bases.Lung function test showed a restrictive pulmonary dysfunction pattern and impaired gas exchange.High-resolution computed tomography scan revealed (predominantly basilar)reticular and ground-glass opacities,accompanied by a loss of lung volume,traction bronchiectasis,and scattered (usually peribronchovascular)areas of consolidation.This distinct pattern was highly suggestive of antisynthetase syndrome-related ILD.Four of 5 anti-PL-12 positive patients had underlying connective tissue disease,including dermatomyositis (2 cases),Sjoegren’s syndrome/rheumatoid arthritis (1 case),and systemic sclerosis (1 case).Only one of five anti-PL-12 positive patients demonstrated clinical myositis,including symmetrical proximal muscle weakness,increased serum creatine phosphokinase,myogenic damage on electromyography,and positive results on biopsy.Two patients had typical rash of dermatomyositis (DM).Mechanic hands occurred in 2 patients,fever in 1 patient,Raynaud’ s phenomenon in 1 patient,and inflammatory polyarthritis with anti-cyclic citrullinated polypeptide antibody in 1 patient.Four patients demonstrated a cytoplasmic pattern when tested for antinuclear antibodies by immunofluorescent assay.Four patients had positive anti-Ro-52 antibodies.Four of 5 patients received high-dose glucocorticosteroid (prednisone 0.8 ~1.5 mg·kg -1 ·d -1 )plus cyclophosphamide (100 mg·d -1 ) therapy.Response was favorable with recovery

  11. Systemic Sclerosis Combined with Rheumatoid Arthritis:Report of Three Cases and Literature Review%系统性硬化症合并类风湿关节炎三例并文献复习

    Institute of Scientific and Technical Information of China (English)

    鲁芙爱; 王慧; 王永福

    2014-01-01

    Objective Toexploretheclinicalfeaturesandlaboratorytestsofpatientswithsystemicsclerosis(SSc) combinedwithrheumatoidarthritis(RA),toimprovetheunderstandingofoverlapsyndrome(OS).Methods Theclinical manifestations,laboratory results and X-ray examinations of the hands of 3 patients with SSc combined with RA were analyzed andliteraturesbothdomesticandabroadreviewed.Results Allpatientswerefemale,startingwithSSc,withtypicalSScclini-cal manifestations,including Raynaud′s phenomenon,mask -like face,limitation of mouth opening,skin sclerosis of finger tips,both-hand contracture deformity,etc. and combined with interstitial lung disease,hypertensive pulmonary vascular dis-ease. None of patients had obvious arthritis manifestations but all or part of rheumatoid factors( RF) were positive;patient 1 had positive anti-perinuclear factor( APF),patient 2 had positive anti-cyclic citrullinated peptide( CCP)antibody,patient 3 had positive APF,anti-CCP antibody,anti-keratin antibody( AKA) and anti-RA33 antibody. The patients′hand X-ray suggested that there were different degrees of narrow or even disappeared joint space in metacarpophalangeal joints or carpal joints,part sclerotin being cystic-degenerated and destructed. Three patients were discharged after treatment of glucocorticoids andcyclophosphamide.Conclusion InOS,SSccombinedwithRAisrare.Patientsmayhavenotypicalarthritismanifesta-tions when SSc combined with RA. Screening RF antibody,anti-CCP antibody,AKA,APF,anti-RA33 antibody and hand X-ray help its diagnoses,to carry out standard treatment and improve patients′outcomes.%目的:探讨系统性硬化症( SSc)合并类风湿关节炎( RA)患者的临床特征和实验室检查特点,提高临床医生对重叠综合征的认识。方法对我科近年收治的3例SSc合并RA患者的临床表现、实验室检查结果及双手X线检查结果进行分析,并结合国内外文献进行复习。结果3例患者均为女性,以SSc起病,有典型的SSc

  12. Risk Factors, Clinical and Radiographic Characteristics of Rheumatoid Arthritis Associated Interstitial Lung Disease%类风湿关节炎相关肺间质病变的危险因素、临床和影像学特点

    Institute of Scientific and Technical Information of China (English)

    赵颖; 李菁; 吴庆军; 唐福林

    2012-01-01

    -citrullinaLed peptide/protein antihodies ( ACPA ) ( including anti-cyclic citrullinated peptide, anti-perinuclear factor, and anti-keratin antihody ) ( all P < 0. 05 ). RA-ILD manifesLed as cough, progressive exertional dyspnea, and cracles at both lung fields clinically. The findings on high-resolution computed Lomography ( 11RCT ) wore fiberoLic lines/reLiculaLion ( 68.75%, 33/48 ), exudalivo infikration/ consolidation ( 21/48, 43.75% ), ground glass opackies ( 15/48, 31.25% ), honeycombing ( 11/48, 22.92% ), and ploural effusion ( 5/48, 10.42% ). Pulmonary funcLion tests ( PFT ) were done in 37 pationts with RA-ILD and 75. 68% of these pationLs were abnormal. DLCO( diffusion oapaoity for carbon monoxide of the lung) was decreased in all 28 pationts, complicated with restrictive vontilation impairment in 7 paLionLs, and 4 pationLs vviLh mixed vonLilaLion disLurbance. Conclusions RA-ILD is an important feature of RA. IL is relaLed vviLh age, course of disease, smoking, serum level of rheumaLoid facLor, positivo ACPA, and disease severiLy. RA-ILD is noL relaLed Lo disease acLiviLy. IIRCT and PFT are valuable examinations for oarly diagnosis and appropriaLed Lhorapy of RA-ILD.

  13. 类风湿关节炎并发肺间质纤维化的相关因素研究%Related Factors of Rheumatoid Arthritis Complicated with Interstitial Pulmonary Fibrosis

    Institute of Scientific and Technical Information of China (English)

    李宗英; 王俊祥

    2013-01-01

    groups. Results Among these 345 HA patients, IPF was identified in 60 patients ( 17. 39% ). Compared with the non - IPF group, the RA - IPF group had significantly more males and smokers; Were significantly older; Had significantly higher levels of rheumatoid factor ( HF ), anti - cyclic citrullinated antibody ( anti - CCP ), and C - reactive protein ( CHP ) ( all P 0.05 ). Comparison of the OR values showed that male gender, smoking history, bone destruction, older age ( ≥60 yrs ), onset age ( ≥50 yrs ), long disease duration (≥5 yrs ), high CHP level ( ≥ 24 mg/L ), high RF titer ( ≥60 U/ml ) and high anti -CCP antibody level (≥75 RU/ml) were independent risk factors of RA - IPF. Conclusion IPF is a common complication of RA, with male gender, old age, smoking, high levels of CRP and high titer of RF or anti - CCP being its independent risk factors. HHCT provides a useful tool for early diagnosis and treatment.

  14. Evaluation of MRI in the diagnosis of rheumatoid arthritis%磁共振成像对类风湿关节炎诊断价值的评估

    Institute of Scientific and Technical Information of China (English)

    宋丁; 刘波; 李萍; 毕黎琦

    2010-01-01

    目的 探讨类风湿关节炎(RA)手关节的磁共振成像(MRI)表现及与临床表现的相关性,从而评价其在RA诊断中的价值.方法 采用对比及相关性分析的方法,分析符合2009年美国风湿病学会/欧洲抗风湿病联盟(ACR/EULAR)诊断标准的25例RA患者双手X线片、MRI及临床特点.结果 25例RA患者在MRI扫描中均显示异常征象,而其中16例患者的X线平片未见异常,18例早期RA患者中有12例未出现X线平片改变.无论患者病程<1年还是>1年,MRI骨侵蚀在抗CCP抗体阳性与阴性组之间差异有统计学意义(P<0.01),RF-IgG阳性组出现滑膜炎的患者明显多于阴性组(P<0.05).血清RF-IgA浓度与骨髓水肿呈正相关(r=0.561,P<0.05).在早期患者中,RF-IgM阳性组出现滑膜炎的患者明显多于阴性组(P<0.01).VAS评分与骨髓水肿评分呈正相关(r=0.562,P<0.01).C反应蛋白(CRP)水平与MRI积液关节的数目呈正相关(r=0.457,P<0.05).结论 MRI对于早期RA的诊断价值要远远大于X线平片.早期RA的MRI改变最常见于腕关节.抗CCP抗体阳性时MRI更易出现骨侵蚀,RF-IgG阳性时MRI更易出现滑膜炎.MRI骨髓水肿评分是VAS评分的重要影响因素.CRP水平可反映有关节腔积液的关节数目.%Objective To study the features of rheumatoid arthritis (RA) on MRI of hands and wrists and compare MRI with clinical manifestations and laboratory tests of RA. Methods A total of 25 patients fulfilling the 2009 ACR/EULAR RA criteria were enrolled. MRI and plain films of hands and wrists and clinical data of swollen joints, tender joints, visual scale (VAS) score, DAS28 score, erythrocyte sedimentation rate ( ESR), C-reactive protein ( CRP), RF classification, anti-cyclic citrullinated peptide antibodies (anti-CCP antibodies) were obtained simultaneously. Results MRI revealed pathologic findings on the wrist and hand joints in all diagnosed RA patients. VAS had a positive correlation with bone marrow edema

  15. ASSOCIATION OF BLOOD GENE EXPRESSIONS IN RHEUMATOID ARTHRITIS PATIENTS WITH CLINICAL AND LABORATORY PARAMETERS BEFORE AND AFTER METHOTREXATE THERAPY

    Directory of Open Access Journals (Sweden)

    E. V. Chetina

    2016-01-01

    Full Text Available The genes, the high basic expression of which indicates the efficiency of methotrexate (MTX therapy in relieving joint inflammation and destruction in patients with rheumatoid arthritis (RA, have been defined.Objective: to find an association between the initial expression of the genes: mTOR (mammalian target of rapamycin, a major regulator of cell growth and proliferation; ULK1 (an autophagy marker 1; p21 (a cyclin-dependent kinase inhibitor; kaspase-3 (an apoptosis activity indicator; MMP-9 (matrix metalloproteinase 9, and cathepsin K, which are involved in joint destruction, and the cytokines: TNF-α (tumor necrosis factor-α, TGFβ1 (transforming growth factor β1 and Runx2 (Runt-related transcription factor 2 in the blood of RA patients with disease activity and joint destruction before and after MTX therapy during 24 months.Subjects and methods. Forty patients (mean age, 47.5 years with RA lasting < 2 years and 26 healthy donors (mean age, 45.1 years were examined. All the patients took MTX for 2 years. A clinical response was assessed with disease activity score (DAS28; erythrocyte sedimentation rate and the serum levels of anti-cyclic citrullinated peptide antibodies (ACCPA, C-reactive protein (CRP, and rheumatoid factor (RF were also estimated. Joint destructive changes were assessed by radiography. Furthermore, blood and knee articular cartilage samples from 21 patients (mean age, 50.4 years with late-stage RA and cartilage samples from 25 healthy individuals were investigated. Gene expression in the cells of peripheral blood and cartilage was determined by real-time reverse transcriptase polymerase chain reaction.Results and discussion. MTX therapy considerably reduced disease activity assessed by DAS28, CRP levels, stiffness, tender and swollen joint counts (TJC and SJC; however, joint space (JS narrowing (JSN substantially increased compared with the baseline values. The expression of the ULK1, p21, MMP-9, cathepsin K genes, and

  16. The clinical significance of peripheral blood hematopoietic stem/progenitor cell in patients with rheumatoid arthritis%类风湿关节炎患者外周血CD34+造血干/祖细胞的检测及其意义

    Institute of Scientific and Technical Information of China (English)

    陈杰; 钱龙; 秦明明; 汪国生; 李向培

    2010-01-01

    目的 探讨类风湿关节炎(RA)外周血造血干/祖细胞(HSC/HPC)的数量及细胞膜CD34平均荧光强度(MFI)变化及其与临床指标的关系,以期阐明其在RA中的意义.方法 收集34例RA患者和16名健康对照者外周血,利用单克隆抗体标记CD34+细胞,流式方法测定CD34+HSC/HPC所占外周血淋巴细胞的比例及膜CD34的MFI,分析其与外周血细胞计数、疾病活动病程和药物应用的关系.数据分析采用t检验和方差分析,相关性分析采用Pearson相关分析.结果 ①RA患者CD34+细胞在外周血中淋巴细胞中所占的比例比健康人偏低[分别为(0.13±0.09)%和(0.38±0.21)%,P<0.05],但两者所占外周血淋巴细胞的比例均低于0.5%;但MFI偏高(分别为57±33和31±11,P<0.05).②RA患者CD34+细胞在外周血淋巴细胞中所占的比例与外周血红细胞计数、血红蛋白浓度呈正相关性,和C反应蛋白呈负相关;其MFI与健康评价问卷表(HAQ)评分、X线分期呈正相关,与血小板计数呈负相关.③RA患者CD34+细胞在外周血淋巴细胞中所占比例的降低程度以及MFI与疾病的活动性、病程和用药情况无明显相关性(P>0.05).结论 造血干细胞可能在RA的发病机制中起作用.%Objective To measure the number of peripheral blood CD34+ hematopoietic stem/progenitor cells (HSC/HPC) expression of CD34 in the peripheral blood of patients with rheumatoid arthritis and exploreits relationship with clinical manifestations. Methods CD34+ HSC/HPCs in the peripheral blood of RA patients (n=32) and healthy controls (n=16) were detected using flow cytometry. The relationship between the frequency of HSC/HPCs, mean fluorescence intensities (MFI) of CD34 and clinical manifestations and rheumatoid factor (RF), anti-cyclic citrullinated peptide (CCP) antibodies, disease activity score (DAS) 28,X rays stages and healthy assessment questionnaire (HAQ) were analyzed. Student's t-test and pearont test were used for

  17. The diagnostic value of antibodies in synovial fluid for rheumatoid arthritis%滑液多种抗体检测诊断类风湿关节炎的临床价值

    Institute of Scientific and Technical Information of China (English)

    徐瑞香; 刘暘; 王文灏; 赵静; 张桂芝; 袁静

    2015-01-01

    目的 通过检测RA患者血清及滑液中抗CCP抗体、RF、免疫球蛋白(Ig)G水平,及AKA、APF的阳性率,探讨滑液多种抗体检测在RA诊断中的价值.方法 入选RA患者30例,常规关节穿刺术抽取关节滑液,静脉采外周血,均离心后-70℃保存.所得数据经受试者工作特征曲线(ROC)分析确定滑液中各抗体诊断RA的阈值,及该阈值的灵敏度、特异度;并对血清及滑液中上述抗体水平进行比较.采用t检验、x2检验及Pearson相关分析进行统计学分析.结果 抗CCP抗体诊断RA滑液有更高的阈值(29.00 U/ml),灵敏度87%;特异度94%.RA患者滑液与血清中抗CCP抗体[(326±313) U/ml,(497±454) U/ml,t=-2.399,P<0.05]、IgG[(57±39) mg/L,(173±86) mg/L,t=-7.4792,P<0.05]、RF[(53±36) U/ml,(149±104) U/ml,t=-2.402,P<0.05]水平差异有统计学意义.结论 RA患者滑液与血清抗CCP抗体、RF、IgG不仅存在数量的不同,同时存在质(分型)的不同.测定滑液抗CCP抗体、RF、IgG较血清学检测能提供更早的信息.%Objective To study the concentration of anti-cyclic citrullinated peptide antibody (CCP),rheumatoid factor (RF),immunoglobulin (Ig) in synovial fluid and serum of patients with rheumatoid arthritis (RA),and the positive rate of anti-keratin antibodies (AKA) and anti-perinuclear factor (APF) were also measured.The aim was to explore the diagnostic value of these antibodies in synovial fluid for RA.Methods Thirty RA patients were collected,synovial fluid were withdrawn from the inflamed joints and peripheral blood were collected at the same time.Anti-CCP antibody,AKA,APF and RF were measured.The cut-off value of CCP in synovial fluid was determined through operating characteristic curve (ROC curves) analysis.The antibody levels in serum and synovial fluid were compared.Statistical analyses were performed using t test,and Pearson's correlation test.Results The cutoff value of CCP in RA synovial was higher,which was 29.00 U/ml,with the

  18. The expression and significance of semaphoring 3A in the serum and peripheral blood mononuclear cell of the patients with rheumatoid arthritis%Semaphorin 3A 在类风湿关节炎患者血清及外周血单个核细胞中的表达及意义

    Institute of Scientific and Technical Information of China (English)

    马晓旭; 胡凡磊; 石连杰; 李英妮; 杜燕; 李春; 郑莉; 张学武

    2013-01-01

    meanwhile, its correlation with the clinical and laboratory indexes of the patients with RA was analyzed. ( 2 ) In addition, the message ribonucleic acid ( mRNA ) expression level of Sema 3A in the PBMC from 45 patients with RA and 35 healthy controls was detected by the real-time quantitative polymerase chain reaction ( PCR ). Results The concentration of Sema 3A in the serum of the patients with RA ( 15.89±8.58 ) ng/ml was significantly higher than that of the healthy controls ( 6.96±2.62 ) ng/ml, ( P<0.01 ) and that of the patients with other rheumatic diseases ( 4.35±3.57 ) ng/ml, ( P<0.01 ). The mRNA expression level of Sema 3A in the PBMC of the patients with RA was ( 1.83±1.47 ), which was significantly higher than ( 1.01±0.85 ) of the healthy controls ( P<0.01 ). There was a positive correlation of the concentration of Sema 3A in the serum and the platelet count ( r=0.229, P<0.01 ), erythrocyte sedimentation rate ( ESR ) ( r=0.174, P<0.05 ), immunoglobulin M ( IgM ) ( r=0.254, P<0.01 ), rheumatoid factor ( RF ) ( r=0.299, P<0.01 ) and histamine-releasing factor-immunoglobulin G ( HRF-IgG ) ( r=0.203, P<0.05 ). In addition, the concentration of Sema 3A in the serum of the RF-, HRF-IgG-, animal protein factor ( APF )-, anti-keratin antibody ( AKA )-, anti-cyclic citrullinated peptide ( CCP ) antibody-positive patients with RA was significantly greater than that of the RF-, HRF-IgG-, APF-, AKA-, anti-CCP antibody-negative patients ( P<0.05 ). Conclusions The expression level of Sema 3A in the serum and PBMC is significantly elevated in the patients with RA, which is closely correlated with the RA disease activity. It is indicated that Sema 3A may become a potentially new target in the treatment of RA.

  19. Serum levels of sTREM-1 and IL-17 in patients with rheumatoid arthritis%类风湿关节炎患者血清可溶性髓样细胞触发受体-1、白介素-17水平分析

    Institute of Scientific and Technical Information of China (English)

    吴志明; 郭健; 黄清水

    2012-01-01

    Rheumatoid arthritis (RA) is an inflammatory autoimmune disease of unknown etiology. Many cytokines have been found to be associated with RA pathogenesis. This study aims to investigate the role of soluble triggering receptors expressed on myeloid cells-1 (sTREM-1) and interleukin 17 (IL-17) in RA. sTREM-1 and IL-17 serum levels were determined in serum samples of 114 RA patients (RA group), 39 other antoimmune disease patients (NO RA group), and 32 healthy subjects (HS group) by enzyme-linked immunosorbent assay (ELISA). Anti-cyclic citrullinated peptide (anti-CCP) antibodies, Rheumatoid factor (RHF), C reactive protein (CRP) and the erythrocyte sedimentation rate (ESR) were measured by routine methods. Disease activity was evaluated using the DAS28 score. Patients were subgrouped according to disease activity and years of disease evolution. Statistical analysis was carried out by SPSS 13.0. Inter-group comparison showed increased levels of sTREM-1 and IL-17 in RA patients versus HS (P = 0.03,0.02); in the RA group, the levels of sTREM-1 was higher in active RA patients than stable RA patients (P = 0.02), and sTREM-1 serum levels positively correlated with IL-17 (r = 0.97, P = 0.001), CRP (r = 0.255, P = 0.006) and ESR (r = 0.442, P = 0.001). Moreover, the serum levels of sTREM-1 and IL-17 were not found to be associated with years of RA evolution (P= 0.64, 0.50); there was no significant difference in the sTREM-1 and IL-17 serum levels between RA group and NO RA group (P = 0.39,0.09). These results suggest that serum sTREM-1 and IL-17 levels are positively correlated in RA, they may play a synergistic effect in RA pathogenesis. Furthermore, sTREM-1 may be associated with RA pathogenesis and disease activity, which could be a potent serum marker for clinical activity of rheumatoid arthritis.%目的 检测类风湿关节炎血清可溶性髓样细胞触发受体1 (soluble triggering receptors expressed on myeloid cells-1,sTREM-1)与白介素-17(interleukin-17,IL-17

  20. A Contra Capture Protein Array Platform for Studying Post-translationally Modified (PTM) Auto-antigenomes.

    Science.gov (United States)

    Karthikeyan, Kailash; Barker, Kristi; Tang, Yanyang; Kahn, Peter; Wiktor, Peter; Brunner, Al; Knabben, Vinicius; Takulapalli, Bharath; Buckner, Jane; Nepom, Gerald; LaBaer, Joshua; Qiu, Ji

    2016-07-01

    Aberrant modifications of proteins occur during disease development and elicit disease-specific antibody responses. We have developed a protein array platform that enables the modification of many proteins in parallel and assesses their immunogenicity without the need to express, purify, and modify proteins individually. We used anticitrullinated protein antibodies (ACPAs) in rheumatoid arthritis (RA) as a model modification and profiled antibody responses to ∼190 citrullinated proteins in 20 RA patients. We observed unique antibody reactivity patterns in both clinical anticyclic citrullinated peptide assay positive (CCP+) and CCP- RA patients. At individual antigen levels, we detected antibodies against known citrullinated autoantigens and discovered and validated five novel antibodies against specific citrullinated antigens (osteopontin (SPP1), flap endonuclease (FEN1), insulin like growth factor binding protein 6 (IGFBP6), insulin like growth factor I (IGF1) and stanniocalcin-2 (STC2)) in RA patients. We also demonstrated the utility of our innovative array platform in the identification of immune-dominant epitope(s) for citrullinated antigens. We believe our platform will promote the study of post-translationally modified antigens at a breadth that has not been achieved before, by both identifying novel autoantigens and investigating their roles in disease development. The developed platforms can potentially be used to study many autoimmune disease-relevant modifications and their immunogenicity. PMID:27141097

  1. The nitric oxide synthase of mouse spermatozoa.

    Science.gov (United States)

    Herrero, M B; Goin, J C; Boquet, M; Canteros, M G; Franchi, A M; Perez Martinez, S; Polak, J M; Viggiano, J M; Gimeno, M A

    1997-07-01

    Nitric oxide synthase (NOS) was evidenced in mature mouse spermatozoa by means of biochemical techniques and Western blot. During 120 min of incubation, 10(7) spermatozoa synthesized 7 +/- 2 pmol of L-[14C]citrulline. Besides, L-citrulline formation depended on the incubation time and on the concentration of L-arginine present in the incubation medium. Different concentrations of N(G)-nitro-L-arginine methyl ester (L-NAME) but not aminoguanidine, inhibited L-[14C]citrulline formation. Western-blot analysis of solubilized sperm proteins revealed a unique band of M(r)=140 kDa with the neural, endothelial and inducible NOS antisera tested. These results provide evidence that mature mouse sperm contains a NOS isoform and that spermatozoa have the potential ability to synthesize NO, suggesting a role for endogenous NO on mammalian sperm function.

  2. Arginine synthesis from enteral glutamine in healthy adults in the fed state.

    Science.gov (United States)

    Tomlinson, Chris; Rafii, Mahroukh; Ball, Ronald O; Pencharz, Paul

    2011-08-01

    Recent studies have documented transfer of labeled nitrogen from [2-(15)N]glutamine to citrulline and arginine in fasting human adults. Conversely, in neonates and piglets we have shown no synthesis of arginine from [2-(15)N]glutamate, and others have shown in mice that glutamine is a nitrogen, but not a carbon donor, for arginine synthesis. Therefore, we performed a multitracer study to determine whether glutamine is a nitrogen and/or carbon donor for arginine in healthy adult men. Two glutamine tracers, 2-(15)N and 1-(13)C, were given enterally to five healthy men fed a standardized milkshake diet. There was no difference in plasma enrichments between the two glutamine tracers. 1-(13)C isotopomers of citrulline and arginine were synthesized from [1-(13)C]glutamine. Three isotopomers each of citrulline and arginine were synthesized from the [2-(15)N]glutamine tracer: 2-(15)N, 5-(15)N, and 2,5-(15)N(2). Significantly greater enrichment was found of both [5-(15)N]arginine (0.75%) and citrulline (3.98%) compared with [2-(15)N]arginine (0.44%) and [2-(15)N]citrulline (2.62%), indicating the amino NH(2) from glutamine is mostly transferred to arginine and citrulline by transamination. Similarly, the enrichment of the 1-(13)C isotopomers was significantly less than the 2-(15)N isotopomers, suggesting rapid formation of α-ketoglutarate and recycling of the nitrogen label. Our results show that the carbon for 50% of newly synthesized arginine comes from dietary glutamine but that glutamine acts primarily as a nitrogen donor for arginine synthesis. Hence, studies using [2-(15)N]glutamine will overestimate arginine synthesis rates.

  3. The interleukin-20 receptor axis in early rheumatoid arthritis

    DEFF Research Database (Denmark)

    Kragstrup, Tue Wenzel; Greisen, Stinne Ravn; Nielsen, Morten Aagaard;

    2016-01-01

    BACKGROUND: Rheumatoid arthritis (RA) is often characterized by the presence of rheumatoid factor, anti-citrullinated protein antibodies, and bone erosions. Current therapies can compromise immunity, leading to risk of infection. The interleukin-20 receptor (IL-20R) axis comprising IL-19, IL-20...... RA synovial fluid compared with monocytes from both RA and healthy control peripheral blood. The plasma concentrations of IL-20 and IL-24 were increased in rheumatoid factor and anti-citrullinated protein antibody positive compared with negative early RA patients (all P < 0.0001). Immune complexes...

  4. 类风湿关节炎患者血清抗鼠科肉瘤病毒癌基因同源物B1抗体检测及临床意义探讨%Detection of serum anti v-raf murine sarcoma viral oncogene homologue B1 antibodies in patients with rheumatoid arthritis and the assessment of its clinical significance

    Institute of Scientific and Technical Information of China (English)

    李扬; 竺红

    2014-01-01

    Objective To investigate the serum levels of anti-BRAF antibody in patients with rheumatoid arthritis (RA) and its clinical significance.Methods Blood samples from 129 patients with RA,71 patients with other rheumatic diseases and 68 healthy controls were measured by enzyme-linked immunosorbent assays (ELISA).The serum levels of anti-BRAF antibody in patients with RA were divided into high,medium and low groups,each group had 43 cases.The clinical symptoms,immunological findings and related imaging data of all cases were recorded.We investigated the relationship between the serum levels of anti-BRAF antibody and the clinical symptoms,immunological findings and related imaging changes of the RA group.Chi-square test,Mann-Whitney U test,Spearman's correlation test were used for statistical analysis.Results ① The serum levels of anti-BRAF antibody were significantly higher in RA group [A:1.093 (0.150-3.210)] than in other rheumatic diseases group [A:0.278 (0.093-0.455)] and healthy controls [A:0.202 (0.121-0.443)] (Z=-11.223,P=0.021 ; Z=-7.839,P=0.008 7).The serum levels of anti-BRAF antibody was not significantly different between patients with other rheumatic diseases group and healthy controls (Z=-0.905,P=0.445).② The difference in positive rate of rheumatoid factor (RF) in different groups of anti-BRAF antibody levels was statistically significant (λx2=10.14,P=0.006),namely the positive rate was significantlyhigher in the low-level group (88%) than in the high-level group (58%) (x2=10.03,P=0.002).Thepositive rate of anti-cyclic citrullinated peptide antibody and anti-keratin antibody of the three groups was not significantly different (x2=0.721,P=0.697; x2=1.735,P=0.188).③ There was significant negative correlation between the levels of anti-BRAF antibody and the course of disease (r=-0.233,P=0.047),RF(r=-2.84,P=0.039),IgM (r=-0.234,P=0.038),and hemoglobin (r=-0.287,P=0.032).There was no significant correlation between the level of anti-BRAF antibody

  5. Survey on Log-Normally Distributed Market-Technical Trend Data

    Directory of Open Access Journals (Sweden)

    René Brenner

    2016-07-01

    Full Text Available In this survey, a short introduction of the recent discovery of log-normally-distributed market-technical trend data will be given. The results of the statistical evaluation of typical market-technical trend variables will be presented. It will be shown that the log-normal assumption fits better to empirical trend data than to daily returns of stock prices. This enables one to mathematically evaluate trading systems depending on such variables. In this manner, a basic approach to an anti-cyclic trading system will be given as an example.

  6. The Euro crisis. Causes and Symptoms

    OpenAIRE

    Weber, Christoph S.

    2015-01-01

    The Euro crisis is mainly a consequence of the international financial crisis of 2008. Thereby, the term Euro crisis is misleading as there is no currency crisis. First, the article shows some of the birth defects of the Euro. Second, it shows that the increase in public debt was caused by rescue measures for banks and anti-cyclical fiscal policy. Third, we argue that the Euro crisis is not just one crisis (a sovereign debt crisis) but it is a combination of several macroeconomic crises inclu...

  7. The Euro crisis. Causes and Symptoms

    Directory of Open Access Journals (Sweden)

    Christoph S. Weber

    2015-06-01

    Full Text Available The Euro crisis is mainly a consequence of the international financial crisis of 2008. Thereby, the term Euro crisis is misleading as there is no currency crisis. First, the article shows some of the birth defects of the Euro. Second, it shows that the increase in public debt was caused by rescue measures for banks and anti-cyclical fiscal policy. Third, we argue that the Euro crisis is not just one crisis (a sovereign debt crisis but it is a combination of several macroeconomic crises including a growth crisis, a labour market crisis, a public debt crisis, and a current account crisis.

  8. High-performance liquid chromatography method with radiochemical detection for measurement of nitric oxide synthase, arginase, and arginine decarboxylase activities.

    Science.gov (United States)

    Volke, A; Wegener, G; Vasar, E; Volke, V

    2006-01-01

    Nitric oxide has been shown to be involved in numerous biological processes, and many studies have aimed to measure nitric oxide synthase (NOS) activity. Recently, it has been demonstrated that arginase and arginine decarboxylase (ADC), two enzymes that also employ arginine as a substrate, may regulate NOS activity. We aimed to develop a HPLC-based method to measure simultaneously the products of these three enzymes. Traditionally, the separation of amino acids and related compounds with HPLC has been carried out with precolumn derivatization and reverse phase chromatography. We describe here a simple and fast HPLC method with radiochemical detection to separate radiolabeled L-arginine, L-citrulline, L-ornithine, and agmatine. 3H-labeled L-arginine, L-citrulline, agmatine, and 14C-labeled L-citrulline were used as standards. These compounds were separated in the normal phase column (Allure Acidix 250 x 4.6 mm i.d.) under isocratic conditions in less than 20 min with good sensitivity. Using the current method, we have shown the formation of L-citrulline and L-ornithine in vitro using brain tissue homogenate of rats and that of agmatine by Escherichia coli ADC. PMID:16541190

  9. Cytotoxicity and cellular uptake of ZnS:Mn nanocrystals biofunctionalized with chitosan and aminoacids

    Digital Repository Service at National Institute of Oceanography (India)

    Augustine, M.S.; Anas, A.; Das, A.V.; Sreekanth, S.; Jayalekshmi, S.

    Highly luminescent, manganese doped, zinc sulphide (ZnS:Mn) nanocrystals biofunctionalized with chitosan and various aminoacids such as l-citrulline, l-lysine, l-arginine, l-serine, l-histidine and glycine were synthesized by chemical capping co...

  10. Combining a Laboratory Practical Class with a Computer Simulation: Studies on the Synthesis of Urea in Isolated Hepatocytes.

    Science.gov (United States)

    Bender, David A.

    1986-01-01

    Describes how a computer simulation is used with a laboratory experiment on the synthesis of urea in isolated hepatocytes. The simulation calculates the amount of urea formed and the amount of ammonium remaining as the concentrations of ornithine, citrulline, argininosuccinate, arginine, and aspartate are altered. (JN)

  11. Fine Mapping Seronegative and Seropositive Rheumatoid Arthritis to Shared and Distinct HLA Alleles by Adjusting for the Effects of Heterogeneity

    NARCIS (Netherlands)

    Han, Buhm; Diogo, Dorothee; Eyre, Steve; Kallberg, Henrik; Zhernakova, Alexandra; Bowes, John; Padyukov, Leonid; Okada, Yukinori; Gonzalez-Gay, Miguel A.; Rantapaa-Dahlqvist, Solbritt; Martin, Javier; Huizinga, Tom W. J.; Plenge, Robert M.; Worthington, Jane; Gregersen, Peter K.; Klareskog, Lars; de Bakker, Paul I. W.; Raychaudhuri, Soumya

    2014-01-01

    Despite progress in defining human leukocyte antigen (HLA) alleles for anti-citrullinated-protein-autoantibody-positive (ACPA(+)) rheumatoid arthritis (RA), identifying HLA alleles for ACPA-negative (ACPA(-)) RA has been challenging because of clinical heterogeneity within clinical cohorts. We imput

  12. Inflammatory response to mucosal barrier injury after myeloablative therapy in allogeneic stem cell transplant recipients.

    NARCIS (Netherlands)

    Blijlevens, N.M.A.; Donnelly, J.P.; Pauw, B.E. de

    2005-01-01

    We noted a significant increase of interleukin-8 (IL-8), LBP and CRP mirroring the pattern of mucosal barrier injury as measured by gut integrity (lactulose/rhamnose ratio), daily mucositis score (DMS) and serum citrulline concentrations of 32 haematopoietic stem cell transplant (HSCT) recipients fo

  13. Dietary supplementation with watermelon pomace juice enhances arginine availability and ameliorates the metabolic syndrome in Zucker diabetic fatty rats

    Science.gov (United States)

    Watermelon is rich in L-citrulline, an effective precursor of L-arginine. This study was conducted to determine whether dietary supplementation with watermelon pomace juice could ameliorate the metabolic syndrome in the Zucker diabetic fatty (ZDF) rat, an animal model of noninsulin-dependent diabet...

  14. Watermelon enhances arginine availability in an animal model of type-II diabetes

    Science.gov (United States)

    Watermelon fruit contain lycopene, a red pigment known for its ability to scavenge free hydroxyl radicals. L-Citrulline, an amino acid that acts as a vasodilator and is a precursor of L-arginine, is found in all cucurbits, but is most plentiful in watermelon. In a study with Zucker diabetic fatty ...

  15. A metabolomics approach to identify and quantify the phytochemicals in watermelons by quantitative (1)HNMR.

    Science.gov (United States)

    Jayaprakasha, G K; Patil, Bhimanagouda S

    2016-06-01

    Watermelon (Citrullus vulgaris) contains many health-promoting compounds, such as ascorbic acid, carotenoids, phenolic acids and amino acids including l-citrulline, arginine, and glutathione. Reported HPLC method for quantification of l-citrulline and sugars in watermelon involves, time-consuming sample preparation, post-column color development and detection with fluorescence and refractive index detectors. The present study describes development of a method to identify and quantify amino acids and sugars simultaneously from watermelon samples using quantitative proton NMR. Lyophilized watermelon samples (30-50mg) were extracted with deuterium oxide (D2O) by sonication and the centrifuged extract was directly used for quantification and identification with (1)HNMR. An external coaxial insert containing a 65µL of 0.012% 3-(trimethylsilyl) propionic-(2,2,3,3-d4) acid sodium salt (TSP-d4) in D2O was used as a quantitative reference. The levels of l-citrulline and sugars were measured in less than 6min. This rapid quantitation method was validated for specificity, linearity, accuracy, precision, reproducibility, and robustness. The limit of detection for l-citrulline was 38µg/mL and the limit of quantification was 71µg/mL; for sugars, the limits were 59-94µg/mL and 120µg/mL, respectively. This method can be used widely for confirmation and rapid quantitation of multiple compounds in large number of biological or breeding samples for routine analysis. PMID:27130118

  16. The microbiome, intestinal function, and arginine metabolism of healthy Indian women are different from those of American and Jamaican women

    Science.gov (United States)

    Indian women have slower arginine flux during pregnancy compared with American and Jamaican women. Arginine is a semi-essential amino acid that becomes essential during periods of rapid lean tissue deposition. It is synthesized only from citrulline, a nondietary amino acid produced mainly in the gut...

  17. Ruthenium(III) readily abstracts NO from L-arginine, the physiological precursor to NO, in the presence of H2O2. A remarkably simple model system for NO synthases.

    Science.gov (United States)

    Marmion, C J; Murphy, T; Nolan, K B

    2001-09-21

    Reaction of [Ru(Hedta)Cl]- with H2O2 in the presence of arginine, produces NO, in the form of an Ru(II)-(NO+) complex and citrulline which is a remarkably simple model system for the physiological NO synthase reaction. PMID:12240355

  18. Short- and long-term efficacy of intra-articular injections with betamethasone as part of a treat-to-target strategy in early rheumatoid arthritis

    DEFF Research Database (Denmark)

    Hetland, Merete Lund; Østergaard, Mikkel; Ejbjerg, Bo;

    2012-01-01

    OBJECTIVE: To investigate the short-term and long-term efficacy of intra-articular betamethasone injections, and the impact of joint area, repeated injections, MRI pathology, anticyclic citrullinated peptide (CCP) and immunoglobulin M rheumatoid factor (IgM-RF) status in patients with early...

  19. Post-translational modifications of the extracellular matrix are key events in cancer progression: opportunities for biochemical marker development

    DEFF Research Database (Denmark)

    Leeming, D J; Bay-Jensen, A C; Vassiliadis, E;

    2011-01-01

    The aim of this review is to discuss the potential usefulness of a novel class of biochemical markers, designated neoepitopes. Neoepitopes are post-translational modifications (PTMs) of proteins and are derived by processes, such as protease cleavage, citrullination, nitrosylation, glycosylation ...

  20. Activity of L-alpha-amino acids at the promiscuous goldfish odorant receptor 5.24

    DEFF Research Database (Denmark)

    Christiansen, Bolette; Wellendorph, Petrine; Bräuner-Osborne, Hans

    2006-01-01

    report the agonist activities of the 20 proteinogenic L-alpha-amino acids, and L-ornithine and L-citrulline, measured in an intracellular calcium release assay in mammalian tsA cells. The results show that receptor 5.24 is broadly activated by 19 of the tested L-alpha-amino acids and displays...

  1. Disease: H00185 [KEGG MEDICUS

    Lifescience Database Archive (English)

    Full Text Available by a deficiency of argininosuccinate synthetase/citrin and characterized primarily by elevated serum and urine citrulline level...H00185 Citrullinemia (CTLN) Citrullinemia is an autosomal recessive disease caused ...s. Inherited metabolic disease; Nervous system disease hsa00330(445) Arginine and proline

  2. Pre-treatment antinuclear antibody positivity, therapeutic efficacy and persistence of biologics in rheumatoid arthritis

    Directory of Open Access Journals (Sweden)

    Codreanu Cătălin

    2016-06-01

    Full Text Available Introduction: Rheumatoid factor (RF and anti-citrullinated protein antibodies (ACPA are poor prognostic factors in rheumatoid arthritis (RA. The therapeutic implication of antinuclear antibody (ANA positivity in RA is still debated. The study aims to evaluate ANA positivity as a prognostic factor for the therapeutic response to biologics in RA.

  3. Perioperative glutamine supplementation restores disturbed renal arginine synthesis after open aortic surgery: a randomized controlled clinical trial.

    Science.gov (United States)

    Brinkmann, Saskia J H; Buijs, Nikki; Vermeulen, Mechteld A R; Oosterink, Efraim; Schierbeek, Henk; Beishuizen, Albertus; de Vries, Jean-Paul P M; Wisselink, Willem; van Leeuwen, Paul A M

    2016-09-01

    Postoperative renal failure is a common complication after open repair of an abdominal aortic aneurysm. The amino acid arginine is formed in the kidneys from its precursor citrulline, and citrulline is formed from glutamine in the intestines. Arginine enhances the function of the immune and cardiovascular systems, which is important for recovery after surgery. We hypothesized that renal arginine production is diminished after ischemia-reperfusion injury caused by clamping of the aorta during open abdominal aortic surgery and that parenteral glutamine supplementation might compensate for this impaired arginine synthesis. This open-label clinical trial randomized patients who underwent clamping of the aorta during open abdominal aortic surgery to receive a perioperative supplement of intravenous alanyl-glutamine (0.5 g·kg(-1)·day(-1); group A, n = 5) or no supplement (group B, n = 5). One day after surgery, stable isotopes and tracer methods were used to analyze the metabolism and conversion of glutamine, citrulline, and arginine. Whole body plasma flux of glutamine, citrulline, and arginine was significantly higher in group A than in group B (glutamine: 391 ± 34 vs. 258 ± 19 μmol·kg(-1)·h(-1), citrulline: 5.7 ± 0.4 vs. 2.8 ± 0.4 μmol·kg(-1)·h(-1), and arginine: 50 ± 4 vs. 26 ± 2 μmol·kg(-1)·h(-1), P glutamine (4.8 ± 0.7 vs. 1.6 ± 0.3 μmol·kg(-1)·h(-1)), citrulline from arginine (2.3 ± 0.3 vs. 0.96 ± 0.1 μmol·kg(-1)·h(-1)), and arginine from glutamine (7.7 ± 0.4 vs. 2.8 ± 0.2 μmol·kg(-1)·h(-1)), respectively (P arginine is severely reduced after clamping during aortic surgery. This study shows that an intravenous supplement of glutamine increases the production of citrulline and arginine and compensates for the inhibitory effect of ischemia-reperfusion injury.

  4. DOMETI FISKALNE POLITIKE U PODSTICANJU PRIVREDNOG RASTA

    Directory of Open Access Journals (Sweden)

    Predrag Mlinarević

    2013-01-01

    Full Text Available The issue of use of expansive fiscal instruments in order to encourage the growth has produced in the past as it does today, a significant division among economists. This division is reflected in the doctrinal and practical aspects. Doctrinal aspect touches different approaches to state interventionism. While its supporters within Keynesian teachings believe that the recession could become a permanent condition if the state does not reach for anti-cyclical policies, the supporters of the infallibility of the market believe that it would result in inflation without affecting growth. This paper presents, with inevitable reference to the genesis of bipolar thinking, the results of empirical studies and our own research on the value of the relevant fiscal multipliers as realistic measures of effectualization of fiscal stimulus to economic growth. Conferring a direct connection between appropriate fiscal stimulus packages with the growth of economic activity will give an answer to the issue about the effectiveness of cyclic measures. It turned out that the results are different for developed countries compared to those who belong to the group of developing countries. It has been proved that for developing countries it is much more useful to rely on public investment in relation to the growth of public spending. It should serve as a guide to the holders of economic power in composing the measures that will form the anti-cyclical economic policies during the current crisis.

  5. Arthritogenic peptide binding to DRB1*01 alleles correlates with susceptibility to rheumatoid arthritis.

    Science.gov (United States)

    Roark, Christina L; Anderson, Kirsten M; Aubrey, Michael T; Rosloniec, Edward F; Freed, Brian M

    2016-08-01

    Genetic susceptibility to rheumatoid arthritis (RA) is often defined by the presence of a shared epitope (QKRAA, QRRAA, or RRRAA) at positions 70-74 in HLA-DRβ1. However, DRβ1*01:01 and 01:02 contain the same QRRAA epitope, but differ considerably in their susceptibility to RA. The purpose of this study was to determine if this difference could be explained by their ability to bind three arthritogenic peptides that we have previously shown to bind to the archetypal RA-susceptible allele, DRβ1*04:01, but not to the resistant DRβ1*08:01 allele. Binding of type II collagen(258-272), citrullinated and native vimentin(66-78), and citrullinated and native α-enolase(11-25) were measured on cell lines expressing either DRβ1*01:01, *01:02 or *01:03 in association with DRα1*01:01. DRβ1*01:01 and *01:02 both exhibited a 6.5-fold preference for citrullinated vimentin(66-78) compared to native vimentin. However, DRβ1*01:01 also exhibited a 1.7-fold preference for citrullinated α-enolase(11-25) and bound collagen(258-272), while DRβ1*01:02 bound neither of these peptides. Consistent with its known resistance to RA, DRβ1*01:03 preferentially bound native vimentin(66-78) and α-enolase(11-25) over the citrullinated forms of these peptides, and also failed to bind collagen(258-272). Site-directed mutagenesis was performed to determine which amino acid residues were responsible for the differences between these alleles. Mutating position 86 in DRβ1*01:01 from glycine to the valine residue found in DRβ1*01:02 eliminated binding of both citrullinated α-enolase(11-25) and collagen(258-272), thereby recapitulating the peptide-binding profile of DRβ1*01:02. The difference in susceptibility to rheumatoid arthritis between DRβ1*01:01 and *01:02 thus correlates with the effect of position 86 on the binding of these arthritogenic peptides. Consistent with their association with RA resistance, positions I67, D70 and E71 all contributed to the inability of DRβ1*01:03 to bind

  6. Evolution of watermelon fruit physicochemical and phytochemical composition during ripening as affected by grafting.

    Science.gov (United States)

    Soteriou, G A; Kyriacou, M C; Siomos, A S; Gerasopoulos, D

    2014-12-15

    Flesh reflectance colorimetry, mechanical texture analysis, pH, titratable acidity (TA), and soluble solid (SS), soluble carbohydrate, lycopene and citrulline content of watermelon fruit were assessed throughout ripening (30-50 days post-anthesis; dpa) in grafted and self-rooted plants. Grafting increased firmness, TA, and lycopene content though it delayed its peak. Lycopene content was mostly ripening-dependant, highly correlated and synchronous with changes in pulp chroma (C) and colour a. The sweetness was affected only by ripening. However, total sugars and SS peaked later in fruit of grafted plants than in non-grafted ones, and significant interaction of ripening with grafting was observed. Citrulline content increased with ripening in fruit of grafted plants, reaching a peak at 45 dpa; whereas in non-grafted ones it was unchanged between 30 and 45 dpa and declined at 50 dpa. As ripening overall was retarded by grafting, fruit quality of grafted watermelon may benefit from belated harvest.

  7. [GABA-NO interaction in the N. Accumbens during danger-induced inhibition of exploratory behavior].

    Science.gov (United States)

    2013-01-01

    In Sprague-Dawley rats by means of in vivo microdialysis combined with HPLC analysis, it was shown that presentation to rats during exploratory activity of a tone previously pared with footshock inhibited the exploration and prevented the exploration-induced increase in extracellular levels of citrulline (an NO co-product) in the medial n. accumbens. Intra-accumbal infusions of 20 μM bicuculline, a GABA(A)-receptor antagonist, firstly, partially restored the exploration-induced increase of extracellular citrulline levels in this brain area, which was inhibited by presentation of the tone, previously paired with foot-shock and, secondly, prevented the inhibition of exploratory behavior produced by this sound signal of danger. The data obtained indicate for the first time that signals of danger inhibit exploratory behavior and exploration-induced activation of the accumbal nitrergic system via GABA(A)-receptor mechanisms. PMID:25508395

  8. Endothelial arginine resynthesis contributes to the maintenance of vasomotor function in male diabetic mice

    DEFF Research Database (Denmark)

    Chennupati, Ramesh; Meens, Merlijn J P M T; Marion, Vincent;

    2014-01-01

    AIM: Argininosuccinate synthetase (ASS) is essential for recycling L-citrulline, the by-product of NO synthase (NOS), to the NOS substrate L-arginine. Here, we assessed whether disturbed arginine resynthesis modulates endothelium-dependent vasodilatation in normal and diabetic male mice. METHODS...... AND RESULTS: Endothelium-selective Ass-deficient mice (Assfl/fl/Tie2Cretg/- = Ass-KOTie2) were generated by crossing Assfl/fl mice ( = control) with Tie2Cre mice. Gene ablation in endothelial cells was confirmed by immunohistochemistry. Blood pressure (MAP) was recorded in 34-week-old male mice. Vasomotor...... of endothelial citrulline recycling to arginine did not affect blood pressure and systemic arterial vasomotor responses in healthy mice. EDNO-mediated vasodilatation was significantly more impaired in diabetic Ass-KOTie2 than in control mice demonstrating that endothelial arginine recycling becomes a limiting...

  9. Inhibition of PAD4 activity is sufficient to disrupt mouse and human NET formation

    Science.gov (United States)

    Lewis, Huw D.; Liddle, John; Coote, Jim E.; Atkinson, Stephen J.; Barker, Michael D.; Bax, Benjamin, D.; Bicker, Kevin L.; Bingham, Ryan P.; Campbell, Matthew; Chen, Yu Hua; Chung, Chun-wa; Craggs, Peter D.; Davis, Rob P.; Eberhard, Dirk; Joberty, Gerard; Lind, Kenneth E.; Locke, Kelly; Maller, Claire; Martinod, Kimberly; Patten, Chris; Polyakova, Oxana; Rise, Cecil E.; Rüdiger, Martin; Sheppard, Robert J.; Slade, Daniel J.; Thomas, Pamela; Thorpe, Jim; Yao, Gang; Drewes, Gerard; Wagner, Denisa D.; Thompson, Paul R.; Prinjha, Rab K.; Wilson, David M.

    2015-01-01

    PAD4 has been strongly implicated in the pathogenesis of autoimmune, cardiovascular and oncological diseases, through clinical genetics and gene disruption in mice. Novel, selective PAD4 inhibitors binding to a calcium-deficient form of the PAD4 enzyme have, for the first time, validated the critical enzymatic role of human and mouse PAD4 in both histone citrullination and neutrophil extracellular trap formation. The therapeutic potential of PAD4 inhibitors can now be explored. PMID:25622091

  10. Comparison of Watermelon and Carbohydrate Beverage on Exercise-Induced Alterations in Systemic Inflammation, Immune Dysfunction, and Plasma Antioxidant Capacity.

    Science.gov (United States)

    Shanely, R Andrew; Nieman, David C; Perkins-Veazie, Penelope; Henson, Dru A; Meaney, Mary P; Knab, Amy M; Cialdell-Kam, Lynn

    2016-01-01

    Consuming carbohydrate- and antioxidant-rich fruits during exercise as a means of supporting and enhancing both performance and health is of interest to endurance athletes. Watermelon (WM) contains carbohydrate, lycopene, l-citrulline, and l-arginine. WM may support exercise performance, augment antioxidant capacity, and act as a countermeasure to exercise-induced inflammation and innate immune changes. Trained cyclists (n = 20, 48 ± 2 years) participated in a randomized, placebo controlled, crossover study. Subjects completed two 75 km cycling time trials after either 2 weeks ingestion of 980 mL/day WM puree or no treatment. Subjects drank either WM puree containing 0.2 gm/kg carbohydrate or a 6% carbohydrate beverage every 15 min during the time trials. Blood samples were taken pre-study and pre-, post-, 1 h post-exercise. WM ingestion versus no treatment for 2-weeks increased plasma l-citrulline and l-arginine concentrations (p 0.05), however, the rating of perceived exertion was greater during the WM trial (p > 0.05). WM puree versus carbohydrate beverage resulted in a similar pattern of increase in blood glucose, and greater increases in post-exercise plasma antioxidant capacity, l-citrulline, l-arginine, and total nitrate (all p < 0.05), but without differences in systemic markers of inflammation or innate immune function. Daily WM puree consumption fully supported the energy demands of exercise, and increased post-exercise blood levels of WM nutritional components (l-citrulline and l-arginine), antioxidant capacity, and total nitrate, but without an influence on post-exercise inflammation and changes in innate immune function. PMID:27556488

  11. Route of administration (enteral or parenteral) affects the contribution of L-glutamine to de novo L-arginine synthesis in mice: a stable-isotope study.

    Science.gov (United States)

    Boelens, Petra G; Melis, Gerdien C; van Leeuwen, Paul A; ten Have, Gabrie A; Deutz, Nicolaas E

    2006-10-01

    A pathway from enteral L-glutamine as substrate for L-arginine synthesis is suggested by previous studies. L-Glutamine and L-glutamine dipeptides exhibit numerous beneficial effects in experimental and clinical studies. In trauma patients, enteral L-glutamine supply increased plasma L-arginine. The present study was designed to quantify the contribution of L-glutamine to the de novo L-citrulline and L-arginine synthesis in mice when L-glutamine is administered in a high dose of labeled L-glutamine or L-alanyl-L-glutamine by the enteral or parenteral route. For this purpose, male Swiss mice (n = 43) underwent a laparotomy, and catheters were inserted for sampling and infusion. A primed, constant, and continuous infusion of L-alanyl-L-[2-(15)N]glutamine (dipeptide groups) or L-[2-(15)N]glutamine (free L-glutamine groups), simultaneously with L-[ureido-(13)C,(2)H(2)]citrulline and L-[guanidino-(15)N(2),(2)H(2)]arginine, was given (steady-state model). Mice received the L-glutamine tracers intravenously (jugular vein) or enterally (duodenum). Enrichments of metabolites were measured by LC-MS. Arterial L-glutamine concentrations were the highest in the intravenous dipeptide group. L-Glutamine was converted to L-citrulline and L-arginine when L-[2-(15)N]glutamine and L-alanyl-L-[2-(15)N]glutamine were given by enteral or parenteral route. The contribution of L-glutamine to the de novo synthesis of L-citrulline and L-arginine was higher in the enteral groups when compared with the intravenous groups (P glutamine, provided as free molecule or dipeptide, to the de novo synthesis of L-arginine in mice.

  12. Fine Mapping Seronegative and Seropositive Rheumatoid Arthritis to Shared and Distinct HLA Alleles by Adjusting for the Effects of Heterogeneity

    OpenAIRE

    Han, Buhm; Diogo, Dorothée; Eyre, Steve; Kallberg, Henrik; Zhernakova, Alexandra; Bowes, John; Padyukov, Leonid; Okada, Yukinori; González-Gay, Miguel A.; Rantapää-Dahlqvist, Solbritt; Martin, Javier; Huizinga, Tom W.J.; Plenge, Robert M.; Worthington, Jane; Gregersen, Peter K.

    2014-01-01

    Despite progress in defining human leukocyte antigen (HLA) alleles for anti-citrullinated-protein-autoantibody-positive (ACPA+) rheumatoid arthritis (RA), identifying HLA alleles for ACPA-negative (ACPA−) RA has been challenging because of clinical heterogeneity within clinical cohorts. We imputed 8,961 classical HLA alleles, amino acids, and SNPs from Immunochip data in a discovery set of 2,406 ACPA− RA case and 13,930 control individuals. We developed a statistical approach to identify and ...

  13. Comparison of Watermelon and Carbohydrate Beverage on Exercise-Induced Alterations in Systemic Inflammation, Immune Dysfunction, and Plasma Antioxidant Capacity

    Directory of Open Access Journals (Sweden)

    R. Andrew Shanely

    2016-08-01

    Full Text Available Consuming carbohydrate- and antioxidant-rich fruits during exercise as a means of supporting and enhancing both performance and health is of interest to endurance athletes. Watermelon (WM contains carbohydrate, lycopene, l-citrulline, and l-arginine. WM may support exercise performance, augment antioxidant capacity, and act as a countermeasure to exercise-induced inflammation and innate immune changes. Trained cyclists (n = 20, 48 ± 2 years participated in a randomized, placebo controlled, crossover study. Subjects completed two 75 km cycling time trials after either 2 weeks ingestion of 980 mL/day WM puree or no treatment. Subjects drank either WM puree containing 0.2 gm/kg carbohydrate or a 6% carbohydrate beverage every 15 min during the time trials. Blood samples were taken pre-study and pre-, post-, 1 h post-exercise. WM ingestion versus no treatment for 2-weeks increased plasma l-citrulline and l-arginine concentrations (p < 0.0125. Exercise performance did not differ between WM puree or carbohydrate beverage trials (p > 0.05, however, the rating of perceived exertion was greater during the WM trial (p > 0.05. WM puree versus carbohydrate beverage resulted in a similar pattern of increase in blood glucose, and greater increases in post-exercise plasma antioxidant capacity, l-citrulline, l-arginine, and total nitrate (all p < 0.05, but without differences in systemic markers of inflammation or innate immune function. Daily WM puree consumption fully supported the energy demands of exercise, and increased post-exercise blood levels of WM nutritional components (l-citrulline and l-arginine, antioxidant capacity, and total nitrate, but without an influence on post-exercise inflammation and changes in innate immune function.

  14. Nitric Oxide: Role in Human Biology

    OpenAIRE

    Nikhil Omer; Ankur Rohilla; Seema Rohilla; Ashok Kushnoor

    2012-01-01

    Nitric oxide (NO), a free radical, possesses various modulatory effects on biological systems. NO is synthesized from L-arginine by converting it to L-citrulline via nitric oxide synthase (NOS) enzymes. Moreover, various precursors of NO have been reported that include arginine, citruline, arginine alphaketoglutarate (A-AKG) and arginineketoisocaproate (A-KIC). NO possess various direct and indirect effects that broadly affect various tissues and organ systems inside the body. The present rev...

  15. Low-load resistance training during step-reduction attenuates declines in muscle mass and strength and enhances anabolic sensitivity in older men

    OpenAIRE

    Devries, Michaela C; Breen, Leigh; Von Allmen, Mark; MacDonald, Maureen J; Moore, Daniel R.; Offord, Elizabeth A; Horcajada, Marie-Noëlle; Breuillé, Denis; Phillips, Stuart M.

    2015-01-01

    Step-reduction (SR) in older adults results in muscle atrophy and an attenuated rise in postprandial muscle protein synthesis (MPS): anabolic resistance. Knowing that resistance exercise (RT) can enhance MPS, we examined whether RT could enhance MPS following 2 weeks of SR. In addition, as we postulated that SR may impair feeding-induced vasodilation limiting nutrient delivery to muscle, we also examined whether citrulline (CIT), as an arginine and nitric oxide precursor, could attenuate musc...

  16. A specific method for measurement of nitric oxide synthase enzymatic activity in peritoneal biopsies.

    OpenAIRE

    Combet, S.; Balligand, Jean-Luc; Lameire, N.; Goffin, Eric; Devuyst, Olivier

    2000-01-01

    A specific method for measurement of nitric oxide synthase enzymatic activity in peritoneal biopsies. BACKGROUND: Nitric oxide (NO) is synthesized by NO synthase (NOS) isoforms that are expressed in the peritoneum. Thus far, NOS activity in the peritoneum has been assessed by nonspecific methods. We describe the application of a specific method for determination of NOS activity in rat and human peritoneal biopsies. METHODS: The L-citrulline assay is based on the stoechiometric production of N...

  17. Relationship between plasmid content and auxotype in Neisseria gonorrhoeae isolates.

    OpenAIRE

    Dillon, J R; Pauzé, M

    1981-01-01

    One hundred and forty strains of Neisseria gonorrhoeae, representing 12 different auxotype groups, were examined for differences in plasmid content. Most auxotype groups harbored a phenotypically cryptic 2,6-megadalton plasmid; a few groups also carried a 24.5-megadalton plasmid which has been previously characterized as a transfer plasmid. However, isolates of the proline-, citrulline-, and uracil-requiring (PCU-) auxotype were consistently free of plasmids. The correlation between auxotype ...

  18. Citrullus lanatus `Sentinel' (Watermelon) Extract Reduces Atherosclerosis in LDL Receptor Deficient Mice

    OpenAIRE

    Poduri, Aruna; Rateri, Debra L.; Saha, Shubin K.; Saha, Sibu; Daugherty, Alan

    2012-01-01

    Watermelon (Citrullus lanatus or C. lanatus) has many potentially bioactive compounds including citrulline, which may influence atherosclerosis. In this study, we determined the effects of C. lanatus, provided as an extract of the cultivar `sentinel', on hypercholesterolemia-induced atherosclerosis in mice. Male LDL receptor deficient mice at 8 weeks old were given either C. lanatus `sentinel' extract (2% vol/vol; n=10) or a mixture of matching carbohydrates (2% vol/vol; n=8) as the control i...

  19. Skrining Fitokimia Dan Uji Aktivitas Antioksidan Sari Daging Buah Semangka (Citrullus Lanatus (Thunb.) Matsum. & Nakai) Dengan Metode Dpph (1,1diphenyl-2-Picrylhydrazil)

    OpenAIRE

    Cristin, Novia

    2015-01-01

    Watermelon (Citrullus lanatus (Thunb.) Matsum. & Nakai ) is the plants fruit of the tropics areas which is generally widely grown in Indonesia; one of those areas that develop these plants is North Sumatra. Watermelon contains various nutrients such as vitamins (A, B and C), amino acid citrulline, lycopene, carotene, fructose, dextrose, and sucrose. Watermelon fruit flesh is composed of layers of white and red layers. The research objective was to determine the phytochemical screening and ant...

  20. Activation of PAD4 in NET formation.

    Science.gov (United States)

    Rohrbach, Amanda S; Slade, Daniel J; Thompson, Paul R; Mowen, Kerri A

    2012-01-01

    Peptidylarginine deiminases, or PADs, convert arginine residues to the non-ribosomally encoded amino acid citrulline in a variety of protein substrates. PAD4 is expressed in granulocytes and is essential for the formation of neutrophil extracellular traps (NETs) via PAD4-mediated histone citrullination. Citrullination of histones is thought to promote NET formation by inducing chromatin decondensation and facilitating the expulsion of chromosomal DNA that is coated with antimicrobial molecules. Numerous stimuli have been reported to lead to PAD4 activation and NET formation. However, how this signaling process proceeds and how PAD4 becomes activated in cells is largely unknown. Herein, we describe the various stimuli and signaling pathways that have been implicated in PAD4 activation and NET formation, including the role of reactive oxygen species generation. To provide a foundation for the above discussion, we first describe PAD4 structure and function, and how these studies led to the development of PAD-specific inhibitors. A comprehensive survey of the receptors and signaling pathways that regulate PAD4 activation will be important for our understanding of innate immunity, and the identification of signaling intermediates in PAD4 activation may also lead to the generation of pharmaceuticals to target NET-related pathogenesis. PMID:23264775

  1. A novel method for simultaneous measurement of concentration and enrichment of NO synthesis-specific amino acids in human plasma using stable isotopes and LC/MS ion trap analysis.

    Science.gov (United States)

    Oosterink, J Efraim; Buijs, Nikki; van Goudoever, Johannes B; Schierbeek, Henk

    2014-05-01

    Stable isotope studies offer the opportunity to study the in-depth metabolic pathway of glutamine, citrulline, and arginine amino acids involved in NO synthesis. The use of multiple stable isotopes can be used to elucidate the exact transformation of glutamine to citrulline and arginine de novo synthesis. This novel method provides a purification step using cation exchange resin in combination with a rapid and easy derivatization procedure for a precise and robust measurement of the concentration and isotopic enrichments of NO synthesis-specific amino acids using a liquid chromatography mass spectrometry (LC/MS) ion trap system with high sensitivity and selectivity. The ethyl chloroformate derivatization procedure is beneficial in terms of robustness, velocity, simplicity, and derivative stability. In addition, the ethyl chloroformate derivatization can be performed at room temperature in an aqueous environment without incubation and the isolation of the derivatives from the reaction mixture also serves as a purification step. The concentration and enrichment of NO synthesis-specific amino acids as well as phenylalanine and tyrosine to determine protein turnover, were measured with good inter-day precision for the concentration (glutamine-citrulline-arginine pathway by using stable isotope studies.

  2. Inhibition of insulin fibrillation by osmolytes: Mechanistic Insights

    Science.gov (United States)

    Choudhary, Sinjan; Kishore, Nand; Hosur, Ramakrishna V.

    2015-11-01

    We have studied here using a number of biophysical tools the effects of osmolytes, betaine, citrulline, proline and sorbitol which differ significantly in terms of their physical characteristics such as, charge distribution, polarity, H-bonding abilities etc, on the fibrillation of insulin. Among these, betaine, citrulline, and proline are very effective in decreasing the extent of fibrillation. Proline also causes a substantial delay in the onset of fibrillation in the concentration range (50-250 mM) whereas such an effect is seen for citrulline only at 250 mM, and in case of betaine this effect is not seen at all in the whole concentration range. The enthalpies of interaction at various stages of fibrillation process have suggested that the preferential exclusion of the osmolyte and its polar interaction with the protein are important in inhibition. The results indicate that the osmolytes are most effective when added prior to the elongation stage of fibrillation. These observations have significant biological implications, since insulin fibrillation is known to cause injection amyloidosis and our data may help in designing lead drug molecules and development of potential therapeutic strategies.

  3. Posttranslational Protein Modification in the Salivary Glands of Sjögren’s Syndrome Patients

    Directory of Open Access Journals (Sweden)

    Rafael Herrera-Esparza

    2013-01-01

    Full Text Available The present study investigated posttranslational reactions in the salivary glands of patients with Sjögren’s syndrome. We analysed the biopsies of primary Sjögren’s patients using immunohistochemistry and a tag-purified anticyclic citrullinated protein (CCP antibody to detect citrullinated peptides, and the presence of peptidylarginine deiminase 2 (PAD2 was assessed simultaneously. The present work demonstrated the weak presence of the PAD2 enzyme in some normal salivary glands, although PAD2 expression was increased considerably in Sjögren’s patients. The presence of citrullinated proteins was also detected in the salivary tissues of Sjögren’s patients, which strongly supports the in situ posttranslational modification of proteins in this setting. Furthermore, the mutual expression of CCP and PAD2 suggests that this posttranslational modification is enzyme dependent. In conclusion, patients with Sjögren’s syndrome expressed the catalytic machinery to produce posttranslational reactions that may result in autoantigen triggering.

  4. HPLC-MS/MS investigation of biochemical markers for the disclosure of erythropoietin abuse in sports

    Science.gov (United States)

    Appolonova, S. A.; Dikunets, M. A.; Rodchenkov, G. M.

    2009-04-01

    The polypeptide hormone erythropoietin (EPO), which is a forbidden doping drug, was determined by high-performance liquid chromatography combined with tandem mass spectrometry (HPLC-MS/MS). The hypothesis about the influence of EPO on the asymmetric dimethylarginine (ADMA)-dimethylargininedime-thylaminohydrolase (DDAH)-NO-synthase system was verified. Changes in this system can serve as indirect biochemical markers of the presence of the forbidden EPO drug in the organism. In the test group, the concentrations of biochemical markers varied from 10 to 40 μg/ml for ADMA and symmetrical DMA (SDMA) and from 0.5 to 10 μg/ml for arginine and citrulline. A single intravenous administration of r-HuEPO (Epocrin, 2000 ME/day) for two volunteers reliably increased ADMA, SDMA, arginine, and citrulline concentrations to 40-270 μg/ml, 40-240μg/ml, 10-60 μg/ml, and 12-140 μg/ml, respectively, with respect to the reference values. The simultaneous increase in arginine, methylarginines, and citrulline contents could be an indirect marker of EPO abuse. The method is recommended for fast screening analysis.

  5. Inhibition of insulin fibrillation by osmolytes: Mechanistic insights.

    Science.gov (United States)

    Choudhary, Sinjan; Kishore, Nand; Hosur, Ramakrishna V

    2015-01-01

    We have studied here using a number of biophysical tools the effects of osmolytes, betaine, citrulline, proline and sorbitol which differ significantly in terms of their physical characteristics such as, charge distribution, polarity, H-bonding abilities etc, on the fibrillation of insulin. Among these, betaine, citrulline, and proline are very effective in decreasing the extent of fibrillation. Proline also causes a substantial delay in the onset of fibrillation in the concentration range (50-250 mM) whereas such an effect is seen for citrulline only at 250 mM, and in case of betaine this effect is not seen at all in the whole concentration range. The enthalpies of interaction at various stages of fibrillation process have suggested that the preferential exclusion of the osmolyte and its polar interaction with the protein are important in inhibition. The results indicate that the osmolytes are most effective when added prior to the elongation stage of fibrillation. These observations have significant biological implications, since insulin fibrillation is known to cause injection amyloidosis and our data may help in designing lead drug molecules and development of potential therapeutic strategies. PMID:26616401

  6. GAP1, a novel selection and counter-selection marker for multiple gene disruptions in Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    Regenberg, Birgitte; Hansen, J.

    2000-01-01

    We report on the use of a new homologous marker for use in multiple gene deletions in S, cerevisiae, the general amino acid permease gene (GAP1), A GAP1 strain can utilize L-citrulline as the sole nitrogen source but cannot grow in the presence of the toxic amino acid D-histidine, L-citrulline as...... well as D-histidine uptake is mediated solely by the general amino acid permease, and a gap1 strain is therefore able to grow in the presence of D-histidine but cannot utilize L-citrulline, Gene disruption is effected by transforming a gap1 strain with a gene cassette generated by PCR, containing GAP1...... the GAP1 gene. This is caused by recombination between two Salmonella typuimurium hisG direct repeats embracing GAP1, and will result in a sub-population of gap1 cells. Such cells are selected on a medium containing D-histidine, and may subsequently be used for a second gene disruption. Hence...

  7. Development of an optofluidic SERS-based biomedical sensor

    Science.gov (United States)

    Walton, Brian; Huang, Po-Jung; Kameoka, Jun; Deutz, Nicolaas; Coté, Gerard L.

    2016-03-01

    Rapid assessment of radiation exposure to sensitive organs like the gut is extremely important for large populations exposed to ionized radiation, for instance during warfare. Recent results have shown that plasma citrulline levels appear to track gut function after irradiation levels in mice and humans. The current ways to monitor blood citrulline levels are bulky, laborious, time-consuming and expensive methods. Therefore, an optofludic point-of-care (POC) system using surface enhanced Raman spectroscopy to measure plasma citrulline as a marker for radiation exposure that overcomes the above issues is being developed. As a first step toward development of this system four colloidal nanoparticles, spherical gold, silver cubes, silica-gold nanoshells, and silver-gold nanocages have been analyzed for use in the POC system. Transmission electron microscopy (TEM) images have been taken of each nanoparticle to visualize the morphology of the nanoparticles, which is vital for SERS. Ultraviolet-visible (UV/Vis) spectroscopy was also collected to verify the extinction spectra for each nanoparticle was in resonance with the excitation wavelength. The nanoparticles were functionalized with mercaptobenzoic acid (MBA), a Raman reporter molecule, and SERS spectra were collected to determine which has better utility in a novel micro-to-nanochannel. The data showed that the silver nanocubes have a larger enhancement factor than the gold nanospheres, nanoshells, or nanocages. Currently, these nanocubes are being functionalized with the citulline for assessing the concentration sensitivity and dynamic range for ultimate use as a marker for radiation.

  8. Four weeks of treatment with liraglutide reduces insulin dose without loss of glycemic control in type 1 diabetic patients with and without residual beta-cell function

    DEFF Research Database (Denmark)

    Kielgast, Urd; Holst, Jens Juul; Madsbad, Sten;

    2011-01-01

    without (C-peptide negative) were studied. All C-peptide-positive patients were treated with liraglutide plus insulin, whereas C-peptide-negative patients were randomly assigned to liraglutide plus insulin or insulin monotherapy. Continuous glucose monitoring with identical food intake and physical...... activity was performed before (week 0) and during (week 4) treatment. Differences in insulin dose; HbA(1c); time spent with blood glucose 10, and 3.9-9.9 mmol/L; and body weight were evaluated. RESULTS Insulin dose decreased from 0.50 ± 0.06 to 0.31 ± 0.08 units/kg per day (P ...-cell function at baseline, and two patients discontinued insulin treatment. In C-peptide-positive patients, time spent with blood glucose mean [range] -2.3 ± 0.3 kg [-0.5 to -5...

  9. Arginine depletion by arginine deiminase does not affect whole protein metabolism or muscle fractional protein synthesis rate in mice.

    Directory of Open Access Journals (Sweden)

    Juan C Marini

    Full Text Available Due to the absolute need for arginine that certain cancer cells have, arginine depletion is a therapy in clinical trials to treat several types of cancers. Arginine is an amino acids utilized not only as a precursor for other important molecules, but also for protein synthesis. Because arginine depletion can potentially exacerbate the progressive loss of body weight, and especially lean body mass, in cancer patients we determined the effect of arginine depletion by pegylated arginine deiminase (ADI-PEG 20 on whole body protein synthesis and fractional protein synthesis rate in multiple tissues of mice. ADI-PEG 20 successfully depleted circulating arginine (<1 μmol/L, and increased citrulline concentration more than tenfold. Body weight and body composition, however, were not affected by ADI-PEG 20. Despite the depletion of arginine, whole body protein synthesis and breakdown were maintained in the ADI-PEG 20 treated mice. The fractional protein synthesis rate of muscle was also not affected by arginine depletion. Most tissues (liver, kidney, spleen, heart, lungs, stomach, small and large intestine, pancreas were able to maintain their fractional protein synthesis rate; however, the fractional protein synthesis rate of brain, thymus and testicles was reduced due to the ADI-PEG 20 treatment. Furthermore, these results were confirmed by the incorporation of ureido [14C]citrulline, which indicate the local conversion into arginine, into protein. In conclusion, the intracellular recycling pathway of citrulline is able to provide enough arginine to maintain protein synthesis rate and prevent the loss of lean body mass and body weight.

  10. Metabolism via arginase or nitric oxide synthase: two competing arginine pathways in macrophages

    Directory of Open Access Journals (Sweden)

    Meera eRath

    2014-10-01

    Full Text Available Macrophages play a major role in the immune system, both as antimicrobial effector cells and as immunoregulatory cells, which induce, suppress or modulate adaptive immune responses. These key aspects of macrophage biology are fundamentally driven by the phenotype of macrophage arginine metabolism that is prevalent in an evolving or ongoing immune response. M1 macrophages express the enzyme nitric oxide synthase (NOS, which metabolizes arginine to nitric oxide (NO and citrulline. NO can be metabolized to further downstream reactive nitrogen species, while citrulline might be reused for efficient NO synthesis via the citrulline-NO cycle. M2 macrophages are characterized by expression of the enzyme arginase, which hydrolyzes arginine to ornithine and urea. The arginase pathway limits arginine availability for NO synthesis and ornithine itself can further feed into the important downstream pathways of polyamine and proline syntheses, which are important for cellular proliferation and tissue repair. M1 versus M2 polarization leads to opposing outcomes of inflammatory reactions, but depending on the context, M1 and M2 macrophages can be both pro- and antiinflammatory. Notably, M1/M2 macrophage polarization can be driven by microbial infection or innate danger signals without any influence of adaptive immune cells, secondarily driving the T helper (Th1/Th2 polarization of the evolving adaptive immune response. Since both arginine metabolic pathways cross-inhibit each other on the level of the respective arginine break-down products and Th1 and Th2 lymphocytes can drive or amplify macrophage M1/M2 dichotomy via cytokine activation, this forms the basis of a self-sustaining M1/M2 polarization of the whole immune response. Understanding the arginine metabolism of M1/M2 macrophage phenotypes is therefore central to find new possibilities to manipulate immune responses in infection, autoimmune diseases, chronic inflammatory conditions and cancer.

  11. A prospective comparison of acute intestinal toxicity following whole pelvic versus small field intensity-modulated radiotherapy for prostate cancer

    Directory of Open Access Journals (Sweden)

    Kim YJ

    2016-03-01

    Full Text Available Yeon Joo Kim, Jin-hong Park, In-Ha Yun, Young Seok KimDepartment of Radiation Oncology, Asan Medical Center, College of Medicine, University of Ulsan, Seoul, Republic of KoreaPurpose: To compare the acute intestinal toxicity of whole pelvic (WP and small field (SF intensity-modulated radiotherapy (IMRT for prostate cancer using dosimetric and metabolic parameters as well as clinical findings.Methods: Patients who received IMRT in either a definitive or postoperative setting were prospectively enrolled. Target volume and organs at risk including intestinal cavity (IC were delineated in every patient by a single physician. The IC volume that received a 10–50 Gy dose at 5-Gy intervals (V10–V50 and the percentage of irradiated volume as a fraction of total IC volume were calculated. Plasma citrulline levels, as an objective biological marker, were checked at three time points: baseline and after exposure to 30 Gy and 60 Gy.Results: Of the 41 patients, only six experienced grade 1 acute intestinal toxicity. Although all dose–volume parameters were significantly worse following WP than SF IMRT, there was no statistically significant relationship between these dosimetric parameters and clinical symptoms. Plasma citrulline levels did not show a serial decrease by radiotherapy volume difference (WP versus SF and were not relevant to the irradiated doses.Conclusion: Given that WP had comparable acute intestinal toxicities to those associated with SF, WP IMRT appears to be a feasible approach for the treatment of prostate cancer despite dosimetric disadvantages.Keywords: prostate cancer, intensity-modulated radiotherapy, intestinal toxicity, citrulline

  12. Increased erythrocytes by-products of arginine catabolism are associated with hyperglycemia and could be involved in the pathogenesis of type 2 diabetes mellitus.

    Science.gov (United States)

    Ramírez-Zamora, Serafín; Méndez-Rodríguez, Miguel L; Olguín-Martínez, Marisela; Sánchez-Sevilla, Lourdes; Quintana-Quintana, Miguel; García-García, Norberto; Hernández-Muñoz, Rolando

    2013-01-01

    Diabetes mellitus (DM) is a worldwide disease characterized by metabolic disturbances, frequently associated with high risk of atherosclerosis and renal and nervous system damage. Here, we assessed whether metabolites reflecting oxidative redox state, arginine and nitric oxide metabolism, are differentially distributed between serum and red blood cells (RBC), and whether significant metabolism of arginine exists in RBC. In 90 patients with type 2 DM without regular treatment for diabetes and 90 healthy controls, paired by age and gender, we measured serum and RBC levels of malondialdehyde (MDA), nitrites, ornithine, citrulline, and urea. In isolated RBC, metabolism of L-[(14)C]-arginine was also determined. In both groups, nitrites were equally distributed in serum and RBC; citrulline predominated in serum, whereas urea, arginine, and ornithine were found mainly in RBC. DM patients showed hyperglycemia and increased blood HbA1C, and increased levels of these metabolites, except for arginine, significantly correlating with blood glucose levels. RBC were observed to be capable of catabolizing arginine to ornithine, citrulline and urea, which was increased in RBC from DM patients, and correlated with an increased affinity for arginine in the activities of putative RBC arginase (Km = 0.23±0.06 vs. 0.50±0.13 mM, in controls) and nitric oxide synthase (Km = 0.28±0.06 vs. 0.43±0.09 mM, in controls). In conclusion, our results suggest that DM alters metabolite distribution between serum and RBC, demonstrating that RBC regulate serum levels of metabolites which affect nitrogen metabolism, not only by transporting them but also by metabolizing amino acids such as arginine. Moreover, we confirmed that urea can be produced also by human RBC besides hepatocytes, being much more evident in RBC from patients with type 2 DM. These events are probably involved in the specific physiopathology of this disease, i.e., endothelial damage and dysfunction.

  13. Amino acid residues important for substrate specificity of the amino acid permeases Can I p and Gnp I p in Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    Regenberg, Birgitte; Kielland-Brandt, M.C.

    2001-01-01

    in the arginine permease gene CAN1. One similar mutation was found in the glutamine-asparagine permease gene GNP1. L-[C-14]citrulline uptake measurements confirmed that suppressor mutations in CAN1 conferred uptake of this amino acid, while none of the mutant permeases had lost the ability to transport L-[C-14......]arginine. Substrate specificity seemed to remain narrow in most cases, and broad substrate specificity was only observed in the cases where mutations affect two proline residues (P148 and P313) that are both conserved in the amino acid-polyamine-choline (APC) transporter superfamily. We found mutations...

  14. Glucagon-like peptide-2 (GLP-2) increases net amino acid utilization by the portal-drained viscera of ruminating calves

    DEFF Research Database (Denmark)

    Taylor-Edwards, C C; Burrin, D G; Kristensen, N B;

    2012-01-01

    , potentially by increased small intestinal epithelial growth and thus energy and amino acid requirements of this tissue. Increased PDV extraction of glutamine and alterations in PDV metabolism of arginine, ornithine and citrulline support the concept that GLP-2 influences intestine-specific amino acid...... periods: baseline (saline infusion), treatment (infusion of bovine serum albumin or 3.76 μg/kg BW per h GLP-2) and recovery (saline infusion). Arterial concentrations and net PDV, hepatic and total splanchnic fluxes of glucose, lactate, glutamate, glutamine, β-hydroxybutyrate and urea-N were measured...

  15. Doxorubicin-Induced Gut Toxicity in Piglets fed Bovine Milk and Colostrum

    DEFF Research Database (Denmark)

    Shen, René Liang; Rathe, Mathias; Jiang, Pingping;

    2016-01-01

    gut permeability and plasma C-reactive protein (CRP) levels were increased (all P enzymes, pro-inflammatory cytokines or tight-junction proteins in the intestine. Increasing BC supplementation to doxorubicin-treated piglets (Study 2) had...... no consistent effects on plasma CRP and citrulline levels, intestinal morphology, digestive enzymes, permeability, or proinflammatory cytokines. However, Only-BC pigs had lower diarrhea severity towards the end of the experiment (P ... gut toxicity. Colostrum supplementation had limited effects on doxorubicin-induced toxicity in milk-fed piglets suggesting that colostrum and a bovine milk diet enriched with whey protein provided similar...

  16. Identification of Secreted Phosphoprotein 1 Gene as a new Rheumatoid Arthritis Susceptibility Gene

    OpenAIRE

    Gazal, Steven; Sacre, Karim; Allanore, Yannick; Teruel, Maria; Goodall, Alison,; Tohma, Shigeto; Alfredsson, Lars; Okada, Yukinori; Xie, Gang; Constantin, Arnaud; Balsa, Alejandro; Kawasaki, Aya; Nicaise, Pascale; Amos, Christopher; Rodriguez-Rodriguez, Luis

    2014-01-01

    International audience Objective To evaluate the contribution of the SPP1 rs11439060 and rs9138 polymorphisms, previously reported as autoimmune risk variants, in the RA genetic background according to anti-citrullinated protein antibodies (ACPAs) status of RA individuals. Methods We analyzed a total of 11,715 RA cases and 26,493 controls from 9 independent cohorts, all individuals were genotyped or had imputed genotypes for SPP1 rs11439060 and rs9138. The effect of the SPP1 rs11439060 and...

  17. 14CO2 fixation pattern of cyanobacteria

    International Nuclear Information System (INIS)

    The 14CO2 fixation pattern of three cyanobacteria in the light and dark were studied. Two different chromatographic methods widely used for separating labelled photosynthetic intermediates were compared. After ethanolic extraction, a rather uniform fixation pattern reflecting mainly the β-carboxylation pathway is obtained for all 3 species. Of the intermediates, glucosylglycerol is specific and high citrulline and low malate contents are fairly specific to cyanobacteria. The composition of the 14CO2 fixation pattern is hardly affected by changes in temperature or light intensity, but it is severely affected by changes in the water potential of the medium. (author)

  18. Metabolic alterations in children with environmental enteric dysfunction.

    Science.gov (United States)

    Semba, Richard D; Shardell, Michelle; Trehan, Indi; Moaddel, Ruin; Maleta, Kenneth M; Ordiz, M Isabel; Kraemer, Klaus; Khadeer, Mohammed; Ferrucci, Luigi; Manary, Mark J

    2016-01-01

    Environmental enteric dysfunction, an asymptomatic condition characterized by inflammation of the small bowel mucosa, villous atrophy, malabsorption, and increased intestinal permeability, is a major contributor to childhood stunting in low-income countries. Here we report the relationship of increased intestinal permeability with serum metabolites in 315 children without acute malnutrition, aged 12-59 months, in rural Malawi. Increased gut permeability was associated with significant differences in circulating metabolites that included lower serum phosphatidylcholines, sphingomyelins, tryptophan, ornithine, and citrulline, and elevated serum glutamate, taurine, and serotonin. Our findings suggest that environmental enteric dysfunction is characterized by alterations in important metabolites involved in growth and differentiation and gut function and integrity. PMID:27294788

  19. 正常及患病心脏中的一氧化氮和一氧化氮合酶亚型(英文)

    Institute of Scientific and Technical Information of China (English)

    李中言; Soban; Urma

    2009-01-01

    We have reviewed the role of nitric oxide synthase(NOS)isoforms and nitric oxide(NO)in the normal heart and in heart failure.NO is a highly reactive siganaling molecule produced normally in the heart and is an important modulator of myocardial function.NOS catalyzes the conversion of L-arginine to L-citrulline and NO.In the heart,three NOS isoforms are present:neuronal NOS(nNOS),and endothelial NOS(eNOS),are constitutively present enzymes in distinct subcellular locations within cardiomyocytes,whereas induc...

  20. The Nongenomic Effects of Progesterone in Repressing iNOS Activation through P38MAPK Pathways in Gonococci-Infected Polymorphonuclear Leukocytes and the Clinical Significance

    Institute of Scientific and Technical Information of China (English)

    陈嵘祎; 涂亚庭; 林加西; 佘惟槟; 李娟; 吴志洪; 许莉; 陈宏翔

    2010-01-01

    Progesterone has nongenomic effects on inducible nitric oxide synthase(iNOS),which is mediated by mitogen activated protein kinase(MAPK)pathways.This effect is supposed to have some potential association with asymptomatic gonococcal infections in women by immunological depression.In this study,polymorphonuclear leukocytes(PMNs)challenged by gonococci were used to study the nongenomic effects of progesterone.The activation of iNOS was assessed by measuring [3H] L-arginine converses to [3H] L-citrulline,and t...

  1. Selective Irreversible Inhibition of Neuronal and Inducible Nitric-oxide Synthase in the Combined Presence of Hydrogen Sulfide and Nitric Oxide*

    OpenAIRE

    Heine, Christian L.; Schmidt, Renate; Geckl, Kerstin; Schrammel, Astrid; Gesslbauer, Bernd; Schmidt, Kurt; Mayer, Bernd; Gorren, Antonius C. F.

    2015-01-01

    Citrulline formation by both human neuronal nitric-oxide synthase (nNOS) and mouse macrophage inducible NOS was inhibited by the hydrogen sulfide (H2S) donor Na2S with IC50 values of ∼2.4·10−5 and ∼7.9·10−5 m, respectively, whereas human endothelial NOS was hardly affected at all. Inhibition of nNOS was not affected by the concentrations of l-arginine (Arg), NADPH, FAD, FMN, tetrahydrobiopterin (BH4), and calmodulin, indicating that H2S does not interfere with substrate or cofactor binding. T...

  2. Smoking is associated with increased levels of extracellular peptidylarginine deiminase 2 (PAD2) in the lungs

    DEFF Research Database (Denmark)

    Damgaard, Dres; Friberg Bruun Nielsen, Michael; Quisgaard Gaunsbaek, Maria;

    2015-01-01

    lavage (BAL) fluid from smokers, but intracellularly located PAD cannot be responsible for citrullination of extracellular self-antigens. We aimed to establish a link between smoking and extracellular PAD2 in the lungs. METHODS: BAL fluid samples were obtained from 13 smokers and 11 nonsmoking controls...... fluids from smokers as compared to non-smokers (p=0.018). The PAD2 content correlated with the overall CRP levels (p=0.009) and cell count (p=0.016). CONCLUSIONS: This first demonstration of increased levels of extracellular PAD2 in the lungs of smokers supports the hypothesis that smoking promotes...

  3. Calcium-independent NO-synthase activity and nitrites/nitrates production in transient focal cerebral ischaemia in mice

    OpenAIRE

    Grandati, M; Verrecchia, C; Revaud, M L; Allix, M.; Boulu, R. G.; Plotkine, M.

    1997-01-01

    The temporal changes in constitutive NO-synthase (cNOS) and in calcium-independent NO-synthase activities were studied in mice subjected to 2 h of transient focal cerebral ischaemia. The changes in brain nitrites/nitrates (NOx) content were also studied.NOS activities were measured by the conversion of L-[14C]-arginine to L-[14C]-citrulline. Brain NOx contents were investigated by the Griess colourimetric method.cNOS activity in the infarcted cortical area was significantly reduced after 6 h ...

  4. 中国上市企业R&D投入的周期性特征研究——来自深圳中小板2008年金融危机期间的证据%Study on Cyclicality of R&D of the Listed Company in China: Evidence from Shenzhen Stock Market in Financial Crisis in 2008

    Institute of Scientific and Technical Information of China (English)

    关勇军; 洪开荣

    2012-01-01

    This paper reviews theory of the cyclicality of R&D, considers the effect of financial crisis on Chinese economy, takes 216 small-medium enterprise of Shenzhen stock market for sample, checks the R&D investment strategy and its affecting factors chosen by the listed company in China and draws the following conclusions: (1) only 22% of sample chosen the strategy of reducing R&D investment (pro-cyclicality behaviors) and back the theo- ry of opportunity cost;(2) the company with rich cash flow would choose the anti-cyclicality behaviors;(3) the com- pany with taking part in R&D coloration would choose the anti-cyclicality behaviors;(4) the high-tech company with high R&D intensity would choose the anti-cyclicality behaviors;(5) the company under high product competi- tive pressure would choose the pro-cyclicality behaviors.%在对研发投资周期性特征相关理论综述的基础上,结合2008年金融危机对中国经济的影响,以2006--2009年深圳中小板的216家企业为样本,考察了我国上市企业在金融危机期间所采取的研发投资策略及其影响因素。得出以下结论:(1)深圳中小板上市公司在金融危机期间仅有22%的企业采取了减少研发投资(顺周期)的策略,这支持了机会成本效应理论;(2)现金流量比较充裕的企业在金融危机期间会采取研发投入的逆周期行为;(3)参与研发合作的企业在金融危机期间会采取研发投入的逆周期行为;(4)研发强度大的高新技术企业在金融危机期间会采取研发投入的逆周期行为;(5)产品竞争激烈的企业在金融危机期间会采取研发投入的顺周期行为。

  5. Aloe vera affects changes induced in pulmonary tissue of mice caused by cigarette smoke inhalation.

    Science.gov (United States)

    Koul, Ashwani; Bala, Shashi; Yasmeen; Arora, Neha

    2015-09-01

    This study was undertaken to determine the influence of Aloe vera (AV) on changes induced in pulmonary tissue of cigarette smoke (CS) inhaling mice. CS inhalation for 4 weeks caused pulmonary damage as evident by histoarchitectural alterations and enhanced serum and tissue lactate dehydrogenase (LDH) activities. CS inhalation also led to increased mucin production as revealed by mucicarmine and Alcian Blue-Periodic Acid Schiff (AB-PAS) staining. Studies on bronchoalveolar lavage fluid (balf) of CS exposed animals revealed structural changes in phospholipids and increase in surface tension when compared with control counterparts. These changes were accompanied by enhanced nitric oxide (NO) levels, citrulline levels, peroxidative damage, and differential modulation of antioxidant defense system. AV administration (seven weeks, 500 mg/kg b.w. daily) to CS inhaling mice led to modulation of CS induced pulmonary changes as revealed by lesser degree of histoarchitectural alterations, lesser mucin production, decreased NO levels, citrulline levels, peroxidative damage, and serum LDH activity. AV treatment to CS inhaling mice was associated with varying response to antioxidant defense system, however balf of CS + AV treated animals did not exhibit appreciable changes when compared with that of CS exposed animals. These observations suggest that AV has the potential to modulate CS induced changes in the pulmonary tissue which could have implications in management of CS associated pulmonary diseases, however, further investigations are required to explore its complete mechanism of action.

  6. Inhibition of peptidyl-arginine deiminases reverses protein-hypercitrullination and disease in mouse models of multiple sclerosis

    Directory of Open Access Journals (Sweden)

    Mario A. Moscarello

    2013-03-01

    Multiple sclerosis (MS is the most common CNS-demyelinating disease of humans, showing clinical and pathological heterogeneity and a general resistance to therapy. We first discovered that abnormal myelin hypercitrullination, even in normal-appearing white matter, by peptidylarginine deiminases (PADs correlates strongly with disease severity and might have an important role in MS progression. Hypercitrullination is known to promote focal demyelination through reduced myelin compaction. Here we report that 2-chloroacetamidine (2CA, a small-molecule, PAD active-site inhibitor, dramatically attenuates disease at any stage in independent neurodegenerative as well as autoimmune MS mouse models. 2CA reduced PAD activity and protein citrullination to pre-disease status. In the autoimmune models, disease induction uniformly induced spontaneous hypercitrullination with citrulline+ epitopes targeted frequently. 2CA rapidly suppressed T cell autoreactivity, clearing brain and spinal cord infiltrates, through selective removal of newly activated T cells. 2CA essentially prevented disease when administered before disease onset or before autoimmune induction, making hypercitrullination, and specifically PAD enzymes, a therapeutic target in MS models and thus possibly in MS.

  7. Comparative aspects of tissue glutamine and proline metabolism.

    Science.gov (United States)

    Bertolo, Robert F; Burrin, Douglas G

    2008-10-01

    The cellular metabolism of glutamine and proline are closely interrelated, because they can be interconverted with glutamate and ornithine via the mitochondrial pathway involving pyrroline-5-carboxylate (P5C). In adults, glutamine and proline are converted via P5C to citrulline in the gut, then citrulline is converted to arginine in the kidney. In neonates, arginine is a semiindispensable amino acid and is synthesized from proline completely in the gut; because of low P5C synthase activity, glutamine is not an important precursor for neonatal arginine synthesis. Thus, splanchnic metabolism of glutamine and proline is important, because both amino acids serve as key precursors for arginine synthesis with some developmental differences. Studies investigating splanchnic extraction demonstrate that about two-thirds of dietary glutamine and almost all dietary glutamate are extracted on first pass and the vast majority is oxidized in the gut. This capacity to extract glutamine and glutamate appears to be very large, so diets high in glutamine or glutamate probably have little impact on circulating concentrations and consequent potential toxicity. In contrast, it appears that very little proline is extracted by the gut and liver, at least in the neonate, which may result in hyperprolinemia and potential toxicity. Therefore, the upper limits of safe dietary intake for glutamine and proline, and other amino acids, appear to be substantially different depending on the extent of first-pass splanchnic extraction and irreversible catabolism.

  8. Influence of phenolic compounds on the growth and arginine deiminase system in a wine lactic acid bacterium

    Directory of Open Access Journals (Sweden)

    María R. Alberto

    2012-03-01

    Full Text Available The influence of seven phenolic compounds, normally present in wine, on the growth and arginine deiminase system (ADI of Lactobacillus hilgardii X1B, a wine lactic acid bacterium, was established. This system provides energy for bacterial growth and produces citrulline that reacts with ethanol forming the carcinogen ethyl carbamate (EC, found in some wines. The influence of phenolic compounds on bacterial growth was compound dependent. Growth and final pH values increased in presence of arginine. Arginine consumption decreased in presence of protocatechuic and gallic acids (31 and 17%, respectively and increased in presence of quercetin, rutin, catechin and the caffeic and vanillic phenolic acids (between 10 and 13%, respectively. ADI enzyme activities varied in presence of phenolic compounds. Rutin, quercetin and caffeic and vanillic acids stimulated the enzyme arginine deiminase about 37-40%. Amounts of 200 mg/L gallic and protocatechuic acids inhibited the arginine deiminase enzyme between 53 and 100%, respectively. Ornithine transcarbamylase activity was not modified at all concentrations of phenolic compounds. As gallic and protocatechuic acids inhibited the arginine deiminase enzyme that produces citrulline, precursor of EC, these results are important considering the formation of toxic compounds.

  9. The role for neutrophil extracellular traps in cystic fibrosis autoimmunity

    Science.gov (United States)

    Skopelja, Sladjana; Hamilton, B. JoNell; Jones, Jonathan D.; Yang, Mei-Ling; Mamula, Mark; Ashare, Alix; Gifford, Alex H.; Rigby, William F.C.

    2016-01-01

    While respiratory failure in cystic fibrosis (CF) frequently associates with chronic infection by Pseudomonas aeruginosa, no single factor predicts the extent of lung damage in CF. To elucidate other causes, we studied the autoantibody profile in CF and rheumatoid arthritis (RA) patients, given the similar association of airway inflammation and autoimmunity in RA. Even though we observed that bactericidal permeability-increasing protein (BPI), carbamylated proteins, and citrullinated proteins all localized to the neutrophil extracellular traps (NETs), which are implicated in the development of autoimmunity, our study demonstrates striking autoantibody specificity in CF. Particularly, CF patients developed anti-BPI autoantibodies but hardly any anti-citrullinated protein autoantibodies (ACPA). In contrast, ACPA-positive RA patients exhibited no reactivity with BPI. Interestingly, anti-carbamylated protein autoantibodies (ACarPA) were found in both cohorts but did not cross-react with BPI. Contrary to ACPA and ACarPA, anti-BPI autoantibodies recognized the BPI C-terminus in the absence of posttranslational modifications. In fact, we discovered that P. aeruginosa–mediated NET formation results in BPI cleavage by P. aeruginosa elastase, which suggests a novel mechanism in the development of autoimmunity to BPI. In accordance with this model, autoantibodies associated with presence of P. aeruginosa on sputum culture. Finally, our results provide a role for autoimmunity in CF disease severity, as autoantibody levels associate with diminished lung function. PMID:27777975

  10. Effects of microalgae chlorella species crude extracts on intestinal adaptation in experimental short bowel syndrome

    Institute of Scientific and Technical Information of China (English)

    Mustafa Kerem; Bulent Salman; Hatice Pasaoglu; Abdulkadir Bedirli; Murat Alper; Hikmet Katircioglu; Tahir Atici; E Ferda Pertoin; Ebru Ofluoglu

    2008-01-01

    AIM: To evaluate the effects of chlorella crude extract (CCE) on intestinal adaptation in rats subjected to short bowel syndrome (SBS).METHODS: Wistar rats weighing 230-260 g were used in the study. After anesthesia a 75% small bowel resection was performed. Rats were randomized and divided into groups. Control group (n = 10): where 5% dextrose was given through a gastrostomy tube, Enteral nutrition (EN) group (n = 10): Isocaloric and isonitrogen EN (Alitraq, Abbott, USA), study group (/7 = 10): CCE was administrated through a gastrostomy tube. Rats were sacrificed on the fifteenth postoperative day and blood and tissue samples were taken. Histopathologic evaluation, intestinal mucosal protein and DNA levels,intestinal proliferation and apoptosis were determined in intestinal tissues, and total protein, albumin and citrulline levels in blood were studied.RESULTS: In rats receiving CCE, villus lengthening,crypt depth, mucosal DNA and protein levels, intestinal proliferation, and serum citrulline, protein and albumin levels were found to be significantly higher than those in control group. Apoptosis in CCE treated rats was significantly reduced when compared to EN group rats.CONCLUSION: CCE has beneficial effects on intestinala daptation in experimental SBS.

  11. Investigation of nutrient sensing in the yeast Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    Eckert-Boulet, Nadine

    2006-01-01

    -citrulline kan ikke optages af disse stammer, da de mangler GAP1 (General Amino acid Permease). Det blev bekræftet, at AAP generne TAT1, BAP2, BAP3 og PTR2 er under kontrol af aminosyre induktionsvejen, mens AGP2 blev identificeret som et nyt gen under kontrol af denne mekanisme. Den globale...... for at aminosyre induktion kan finde sted. Hel-genom transkriptionsanalyser af en vildtype stamme og en grr1 stamme med og uden L-citrullin blev udført. Dataanalyse viste at aminosyre induktion af AAP generne AGP1, BAP2, BAP3, DIP5, TAT1, og GNP1 er helt afhængig af tilstedeværelsen af GRR1. Sammenligningen af de....... RTS1 blev slået ud, og dette medførte konstitutiv aktivering af AGP1 og BAP2 promotorer. Dette var afhængigt af SSY1, PTR3, SSY5, GRR1 eller STP1, STP2 og deres homolog STP3. Den forøgede transkription fra AGP1 og BAP2 promotorer i rts1 celler ser ud til at skyldes forøget processering af Stp1p. Med...

  12. Rheumatoid Arthritis-Associated Interstitial Lung Disease and Idiopathic Pulmonary Fibrosis: Shared Mechanistic and Phenotypic Traits Suggest Overlapping Disease Mechanisms.

    Science.gov (United States)

    Paulin, Francisco; Doyle, Tracy J; Fletcher, Elaine A; Ascherman, Dana P; Rosas, Ivan O

    2015-01-01

    The prevalence of clinically evident interstitial lung disease in patients with rheumatoid arthritis is approximately 10%. An additional 33% of undiagnosed patients have interstitial lung abnormalities that can be detected with high-resolution computed tomography. Rheumatoid arthritis-interstitial lung disease patients have three times the risk of death compared to those with rheumatoid arthritis occurring in the absence of interstitial lung disease, and the mortality related to interstitial lung disease is rising. Rheumatoid arthritis-interstitial lung disease is most commonly classified as the usual interstitial pneumonia pattern, overlapping mechanistically and phenotypically with idiopathic pulmonary fibrosis, but can occur in a non-usual interstitial pneumonia pattern, mainly nonspecific interstitial pneumonia. Based on this, we propose two possible pathways to explain the coexistence of rheumatoid arthritis and interstitial lung disease: (i) Rheumatoid arthritis-interstitial lung disease with a non-usual interstitial pneumonia pattern may come about when an immune response against citrullinated peptides taking place in another site (e.g. the joints) subsequently affects the lungs; (ii) Rheumatoid arthritis-interstitial lung disease with a usual interstitial pneumonia pattern may represent a disease process in which idiopathic pulmonary fibrosis-like pathology triggers an immune response against citrullinated proteins that promotes articular disease indicative of rheumatoid arthritis. More studies focused on elucidating the basic mechanisms leading to different sub-phenotypes of rheumatoid arthritis-interstitial lung disease and the overlap with idiopathic pulmonary fibrosis are necessary to improve our understanding of the disease process and to define new therapeutic targets.

  13. Inducible nitric oxide synthase is expressed in synovial fluid granulocytes

    Science.gov (United States)

    CEDERGREN, J; FORSLUND 2, T; SUNDQVIST 2, T; SKOGH 1, T

    2002-01-01

    The objective of the study was to evaluate the NO-producing potential of synovial fluid (SF) cells. SF from 15 patients with arthritis was compared with blood from the same individuals and with blood from 10 healthy controls. Cellular expression of inducible nitric oxide synthase (iNOS) was analysed by flow cytometry. High-performance liquid chromatography was used to measure l-arginine and l-citrulline. Nitrite and nitrate were measured colourimetrically utilizing the Griess’ reaction. Compared to whole blood granulocytes in patients with chronic arthritis, a prominent iNOS expression was observed in SF granulocytes (P < 0·001). A slight, but statistically significant, increase in iNOS expression was also recorded in lymphocytes and monocytes from SF. l-arginine was elevated in SF compared to serum (257 ± 78 versus 176 ± 65 µmol/l, P = 0·008), whereas a slight increase in l-citrulline (33 ± 11 versus 26 ± 9 µmol/l), did not reach statistical significance. Great variations but no significant differences were observed comparing serum and SF levels of nitrite and nitrate, respectively, although the sum of nitrite and nitrate tended to be elevated in SF (19·2 ± 20·7 versus 8·6 ± 6·5 µmol/l, P = 0·054). Synovial fluid leucocytes, in particular granulocytes, express iNOS and may thus contribute to intra-articular NO production in arthritis. PMID:12296866

  14. A Comparative Metabolomic Evaluation of Behcet's Disease with Arthritis and Seronegative Arthritis Using Synovial Fluid.

    Directory of Open Access Journals (Sweden)

    Joong Kyong Ahn

    Full Text Available Behcet's disease (BD with arthritis is often confused with seronegative arthritis (SNA because of shared clinical symptoms and the lack of definitive biomarkers for BD. To investigate possible metabolic patterns and potential biomarkers of BD with arthritis, metabolomic profiling of synovial fluid (SF from 6 patients with BD with arthritis and 18 patients with SNA was performed using gas chromatography/time-of-flight mass spectrometry in conjunction with univariate and multivariate statistical analyses. A total of 123 metabolites were identified from samples. Orthogonal partial least square-discriminant analysis showed clear discrimination between BD with arthritis and SNA. A set of 11 metabolites were identified as potential biomarkers for BD using variable importance for projection values and the Wilcoxon-Mann-Whitney test. Compared with SNA, BD with arthritis exhibited relatively high levels of glutamate, valine, citramalate, leucine, methionine sulfoxide, glycerate, phosphate, lysine, isoleucine, urea, and citrulline. There were two markers identified, elevated methionine sulfoxide and citrulline, that were associated with increased oxidative stress, providing a potential link to BD-associated neutrophil hyperactivity. Glutamate, citramalate, and valine were selected and validated as putative biomarkers for BD with arthritis (sensitivity, 100%; specificity, 61.1%. This is the first report to present potential biomarkers from SF for discriminating BD with arthritis from SNA. The metabolomics of SF may be helpful in searching for potential biomarkers and elucidating the clinicopathogenesis of BD with arthritis.

  15. Arginine depletion by arginine deiminase does not affect whole protein metabolism or muscle fractional protein synthesis rate in mice.

    Science.gov (United States)

    Marini, Juan C; Didelija, Inka Cajo

    2015-01-01

    Due to the absolute need for arginine that certain cancer cells have, arginine depletion is a therapy in clinical trials to treat several types of cancers. Arginine is an amino acids utilized not only as a precursor for other important molecules, but also for protein synthesis. Because arginine depletion can potentially exacerbate the progressive loss of body weight, and especially lean body mass, in cancer patients we determined the effect of arginine depletion by pegylated arginine deiminase (ADI-PEG 20) on whole body protein synthesis and fractional protein synthesis rate in multiple tissues of mice. ADI-PEG 20 successfully depleted circulating arginine (pancreas) were able to maintain their fractional protein synthesis rate; however, the fractional protein synthesis rate of brain, thymus and testicles was reduced due to the ADI-PEG 20 treatment. Furthermore, these results were confirmed by the incorporation of ureido [14C]citrulline, which indicate the local conversion into arginine, into protein. In conclusion, the intracellular recycling pathway of citrulline is able to provide enough arginine to maintain protein synthesis rate and prevent the loss of lean body mass and body weight. PMID:25775142

  16. Arginase activity in mitochondria - An interfering factor in nitric oxide synthase activity assays

    International Nuclear Information System (INIS)

    Previously, in tightly controlled studies, using three independent, yet complementary techniques, we refuted the claim that a mitochondrial nitric oxide synthase (mtNOS) isoform exists within pure, rat liver mitochondria (MT). Of those techniques, the NOS-catalyzed [14C]-L-arginine to [14C]-L-citrulline conversion assay (NOS assay) with MT samples indicated a weak, radioactive signal that was NOS-independent . Aliquots of samples from the NOS assays were then extracted with acetone, separated by high performance thin-layer chromatography (HPTLC) and exposed to autoradiography. Results obtained from these samples showed no radioactive band for L-citrulline. However, a fast-migrating, diffuse, radioactive band was observed in the TLC lanes loaded with MT samples. In this manuscript, we identify and confirm that this radioactive signal in MT samples is due to the arginase-catalyzed conversion of [14C]-L-arginine to [14C]-urea. The current results, in addition to reconfirming the absence of NOS activity in rat liver MT, also show the need to include arginase inhibitors in studies using MT samples in order to avoid confounding results when using NOS activity assays.

  17. Cigarette Smoke Induces Immune Responses to Vimentin in both, Arthritis-Susceptible and -Resistant Humanized Mice.

    Science.gov (United States)

    Bidkar, Mitali; Vassallo, Robert; Luckey, David; Smart, Michele; Mouapi, Kelly; Taneja, Veena

    2016-01-01

    Rheumatoid arthritis (RA) is an autoimmune disease marked by chronic synovial inflammation and both, genetic and environmental factors are involved in its pathogenesis. Human leukocyte antigen (HLA) DRB1*0401 is associated with susceptibility to develop RA, while cigarette smoke (CS) exposure promotes seropositive disease with increased severity in DRB1*0401+ individuals. Smokers have higher levels of antibodies against citrullinated peptides. In this study, we determined whether the response to a known autoantigen, Vimentin (Vim) is shared epitope specific and how CS influences this response using transgenic-mice carrying RA-susceptible,*0401, and -resistant, *0402, genes. Following relatively brief exposure to CS, peptidyl arginine deiminase (PAD) enzyme expression was increased in murine lungs. Cigarette smoking led to production of Interferon (IFN)-γ with reduced levels of Interleukin (IL)-10 by splenocytes of *0401 mice. In contrast, CS augmented Th2 cytokines along with T-regulatory cells in *0402 mice. An increase in levels of antibodies to native and citrullinated Vim was observed in naïve mice of both strains following CS exposure. Our data showed that both arthritis-susceptible and -resistant mice can generate cellular and humoral immunity to Vim; however CS-induced modulation of host immunity is dependent on the interaction with the host HLA genes. PMID:27602574

  18. Enzymatic Modification of Soluble Cyanophycin Using the Type II Peptidyl Arginine Deiminase from Oryctolagus cuniculus.

    Science.gov (United States)

    Wiefel, Lars; Steinbüchel, Alexander

    2016-07-01

    An increased structural variety expands the number of putative applications for cyanophycin (multi-l-arginyl-poly-[l-aspartic acid], CGP). Therefore, structural modifications of CGP are of major interest; these are commonly obtained by modification and optimization of the bacterial producing strain or by chemical modification. In this study, an enzymatic modification of arginine side chains from lysine-rich CGP is demonstrated using the peptidyl arginine deiminase from Oryctolagus cuniculus, purified from Escherichia coli after heterologous expression. About 10% of the arginine side chains are converted to citrulline which corresponds to 4% of the polymer's total side chains. An inhibition of the reaction in the presence of small amounts of l-citrulline is observed, thereby explaining the low conversion rate. CGP dipeptides can be modified with about 7.5 mol% of the Asp-Arg dipeptides being converted to Asp-Cit. These results show that the enzymatic modification of CGP is feasible, opening up a whole new area of possible CGP modifications for further research. PMID:26953800

  19. Rheumatoid Arthritis: The Stride from Research to Clinical Practice

    Directory of Open Access Journals (Sweden)

    Ill-Min Chung

    2016-06-01

    Full Text Available Over 70 different genetic variants with a significant association with rheumatoid arthritis (RA have been discovered. Anti-citrullination protein antibodies (ACPA-positive RA variants are more well-defined than their ACPA-negative counterparts. The human leukocyte antigen, HLA-DRB1 locus remains the prime suspect in anti-citrullination protein antibodies (ACPA—positive RA. Different HLA-DRB1 alleles are linked to RA susceptibility across different ethnicities. With evolving techniques, like genome-wide association studies (GWAS and single nucleotide polymorphism (SNP arrays, more non-HLA susceptibility loci have been identified for both types of RA. However, the functional significance of only a handful of these variants is known. Their roles include increasing susceptibility to RA or in determining the speed at which the disease progresses. Additionally, a couple of variations are associated with protection from RA. Defining such clear-cut biological functions can aid in the clinical diagnosis and treatment of RA. Recent research has focused on the implication of microRNAs, with miR-146a widely studied. In addition to disease susceptibility, genetic variations that influence the efficacy and toxicity of anti-RA agents have also been identified. Polymorphisms in the MTHFR gene influence the effectiveness of methotrexate, the first line of therapy in RA. Larger studies are, however, needed to identify potential biomarkers for early disease identification and monitoring disease progression.

  20. Nitric oxide synthase expression in the opossum superior colliculus: a histochemical, immunohistochemical and biochemical study.

    Science.gov (United States)

    Giraldi-Guimarães, A; Tenório, F; Brüning, G; Mayer, B; Mendez-Otero, R; Cavalcante, L A

    1999-12-01

    The expression of neuronal nitric oxide synthase (nNOS) in the superior colliculus (SC) of the opossum Didelphis marsupialis was studied by NADPH diaphorase (NADPH-d) histochemistry and nNOS immunohistochemistry. In addition, the activity of nNOS was quantified by measurement of [(3)H]-L-arginine conversion to [(3)H]-L-citrulline in tissue extracts from SC superficial layers in opossums and rats. Our results show that the number of NADPH-d stained cells was small and virtually identical in stratum opticum (SO) and stratum griseum superficiale (SGS) and their staining was very light, particularly in SGS. Neuropil staining was heavier in the stratum zonale (SZ) than in SGS or SO. The intermediate and deep layers contained heavily stained cells and moderate neuropil staining. Surprisingly, nNOS-immunoreactive cells were far more numerous than NADPH-d+ cells in every layer. The production of [(3)H]-L-citrulline from [(3)H]-L-arginine in tissue extracts enriched in superficial layers indicated that nNOS specific activity is as high in the opossum as in the rat. Our results suggest that the location of nNOS-expressing neurons in retino-receptive layers may be related to inter-specific differences in the processing of visual information. PMID:10681601

  1. Mutation of Glu-361 in Human Endothelial Nitric-oxide Synthase Selectively Abolishes L-Arginine Binding without Perturbing the Behavior of Heme and Other Redox Centers

    Science.gov (United States)

    Chen, Pei-Feng; Tsai, Ah-Lim; Berka, Vladimir; Wu, Kenneth K.

    2010-01-01

    Nitric oxide (NO) and L-citrulline are formed from the oxidation of L-arginine by three different isoforms of NO synthase (NOS). Defining amino acid residues responsible for L-arginine binding and oxidation is a primary step toward a detailed understanding of the NOS reaction mechanisms and designing strategies for the selective inhibition of the individual isoform. We have altered Glu-361 in human endothelial NOS to Gln or Leu by site-directed mutagenesis and found that these mutations resulted in a complete loss of L-citrulline formation without disruption of the cytochrome c reductase and NADPH oxidase activities. Optical and EPR spectroscopic studies demonstrated that the Glu-361 mutants had similar spectra either in resting state or reduced CO-complex as the wild type. The heme ligand, imidazole, could induce a low spin state in both wild-type and Glu-361 mutants. However, unlike the wild-type enzyme, the low spin imidazole complex of Glu-361 mutants was not reversed to a high spin state by addition of either L-arginine, acetylguanidine, or 2-aminothiazole. Direct L-arginine binding could not be detected in the mutants either. These results strongly indicate that Glu-361 in human endothelial NOS is specifically involved in the interaction with L-arginine. Mutation of this residue abolished the L-arginine binding without disruption of other functional characteristics. PMID:9045621

  2. A prospective comparison of acute intestinal toxicity following whole pelvic versus small field intensity-modulated radiotherapy for prostate cancer

    Science.gov (United States)

    Kim, Yeon Joo; Park, Jin-hong; Yun, In-Ha; Kim, Young Seok

    2016-01-01

    Purpose To compare the acute intestinal toxicity of whole pelvic (WP) and small field (SF) intensity-modulated radiotherapy (IMRT) for prostate cancer using dosimetric and metabolic parameters as well as clinical findings. Methods Patients who received IMRT in either a definitive or postoperative setting were prospectively enrolled. Target volume and organs at risk including intestinal cavity (IC) were delineated in every patient by a single physician. The IC volume that received a 10–50 Gy dose at 5-Gy intervals (V10–V50) and the percentage of irradiated volume as a fraction of total IC volume were calculated. Plasma citrulline levels, as an objective biological marker, were checked at three time points: baseline and after exposure to 30 Gy and 60 Gy. Results Of the 41 patients, only six experienced grade 1 acute intestinal toxicity. Although all dose–volume parameters were significantly worse following WP than SF IMRT, there was no statistically significant relationship between these dosimetric parameters and clinical symptoms. Plasma citrulline levels did not show a serial decrease by radiotherapy volume difference (WP versus SF) and were not relevant to the irradiated doses. Conclusion Given that WP had comparable acute intestinal toxicities to those associated with SF, WP IMRT appears to be a feasible approach for the treatment of prostate cancer despite dosimetric disadvantages. PMID:27022287

  3. Markers of Perioperative Bowel Complications in Colorectal Surgery Patients

    Directory of Open Access Journals (Sweden)

    Radomír Hyšpler

    2015-01-01

    Full Text Available Colorectal cancer is a clinical condition whose treatment often involves intestinal resection. Such treatment frequently results in two major gastrointestinal complications after surgery: anastomotic leakage and prolonged ileus. Anastomotic leakage is a serious complication which, more often than not, is diagnosed late; to date, C-reactive protein is the only available diagnostic marker. A monocentric, prospective, open case-control study was performed in patients (n=117 undergoing colorectal surgery. Intestinal fatty acid binding protein (i-FABP, citrulline, D-lactate, exhaled hydrogen, Escherichia coli genomic DNA, and ischemia modified albumin (IMA were determined preoperatively, postoperatively, and on the following four consecutive days. Bacterial DNA was not detected in any sample, and i-FABP and D-lactate lacked any distinct potential to detect postoperative bowel complications. Exhaled breath hydrogen content showed unacceptably low sensitivity. However, citrulline turned out to be a specific marker for prolonged ileus on postoperative days 3-4. Using a cut-off value of 20 μmol/L, a sensitivity and specificity of ~75% was achieved on postoperative day 4. IMA was found to be an efficient predictor of anastomosis leak by calculating the difference between preoperative and postoperative values. This test had 100% sensitivity and 80% specificity and 100% negative and 20% positive predictive value.

  4. The T1405N carbamoyl phosphate synthetase polymorphism does not affect plasma arginine concentrations in preterm infants.

    Directory of Open Access Journals (Sweden)

    Rob M J Moonen

    Full Text Available BACKGROUND: A C-to-A nucleotide transversion (T1405N in the gene that encodes carbamoyl-phosphate synthetase 1 (CPS1 has been associated with changes in plasma concentrations of L-arginine in term and near term infants but not in adults. In preterm infants homozygosity for the CPS1 Thr1405 variant (CC genotype was associated with an increased risk of having necrotizing enterocolitis (NEC. Plasma L-arginine concentrations are decreased in preterm infants with NEC. AIM: To examine the putative association between the CPS1 T1405N polymorphism and plasma arginine concentrations in preterm infants. METHODS: Prospective multicenter cohort study. Plasma and DNA samples were collected from 128 preterm infants (<30 weeks between 6 and 12 hours after birth. Plasma amino acid and CPS1 T1405N polymorphism analysis were performed. RESULTS: Distribution of genotypes did not differ between the preterm (CC:CA:AA = 55.5%:33.6%:10.9%, n = 128 and term infants (CC:CA:AA = 54.2%:35.4%:10.4%, n = 96. There was no association between the CPS1 genotype and plasma L-arginine or L-citrulline concentration, or the ornithine to citrulline ratio, which varies inversely with CPS1 activity. Also the levels of asymmetric dimethylarginine, and symmetric dimethylarginine were not significantly different among the three genotypes. CONCLUSIONS: The present study in preterm infants did not confirm the earlier reported association between CPS1 genotype and L-arginine levels in term infants.

  5. Aloe vera affects changes induced in pulmonary tissue of mice caused by cigarette smoke inhalation.

    Science.gov (United States)

    Koul, Ashwani; Bala, Shashi; Yasmeen; Arora, Neha

    2015-09-01

    This study was undertaken to determine the influence of Aloe vera (AV) on changes induced in pulmonary tissue of cigarette smoke (CS) inhaling mice. CS inhalation for 4 weeks caused pulmonary damage as evident by histoarchitectural alterations and enhanced serum and tissue lactate dehydrogenase (LDH) activities. CS inhalation also led to increased mucin production as revealed by mucicarmine and Alcian Blue-Periodic Acid Schiff (AB-PAS) staining. Studies on bronchoalveolar lavage fluid (balf) of CS exposed animals revealed structural changes in phospholipids and increase in surface tension when compared with control counterparts. These changes were accompanied by enhanced nitric oxide (NO) levels, citrulline levels, peroxidative damage, and differential modulation of antioxidant defense system. AV administration (seven weeks, 500 mg/kg b.w. daily) to CS inhaling mice led to modulation of CS induced pulmonary changes as revealed by lesser degree of histoarchitectural alterations, lesser mucin production, decreased NO levels, citrulline levels, peroxidative damage, and serum LDH activity. AV treatment to CS inhaling mice was associated with varying response to antioxidant defense system, however balf of CS + AV treated animals did not exhibit appreciable changes when compared with that of CS exposed animals. These observations suggest that AV has the potential to modulate CS induced changes in the pulmonary tissue which could have implications in management of CS associated pulmonary diseases, however, further investigations are required to explore its complete mechanism of action. PMID:24615921

  6. Determination of homocitrulline in urine of patients with HHH syndrome by liquid chromatography tandem mass spectrometry.

    Science.gov (United States)

    Al-Dirbashi, Osama Y; Al-Hassnan, Zuhair N; Rashed, Mohamed S

    2006-12-01

    A liquid chromatography tandem mass spectrometric method is described for the analysis of homocitrulline in human urine, a key metabolite in the differential diagnosis of hyperammonemia, hyperornithinemia, homocitrullinuria (HHH) syndrome. Urine samples were prepared by mere five-fold dilution with a mixture of internal standards (2H2-citrulline and 2H3-creatinine) used for the simultaneous quantification of creatinine. Analytes were separated on a cyano column and eluted isocratically within seven min. Detection was achieved by monitoring transitions of 190 > 84 and 190 > 127 for homocitrulline, 178 > 115 for 2H2-citrulline, 114 > 44 for creatinine and 117 > 47 for 2H3-creatinine. Calibration curves were linear up to 100 micromol/L. Intraday (n = 7) and interday (n = 6) variations were less than 10%. In urine samples from three siblings confirmed to have HHH syndrome, homocitrulline levels were at 13.3 (74), 21.1 (50) and 108.2 (103) mmol/mol creatinine (micromol/L). Control values were 0-9 mmol/mol creatinine (n = 120). The current method solves specificity issues in homocitrulline determination often encountered with some ninhydrin-based systems (coelution with methionine) and some o-phthalaldehyde-based ones (coelution with taurine), and presents an attractive alternative with a relatively high throughput.

  7. Regulation of [15N]urea synthesis from [5-15N]glutamine. Role of pH, hormones, and pyruvate.

    Science.gov (United States)

    Nissim, I; Yudkoff, M; Brosnan, J T

    1996-12-01

    We have utilized both [5-15N]glutamine and [3-13C] pyruvate as metabolic tracers in order to: (i) examine the effect of pH, glucagon (GLU), or insulin on the precursor-product relationship between 15NH3, [15N]citrulline, and, thereby, [15N]urea synthesis and (ii) elucidate the mechanism(s) by which pyruvate stimulates [15N] urea synthesis. Hepatocytes isolated from rat were incubated at pH 6.8, 7.4, or 7.6 with 1 mM [5-15N]glutamine and 0.1 mM 14NH4Cl in the presence or the absence of [3-13C] pyruvate (2 mM). A separate series of experiments was performed at pH 7.4 in the presence of insulin or GLU. 15NH3 enrichment exceeded or was equal to that of [15N]citrulline under all conditions except for pH 7.6, when the 15N enrichment in citrulline exceeded that in ammonia. The formation of [15N]citrulline (atom % excess) was increased with higher pH. Flux through phosphate-dependent glutaminase (PDG) and [15N]urea synthesis were stimulated (p < 0.05) at pH 7.6 or with GLU and decreased (p < 0.05) at pH 6.8. Insulin had no significant effect on flux through PDG or on [15N]urea synthesis. Decreased [15N]urea production at pH 6.8 was associated with depleted aspartate and glutamate levels. Pyruvate attenuated this decrease in the aspartate and glutamate pools and stimulated [15N]urea synthesis. Production of Asp from pyruvate was increased with increasing medium pH. Approximately 80% of Asp was derived from [3-13C]pyruvate regardless of incubation pH or addition of hormone. Furthermore, approximately 20, 40, and 50% of the mitochondrial N-acetylglutamate (NAG) pool was derived from [3-13C]pyruvate at pH 6.8, 7.4, and 7.6, respectively. Both the concentration and formation of [13C]NAG from [3-13C]pyruvate were increased (p < 0.05) with glucagon and decreased (p < 0.05) with insulin or at pH 6.8. The data suggest a correlation between changes in [15N]urea synthesis and alterations in the level and synthesis of [13C]NAG from pyruvate. The current observations suggest that the

  8. BEHAVIOR OF THE TEN LARGEST BRAZILIAN BANKS DURING THE SUBPRIME CRISIS: AN ANALYSIS BASED ON FINANCIAL INDICATORS

    Directory of Open Access Journals (Sweden)

    Rosane Maria Pio da Silva

    2012-06-01

    Full Text Available The aim of this paper is to demonstrate the behavior of the ten largest Brazilian banks between June 2008 and September 2009, based on the analysis of financial indicators. Therefore, 16 three-monthly indices were calculated, extracted from financial statement information, which characterizes a documentary research. The indices were separated in five categories: liquidity, capital, profitability, income and market. The obtained results appointed that most financial institutions in the sample were able to manage their resources so as to gain conditions to maintain credit initially. Then, as from the first term of 2009, driven by public banks, they increased their credit operations. In addition, most banks revealed an anti-cyclical trend to encourage productive activities, preferably activities with higher liquidity levels, to the detriment of profitability, which reveals a more conservative attitude. Finally, it was verified that government initiatives, the Brazilian economic balance and the resources the banks offered helped to produce an environment to reactivate business activities during the most acute period of the subprime crisis.

  9. Brazilian economic performance since the emergence of the great recession: The effects of income distribution on consumption

    Directory of Open Access Journals (Sweden)

    Arestis Philip

    2016-01-01

    Full Text Available After a long period of unstable and low economic activity, Brazil achieved a relatively high economic growth with low inflation from 2004 to 2008, when the world scenario was favourable for the Brazilian trade balance. An incomes policy, focused on real increases in the minimum wage along with a credit boom, led to a decade of high consumption growth rates. High levels of consumption and exports, in turn, induced investment and stimulated manufacturing production, despite the real appreciation of the national currency. However, the Great Recession that emerged after the global financial crisis of 2007/2008 brought challenges to the Brazilian economic performance, with unpleasant consequences for the country’s GDP growth. Consumption, investment and exports have decelerated, despite anti-cyclical macroeconomic policies. In this setting, manufacturing production stagnated and GDP growth slowed down substantially, while imports continued rising considerably. The aim of this paper is to provide an explanation to the slowdown of Brazilian growth rates after the Great Recession. The main hypothesis is that consumption was the main source of effective demand in the country since 2003. However, Brazil has not yet been able to sustain manufacturing and economic growth without a more active government policy to stimulate productive investment.

  10. Legal Instruments of Regulation of Development of Banking Activity in Ukraine

    Directory of Open Access Journals (Sweden)

    Senyshch Pavlo M.

    2014-03-01

    Full Text Available The article considers main approaches to identification of essence of legal instruments of regulation of development of the banking activity, identifies the mechanism of legal regulation of the banking activity and its elements and justifies the system and form of legal regulation of the banking activity in Ukraine. It describes subjects of legal regulation of the banking activity at the international level, which are the Basel Committee on Banking Supervision, European Central Bank, IMF, International Financial Reporting Standards Foundation and others. The article considers specific features of the regulatory requirements of Basel II and Basel III and specific features of their introduction into the banking activity. It describes anti-cyclic measures offered by the Basel Committee, which should facilitate formation of such conditions, under which the banking sector could have a lower level of leverage and stability with respect to influence of system risks. Significant attention is paid to international instruments of regulation of the banking activity, which include the following legal acts: Uniform Rules for Collections, Uniform Customs and Practice for Documentary Credits, and Unified Rules for Loan Guarantees. The article shows that the share of subordinate legal acts is significant in the Ukrainian system of banking regulatory and legal acts since the state cannot operatively react to the changing processes in banking at the legislative level and, that is why, basic provisions on carrying out banking activity should be fixed in law.

  11. DIAGNOSTIC VALUE OF SEROLOGICAL MARKERS OF RHEUMATOID ARTHRITIS

    Directory of Open Access Journals (Sweden)

    Aleksei Leonidovich Maslaynski

    2012-01-01

    Full Text Available Rheumatoid arthritis (RA is a classic autoimmune disease associated with the production of wide range of autoantibodies, and their detection has diagnostic and prognostic implication. The objective of this study was to estimate the diagnostic value of antibodies against modified citrullinated vimentin (AMCV and nuclear antigen RA33 of the IgA rheumatoid factor (RF versus the value of routinely used profile of autoantibodies in diagnostic work-up of RA. Material and methods. 253 patients with RA prehistory of varying duration were included into the study group. The control group was comprised of 92 patients, including patients with seronegative spondyloarthropathies and diffuse connective tissue diseases, as well as sex and age matched healthy controls. Serum levels of IgM and IgA RF, antibodies against cyclic citrullinated peptide (ACCP, ACMV, anti-keratin antibodies (AKA, antibodies against RA33 antigen (ARA33 and antinuclear factor (ANF were measured in all patients and controls. Results and discussion. Diagnostic sensitivity of AMCV equaled 78%, ACCP — 77%, IgM RF — 71%, IgA RF — 43%, AKA — 43%, ARA33 — 31% and ANF — 31%. All anti-citrullinic antibodies (AKA, ACCP, ACMV were significantly more commonly associated with IgM RF. Among RF and ACCP seronegative patients ACMV were found in 24% cases with 20 IU/Ml detection threshold, and in 21% — with 30 IU/Ml, allowing to increase diagnostic specificity of the test up to 91% with the increment of diagnostic threshold. Incidence of ARA33 was not significantly different among the RF and ACCP positive or negative subgroups, thus making ARA33 an independent RA marker. Specificity of this marker was 87,9%, thus making it inferior to RF and ACCP by a composite of diagnostic characteristics. Conclusions. Integrated measurement of ACMV and ARA33 is a rational approach at the second stage of serologic testing work-up in suspected cases of RA onset, when initial RF and ACCP tests were negative.

  12. PADI2 is significantly associated with rheumatoid arthritis.

    Directory of Open Access Journals (Sweden)

    Xiaotian Chang

    Full Text Available Citrullination, a posttranslational modification of peptidyl arginine to citrulline, plays an essential role in rheumatoid arthritis (RA. Citrullination is catalyzed by a group of peptidylarginine deiminases (PADs including PADI 1, 2, 3, 4 and 6. Many studies have indicated that the gene encoding PADI4 is a factor in susceptibility to RA. Some studies have detected PADI2 expression in RA synovial tissues, suggesting that PADI2 also plays an important role in the disease. This study evaluated the possible association between the PADI2-encoding gene and RA. Seventeen tag SNPs across the PAD locus were genotyped using a custom-designed Illumina 96-SNP VeraCode microarray. Peripheral blood samples were collected from patients with RA (n = 267, ankylosing spondylitis (AS, n = 51 and healthy controls (n = 160. The results of genotyping were verified using Sequenom MassARRAY in an independent cohort of 307 patients with RA, 324 patients with AS and 509 healthy controls. A western blot analysis was performed using synovial tissue from patients with RA (n = 7, osteoarthritis (OA, n = 7 and AS (n = 5 to determine the levels of expression of PADI2. A microarray analysis revealed a significant association between three selected PADI2 SNPs (rs2235926, rs2057094, rs2076616 and the presence of RA. The increased susceptibility to RA associated with rs2235926 (OR = 1.706733, 95% CI = [1.576366-1.866587], p = 0.000839 and rs2057094 (OR = 1.360432, 95% CI = [1.065483-1.869482], p = 0.003291 was further confirmed by the Sequenom MassARRAY. No tag SNPs in the PADI2 locus showed a significant association with AS. Increased expression of PADI2 was detected in RA synovial tissues compared with samples from patients with OA and AS. PADI2 is significantly associated with RA and may be involved in the pathogenesis of the disease.

  13. Increased PADI4 expression in blood and tissues of patients with malignant tumors

    Science.gov (United States)

    2009-01-01

    Background Peptidylarginine deiminase type 4 (PAD4/PADI4) post-translationally converts peptidylarginine to citrulline. Recent studies suggest that PADI4 represses expression of p53-regulated genes via citrullination of histones at gene promoters. Methods Expression of PADI4 was investigated in various tumors and non-tumor tissues (n = 1673) as well as in A549, SKOV3 and U937 tumor cell lines by immunohistochemistry, real-time PCR, and western blot. Levels of PADI4 and citrullinated antithrombin (cAT) were investigated in the blood of patients with various tumors by ELISA (n = 1121). Results Immunohistochemistry detected significant PADI4 expression in various malignancies including breast carcinomas, lung adenocarcinomas, hepatocellular carcinomas, esophageal squamous cancer cells, colorectal adenocarcinomas, renal cancer cells, ovarian adenocarcinomas, endometrial carcinomas, uterine adenocarcinomas, bladder carcinomas, chondromas, as well as other metastatic carcinomas. However, PADI4 expression was not observed in benign leiomyomas of stomach, uterine myomas, endometrial hyperplasias, cervical polyps, teratomas, hydatidiform moles, trophoblastic cell hyperplasias, hyroid adenomas, hemangiomas, lymph hyperplasias, schwannomas, neurofibromas, lipomas, and cavernous hemangiomas of the liver. Additionally, PADI4 expression was not detected in non-tumor tissues including cholecystitis, cervicitis and synovitis of osteoarthritis, except in certain acutely inflamed tissues such as in gastritis and appendicitis. Quantitative PCR and western blot analysis showed higher PADI4 expression in gastric adenocarcinomas, lung adenocarcinomas, hepatocellular carcinomas, esophageal squamous cell cancers and breast cancers (n = 5 for each disease) than in the surrounding healthy tissues. Furthermore, western blot analysis detected PADI4 expression in cultured tumor cell lines. ELISA detected increased PADI4 and cAT levels in the blood of patients with various malignant tumors

  14. Role of the Cationic C-Terminal Segment of Melittin on Membrane Fragmentation.

    Science.gov (United States)

    Therrien, Alexandre; Fournier, Alain; Lafleur, Michel

    2016-05-01

    The widespread distribution of cationic antimicrobial peptides capable of membrane fragmentation in nature underlines their importance to living organisms. In the present work, we determined the impact of the electrostatic interactions associated with the cationic C-terminal segment of melittin, a 26-amino acid peptide from bee venom (net charge +6), on its binding to model membranes and on the resulting fragmentation. In order to detail the role played by the C-terminal charges, we prepared a melittin analogue for which the four cationic amino acids in positions 21-24 were substituted with the polar residue citrulline, providing a peptide with the same length and amphiphilicity but with a lower net charge (+2). We compared the peptide bilayer affinity and the membrane fragmentation for bilayers prepared from 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC)/1,2-dipalmitoyl-sn-glycero-3-phospho-l-serine (DPPS) mixtures. It is shown that neutralization of the C-terminal considerably increased melittin affinity for zwitterionic membranes. The unfavorable contribution associated with transferring the cationic C-terminal in a less polar environment was reduced, leaving the hydrophobic interactions, which drive the peptide insertion in bilayers, with limited counterbalancing interactions. The presence of negatively charged lipids (DPPS) in bilayers increased melittin binding by introducing attractive electrostatic interactions, the augmentation being, as expected, greater for native melittin than for its citrullinated analogue. The membrane fragmentation power of the peptide was shown to be controlled by electrostatic interactions and could be modulated by the charge carried by both the membrane and the lytic peptide. The analysis of the lipid composition of the extracted fragments from DPPC/DPPS bilayers revealed no lipid specificity. It is proposed that extended phase separations are more susceptible to lead to the extraction of a lipid species in a specific manner

  15. Increased PADI4 expression in blood and tissues of patients with malignant tumors

    Directory of Open Access Journals (Sweden)

    Zhao Yan

    2009-01-01

    Full Text Available Abstract Background Peptidylarginine deiminase type 4 (PAD4/PADI4 post-translationally converts peptidylarginine to citrulline. Recent studies suggest that PADI4 represses expression of p53-regulated genes via citrullination of histones at gene promoters. Methods Expression of PADI4 was investigated in various tumors and non-tumor tissues (n = 1673 as well as in A549, SKOV3 and U937 tumor cell lines by immunohistochemistry, real-time PCR, and western blot. Levels of PADI4 and citrullinated antithrombin (cAT were investigated in the blood of patients with various tumors by ELISA (n = 1121. Results Immunohistochemistry detected significant PADI4 expression in various malignancies including breast carcinomas, lung adenocarcinomas, hepatocellular carcinomas, esophageal squamous cancer cells, colorectal adenocarcinomas, renal cancer cells, ovarian adenocarcinomas, endometrial carcinomas, uterine adenocarcinomas, bladder carcinomas, chondromas, as well as other metastatic carcinomas. However, PADI4 expression was not observed in benign leiomyomas of stomach, uterine myomas, endometrial hyperplasias, cervical polyps, teratomas, hydatidiform moles, trophoblastic cell hyperplasias, hyroid adenomas, hemangiomas, lymph hyperplasias, schwannomas, neurofibromas, lipomas, and cavernous hemangiomas of the liver. Additionally, PADI4 expression was not detected in non-tumor tissues including cholecystitis, cervicitis and synovitis of osteoarthritis, except in certain acutely inflamed tissues such as in gastritis and appendicitis. Quantitative PCR and western blot analysis showed higher PADI4 expression in gastric adenocarcinomas, lung adenocarcinomas, hepatocellular carcinomas, esophageal squamous cell cancers and breast cancers (n = 5 for each disease than in the surrounding healthy tissues. Furthermore, western blot analysis detected PADI4 expression in cultured tumor cell lines. ELISA detected increased PADI4 and cAT levels in the blood of patients with

  16. Antepartum ornithine transcarbamylase deficiency.

    Science.gov (United States)

    Nakajima, Hitoshi; Sasaki, Yosuke; Maeda, Tadashi; Takeda, Masako; Hara, Noriko; Nakanishi, Kazushige; Urita, Yoshihisa; Hattori, Risa; Miura, Ken; Taniguchi, Tomoko

    2014-01-01

    Ornithine transcarbamylase deficiency (OTCD) is the most common type urea cycle enzyme deficiencies. This syndrome results from a deficiency of the mitochondrial enzyme ornithine transcarbamylase, which catalyzes the conversion of ornithine and carbamoyl phosphate to citrullin. Our case was a 28-year-old female diagnosed with OTCD following neurocognitive deficit during her first pregnancy. Although hyperammonemia was suspected as the cause of the patient's mental changes, there was no evidence of chronic liver disease. Plasma amino acid and urine organic acid analysis revealed OTCD. After combined modality treatment with arginine, sodium benzoate and hemodialysis, the patient's plasma ammonia level stabilized and her mental status returned to normal. At last she recovered without any damage left. PMID:25759629

  17. Potential biological indicators of multi-organ damage: Application to radiation accident victims; Bio-indicateurs potentiels d'atteinte multi-organe: application au cas des victimes d'irradiation accidentelles

    Energy Technology Data Exchange (ETDEWEB)

    Bertho, J.M.; Souidi, M.; Gourmelon, P. [Institut de Radioprotection et de Surete Nucleaire (IRSN), Dir. de la Radioprotection de l' Homme 92 - Fontenay-aux-Roses (France)

    2009-09-15

    Accidental irradiations induce a complex pathological situation, difficult to assess and to treat. However, recent results describing new biological indicators of radiation-induced damages such as Flt3-ligand, citrulline and oxy-sterol concentration in the plasma, together with results obtained in large animal models of high dose irradiation, allowed a better understanding of pathophysiological mechanisms induced by uncontrolled irradiations. This conducted to leave the classical paradigm of the acute radiation syndrome, described as the association of three individual syndromes, the hematopoietic syndrome, the gastro-intestinal syndrome and the cerebrovascular syndrome, in favour of a multiple organ dysfunction syndrome, with the implication of other organs and systems. Follow-up of victims from two recent radiation accidents brings a confirmation of the usefulness of the newly described biological indicators, and also a partial confirmation of this new concept of a multiple organ dysfunction syndrome. (authors)

  18. Azospirillum lectin – induced changes in the content of nitric oxide in wheat seedling roots

    Directory of Open Access Journals (Sweden)

    Alen’kina S.A.

    2010-11-01

    Full Text Available The lectin of Azospirillum brasilense Sp7 at 40 μg ml-1 elicited two peaks of induction of nitric oxide synthesis in the roots of wheat seedlings after 3 and 26 h of coincubation. The lectin of A. brasilense Sp7.2.3, a mutant defective in lectin activity, produced the same effect, but the activation of nitric oxide synthesis in the roots was less in the case of 26-h incubation. Exposure to the lectins for 3 h increased citrulline synthesis in the plant cell to the same extent. This finding indicated that the Azospirillum lectins activate nitric oxide production through the NO signal system of plants, thereby acting as inducers of adaptation processes in the roots of wheat seedlings.

  19. PACAP-38 but not VIP induces release of CGRP from trigeminal nucleus caudalis via a receptor distinct from the PAC1 receptor

    DEFF Research Database (Denmark)

    Jansen-Olesen, Inger; Baun, Michael; Amrutkar, Dipak V;

    2014-01-01

    nucleus caudalis (TNC) was quantified by EIA. Regulation of NOS-enzymes caused by VIP and PACAP was investigated in dura mater, TG and TNC by measuring the conversion of L-[3H]arginine to L-[3H]citrulline. Co-expression of PACAP, neuronal nitric oxide synthase (nNOS) and CGRP was explored....... Immunohistochemistry of PACAP and CGRP showed co-expression in TG and TNC. PACAP and nNOS were co-localized in TG, but not in TNC. PACAP was found to co-localize with glutamine synthetase in TG satellite glial cells. CONCLUSION: PACAP-38 cause release of CGRP from TNC but not from TG. We suggest that the release...

  20. Identification of amino acids in Securigera securidaca, a popular medicinal herb in Iranian folk medicine

    Directory of Open Access Journals (Sweden)

    S.E. Sadat-Ebrahimi

    2014-01-01

    Full Text Available Securigera securidaca (L. Degen & Dorfl grows in different parts of Iran. The seeds of the species are used in Iranian folk medicine as an anti-diabetic agent. Many studies have established hypoglycemic effects of amino acids and in the present investigation, amino acids of Securigera securidaca seeds have been evaluated. The ground seeds were extracted using petroleum ether, hot ethanol and ethanol 50%, respectively. ethanol 50% extract was chromatographed over cation exchanging resin and the resulting amino acid fraction was subjected to HPLC after OPA derivatization and the amino acids were identified by comparing to standards. The results evidenced the presence of 19 amino acids in the plant extract including alanine, arginine, asparagine, aspartic acid, citrulline, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, serine, threonine, tyrosine and valine. Considering the role of some amino acids in diabetes the above amino acids could be noted as hypoglycemic agents of the plant seeds but further studies are necessary.

  1. Sialylation converts arthritogenic IgG into inhibitors of collagen-induced arthritis.

    Science.gov (United States)

    Ohmi, Yuhsuke; Ise, Wataru; Harazono, Akira; Takakura, Daisuke; Fukuyama, Hidehiro; Baba, Yoshihiro; Narazaki, Masashi; Shoda, Hirofumi; Takahashi, Nobunori; Ohkawa, Yuki; Ji, Shuting; Sugiyama, Fumihiro; Fujio, Keishi; Kumanogoh, Atsushi; Yamamoto, Kazuhiko; Kawasaki, Nana; Kurosaki, Tomohiro; Takahashi, Yoshimasa; Furukawa, Koichi

    2016-01-01

    Rheumatoid arthritis (RA)-associated IgG antibodies such as anti-citrullinated protein antibodies (ACPAs) have diverse glycosylation variants; however, key sugar chains modulating the arthritogenic activity of IgG remain to be clarified. Here, we show that reduced sialylation is a common feature of RA-associated IgG in humans and in mouse models of arthritis. Genetically blocking sialylation in activated B cells results in exacerbation of joint inflammation in a collagen-induced arthritis (CIA) model. On the other hand, artificial sialylation of anti-type II collagen antibodies, including ACPAs, not only attenuates arthritogenic activity, but also suppresses the development of CIA in the antibody-infused mice, whereas sialylation of other IgG does not prevent CIA. Thus, our data demonstrate that sialylation levels control the arthritogenicity of RA-associated IgG, presenting a potential target for antigen-specific immunotherapy. PMID:27046227

  2. Autoantibodies to Posttranslational Modifications in Rheumatoid Arthritis

    Directory of Open Access Journals (Sweden)

    Agata N. Burska

    2014-01-01

    Full Text Available Autoantibodies have been associated with human pathologies for a long time, particularly with autoimmune diseases (AIDs. Rheumatoid factor (RF is known since the late 1930s to be associated with rheumatoid arthritis (RA. The discovery of anticitrullinated protein antibodies in the last century has changed this and other posttranslational modifications (PTM relevant to RA have since been described. Such PTM introduce neoepitopes in proteins that can generate novel autoantibody specificities. The recent recognition of these novel specificities in RA provides a unique opportunity to understand human B-cell development in vivo. In this paper, we will review the three of the main classes of PTMs already associated with RA: citrullination, carbamylation, and oxidation. With the advancement of research methodologies it should be expected that other autoantibodies against PTM proteins could be discovered in patients with autoimmune diseases. Many of such autoantibodies may provide significant biomarker potential.

  3. Smoking is associated with increased levels of extracellular peptidylarginine deiminase 2 (PAD2) in the lungs

    DEFF Research Database (Denmark)

    Damgaard, Dres; Friberg Bruun Nielsen, Michael; Quisgaard Gaunsbaek, Maria;

    2015-01-01

    OBJECTIVES: Smoking is a well-established risk factor in rheumatoid arthritis (RA), and citrullination of self-antigens plays a pathogenic role in the majority of patients. Increased numbers of peptidylarginine deiminase 2 (PAD2)-containing macrophages have been demonstrated in bronchoalveolar....... Total protein content and C-reactive protein (CRP) concentration were determined after separating cells from the samples. PAD2 content in cell-free BAL fluids was measured by means of a PAD2-specific sandwich ELISA. RESULTS: Significantly increased levels of soluble PAD2 were detected in cell-free BAL...... fluids from smokers as compared to non-smokers (p=0.018). The PAD2 content correlated with the overall CRP levels (p=0.009) and cell count (p=0.016). CONCLUSIONS: This first demonstration of increased levels of extracellular PAD2 in the lungs of smokers supports the hypothesis that smoking promotes...

  4. Inhibition of MMP-13 with modified polymer particles

    Science.gov (United States)

    Tran, Hai; Bratlie, Kaitlin M.

    2016-06-01

    Matrix metalloproteinases (MMPs) are proteases that destroy the extracellular matrix and have important roles in the foreign body response, wound healing, and disease. Of particular importance is the chronic wound environment in which MMP activity is increased, resulting in destruction of the de novo extracellular matrix. One potential treatment of these wounds would be to use dressings that are capable of inhibiting MMP activity. In this study, we examined the effect of seven polymer modifiers (2-amino-3-guanidinopropionic acid, arginine, carnitine, citrulline, creatine, 3-guanidino propionic acid, and Nw-nitro-L-arginine) on MMP-13 activity. MMP-13 is a collagenase that is present in chronic wounds and is zinc dependent. Our results showed that these polymer modifiers were able to inhibit MMP-13 activity to varying degrees. The mechanism of inhibition appears to be binding zinc to the modifiers.

  5. Smoking and polymorphisms of genes encoding mannose-binding lectin and surfactant protein-D in patients with rheumatoid arthritis

    DEFF Research Database (Denmark)

    Kristiansen, Malthe; Frisch, Morten; Madsen, Hans Ole;

    2014-01-01

    To investigate whether polymorphisms in genes coding for mannose-binding lectin (MBL) and surfactant protein-D (SP-D) are associated directly or by interaction with smoking with rheumatoid arthritis (RA), anti-citrullinated peptide antibody (ACPA) positive RA, and erosive RA. MBL2 genotypes, SFTPD...... genotype at codon 11, and HLA-shared epitope were determined in 456 patients with rheumatoid arthritis and 533 sex- and age-matched controls. Patients were grouped according to the presence of ACPA antibodies and RA-associated bone erosions and sub-stratified according to smoking status as never or ever...... smokers. Odds ratios with 95% confidence interval (OR, 95% CI) were calculated using multiple logistic regression analyses controlling for shared epitope. The low-producing SFTPD genotype was not associated with risk of RA or ACPA positive RA, but with erosive disease in the RA patients (OR = 1.8; 95% CI...

  6. Regulatory effect of arginine metabolism enzymes on ethyl carbamate formation during the fermentation of Chinese yellow rice wine%精氨酸代谢酶对传统黄酒发酵氨基甲酸乙酯产生的调控作用

    Institute of Scientific and Technical Information of China (English)

    方若思; 董亚晨; 徐腾洋; 范琳琳; 陈启和

    2013-01-01

    Summary Ethyl carbamate ( EC) is a potential carcinogenic compound in most of fermented products . It widely exists in the fermented foods , especially alcohol beverages , such as grape wine , Japanese sake and Chinese yellow rice wine , etc . Thus , the search for effective methods to reduce EC level in final products must be carried on and considered . It has been reported that the optimal production methods for grape wine and high quality strains were capable of reducing the level of EC in grape wine product . Arginine is one of the major amino acids found in grape wine , which can be decomposed to the precursors of EC by yeast and malolactic fermentation bacteria . The degradation of arginine by lactic acid bacteria via the arginine deiminase ( ADI) pathway has been reported . It involves three enzymes: ADI , ornithine transcarbamoylase ( OTC ) and carbamate kinase . The degradation of arginine metabolism produced ammonia , adenosine triphosphate , ornithine , carbamyl phosphate and some citrullines . Studies have shown that citrulline is a typical indicator of EC level in grape wine , and a correlation between citrulline production and ethyl carbamate formation by L actobacillus hilgardii isolated from grape wines is reported . As generally acknowledged , urea is considered as the major precursor of EC production in grape wine fermentation which can be degraded by urease .In this study , different disposals were taken during the brewing process . The control group was designed without adding any inhibitor , whereas in the experimental groups , 0 .001 , 0 .01 , 0 .2 mol/L L‐ornithine hydrochlorides were added , and 150 , 750 U ureases per 1 .5 kg solid at the eighth day were added as inhibitors , respectively . As a result , both the inhibitors acted well . The highest level of EC content in control group was 169 mg/L , while in the experimental groups with 0 .01mol/L L‐ornithine hydrochloride and 150 U urease , the EC contents were 78 .81 and 58 .55 mg

  7. Urinary Metabolomic Approach Provides New Insights into Distinct Metabolic Profiles of Glutamine and N-Carbamylglutamate Supplementation in Rats.

    Science.gov (United States)

    Liu, Guangmang; Cao, Wei; Fang, Tingting; Jia, Gang; Zhao, Hua; Chen, Xiaoling; Wu, Caimei; Wang, Jing

    2016-01-01

    Glutamine and N-carbamylglutamate can enhance growth performance and health in animals, but the underlying mechanisms are not yet elucidated. This study aimed to investigate the effect of glutamine and N-carbamylglutamate supplementation in rat metabolism. Thirty rats were fed a control, glutamine, or N-carbamylglutamate diet for four weeks. Urine samples were analyzed by nuclear magnetic resonance (NMR)-based metabolomics, specifically high-resolution ¹H NMR metabolic profiling combined with multivariate data analysis. Glutamine significantly increased the urine levels of acetamide, acetate, citrulline, creatinine, and methymalonate, and decreased the urine levels of ethanol and formate (p glutamine and N-carbamylglutamate could modify urinary metabolome related to nitrogen metabolism and gut microbiota metabolism. Moreover, N-carbamylglutamate could alter energy and lipid metabolism. These findings indicate that different arginine precursors may lead to differences in the biofluid profile in rats. PMID:27527211

  8. The role of immunomodulators on intestinal barrier homeostasis in experimental models.

    Science.gov (United States)

    Andrade, Maria Emília Rabelo; Araújo, Raquel Silva; de Barros, Patrícia Aparecida Vieira; Soares, Anne Danieli Nascimento; Abrantes, Fernanda Alves; Generoso, Simone de Vasconcelos; Fernandes, Simone Odília Antunes; Cardoso, Valbert Nascimento

    2015-12-01

    The intestinal epithelium is composed of specialized epithelial cells that form a physical and biochemical barrier to commensal and pathogenic microorganisms. However, dysregulation of the epithelial barrier function can lead to increased intestinal permeability and bacterial translocation across the intestinal mucosa, which contributes to local and systemic immune activation. The increase in these parameters is associated with inflammatory bowel disease, physical exercise under heat stress, intestinal obstruction, ischemia, and mucositis, among other conditions. Lately, there has been growing interest in immunomodulatory nutrients and probiotics that can regulate host immune and inflammatory responses and possibly restore the intestinal barrier. Immunomodulators such as amino acids (glutamine, arginine, tryptophan, and citrulline), fatty acids (short-chain and omega-3 fatty acids and conjugated linoleic acids), and probiotics (Bifidobacterium, Saccharomyces, and Lactobacillus) have been reported in the literature. Here, we review the critical roles of immunomodulatory nutrients in supporting gut barrier integrity and function. PMID:25660317

  9. Amino acid profile during exercise and training in Standardbreds.

    Science.gov (United States)

    Westermann, C M; Dorland, L; Wijnberg, I D; de Sain-van der Velden, M G M; van Breda, E; Barneveld, A; de Graaf-Roelfsema, E; Keizer, H A; van der Kolk, J H

    2011-08-01

    The objective of this study is to assess the influence of acute exercise, training and intensified training on the plasma amino acid profile. In a 32-week longitudinal study using 10 Standardbred horses, training was divided into four phases, including a phase of intensified training for five horses. At the end of each phase, a standardized exercise test, SET, was performed. Plasma amino acid concentrations before and after each SET were measured. Training significantly reduced mean plasma aspartic acid concentration, whereas exercise significantly increased the plasma concentrations of alanine, taurine, methionine, leucine, tyrosine and phenylalanine and reduced the plasma concentrations of glycine, ornithine, glutamine, citrulline and serine. Normally and intensified trained horses differed not significantly. It is concluded that amino acids should not be regarded as limiting training performance in Standardbreds except for aspartic acid which is the most likely candidate for supplementation. PMID:20863542

  10. [Basic research overview in rheumatoid arthritis].

    Science.gov (United States)

    Iwasaki, Yukiko; Yamamoto, Kazuhiko

    2016-06-01

    Rheumatoid arthritis (RA) is a common autoimmune disease with a prevalence of 0.5-1.0% worldwide. Although advances in understanding the pathogenesis of RA have led to new therapeutics with good outcomes, the real cause of the disease is still unknown. RA is characterized by synovial inflammation and hyperplasia, which erodes cartilage and bone, and autoantibody production (rheumatoid factor (RF) and anti-citrullinated protein antibody (ACPA)). There are many critical questions on the mechanism of the disease onset and progression: How genetic and environmental factors interact with each other? Why does the inflammatory response localize in joints? What are the key players to perpetuate synovial inflammation? In this review, we summarize pathogenetic advances in these issues especially from the point of view of basic research.

  11. Medical immunology: two-way bridge connecting bench and bedside.

    Science.gov (United States)

    Rijkers, Ger T; Damoiseaux, Jan G M C; Hooijkaas, Herbert

    2014-12-01

    Medical immunology in The Netherlands is a laboratory specialism dealing with immunological analyses as well as pre- and post-analytical consultation to clinicians (clinical immunologists and other specialists) involved in patients with immune mediated diseases. The scope of medical immunology includes immunodeficiencies, autoimmune diseases, allergy, transfusion and transplantation immunology, and lymphoproliferative disorders plus the monitoring of these patients. The training, professional criteria, quality control of procedures and laboratories is well organized. As examples of the bridge function of medical immunology between laboratory (bench) and patient (bedside) the contribution of medical immunologists to diagnosis and treatment of primary immunodeficiency diseases (in particular: humoral immunodeficiencies) as well as autoantibodies (anti-citrullinated proteins in rheumatoid arthritis) are given.

  12. Autoantigens as Partners in Initiation and Propagation of Autoimmune Rheumatic Diseases.

    Science.gov (United States)

    Rosen, Antony; Casciola-Rosen, Livia

    2016-05-20

    Systemic autoimmune diseases are characterized by specific targeting of a limited group of ubiquitously expressed autoantigens by the immune system. This review examines the mechanisms underlying their selection as immune targets. Initiation of autoimmune responses likely reflects the presentation of antigens with a distinct structure not previously encountered by the immune system, in a proimmune context (injury, malignancy, or infection). Causes of modified structure include somatic mutation and posttranslational modifications (including citrullination and proteolysis). Many autoantigens are components of multimolecular complexes, and some of the other components may provide adjuvant activity. Propagation of autoimmune responses appears to reflect a bidirectional interaction between the immune response and the target tissues in a mutually reinforcing cycle: Immune effector pathways generate additional autoantigen, which feeds further immune response. We propose that this resonance may be a critical principle underlying disease propagation, with specific autoantigens functioning as the hubs around which amplification occurs. PMID:26907212

  13. (1)H-Nuclear Magnetic Resonance-Based Plasma Metabolic Profiling of Dairy Cows with Fatty Liver.

    Science.gov (United States)

    Xu, Chuang; Sun, Ling-Wei; Xia, Cheng; Zhang, Hong-You; Zheng, Jia-San; Wang, Jun-Song

    2016-02-01

    Fatty liver is a common metabolic disorder of dairy cows during the transition period. Historically, the diagnosis of fatty liver has involved liver biopsy, biochemical or histological examination of liver specimens, and ultrasonographic imaging of the liver. However, more convenient and noninvasive methods would be beneficial for the diagnosis of fatty liver in dairy cows. The plasma metabolic profiles of dairy cows with fatty liver and normal (control) cows were investigated to identify new biomarkers using (1)H nuclear magnetic resonance. Compared with the control group, the primary differences in the fatty liver group included increases in β-hydroxybutyric acid, acetone, glycine, valine, trimethylamine-N-oxide, citrulline, and isobutyrate, and decreases in alanine, asparagine, glucose, γ-aminobutyric acid glycerol, and creatinine. This analysis revealed a global profile of endogenous metabolites, which may present potential biomarkers for the diagnosis of fatty liver in dairy cows. PMID:26732447

  14. (1)H-Nuclear Magnetic Resonance-Based Plasma Metabolic Profiling of Dairy Cows with Fatty Liver.

    Science.gov (United States)

    Xu, Chuang; Sun, Ling-Wei; Xia, Cheng; Zhang, Hong-You; Zheng, Jia-San; Wang, Jun-Song

    2016-02-01

    Fatty liver is a common metabolic disorder of dairy cows during the transition period. Historically, the diagnosis of fatty liver has involved liver biopsy, biochemical or histological examination of liver specimens, and ultrasonographic imaging of the liver. However, more convenient and noninvasive methods would be beneficial for the diagnosis of fatty liver in dairy cows. The plasma metabolic profiles of dairy cows with fatty liver and normal (control) cows were investigated to identify new biomarkers using (1)H nuclear magnetic resonance. Compared with the control group, the primary differences in the fatty liver group included increases in β-hydroxybutyric acid, acetone, glycine, valine, trimethylamine-N-oxide, citrulline, and isobutyrate, and decreases in alanine, asparagine, glucose, γ-aminobutyric acid glycerol, and creatinine. This analysis revealed a global profile of endogenous metabolites, which may present potential biomarkers for the diagnosis of fatty liver in dairy cows.

  15. Integrated analysis of COX-2 and iNOS derived inflammatory mediators in LPS-stimulated RAW macrophages pre-exposed to Echium plantagineum L. bee pollen extract.

    Directory of Open Access Journals (Sweden)

    Eduarda Moita

    Full Text Available Oxidative stress and inflammation play important roles in disease development. This study intended to evaluate the anti-inflammatory and antioxidant potential of Echium plantagineum L. bee pollen to support its claimed health beneficial effects. The hydromethanol extract efficiently scavenged nitric oxide ((•NO although against superoxide (O2(•- it behaved as antioxidant at lower concentrations and as pro-oxidant at higher concentrations. The anti-inflammatory potential was evaluated in LPS-stimulated macrophages. The levels of (•NO and L-citrulline decreased for all extract concentrations tested, while the levels of prostaglandins, their metabolites and isoprostanes, evaluated by UPLC-MS, decreased with low extract concentrations. So, E. plantagineum bee pollen extract can exert anti-inflammatory activity by reducing (•NO and prostaglandins. The extract is able to scavenge the reactive species (•NO and O2(•- and reduce markers of oxidative stress in cells at low concentrations.

  16. Musculoskeletal involvement in systemic sclerosis.

    Science.gov (United States)

    Randone, Silvia Bellando; Guiducci, Serena; Cerinic, Marco Matucci

    2008-04-01

    Musculoskeletal involvement is more frequent than expected in patients with systemic sclerosis (SSc) and is a major cause of disability, even if the prognosis of the disease largely depends on visceral involvement. The most common clinical feature of musculoskeletal involvement is arthralgia; less frequent features are arthritis, flexion contractures, stiffness (affecting predominantly fingers, wrists and ankles), proximal muscle weakness (mainly of the shoulder and hip) and tendon sheath involvement. Tendon friction rubs are predictive of poor prognosis. If musculoskeletal involvement is suspected, serum creatinine phosphokinase, aldolase, lactate dehydrogenase, alkaline phosphate, rheumatoid factor and anticyclic citrullinated peptide autoantibodies should be checked routinely. Treatment for muscle involvement has not yet been considered adequately and, in the future, it is to be hoped that clinical trials will identify new drugs to control this aspect of SSc, which seriously compromises patients' quality of life. PMID:18455689

  17. Utilization of ornithine and arginine as specific precursors of clavulanic acid.

    Science.gov (United States)

    Romero, J; Liras, P; Martín, J F

    1986-01-01

    Ornithine and arginine (5 to 20 mM), but not glutamic acid or proline, exerted a concentration-dependent stimulatory effect on the biosynthesis of clavulanic acid in both resting-cell cultures and long-term fermentations of Streptomyces clavuligerus. Ornithine strongly inhibited cephamycin biosynthesis in the same strain. [1-14C]-, [5-14C]-, or [U-14 C] ornithine was efficiently incorporated into clavulanic acid, whereas the incorporation of uniformly labeled glutamic acid was very poor. [U-14C] citrulline were not incorporated at all. Mutant nca-1, a strain that is blocked in clavulanic acid biosynthesis, did not incorporate arginine into clavulanic acid. S. clavuligerus showed arginase activity, converting arginine into ornithine, but not amidinotransferase activity. Both arginase activity and clavulanic acid formation were enhanced simultaneously by supplementing the production medium with 10 mM arginine. PMID:2877616

  18. Plasma levels of nitric oxide related amino acids in demented subjects with Down syndrome are related to neopterin concentrations.

    Science.gov (United States)

    Coppus, A M W; Fekkes, D; Verhoeven, W M A; Tuinier, S; van Duijn, C M

    2010-03-01

    Subjects with Down syndrome (DS) have abnormalities in virtually all aspects of the immune system and almost all will be affected with Alzheimer's disease (AD). It is thought that nitric oxide (NO) is involved in the pathophysiology of AD. In the present study, including a total of 401 elderly DS subjects, the spectrum of plasma amino acids and neopterin was investigated and related to development of AD. Concentrations of nearly all amino acids in DS subjects differed significantly from those of healthy controls. Neopterin was increased in DS subjects, especially in dementia. The production of NO as reflected by an increased citrulline/arginine ratio (Cit/Arg ratio) was enhanced during development of clinical dementia. Neopterin concentrations correlated to the Cit/Arg ratio only in the group of prevalent demented subjects (rho = 0.48, P = 0.006). The results of this study are suggestive for an increase in oxidative processes in DS subjects with AD.

  19. Left ventricular deformation by speckle tracking echocardiography at 2-year follow-up in treatment naive rheumatoid arthritis patients

    DEFF Research Database (Denmark)

    Logstrup, B. B.; Masic, D.; Laurberg, T. B.;

    2015-01-01

    Background: Patients with rheumatoid arthritis (RA) are at increased risk of cardiovascular disease (CVD) including heart failure. The role of autoimmunity and especially citrullination in the pathogenesis of the increased risk of CVD and the time-course of left ventricular (LV) dysfunction is...... unknown. Objectives: To assess the LV function according to the amount of inflammation during a 2-year follow-up period in a cohort of treatment naive RA patients without known CVD. Methods: Thirty-nine patients (21 women) with mean age 59+/-10.7 years and steroid- and disease-modifying antirheumatic drug...... (DMARD)-naive early RA were included and followed for 2 years. All patients were treated with methotrexate and followed the National treatment algoritm. Disease activity was assessed at baseline and at 2-year follow-up by the use of the Danish national DANBIO registry. IgM rheumafactor (IgM-RF) and anti...

  20. Hyperornithinemia-hyperammonemia-homocitrullinuria syndrome (HHH) presenting with acute fulminant hepatic failure.

    Science.gov (United States)

    Mhanni, A A; Chan, A; Collison, M; Seifert, B; Lehotay, D C; Sokoro, Ah; Huynh, H Q; Greenberg, C R

    2008-03-01

    We report on two Aboriginal patients with the hyperornithinemia-hyperammonemia-homocitrullinuria (HHH) syndrome. Both presented with acute hepatic failure with severe hypertransaminasemia and coagulopathy, prompting evaluation for emergent liver transplantation. The diagnosis of HHH syndrome was based on the presence of typical metabolic abnormalities. A protein-restricted diet and L-arginine or L-citrulline supplementation were immediately started, with rapid normalization of liver function test results and other biochemical abnormalities. Molecular analysis of the SLC25A15 gene showed that the two patients were homozygous for the common French Canadian mutation (F188Delta). The diagnosis of HHH syndrome should be considered in patients with unexplained fulminant hepatic failure. There does not appear to be a genotype-phenotype correlation for this presentation, inasmuch as the only other reported patient presenting with this picture had two different point mutations. Early identification and prompt treatment of these patients is crucial to avoid liver transplantation and can be life saving.

  1. Isotypes of Epstein-Barr virus antibodies in rheumatoid arthritis

    DEFF Research Database (Denmark)

    Westergaard, Marie Wulff; Draborg, Anette Holck; Troelsen, Lone;

    2015-01-01

    In order to study the humoral immune response against Epstein-Barr virus (EBV) in patients with rheumatoid arthritis (RA) and to compare it with the two major autoantibody types in RA, plasma samples from 77 RA patients, 28 patients with systemic lupus erythematosus (SLE), and 28 healthy controls...... (HCs) were investigated by enzyme-linked immunosorbent assays (ELISA). Increased percentages of positives and concentrations of IgG/IgA/IgM antibodies against the latent EBV nuclear antigen-1 (EBNA-1) were observed in RA patients compared to SLE patients and HCs. Increased concentrations......) and anti-citrullinated protein antibodies (ACPAs, IgG) and between elevated IgA concentrations against EAD and the presence of RFs and ACPAs in RA patients were found. Thus, RA patients had elevated antibodies of all isotypes characteristic of latent EBV infection (whereas SLE patients had elevated...

  2. Identification of macrophage extracellular trap-like structures in mammary gland adipose tissue: a preliminary study.

    Directory of Open Access Journals (Sweden)

    Sunish eMohanan

    2013-03-01

    Full Text Available PAD4-mediated hypercitrullination of histone H4 arginine 3 (H4R3 has been previously found to promote the formation of Neutrophil Extracellular Traps (NET in inflamed tissues and the resulting histone H4 citrulline 3 (H4Cit3 modification is thought to play a key role in extracellular trap (ET formation by promoting chromatin decondensation. In addition to neutrophils, macrophages have also recently been found to generate functional extracellular traps (METs. However, a role for PADs in ET formation in macrophages has not been previously described. Transcripts for PAD2 and PAD4 are found in mature macrophages and these cells can be induced to citrullinate proteins, thus raising the possibility that PADs may play a direct role in ET formation in macrophages via histone hypercitrullination. In breast and visceral white adipose tissue from obese patients, infiltrating macrophages are often seen to surround dead adipocytes forming characteristic crown-like structures (CLS and the presence of these lesions is associated with increased levels of inflammatory mediators. In light of these observations, we have initiated studies to test whether PADs are expressed in CLS macrophages and whether these macrophages might form METs. Our preliminary findings show that PAD2 (and to a lesser extent, PAD4 is expressed in both in the macrophage cell line (RAW 264.7 and in CLS lesions. Additionally, we provide evidence that macrophage-derived extracellular histones are seen around presumptive macrophages within CLS lesions and that these histones contain the H4Cit3 modification. These initial findings support our hypothesis that obesity-induced adipose tissue inflammation promotes the formation of METs within CLS lesions via PAD-mediated histone hypercitrullination. Subsequent studies are underway to further validate these findings and to investigate the role in PAD-mediated MET formation in CLS function in the mammary gland.

  3. An enzymatic deconjugation method for the analysis of small molecule active drugs on antibody-drug conjugates.

    Science.gov (United States)

    Li, Yi; Gu, Christine; Gruenhagen, Jason; Yehl, Peter; Chetwyn, Nik P; Medley, Colin D

    2016-01-01

    Antibody-drug conjugates (ADCs) are complex therapeutic agents that use the specific targeting properties of antibodies and the highly potent cytotoxicity of small molecule drugs to selectively eliminate tumor cells while limiting the toxicity to normal healthy tissues. Two critical quality attributes of ADCs are the purity and stability of the active small molecule drug linked to the ADC, but these are difficult to assess once the drug is conjugated to the antibody. In this study, we report a enzyme deconjugation approach to cleave small molecule drugs from ADCs, which allows the drugs to be subsequently characterized by reversed-phase high performance liquid chromatography. The model ADC we used in this study utilizes a valine-citrulline linker that is designed to be sensitive to endoproteases after internalization by tumor cells. We screened several proteases to determine the most effective enzyme. Among the 3 cysteine proteases evaluated, papain had the best efficiency in cleaving the small molecule drug from the model ADC. The deconjugation conditions were further optimized to achieve complete cleavage of the small molecule drug. This papain deconjugation approach demonstrated excellent specificity and precision. The purity and stability of the active drug on an ADC drug product was evaluated and the major degradation products of the active drug were identified. The papain deconjugation method was also applied to several other ADCs, with the results suggesting it could be applied generally to ADCs containing a valine-citrulline linker. Our results indicate that the papain deconjugation method is a powerful tool for characterizing the active small molecule drug conjugated to an ADC, and may be useful in ensuring the product quality, efficacy and the safety of ADCs. PMID:26891281

  4. A novel biochemically salvageable animal model of hyperammonemia devoid of N-acetylglutamate synthase.

    Science.gov (United States)

    Senkevitch, Emilee; Cabrera-Luque, Juan; Morizono, Hiroki; Caldovic, Ljubica; Tuchman, Mendel

    2012-06-01

    All knockout mouse models of urea cycle disorders die in the neonatal period or shortly thereafter. Since N-acetylglutamate synthase (NAGS) deficiency in humans can be effectively treated with N-carbamyl-l-glutamate (NCG), we sought to develop a mouse model of this disorder that could be rescued by biochemical intervention, reared to adulthood, reproduce, and become a novel animal model for hyperammonemia. Founder NAGS knockout heterozygous mice were obtained from the trans-NIH Knock-Out Mouse Project. Genotyping of the mice was performed by PCR and confirmed by Western blotting of liver and intestine. NCG and L-citrulline (Cit) were used to rescue the NAGS knockout homozygous (Nags(-/-)) pups and the rescued animals were characterized. We observed an 85% survival rate of Nags(-/-) mice when they were given intraperitoneal injections with NCG and Cit during the newborn period until weaning and supplemented subsequently with both compounds in their drinking water. This regimen has allowed for normal development, apparent health, and reproduction. Interruption of this rescue intervention resulted in the development of severe hyperammonemia and death within 48 h. In addition to hyperammonemia, interruption of rescue supplementation was associated with elevated plasma glutamine, glutamate, and lysine, and reduced citrulline, arginine, ornithine and proline levels. We conclude that NAGS deprived mouse model has been developed which can be rescued by NCG and Cit and reared to reproduction and beyond. This biochemically salvageable mouse model recapitulates the clinical phenotype of proximal urea cycle disorders and can be used as a reliable model of induced hyperammonemia by manipulating the administration of the rescue compounds. PMID:22503289

  5. Overexpression of arginase in the aged mouse penis impairs erectile function and decreases eNOS activity: influence of in vivo gene therapy of anti-arginase.

    Science.gov (United States)

    Bivalacqua, Trinity J; Burnett, Arthur L; Hellstrom, Wayne J G; Champion, Hunter C

    2007-03-01

    Since both increased nitric oxide (NO) synthase (NOS) abundance and diminished NO signaling have been reported in the aging penis, the role of NO in the adaptations of aging remains controversial. Here we tested the hypothesis that arginase, an enzyme that competes with NOS for the substrate l-arginine, contributes to erectile dysfunction with advanced age in the B6/129 mouse strain. Arginase protein abundance, mRNA expression, and enzyme activity were elevated in aged compared with young penile endothelial cells. In addition, endothelial NOS (NOS3) protein abundance was greater in aged versus young penile endothelial cells, whereas NOS activity and cGMP levels were reduced. Calcium-dependent l-arginine-to-l-citrulline conversion and cGMP formation increased significantly in aged mouse penes in the presence of the arginase inhibitor 2(S)-amino-6-boronohexanoic acid (ABH). However, there was no effect on l-arginine-to-l-citrulline conversion or cGMP accumulation in the endothelium from young mouse penes. To assess the functional role of arginase in the inhibition of NOS pathway responsiveness in the penis, we evaluated the effects of ABH and an adeno-associated virus encoding an antisense sequence to arginase I (AAVanti-arginase) on erectile function in vivo. ABH and AAVanti-arginase enhanced endothelium-dependent erectile responses in the aged mice without altering endothelium-independent responses. Paralleling our in vitro observations, ABH or AAVanti-arginase did not affect vascular responses in the young mice. Inhibition of the arginase pathway improves endothelial function in the aging penile circulation, suggesting that the arginase pathway may be exploited to improve erectile dysfunction associated with aging. PMID:17071735

  6. Toxoplasma gondii lacks the enzymes required for de novo arginine biosynthesis and arginine starvation triggers cyst formation.

    Science.gov (United States)

    Fox, Barbara A; Gigley, Jason P; Bzik, David J

    2004-03-01

    Two separate carbamoyl phosphate synthetase activities are required for the de novo synthesis of pyrimidines and arginine in most eukaryotes. Toxoplasma gondii is novel in possessing a single carbamoyl phosphate synthetase II gene that corresponds to a glutamine-dependent form required for pyrimidine biosynthesis. We therefore examined arginine acquisition in T. gondii to determine whether the single carbamoyl phosphate synthetase II activity could provide both pyrimidine and arginine biosynthesis. We found that arginine deprivation efficiently blocks the replication of intracellular T. gondii, yet has little effect on long-term parasite viability. Addition of citrulline, but not ornithine, rescues the growth defect observed in the absence of exogenous arginine. This rescue with citrulline is ablated when parasites are cultured in a human citrullinemia fibroblast cell line that is deficient in argininosuccinate synthetase activity. These results reveal the absence of genes and activities of the arginine biosynthetic pathway and demonstrate that T. gondii is an arginine auxotroph. Arginine starvation was also found to efficiently trigger differentiation of replicative tachyzoites into bradyzoites contained within stable cyst-like structures. These same parasites expressing bradyzoite antigens can be efficiently switched back to rapidly proliferating tachyzoites several weeks after arginine starvation. We hypothesise that the absence of gene activities that are essential for the biosynthesis of arginine from carbamoyl phosphate confers a selective advantage by increasing bradyzoite switching during the host response to T. gondii infection. These findings are consistent with a model of host-parasite evolution that allowed host control of bradyzoite induction by trading off virulence for increased transmission. PMID:15003493

  7. Hepatic adaptation compensates inactivation of intestinal arginine biosynthesis in suckling mice.

    Directory of Open Access Journals (Sweden)

    Vincent Marion

    Full Text Available Suckling mammals, including mice, differ from adults in the abundant expression of enzymes that synthesize arginine from citrulline in their enterocytes. To investigate the importance of the small-intestinal arginine synthesis for whole-body arginine production in suckling mice, we floxed exon 13 of the argininosuccinate synthetase (Ass gene, which codes for a key enzyme in arginine biosynthesis, and specifically and completely ablated Ass in enterocytes by crossing Ass (fl and Villin-Cre mice. Unexpectedly, Ass (fl/fl /VilCre (tg/- mice showed no developmental impairments. Amino-acid fluxes across the intestine, liver, and kidneys were calculated after determining the blood flow in the portal vein, and hepatic and renal arteries (86%, 14%, and 33%, respectively, of the transhepatic blood flow in 14-day-old mice. Relative to control mice, citrulline production in the splanchnic region of Ass (fl/fl /VilCre (tg/- mice doubled, while arginine production was abolished. Furthermore, the net production of arginine and most other amino acids in the liver of suckling control mice declined to naught or even changed to consumption in Ass (fl/fl /VilCre (tg/- mice, and had, thus, become remarkably similar to that of post-weaning wild-type mice, which no longer express arginine-biosynthesizing enzymes in their small intestine. The adaptive changes in liver function were accompanied by an increased expression of genes involved in arginine metabolism (Asl, Got1, Gpt2, Glud1, Arg1, and Arg2 and transport (Slc25a13, Slc25a15, and Slc3a2, whereas no such changes were found in the intestine. Our findings suggest that the genetic premature deletion of arginine synthesis in enterocytes causes a premature induction of the post-weaning pattern of amino-acid metabolism in the liver.

  8. Expression pattern of a nuclear encoded mitochondrial arginine-ornithine translocator gene from Arabidopsis

    Directory of Open Access Journals (Sweden)

    Schneider Anja

    2003-01-01

    Full Text Available Abstract Background Arginine and citrulline serve as nitrogen storage forms, but are also involved in biosynthetic and catabolic pathways. Metabolism of arginine, citrulline and ornithine is distributed between mitochondria and cytosol. For the shuttle of intermediates between cytosol and mitochondria transporters present on the inner mitochondrial membrane are required. Yeast contains a mitochondrial translocator for ornithine and arginine, Ort1p/Arg11p. Ort1p/Arg11p is a member of the mitochondrial carrier family (MCF essential for ornithine export from mitochondria. The yeast arg11 mutant, which is deficient in Ort1p/Arg11p grows poorly on media lacking arginine. Results High-level expression of a nuclear encoded Arabidopsis thaliana homolog (AtmBAC2 of Ort1p/Arg11p was able to suppress the growth deficiency of arg11. RT-PCR analysis demonstrated expression of AtmBAC2 in all tissues with highest levels in flowers. Promoter-GUS fusions showed preferential expression in flowers, i.e. pollen, in the vasculature of siliques and in aborted seeds. Variable expression was observed in leaf vasculature. Induction of the promoter was not observed during the first two weeks in seedlings grown on media containing NH4NO3, arginine or ornithine as sole nitrogen sources. Conclusion AtmBAC2 was isolated as a mitochondrial transporter for arginine in Arabidopsis. The absence of expression in developing seeds and in cotyledons of seedlings indicates that other transporters are responsible for storage and mobilization of arginine in seeds.

  9. Pneumolysin activates neutrophil extracellular trap formation.

    Science.gov (United States)

    G Nel, J; Theron, A J; Durandt, C; Tintinger, G R; Pool, R; Mitchell, T J; Feldman, C; Anderson, R

    2016-06-01

    The primary objective of the current study was to investigate the potential of the pneumococcal toxin, pneumolysin (Ply), to activate neutrophil extracellular trap (NET) formation in vitro. Isolated human blood neutrophils were exposed to recombinant Ply (5-20 ng ml(-1) ) for 30-90 min at 37°C and NET formation measured using the following procedures to detect extracellular DNA: (i) flow cytometry using Vybrant® DyeCycle™ Ruby; (ii) spectrofluorimetry using the fluorophore, Sytox(®) Orange (5 μM); and (iii) NanoDrop(®) technology. These procedures were complemented by fluorescence microscopy using 4', 6-diamino-2-phenylindole (DAPI) (nuclear stain) in combination with anti-citrullinated histone monoclonal antibodies to visualize nets. Exposure of neutrophils to Ply resulted in relatively rapid (detected within 30-60 min), statistically significant (P DNA impregnated with both citrullinated histone and myeloperoxidase. Microscopy revealed that NETosis appeared to be restricted to a subpopulation of neutrophils, the numbers of NET-forming cells in the control and Ply-treated systems (10 and 20 ng ml(-1) ) were 4·3 (4·2), 14.3 (9·9) and 16·5 (7·5), respectively (n = 4, P < 0·0001 for comparison of the control with both Ply-treated systems). Ply-induced NETosis occurred in the setting of retention of cell viability, and apparent lack of involvement of reactive oxygen species and Toll-like receptor 4. In conclusion, Ply induces vital NETosis in human neutrophils, a process which may either contribute to host defence or worsen disease severity, depending on the intensity of the inflammatory response during pneumococcal infection. PMID:26749379

  10. New aspects on the structure of neutrophil extracellular traps from chronic obstructive pulmonary disease and in vitro generation.

    Directory of Open Access Journals (Sweden)

    Astrid Obermayer

    Full Text Available Polymorphonuclear neutrophils have in recent years attracted new attention due to their ability to release neutrophil extracellular traps (NETs. These web-like extracellular structures deriving from nuclear chromatin have been depicted in ambiguous roles between antimicrobial defence and host tissue damage. NETs consist of DNA strands of varying thickness and are decorated with microbicidal and cytotoxic proteins. Their principal structure has in recent years been characterised at molecular and ultrastructural levels but many features that are of direct relevance to cytotoxicity are still incompletely understood. These include the extent of chromatin decondensation during NET formation and the relative amounts and spatial distribution of the microbicidal components within the NET. In the present work, we analyse the structure of NETs found in induced sputum of patients with acutely exacerbated chronic obstructive pulmonary disease (COPD using confocal laser microscopy and electron microscopy. In vitro induced NETs from human neutrophils serve for purposes of comparison and extended analysis of NET structure. Results demonstrate that COPD sputa are characterised by the pronounced presence of NETs and NETotic neutrophils. We provide new evidence that chromatin decondensation during NETosis is most extensive and generates substantial amounts of double-helix DNA in 'beads-on-a-string' conformation. New information is also presented on the abundance and location of neutrophil elastase (NE and citrullinated histone H3 (citH3. NE occurs in high densities in nearly all non-fibrous constituents of the NETs while citH3 is much less abundant. We conclude from the results that (i NETosis is an integral part of COPD pathology; this is relevant to all future research on the etiology and therapy of the disease; and that (ii release of 'beads-on-a-string' DNA studded with non-citrullinated histones is a common feature of in vivo NETosis; this is of relevance to

  11. Citrullus lanatus 'sentinel' (watermelon) extract reduces atherosclerosis in LDL receptor-deficient mice.

    Science.gov (United States)

    Poduri, Aruna; Rateri, Debra L; Saha, Shubin K; Saha, Sibu; Daugherty, Alan

    2013-05-01

    Watermelon (Citrullus lanatus or C. lanatus) has many potentially bioactive compounds including citrulline, which may influence atherosclerosis. In this study, we determined the effects of C. lanatus, provided as an extract of the cultivar 'sentinel,' on hypercholesterolemia-induced atherosclerosis in mice. Male low-density lipoprotein receptor-deficient mice at 8 weeks old were given either C. lanatus 'sentinel' extract (2% vol/vol; n=10) or a mixture of matching carbohydrates (2% vol/vol; n=8) as the control in drinking water while being fed a saturated fat-enriched diet for 12 weeks ad libitum. Mice consuming C. lanatus 'sentinel' extract had significantly increased plasma citrulline concentrations. Systolic blood pressure was comparable between the two groups. Consumption of C. lanatus 'sentinel' extract led to lower body weight and fat mass without influencing lean mass. There were no differences in food and water intake and in urine output between the two groups. C. lanatus 'sentinel' extract administration decreased plasma cholesterol concentrations that were attributed to reductions of intermediate-/low-density lipoprotein cholesterol. Plasma concentrations of monocyte chemoattractant protein-1 and interferon-gamma were decreased and those of interleukin-10 were increased in mice consuming C. lanatus 'sentinel' extract. Intake of C. lanatus 'sentinel' extract resulted in reductions of atherosclerosis in both aortic arch and thoracic regions. In conclusion, consumption of C. lanatus 'sentinel' extract led to reduced body weight gain, decreased plasma cholesterol concentrations, improved homeostasis of pro- and anti-inflammatory cytokines, and attenuated development of atherosclerosis without affecting systolic blood pressure in hypercholesterolemic mice. PMID:22902326

  12. Neutrophil extracellular traps in sheep mastitis.

    Science.gov (United States)

    Pisanu, Salvatore; Cubeddu, Tiziana; Pagnozzi, Daniela; Rocca, Stefano; Cacciotto, Carla; Alberti, Alberto; Marogna, Gavino; Uzzau, Sergio; Addis, Maria Filippa

    2015-01-01

    Neutrophil extracellular traps (NETs) are structures composed of DNA, histones, and antimicrobial proteins that are released extracellularly by neutrophils and other immune cells as a means for trapping and killing invading pathogens. Here, we describe NET formation in milk and in mammary alveoli of mastitic sheep, and provide a dataset of proteins found in association to these structures. Nucleic acid staining, immunomicroscopy and fluorescent in-situ hybridization of mastitic mammary tissue from sheep infected with Streptococcus uberis demonstrated the presence of extranuclear DNA colocalizing with antimicrobial proteins, histones, and bacteria. Then, proteomic analysis by LTQ-Orbitrap Velos mass spectrometry provided detailed information on protein abundance changes occurring in milk upon infection. As a result, 1095 unique proteins were identified, of which 287 being significantly more abundant in mastitic milk. Upon protein ontology classification, the most represented localization classes for upregulated proteins were the cytoplasmic granule, the nucleus, and the mitochondrion, while function classes were mostly related to immune defence and inflammation pathways. All known NET markers were massively increased, including histones, granule proteases, and antimicrobial proteins. Of note was the detection of protein arginine deiminases (PAD3 and PAD4). These enzymes are responsible for citrullination, the post-translational modification that is known to trigger NET formation by inducing chromatin decondensation and extracellular release of NETs. As a further observation, citrullinated residues were detected by tandem mass spectrometry in histones of samples from mastitic animals. In conclusion, this work provides novel microscopic and proteomic information on NETs formed in vivo in the mammary gland, and reports the most complete database of proteins increased in milk upon bacterial mastitis. PMID:26088507

  13. Protection of DFP-induced oxidative damage and neurodegeneration by antioxidants and NMDA receptor antagonist

    International Nuclear Information System (INIS)

    Prophylactic agents acutely administered in response to anticholinesterases intoxication can prevent toxic symptoms, including fasciculations, seizures, convulsions and death. However, anticholinesterases also have long-term unknown pathophysiological effects, making rational prophylaxis/treatment problematic. Increasing evidence suggests that in addition to excessive cholinergic stimulation, organophosphate compounds such as diisopropylphosphorofluoridate (DFP) induce activation of glutamatergic neurons, generation of reactive oxygen (ROS) and nitrogen species (RNS), leading to neurodegeneration. The present study investigated multiple affectors of DFP exposure critical to cerebral oxidative damage and whether antioxidants and NMDA receptor antagonist memantine provide neuroprotection by preventing DFP-induced biochemical and morphometric changes in rat brain. Rats treated acutely with DFP (1.25 mg/kg, s.c.) developed onset of toxicity signs within 7-15 min that progressed to maximal severity of seizures and fasciculations within 60 min. At this time point, DFP caused significant (p 2-isoprostanes, F2-IsoPs; and F4-neuroprostanes, F4-NeuroPs), RNS (citrulline), and declines in high-energy phosphates (HEP) in rat cerebrum. At the same time, quantitative morphometric analysis of pyramidal neurons of the hippocampal CA1 region revealed significant (p 2-IsoPs, F4-NeuroPs, citrulline, and depletion of HEP were noted. Furthermore, attenuation in oxidative damage following antioxidants or memantine pretreatment was accompanied by rescue from dendritic degeneration of pyramidal neurons in the CA1 hippocampal area. These findings closely associated DFP-induced lipid peroxidation with dendritic degeneration of pyramidal neurons in the CA1 hippocampal area and point to possible interventions to limit oxidative injury and dendritic degeneration induced by anticholinesterase neurotoxicity.

  14. Analysis of the Expression and Activity of Nitric Oxide Synthase from Marine Photosynthetic Microorganisms.

    Science.gov (United States)

    Foresi, Noelia; Correa-Aragunde, Natalia; Santolini, Jerome; Lamattina, Lorenzo

    2016-01-01

    Nitric oxide (NO) functions as a signaling molecule in many biological processes in species belonging to all kingdoms of life. In animal cells, NO is synthesized primarily by NO synthase (NOS), an enzyme that catalyze the NADPH-dependent oxidation of L-arginine to NO and L-citrulline. Three NOS isoforms have been identified, the constitutive neuronal NOS (nNOS) and endothelial NOS (eNOS) and one inducible (iNOS). Plant NO synthesis is complex and is a matter of ongoing investigation and debate. Despite evidence of an Arg-dependent pathway for NO synthesis in plants, no plant NOS homologs to animal forms have been identified to date. In plants, there is also evidence for a nitrate-dependent mechanism of NO synthesis, catalyzed by cytosolic nitrate reductase. The existence of a NOS enzyme in the plant kingdom, from the tiny single-celled green alga Ostreococcus tauri was reported in 2010. O. tauri shares a common ancestor with higher plants and is considered to be part of an early diverging class within the green plant lineage.In this chapter we describe detailed protocols to study the expression and characterization of the enzymatic activity of NOS from O. tauri. The most used methods for the characterization of a canonical NOS are the analysis of spectral properties of the oxyferrous complex in the heme domain, the oxyhemoglobin (oxyHb) and citrulline assays and the NADPH oxidation for in vitro analysis of its activity or the use of fluorescent probes and Griess assay for in vivo NO determination. We further discuss the advantages and drawbacks of each method. Finally, we remark factors associated to the measurement of NOS activity in photosynthetic organisms that can generate misunderstandings in the interpretation of results. PMID:27094418

  15. Immunocytochemical localization and identification of prosomatostatin gene products in medullary carcinoma of human thyroid gland.

    Science.gov (United States)

    Scopsi, L; Ferrari, C; Pilotti, S; Holst, J J; Rehfeld, J F; Fossati, G; Rilke, F

    1990-08-01

    Thirty-three cases of histologically proven calcitonin-positive medullary thyroid carcinoma were studied immunocytochemically for the occurrence of prosomatostatin-related peptides. Positive cells, identified with a panel of antisera raised against four different regions of the prosomatostatin molecule, were found in 100% of the tumors. Most but not all somatostatin-positive cells were also immunoreactive for calcitonin. Notably, seven patients harboring somatostatin-rich tumors revealed a more favorable clinical course. The results (1) indicate that somatostatin production is a universal concomitant of thyroid medullary carcinoma, (2) suggest that these cells are likely to produce a somatostatin precursor molecule similar to mammalian prosomatostatin, and (3) imply that somatostatin-reactive cells may have as yet unknown roles in these tumors, possibly in the realm of paracrine and autocrine regulation of cell growth.

  16. Early arthritis in children – first results of prospective follow up

    Directory of Open Access Journals (Sweden)

    S. O. Salugina

    2007-01-01

    Full Text Available Objective. To perform a prospective investigation of clinico-laboratory and sonographic picture of juvenile arthritis (JA onset, to follow up its evolution during the first years of the disease, to prove the program of appropriate treatment.Material and methods. 128 pts with early JA (61% girls and 39% boys aged 1,5 to 16 years were included. Disease duration varied from 2 weeks to 6 months. General health, joint status, disease activity, functional class (Steinbroker, systemic and organ disturbances, laboratory activity indices, immunologic parameters (ANF, RF were assessed. Antibodies to cyclic citrullinated peptide (ACCP were evaluated in 38 pts by immunofluorescence method (IFM with commercial kits “Axis Shild Diagnostics Limited, UK”, antibodies to modified citrullinated vimentin (MCV were assessed in 60 pts by IFM with commercial kits “Orgentec” (Germany. Pts were examined every 3 months, the main investigations were repeated after 6, 12 and 24 months. Accuracy of the diagnosis was checked at every examination.Results. Mono- oligoarticular variant of onset was found in 68,6% polyarticular – in 21,4%, systemic – in 10% of pts. Knee and ankle joints were involved first (61% and 24% respectively, later – wrist (13% and hand joints (17%. Morning stiffness was absent in 85% of pts. 37% of pts had ANF and 13,2% - RF. 10,5% were ACCP positive and 28,3% – MCV positive. Antibodies correlated with polyarticular variant of JA and RF. Reactive arthritis was the preliminary diagnosis in 39% and JRA – in 21% of pts at inclusion. Later Juvenile chronic arthritis was the most frequent diagnosis (73-80%. Disease modifying antirheumatic drugs were prescribed early (before 6 months in 53% of pts. Exacerbations of the disease appeared in 1/3 of pts, more frequently – in pts with polyarthritis. After 12 months functional class 1 remained in 71,4% of pts.Conclusion. Stabilization of pathological process or remission can

  17. Metabolic and clinical response to Escherichia coli lipopolysaccharide in layer pullets of different genetic backgrounds supplied with graded dietary L-arginine.

    Science.gov (United States)

    Lieboldt, M A; Frahm, J; Halle, I; Görs, S; Schrader, L; Weigend, S; Preisinger, R; Metges, C C; Breves, G; Dänicke, S

    2016-03-01

    L-arginine (Arg) is an essential amino acid in birds that plays a decisive role in avian protein synthesis and immune response. Effects of graded dietary Arg supply on metabolic and clinical response to Escherichia coli lipopolysaccharide (LPS) were studied over 48 hours after a single intramuscular LPS injection in 18-week-old genetically diverse purebred pullets. LPS induced a genotype-specific fever response within 4 hours post injectionem. Whereas brown genotypes showed an initial hypothermia followed by longer-lasting moderate hyperthermia, white genotypes exhibited a biphasic hyperthermia without initial hypothermia. Furthermore, within 2 hours after LPS injection, sickness behavior characterized by lethargy, anorexia, intensified respiration, and ruffled feathers appeared, persisted for 3 to 5 hours and recovered 12 hours post injectionem. The varying grades of Arg did not alter the examined traits named above, whereas insufficient Arg reduced body growth and increased relative weights of liver and pancreas significantly. At 48 hours post injectionem, increased relative weights of liver and spleen were also found in LPS treated pullets, whereas LPS decreased those of pancreas, bursa, thymus, and cecal tonsils. Moreover, LPS lowered the sum of plasma amino acids and decreased plasma concentrations of Arg, citrulline, glutamate, methionine, ornithine, phenylalanine, proline, tryptophan, and tyrosine, and increased those of aspartate, glutamine, lysine, 1- and 3-methyl-histidine. Elevating concentrations of dietary Arg led to increasing plasma concentrations of Arg, citrulline, ornithine, and 3-methyl-histidine subsequently. As quantitative expression of LPS-induced anorexia, proteolysis, and the following changes in plasma amino acids, pullets showed a significant decrease of feed and nitrogen intake and catabolic metabolism characterized by negative nitrogen balance and body weight loss in the first 24 hours post injectionem. Pullets recovered from the

  18. Myelin basic protein as a novel genetic risk factor in rheumatoid arthritis--a genome-wide study combined with immunological analyses.

    Directory of Open Access Journals (Sweden)

    Chikashi Terao

    Full Text Available Rheumatoid arthritis (RA is a major cause of adult chronic inflammatory arthritis and a typical complex trait. Although several genetic determinants have been identified, they account for only a part of the genetic susceptibility. We conducted a genome-wide association study of RA in Japanese using 225,079 SNPs genotyped in 990 cases and 1,236 controls from two independent collections (658 cases and 934 controls in collection1; 332 cases and 302 controls in collection2, followed by replication studies in two additional collections (874 cases and 855 controls in collection3; 1,264 cases and 948 controls in collection4. SNPs showing p<0.005 in the first two collections and p<10(-4 by meta-analysis were further genotyped in the latter two collections. A novel risk variant, rs2000811, in intron2 of the myelin basic protein (MBP at chromosome 18q23 showed strong association with RA (p = 2.7×10(-8, OR 1.23, 95% CI: 1.14-1.32. The transcription of MBP was significantly elevated with the risk allele compared to the alternative allele (p<0.001. We also established by immunohistochemistry that MBP was expressed in the synovial lining layer of RA patients, the main target of inflammation in the disease. Circulating autoantibody against MBP derived from human brain was quantified by ELISA between patients with RA, other connective tissue diseases and healthy controls. As a result, the titer of anti-MBP antibody was markedly higher in plasma of RA patients compared to healthy controls (p<0.001 and patients with other connective tissue disorders (p<0.001. ELISA experiment using citrullinated recombinant MBP revealed that a large fraction of anti-MBP antibody in RA patients recognized citrullinated MBP. This is the first report of a genetic study in RA implicating MBP as a potential autoantigen and its involvement in pathogenesis of the disease.

  19. Investigating the Pathogenic Role of PADI4 in Oesophageal Cancer

    Directory of Open Access Journals (Sweden)

    Xiaotian Chang, Xiuli Hou, Jihong Pan, Kehua Fang, Lin Wang, Jinxiang Han

    2011-01-01

    Full Text Available PADI4 post-translationally converts peptidylarginine to citrulline. PADI4 can disrupt the apoptotic process via the citrullination of histone H3 in the promoter of p53-target genes. The current study focused on PADI4 expression in various subtypes of oesophageal carcinoma (EC by immunohistochemistry, western blotting and real time PCR. The study also investigated the effect of bile acid deoxycholate (DCA on PADI4 expression in Eca-109 cells that originated from EC. Apoptosis and DCA-induced toxicity were analyzed by TUNEL, MTT assay and flow cytometry. Additionally, the present study investigated the correlation between single nucleotide polymorphism (SNP in PADI4 gene and EC risk in Chinese population using Illumina GoldenGate assay. Compared with paraneoplastic tissues, the transcriptional and translational levels of PADI4 were significantly elevated in oesophageal squamous cell carcinoma (ESCC, n=9 and oesophageal adenocarcinoma (EAC, n=5 tissues. Immunolabeling detected expression of PADI4 in ESCC tissues (98.56%, n=139, EAC samples (87.5%, n=16 and oesophageal small cell undifferentiated carcinoma (91.7%, n=12 but not in normal tissues (0%, n=16. Furthermore, PADI4 levels is positively correlated with the pathological classification of ESCC (p=0.009. PADI4 expression levels were consistent with the number of apoptotic cells in the induced Eca-109 cells. rs10437048 [OR= 0.012831; 95% CI, 0.001746~0.094278; p=1.556×10-12] were significantly associated with decreased risk of EC, whereas rs41265997 [OR=12.7; 95% CI, 0.857077~33.207214; p=3.896×10-8] were significantly associated with increased risk of EC. rs41265997 in exon 3 of PADI4 gene is non-synonymous and converts ACG to ATG resulting in a threonine /methionine conversion at position 274 of the protein. Haplotypes GC that carries the variant alleles for rs2501796 and rs2477134 was significantly associated with increased risk of EC (frequency=0.085, p=0.0256, OR=2.7. The results suggest

  20. A new metabolomic assay to examine inflammation and redox pathways following LPS challenge

    Directory of Open Access Journals (Sweden)

    Suh Jung H

    2012-10-01

    Full Text Available Abstract Background Shifts in intracellular arginine (Arg and sulfur amino acid (SAA redox metabolism modulate macrophage activation, polarization and phenotype. Despite their importance in inflammation and redox regulatory pathways, comprehensive analysis of these metabolic networks was not previously possible with existing analytical methods. Methods The Arg/thiol redox LC-MS/MS metabolomics assay permits simultaneous assessment of amino acids and derivative products generated from Arg and SAA metabolism. Using this assay, LPS-induced changes in macrophage amino acid metabolism were monitored to identify pathway shifts during activation and their linkage to cellular redox regulation. Results Metabolite concentrations most significantly changed after treatment of a macrophage-like cell line (RAW with LPS for 24 hrs were citrulline (Cit (48-fold increase, ornithine (Orn (8.5-fold increase, arginine (Arg (66% decrease, and aspartic acid (Asp (73% decrease. The ratio Cit + Orn/Arg + Asp (CO/AA was more sensitive to LPS stimulation than other amino acid ratios commonly used to measure LPS-dependent inflammation (e.g., SAM/SAH, GSH/GSSG and total media NOx. The CO/AA ratio was also the first ratio to change significantly after LPS treatment (4 hrs. Changes in the overall metabolomic profile over time indicated that metabolic pathways shifted from Arg catabolism to thiol oxidation. Conclusions Simultaneous quantification of Arg and SAA metabolic pathway shifts following LPS challenge of macrophage indicate that, in this system, the Arg-Citrulline/NO cycle and arginase pathways are the amino acid metabolic pathways most sensitive to LPS-challenge. The cellular (Cit + Orn/(Arg + Asp ratio, which summarizes this pathway, was more responsive to lower concentrations of LPS and responded earlier than other metabolic biomarkers of macrophage activation including GSH redox. It is suggested that the CO/AA ratio is a redox- independent early biomarker of

  1. RF及抗CCP抗体与类风湿关节炎骨侵蚀及肺间质纤维化的关系分析%The Relationship Analysis of RF and Anti-CCP Antibodies and Rheumatoid Arthritis with Bone Erosion and Pulmonary Fibrosis

    Institute of Scientific and Technical Information of China (English)

    裴冰

    2014-01-01

    Objective:To investigate the rheumatoid factor (RF),anti cyclic citrul inated peptide (CCP) antibody and rheumatoid arthritis (RA) interstitial fibrosis and the relationship between disease activity score and knee injury,lung. Methods:analysis of anti CCP antibodies,RF and rheumatoid arthritis activity score (DAS28),fibrosis,lung relationship between joint bone erosion. Results:pulmonary interstitial fibrosis in patients with RA,RF high titer positive (19.2%) percentage higher than the negative ones (11.5%),anti CCP antibody with high titer was positive (66.7%)percentage higher than the negative ones (14.8%),the dif erence was statistical y significant (P<0.05). Conclusion:high titer of RF and anti CCP antibody may be associated with RA patients with pulmonary interstitial fibrosis related.%目的:探讨类风湿因子(RF)、抗环瓜氨酸(CCP)抗体与类风湿关节炎(RA)疾病活动性评分及关节损害、肺间质纤维化的关系。方法分析抗 CCP抗体、RF与类风湿关节炎活动性评分(DAS28)、关节骨侵蚀、肺间质纤维化的关系。结果合并肺间质纤维化的 RA患者中,RF高滴定度阳性(19.2%)所占百分比高于阴性者(11.5%),抗CCP抗体高滴定度阳性(66.7%)所占百分比高于阴性者(14.8%),差异有统计学意义(P<0.05)。结论高滴定度RF和抗CCP抗体可能与RA患者合并肺间质纤维化相关。

  2. Competition between roots and microorganisms for phosphorus: A novel 33P labeling approach

    Science.gov (United States)

    Zilla, Thomas; Kuzyakov, Yakov; Zavišiæ, Aljoša; Polle, Andrea

    2015-04-01

    While organic N mineralization exhibits clear seasonal uptake dynamics, knowledge about seasonal variation in microbial P uptake and mineralization is scarce. We hypothesize that the dynamics of P uptake and mineralization by microorganisms in temperate forest soils exhibit a seasonality anti-cyclic to plant P uptake. Therefore, the ratio of microbial P to labile P increases by the transition from acquiring ecosystems (in spring) to recycling ones (in fall). To investigate this, intact soil-plant mesocosms containing Ah horizon with 1 year old F. sylvatica were removed from the P-rich field site Bad Brueckenau and the P-depleted field site Luess in Germany. During incubation under controlled conditions, seasonal pulse labeling by 33P-orthophosphate was performed at 5 time points over the course of one year. 33P recovery in microbial compounds of organic and mineral soil horizons was determined 7 and 30 days after the labeling. This procedure will account for temporal changes in P allocation and also considers the rather slow P transport from the mycorrhiza into the plants and other microorganisms. For the first time we analyzed the 33P incorporation into total PLFA and consequently provide a new technique for the analysis of P uptake by microorganisms, which has clear advantages compared to P quantification after chloroform fumigation. Polar lipids are hereby extracted with a Frostegård-modified Bligh-and-Dyer buffer, i.e. a single phase mixture of chloroform, methanol and citrate buffer (0.8:1:2, v:v:v). Phospholipids (PLFA) are isolated and purified by solid phase extraction via a silica gel column chromatography. Subsequently, PLFA are hydrolyzed and the resulting fatty acids derivatized by methylation. The fatty acid methyl esters were extracted with n-hexane and measured by GC/MS to investigate the composition of the microbial community. The remaining extract, containing head groups, phosphate units and glycerol backbones, was used to determine 33P activity

  3. A crise internacional desafia o modelo brasileiro de abertura e liberalização The international crisis challenges the Brazilian model of economic opening and liberalization

    Directory of Open Access Journals (Sweden)

    Carlos Eduardo Carvalho

    2009-01-01

    Full Text Available É surpreendente a resistência da economia brasileira à crise internacional nos primeiros meses de 2009. A queda da produção e dos investimentos foi intensa, mas não houve crise cambial, nem crise fiscal ou abalos no sistema financeiro. O governo conseguiu adotar medidas anticíclicas, com corte de impostos e redução dos juros, favorecido pela arrecadação fiscal elevada e pelo fortalecimento dos bancos, depois de anos de juros elevados, além de dispor dos bancos públicos e dos fundos públicos, instrumentos poderosos herdados do passado. No lado externo, a entrada de capitais é estimulada pela ampla liquidez gerada pelas políticas de expansão monetária dos Estados Unidos, enquanto as exportações refletem a sustentação da demanda chinesa, com forte aumento da participação de produtos primários e redução das vendas de produtos industrializados. A capacidade de reagir à crise é um grande teste para o modelo brasileiro de abertura e liberalização, da mesma forma que as dificuldades para sustentar as exportações de industrializados e para a recuperação dos investimentos e da atividade produtiva são também desafios consideráveis.Reviewing the first months of 2009, it's surprising to see the Brazilian economy resistance over the international crisis. The decline on production and investment were intense, however, there has not been neither a foreign exchange crisis nor fiscal and financial system breakthrough. The government could apply anti-cyclic policies, as tax cuts and interest reduction, helped by the actual large extend of tax collection and banks strength, after years of high interest rates, besides using the public banks and the public funds, valuable legacy instruments. Regarding the external side, the capital inflow is stimulated by the wider liquidity generate by the US recent monetary expansion policy, while the exports are due to China demand maintenance, with significant increase in basic commodities

  4. A novel PAD4/SOX4/PU.1 signaling pathway is involved in the committed differentiation of acute promyelocytic leukemia cells into granulocytic cells.

    Science.gov (United States)

    Song, Guanhua; Shi, Lulu; Guo, Yuqi; Yu, Linchang; Wang, Lin; Zhang, Xiaoyu; Li, Lianlian; Han, Yang; Ren, Xia; Guo, Qiang; Bi, Kehong; Jiang, Guosheng

    2016-01-19

    All-trans retinoic acid (ATRA) treatment yields cure rates > 80% through proteasomal degradation of the PML-RARα fusion protein that typically promotes acute promyelocytic leukemia (APL). However, recent evidence indicates that ATRA can also promote differentiation of leukemia cells that are PML-RARα negative, such as HL-60 cells. Here, gene expression profiling of HL-60 cells was used to investigate the alternative mechanism of impaired differentiation in APL. The expression of peptidylarginine deiminase 4 (PADI4), encoding PAD4, a protein that post-translationally converts arginine into citrulline, was restored during ATRA-induced differentiation. We further identified that hypermethylation in the PADI4 promoter was associated with its transcriptional repression in HL-60 and NB4 (PML-RARα positive) cells. Functionally, PAD4 translocated into the nucleus upon ATRA exposure and promoted ATRA-mediated differentiation. Mechanistic studies using RNAi knockdown or electroporation-mediated delivery of PADI4, along with chromatin immunoprecipitation, helped identify PU.1 as an indirect target and SOX4 as a direct target of PAD4 regulation. Indeed, PAD4 regulates SOX4-mediated PU.1 expression, and thereby the differentiation process, in a SOX4-dependent manner. Taken together, our results highlight an association between PAD4 and DNA hypermethylation in APL and demonstrate that targeting PAD4 or regulating its downstream effectors may be a promising strategy to control differentiation in the clinic. PMID:26673819

  5. Involvement of Nitric Oxide, Neurotrophins and HPA Axis in Neurobehavioural Alterations Induced by Prenatal Stress.

    Science.gov (United States)

    Maur, Damian G; Pascuan, Cecilia G; Genaro, Ana M; Zorrilla-Zubilete, Maria A

    2015-01-01

    Several studies suggest that negative emotions during pregnancy generate adverse effects on the cognitive, behavioural and emotional development of the descendants. The psychoneuroendocrine pathways involve the transplacentary passage of maternal glucocorticoids in order to influence directly on fetal growth and brain development.Nitric oxide is a gaseous neurotransmitter that plays an important role in the control of neural activity by diffusing into neurons and participates in learning and memory processes. It has been demonstrated that nitric oxide is involved in the regulation of corticosterone secretion. Thus, it has been found that the neuronal isoform of nitric oxide synthase (nNOS) is an endogenous inhibitor of glucocorticoid receptor (GR) in the hippocampus and that nNOS in the hippocampus may participate in the modulation of hypothalamic-pituitary-adrenal axis activity via GR.Neurotrophins are a family of secreted growth factors consisting of nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), neurotrophin 3 (NT3) and NT4. Although initially described in the nervous system, they regulate processes such as cell survival, proliferation and differentiation in several other compartments. It has been demonstrated that the NO-citrulline cycle acts together with BDNF in maintaining the progress of neural differentiation.In the present chapter, we explore the interrelation between nitric oxide, glucocorticoids and neurotrophins in brain areas that are key structures in learning and memory processes. The participation of this interrelation in the behavioural and cognitive alterations induced in the offspring by maternal stress is also addressed. PMID:25287536

  6. Anti-CCP Antibodies Are Not Associated with Familial Mediterranean Fever in Childhood

    Directory of Open Access Journals (Sweden)

    Hatice Onur

    2013-01-01

    Full Text Available Objective. Anticyclic citrullinated peptide antibodies (anti-CCP testing is useful in the diagnosis of rheumatoid arthritis (RA with high specificity. Arthritis is a very common clinical manifestation in children with familial Mediterranean fever (FMF. The aim of the study was to show the presence of anti-CCP antibodies in child individuals diagnosed with FMF. Material and Methods. The study groups comprised one hundred and twenty-six patients (126 diagnosed with FMF (female/male (n: 66/60 and 50 healthy controls (female/male (n: 25/25. Clinical and laboratory assessments of the FMF patients were performed during attack-free periods. Erythrocyte sedimentation rate (ESR, serum C-reactive protein (CRP, fibrinogen, and anti-CCP antibody levels were measured. Results. Anti-CCP was negative in healthy controls and also in all FMF patients. There was not a significant difference in anti-CCP between the patient and the control groups. Our study has shown that anti-CCP was correlated moderately with age (rs=0.271; P=0.0020, duration of illness (rs=0.331; P<0.0001, and colchicine therapy (rs=0.259; P=0.004. Conclusion. Our data show that anti-CCP antibodies are not associated with FMF. Anti-CCP does not have a priority for identifying FMF arthritis from the other inflammatory arthritis.

  7. Expression of the inducible nitric oxide synthase gene in diaphragm and skeletal muscle.

    Science.gov (United States)

    Thompson, M; Becker, L; Bryant, D; Williams, G; Levin, D; Margraf, L; Giroir, B P

    1996-12-01

    Nitric oxide (NO) is a pluripotent molecule that can be secreted by skeletal muscle through the activity of the neuronal constitutive isoform of NO synthase. To determine whether skeletal muscle and diaphragm might also express the macrophage-inducible form of NO synthase (iNOS) during provocative states, we examined tissue from mice at serial times after intravenous administration of Escherichia coli endotoxin. In these studies, iNOS mRNA was strongly expressed in the diaphragm and skeletal muscle of mice 4 h after intravenous endotoxin and was significantly diminished by 8 h after challenge. Induction of iNOS mRNA was followed by expression of iNOS immunoreactive protein on Western immunoblots. Increased iNOS activity was demonstrated by conversion of arginine to citrulline. Immunochemical analysis of diaphragmatic explants exposed to endotoxin in vitro revealed specific iNOS staining in myocytes, in addition to macrophages and endothelium. These results may be important in understanding the pathogenesis of respiratory pump failure during septic shock, as well as skeletal muscle injury during inflammation or metabolic stress.

  8. A Combined Metabolomic and Proteomic Analysis of Gestational Diabetes Mellitus

    Directory of Open Access Journals (Sweden)

    Joanna Hajduk

    2015-12-01

    Full Text Available The aim of this pilot study was to apply a novel combined metabolomic and proteomic approach in analysis of gestational diabetes mellitus. The investigation was performed with plasma samples derived from pregnant women with diagnosed gestational diabetes mellitus (n = 18 and a matched control group (n = 13. The mass spectrometry-based analyses allowed to determine 42 free amino acids and low molecular-weight peptide profiles. Different expressions of several peptides and altered amino acid profiles were observed in the analyzed groups. The combination of proteomic and metabolomic data allowed obtaining the model with a high discriminatory power, where amino acids ethanolamine, l-citrulline, l-asparagine, and peptide ions with m/z 1488.59; 4111.89 and 2913.15 had the highest contribution to the model. The sensitivity (94.44% and specificity (84.62%, as well as the total group membership classification value (90.32% calculated from the post hoc classification matrix of a joint model were the highest when compared with a single analysis of either amino acid levels or peptide ion intensities. The obtained results indicated a high potential of integration of proteomic and metabolomics analysis regardless the sample size. This promising approach together with clinical evaluation of the subjects can also be used in the study of other diseases.

  9. Quantitative proteomics and terminomics to elucidate the role of ubiquitination and proteolysis in adaptive immunity.

    Science.gov (United States)

    Klein, Theo; Viner, Rosa I; Overall, Christopher M

    2016-10-28

    Adaptive immunity is the specialized defence mechanism in vertebrates that evolved to eliminate pathogens. Specialized lymphocytes recognize specific protein epitopes through antigen receptors to mount potent immune responses, many of which are initiated by nuclear factor-kappa B activation and gene transcription. Most, if not all, pathways in adaptive immunity are further regulated by post-translational modification (PTM) of signalling proteins, e.g. phosphorylation, citrullination, ubiquitination and proteolytic processing. The importance of PTMs is reflected by genetic or acquired defects in these pathways that lead to a dysfunctional immune response. Here we discuss the state of the art in targeted proteomics and systems biology approaches to dissect the PTM landscape specifically regarding ubiquitination and proteolysis in B- and T-cell activation. Recent advances have occurred in methods for specific enrichment and targeted quantitation. Together with improved instrument sensitivity, these advances enable the accurate analysis of often rare PTM events that are opaque to conventional proteomics approaches, now rendering in-depth analysis and pathway dissection possible. We discuss published approaches, including as a case study the profiling of the N-terminome of lymphocytes of a rare patient with a genetic defect in the paracaspase protease MALT1, a key regulator protease in antigen-driven signalling, which was manifested by elevated linear ubiquitination.This article is part of the themed issue 'Quantitative mass spectrometry'. PMID:27644975

  10. A new mouse model of mild ornithine transcarbamylase deficiency (spf-j displays cerebral amino acid perturbations at baseline and upon systemic immune activation.

    Directory of Open Access Journals (Sweden)

    Tatyana N Tarasenko

    Full Text Available Ornithine transcarbamylase deficiency (OTCD, OMIM# 311250 is an inherited X-linked urea cycle disorder that is characterized by hyperammonemia and orotic aciduria. In this report, we describe a new animal model of OTCD caused by a spontaneous mutation in the mouse Otc gene (c.240T>A, p.K80N. This transversion in exon 3 of ornithine transcarbamylase leads to normal levels of mRNA with low levels of mature protein and is homologous to a mutation that has also been described in a single patient affected with late-onset OTCD. With higher residual enzyme activity, spf-J were found to have normal plasma ammonia and orotate. Baseline plasma amino acid profiles were consistent with mild OTCD: elevated glutamine, and lower citrulline and arginine. In contrast to WT, spf-J displayed baseline elevations in cerebral amino acids with depletion following immune challenge with polyinosinic:polycytidylic acid. Our results indicate that the mild spf-J mutation constitutes a new mouse model that is suitable for mechanistic studies of mild OTCD and the exploration of cerebral pathophysiology during acute decompensation that characterizes proximal urea cycle dysfunction in humans.

  11. Adaptations of Arginine's Intestinal-Renal Axis in Cachectic Tumor-Bearing Rats.

    Science.gov (United States)

    Buijs, Nikki; Vermeulen, Mechteld A R; Weeda, Viola B; Bading, James R; Houdijk, Alexander P J; van Leeuwen, Paul A M

    2015-01-01

    Malignancies induce disposal of arginine, an important substrate for the immune system. To sustain immune function, the tumor-bearing host accelerates arginine's intestinal-renal axis by glutamine mobilization from skeletal muscle and this may promote cachexia. Glutamine supplementation stimulates argi-nine production in healthy subjects. Arginine's intestinal-renal axis and the effect of glutamine supplementation in cancer cach-exia have not been investigated. This study evaluated the long-term adaptations of the interorgan pathway for arginine production following the onset of cachexia and the metabolic effect of glutamine supplementation in the cachectic state. Fischer-344 rats were randomly divided into a tumor-bearing group (n = 12), control group (n = 7) and tumor-bearing group receiving a glutamine-enriched diet (n = 9). Amino acid fluxes and net fractional extractions across intestine, kidneys, and liver were studied. Compared to controls, the portal-drained viscera of tumor-bearing rats took up significantly more glutamine and released significantly less citrulline. Renal metabolism was unchanged in the cachectic tumor-bearing rats compared with controls. Glutamine supplementation had no effects on intestinal and renal adaptations. In conclusion, in the cachectic state, an increase in intestinal glutamine uptake is not accompanied by an increase in renal arginine production. The adaptations found in the cachectic, tumor-bearing rat do not depend on glutamine availability.

  12. The Role of Immunonutrients in the Prevention of Necrotizing Enterocolitis in Preterm Very Low Birth Weight Infants

    Directory of Open Access Journals (Sweden)

    Ping Zhou

    2015-08-01

    Full Text Available Necrotizing enterocolitis (NEC is a critical intestinal emergency condition, which mainly occurs in preterm very low birth weight (PVLBW infants. Despite remarkable advances in the care of PVLBW infants, with considerable improvement of the survival rate in recent decades, the incidence of NEC and NEC-related mortality have not declined accordingly. The fast progression from nonspecific signs to extensive necrosis also makes primary prevention the first priority. Recently, increasing evidence has indicated the important role of several nutrients in primary prevention of NEC. Therefore, the aim of this review is to summarize some potential immunomodulatory nutrients in the prevention of NEC, including bovine colostrum, probiotics, prebiotics (e.g., human milk oligosaccharides, long chain polyunsaturated fatty acids, and amino acids (glutamine, cysteine and N-acetylcysteine, l-arginine and l-citrulline. Based on current research evidence, probiotics are the most documented effective method to prevent NEC, while others still require further investigation in animal studies and clinical randomized controlled trials.

  13. A Simple Enzymatic Method for Production of a Wide Variety of D-Amino Acids Using L-Amino Acid Oxidase from Rhodococcus sp. AIU Z-35-1

    Directory of Open Access Journals (Sweden)

    Kimiyasu Isobe

    2010-01-01

    Full Text Available A simple enzymatic method for production of a wide variety of D-amino acids was developed by kinetic resolution of DL-amino acids using L-amino acid oxidase (L-AAO with broad substrate specificity from Rhodococcus sp. AIU Z-35-1. The optimum pH of the L-AAO reaction was classified into three groups depending on the L-amino acids as substrate, and their respective activities between pH 5.5 and 8.5 accounted for more than 60% of the optimum activity. The enzyme was stable in the range from pH 6.0 to 8.0, and approximately 80% of the enzyme activity remained after incubation at 40∘C for 60 min at pH 7.0. D-Amino acids such as D-citrulline, D-glutamine, D-homoserine or D-arginine, which are not produced by D-aminoacylases or D-hydantoinases, were produced from the racemic mixture within a 24-hr reaction at 30∘C and pH 7.0. Thus, the present method using L-AAO was versatile for production of a wide variety of D-amino acids.

  14. Filaggrin in the frontline: role in skin barrier function and disease.

    Science.gov (United States)

    Sandilands, Aileen; Sutherland, Calum; Irvine, Alan D; McLean, W H Irwin

    2009-05-01

    Recently, loss-of-function mutations in FLG, the human gene encoding profilaggrin and filaggrin, have been identified as the cause of the common skin condition ichthyosis vulgaris (which is characterised by dry, scaly skin). These mutations, which are carried by up to 10% of people, also represent a strong genetic predisposing factor for atopic eczema, asthma and allergies. Profilaggrin is the major component of the keratohyalin granules within epidermal granular cells. During epidermal terminal differentiation, the approximately 400 kDa profilaggrin polyprotein is dephosphorylated and rapidly cleaved by serine proteases to form monomeric filaggrin (37 kDa), which binds to and condenses the keratin cytoskeleton and thereby contributes to the cell compaction process that is required for squame biogenesis. Within the squames, filaggrin is citrullinated, which promotes its unfolding and further degradation into hygroscopic amino acids, which constitute one element of natural moisturising factor. Loss of profilaggrin or filaggrin leads to a poorly formed stratum corneum (ichthyosis), which is also prone to water loss (xerosis). Recent human genetic studies strongly suggest that perturbation of skin barrier function as a result of reduction or complete loss of filaggrin expression leads to enhanced percutaneous transfer of allergens. Filaggrin is therefore in the frontline of defence, and protects the body from the entry of foreign environmental substances that can otherwise trigger aberrant immune responses.

  15. Arginase activities and global arginine bioavailability in wild-type and ApoE-deficient mice: responses to high fat and high cholesterol diets.

    Directory of Open Access Journals (Sweden)

    Aaron Erdely

    Full Text Available Increased catabolism of arginine by arginase is increasingly viewed as an important pathophysiological factor in cardiovascular disease, including atherosclerosis induced by high cholesterol diets. Whereas previous studies have focused primarily on effects of high cholesterol diets on arginase expression and arginine metabolism in specific blood vessels, there is no information regarding the impact of lipid diets on arginase activity or arginine bioavailability at a systemic level. We, therefore, evaluated the effects of high fat (HF and high fat-high cholesterol (HC diets on arginase activity in plasma and tissues and on global arginine bioavailability (defined as the ratio of plasma arginine to ornithine + citrulline in apoE(-/- and wild-type C57BL/6J mice. HC and HF diets led to reduced global arginine bioavailability in both strains. The HC diet resulted in significantly elevated plasma arginase in both strains, but the HF diet increased plasma arginase only in apoE(-/- mice. Elevated plasma arginase activity correlated closely with increased alanine aminotransferase levels, indicating that liver damage was primarily responsible for elevated plasma arginase. The HC diet, which promotes atherogenesis, also resulted in increased arginase activity and expression of the type II isozyme of arginase in multiple tissues of apoE(-/- mice only. These results raise the possibility that systemic changes in arginase activity and global arginine bioavailability may be contributing factors in the initiation and/or progression of cardiovascular disease.

  16. Contribution of a Non-classical HLA Gene, HLA-DOA, to the Risk of Rheumatoid Arthritis.

    Science.gov (United States)

    Okada, Yukinori; Suzuki, Akari; Ikari, Katsunori; Terao, Chikashi; Kochi, Yuta; Ohmura, Koichiro; Higasa, Koichiro; Akiyama, Masato; Ashikawa, Kyota; Kanai, Masahiro; Hirata, Jun; Suita, Naomasa; Teo, Yik-Ying; Xu, Huji; Bae, Sang-Cheol; Takahashi, Atsushi; Momozawa, Yukihide; Matsuda, Koichi; Momohara, Shigeki; Taniguchi, Atsuo; Yamada, Ryo; Mimori, Tsuneyo; Kubo, Michiaki; Brown, Matthew A; Raychaudhuri, Soumya; Matsuda, Fumihiko; Yamanaka, Hisashi; Kamatani, Yoichiro; Yamamoto, Kazuhiko

    2016-08-01

    Despite the progress in human leukocyte antigen (HLA) causal variant mapping, independent localization of major histocompatibility complex (MHC) risk from classical HLA genes is challenging. Here, we conducted a large-scale MHC fine-mapping analysis of rheumatoid arthritis (RA) in a Japanese population (6,244 RA cases and 23,731 controls) population by using HLA imputation, followed by a multi-ethnic validation study including east Asian and European populations (n = 7,097 and 23,149, respectively). Our study identified an independent risk of a synonymous mutation at HLA-DOA, a non-classical HLA gene, on anti-citrullinated protein autoantibody (ACPA)-positive RA risk (p = 1.4 × 10(-9)), which demonstrated a cis-expression quantitative trait loci (cis-eQTL) effect on HLA-DOA expression. Trans-ethnic comparison revealed different linkage disequilibrium (LD) patterns in HLA-DOA and HLA-DRB1, explaining the observed HLA-DOA variant risk heterogeneity among ethnicities, which was most evident in the Japanese population. Although previous HLA fine-mapping studies have identified amino acid polymorphisms of the classical HLA genes as driving genetic susceptibility to disease, our study additionally identifies the dosage contribution of a non-classical HLA gene to disease etiology. Our study contributes to the understanding of HLA immunology in human diseases and suggests the value of incorporating additional ancestry in MHC fine-mapping. PMID:27486778

  17. Global metabolite profiling of synovial fluid for the specific diagnosis of rheumatoid arthritis from other inflammatory arthritis.

    Directory of Open Access Journals (Sweden)

    Sooah Kim

    Full Text Available Currently, reliable biomarkers that can be used to distinguish rheumatoid arthritis (RA from other inflammatory diseases are unavailable. To find possible distinctive metabolic patterns and biomarker candidates for RA, we performed global metabolite profiling of synovial fluid samples. Synovial fluid samples from 38 patients with RA, ankylosing spondylitis, Behçet's disease, and gout were analyzed by gas chromatography/time-of-flight mass spectrometry (GC/TOF MS. Orthogonal partial least-squares discriminant and hierarchical clustering analyses were performed for the discrimination of RA and non-RA groups. Variable importance for projection values were determined, and the Wilcoxon-Mann-Whitney test and the breakdown and one-way analysis of variance were conducted to identify potential biomarkers for RA. A total of 105 metabolites were identified from synovial fluid samples. The score plot of orthogonal partial least squares discriminant analysis showed significant discrimination between the RA and non-RA groups. The 20 metabolites, including citrulline, succinate, glutamine, octadecanol, isopalmitic acid, and glycerol, were identified as potential biomarkers for RA. These metabolites were found to be associated with the urea and TCA cycles as well as fatty acid and amino acid metabolism. The metabolomic analysis results demonstrated that global metabolite profiling by GC/TOF MS might be a useful tool for the effective diagnosis and further understanding of RA.

  18. Analysis of the Citrullus colocynthis transcriptome during water deficit stress.

    Science.gov (United States)

    Wang, Zhuoyu; Hu, Hongtao; Goertzen, Leslie R; McElroy, J Scott; Dane, Fenny

    2014-01-01

    Citrullus colocynthis is a very drought tolerant species, closely related to watermelon (C. lanatus var. lanatus), an economically important cucurbit crop. Drought is a threat to plant growth and development, and the discovery of drought inducible genes with various functions is of great importance. We used high throughput mRNA Illumina sequencing technology and bioinformatic strategies to analyze the C. colocynthis leaf transcriptome under drought treatment. Leaf samples at four different time points (0, 24, 36, or 48 hours of withholding water) were used for RNA extraction and Illumina sequencing. qRT-PCR of several drought responsive genes was performed to confirm the accuracy of RNA sequencing. Leaf transcriptome analysis provided the first glimpse of the drought responsive transcriptome of this unique cucurbit species. A total of 5038 full-length cDNAs were detected, with 2545 genes showing significant changes during drought stress. Principle component analysis indicated that drought was the major contributing factor regulating transcriptome changes. Up regulation of many transcription factors, stress signaling factors, detoxification genes, and genes involved in phytohormone signaling and citrulline metabolism occurred under the water deficit conditions. The C. colocynthis transcriptome data highlight the activation of a large set of drought related genes in this species, thus providing a valuable resource for future functional analysis of candidate genes in defense of drought stress.

  19. Lysozyme affects the microbial catabolism of free arginine in raw-milk hard cheeses.

    Science.gov (United States)

    D'Incecco, P; Gatti, M; Hogenboom, J A; Bottari, B; Rosi, V; Neviani, E; Pellegrino, L

    2016-08-01

    Lysozyme (LZ) is used in several cheese varieties to prevent late blowing which results from fermentation of lactate by Clostridium tyrobutyricum. Side effects of LZ on lactic acid bacteria population and free amino acid pattern were studied in 16 raw-milk hard cheeses produced in eight parallel cheese makings conducted at four different dairies using the same milk with (LZ+) or without (LZ-) addition of LZ. The LZ-cheeses were characterized by higher numbers of cultivable microbial population and lower amount of DNA arising from lysed bacterial cells with respect to LZ + cheeses. At both 9 and 16 months of ripening, Lactobacillus delbrueckii and Lactobacillus fermentum proved to be the species mostly affected by LZ. The total content of free amino acids indicated the proteolysis extent to be characteristic of the dairy, regardless to the presence of LZ. In contrast, the relative patterns showed the microbial degradation of arginine to be promoted in LZ + cheeses. The data demonstrated that the arginine-deiminase pathway was only partially adopted since citrulline represented the main product and only trace levels of ornithine were found. Differences in arginine degradation were considered for starter and non-starter lactic acid bacteria, at different cheese ripening stages. PMID:27052697

  20.  NET and NETosis – new phenomenon in immunology

    Directory of Open Access Journals (Sweden)

    Natalia Matoszka

    2012-06-01

    Full Text Available  Neutrophils are one of the first cells of the immune system recruited to the site of infection, representing the host’s most effective and numerous front-line defenders. Recently, a novel antimicrobial mechanism of neutrophils has been described: upon activation, they release DNA and a subset of their granule content, forming neutrophil extracellular traps (NETs. These extracellular, chromatin structures, which contain histones and neutrophil granule proteins, can trap and kill a broad spectrum of microbes, including Gram-positive and Gram-negative bacteria, fungi, protozoa and viruses. Some of the pathogens, which are trapped and exposed to high local concentrations of antimicrobial compounds, employ strategies against NET binding, including surface modification and/or degradation of NET by DNases. It has been suggested that NETs are formed during active cell death, recently named NETosis. New data indicate that this novel mechanism of cell death requires interaction between three processes – reactive oxygen species generation, histone citrullination and autophagy – and significantly differs from previously known types of cell death, including apoptosis and necrosis. Moreover, the release of nuclear chromatin was also described for other types of cells – mast cells and eosinophils. Mast cells, like neutrophils, under certain conditions release nuclear chromatin and may undergo a similar active cell death program, while eosinophils release only mitochondrial chromatin, and its release does not lead to the death of these cells.

  1. Peroxynitrite inhibits inducible (type 2) nitric oxide synthase in murine lung epithelial cells in vitro.

    Science.gov (United States)

    Robinson, V K; Sato, E; Nelson, D K; Camhi, S L; Robbins, R A; Hoyt, J C

    2001-05-01

    Peroxynitrite, formed by nitric oxide (NO) and superoxide, can alter protein function by nitrating amino acids such as tyrosine, cysteine, trytophan, or methionine. Inducible nitric oxide synthase (Type 2 NOS or iNOS) converts arginine to citrulline, releasing NO. We hypothesized that peroxynitrite could function as a negative feedback modulator of NO production by nitration of iNOS. Confluent cultures of the murine lung epithelial cell line, LA-4 were stimulated with cytokines to express iNOS, peroxynitrite was added, and the flasks sealed. After 3 h, NO in the headspace above the culture was sampled. Peroxynitrite caused a concentration-dependent decrease in NO. Similar results were obtained when 3-morpholinosydnonimine (SIN-1), a peroxynitrite generator, was added to the flasks. PAPA-NONOate, the NO generator, did not affect the headspace NO. Nitration of the iNOS was confirmed by detection of 3-nitrotyrosine by Western blotting. These data suggest a mechanism for inhibition of NO synthesis at inflammatory sites where iNOS, NO, and superoxide would be expected.

  2. Genome-wide analysis reveals PADI4 cooperates with Elk-1 to activate c-Fos expression in breast cancer cells.

    Directory of Open Access Journals (Sweden)

    Xuesen Zhang

    2011-06-01

    Full Text Available Peptidylarginine deiminase IV (PADI4 catalyzes the conversion of positively charged arginine and methylarginine residues to neutrally charged citrulline, and this activity has been linked to the repression of a limited number of target genes. To broaden our knowledge of the regulatory potential of PADI4, we utilized chromatin immunoprecipitation coupled with promoter tiling array (ChIP-chip analysis to more comprehensively investigate the range of PADI4 target genes across the genome in MCF-7 breast cancer cells. Results showed that PADI4 is enriched in gene promoter regions near transcription start sites (TSSs; and, surprisingly, this pattern of binding is primarily associated with actively transcribed genes. Computational analysis found potential binding sites for Elk-1, a member of the ETS oncogene family, to be highly enriched around PADI4 binding sites; and coimmunoprecipitation analysis then confirmed that Elk-1 physically associates with PADI4. To better understand how PADI4 may facilitate gene transactivation, we then show that PADI4 interacts with Elk-1 at the c-Fos promoter and that, following Epidermal Growth Factor (EGF stimulation, PADI4 catalytic activity facilitates Elk-1 phosphorylation, histone H4 acetylation, and c-Fos transcriptional activation. These results define a novel role for PADI4 as a transcription factor co-activator.

  3. Inhibition of Cryptococcus neoformans replication by nitrogen oxides supports the role of these molecules as effectors of macrophage-mediated cytostasis

    Energy Technology Data Exchange (ETDEWEB)

    Alspaugh, J.A.; Granger, D.L. (Department of Medicine, Duke University School of Medicine, Durham, NC (USA))

    1991-07-01

    Activated macrophages are able to inhibit the replication of intracellular microbes and tumor cells. In the murine system, this cytostatic effect is associated with the oxidation of L-arginine to L-citrulline, nitrite, and nitrate and is thought to be mediated by an intermediate of this reaction, possibly nitric oxide (NO.). By exposing replicating Cryptococcus neoformans cells to conditions under which NO. is chemically generated, we have observed a cytostatic effect similar to that caused by activated murine macrophages. Nitric oxide is formed as a decomposition product of nitrite salts in acidic, aqueous solutions. Although C. neoformans replicates well in the presence of high nitrite concentrations at physiologic pH, its growth in acidic media can be inhibited by the addition of low concentrations of sodium nitrite. The degree of cytostasis is dependent on both the pH and the nitrite concentration of the NO. generating solution. The cytostatic effector molecule appears to be a gas since, in addition to inhibiting C. neoformans replication in solution, it is able to exert its inhibitory effect across a gas-permeable but ion-impermeable membrane. At high nitrite concentrations, a fungicidal effect occurs. We propose that the growth inhibition of C. neoformans upon exposure to chemically generated NO. or some related oxide of nitrogen represents a cell-free system simulating the cytostatic effect of activated murine macrophages.

  4. In vivo measurement of nitric oxide production in porcine gut, liver and muscle during hyperdynamic endotoxaemia

    Science.gov (United States)

    Bruins, Maaike J; Lamers, Wouter H; Meijer, Alfred J; Soeters, Peter B; Deutz, Nicolaas E P

    2002-01-01

    During prolonged endotoxaemia, an increase in arginine catabolism may result in limiting substrate availability for nitric oxide (NO) production. These effects were quantitated in a chronically instrumented porcine endotoxaemia model. Ten days prior to the beginning of the experiments, pigs were catheterized. On day 0, pigs received a continuous infusion of endotoxin (3 μg kg−1 h−1) over 24 h and were saline resuscitated. Blood was drawn from the catheters at 0 and 24 h during primed-infusion of 15N2-arginine and P-aminohippurate to assess 15N2-arginine to 15N-citrulline conversion and plasma flow rates, respectively, across the portal-drained viscera, liver and hindquarter. During endotoxin infusion a hyperdynamic circulation with elevated heart rate, cardiac index and decreased mean arterial pressure was achieved, characteristic of the human septic condition. Endotoxin induced NO production by the portal-drained viscera and the liver. The increased NO production was quantitatively matched by an increase in arginine disposal. Nitrite/nitrate levels remained unchanged during endotoxaemia. Despite an increased arginine production from the hindquarter and an increased whole-body arginine appearance rate during endotoxin infusion, the plasma arginine concentration was lower in endotoxin-treated animals than in controls. On a whole-body level, the muscle was found to serve as a major arginine supplier and, considering the lowered arginine plasma levels, seems critical in providing arginine as precursor for NO synthesis in the splanchnic region. PMID:12466232

  5. Effects of Egg White Protein Supplementation on Muscle Strength and Serum Free Amino Acid Concentrations

    Directory of Open Access Journals (Sweden)

    Yukari Kawano

    2012-10-01

    Full Text Available The aim of this study was to evaluate the effects of egg white protein compared to carbohydrate intake prior to exercise on fat free mass (FFM, one repetition maximum (1RM muscle strength and blood biochemistry in female athletes. Thirty healthy female collegiate athletes were recruited for this study and matched by sport type, body fat percentage and 1RM leg curl muscle strength. Participants were randomly divided into two groups: protein group (15.0 g egg white protein; 75 kcal and carbohydrate group (17.5 g maltodextrin, 78 kcal. Supplements were administered daily at the same time in a double-blind manner prior to training during an 8-week period. Measurements were performed before and after the 8-week regimen. The mean dietary energy intake did not change throughout the study period. FFM and 1RM assessments (i.e., leg curl, leg extension, squat, and bench press increased in both groups. Furthermore, serum urea and serum citrulline levels after the 8-week regimen increased significantly only in the protein group. Our findings indicated that compared to the carbohydrate supplement, the protein supplement was associated with some changes in protein metabolites but not with changes in body composition or muscle strength.

  6. Changes in intramuscular amino acid levels in submaximally exercised horses - a pilot study.

    Science.gov (United States)

    van den Hoven, R; Bauer, A; Hackl, S; Zickl, M; Spona, J; Zentek, J

    2010-08-01

    The time-dependent changes in intramuscular amino acid (AA) levels caused by exercise and by feeding a protein/AA supplement were analysed in nine horses. Horses were submitted to a total of four standardized exercise tests (SETs). Amino acid concentrations were determined prior to, immediately after, 4 and 18 h after exercise. The experiment was subdivided into two consecutive periods of 3 weeks. In each period two SETs were performed. In the second period, horses were given a protein/AA supplement within 1 h after exercise. Significant changes in mean plasma AA levels similar to previous studies were noted to be time-dependent and to be associated with feeding the supplement. The intramuscular concentrations of the free AA in relation to pre-exercise levels showed significant time-dependent changes for alanine, asparagine, aspartate, citrulline, glutamine, glycine, isoleucine, leucine, methionine, serine, taurine, threonine, tyrosine and valine. Feeding the supplement significantly increased the 4 h post-exercise intramuscular concentration of alanine, isoleucine, methionine and tyrosine. At 18 h after exercise, apart from isoleucine and methionine, levels were still increased and also those of asparagine, histidine and valine in relation to none treatment. Hence, it was concluded that AA mixtures administered orally to horses within 1 h after exercise increased intramuscular AA pool. PMID:19663973

  7. The effects of short intensive exercise on plasma free amino acids in standardbred trotters.

    Science.gov (United States)

    Hackl, S; van den Hoven, R; Zickl, M; Spona, J; Zentek, J

    2009-04-01

    The aim of this study was to investigate the effect of short intense exercise on plasma amino acid concentrations in trotters and to test the repeatability of plasma amino acids concentration in samples obtained on two independent days under field conditions. Plasma amino acid concentrations were analysed in blood samples of 36 standardbred trotters before and after intense exercise over a distance of 2000 m. Sampling was repeated in 20 horses after 35 days. Exercise intensity was estimated from post-exercise lactate levels. Horses were divided in two groups according to a cut-off lactate concentration at 15 mmol/l. The plasma concentrations of alanine, aspartate, glutamate, isoleucine, leucine, lysine and taurine increased and arginine, asparagine, citrulline, glutamine, glycine, histidine, methionine, serine, tryptophan and 3-methylhistidine decreased after exercise. Ornithine, threonine, tyrosine, phenylalanine and valine concentrations remained constant. Higher intensity of exercise significantly decreased tryptophan and increased taurine concentrations. Sampling day had a significant effect on the absolute pre- and post-exercise amino acid concentrations. Exercise had a significant influence on the concentrations of most plasma amino acids in trotters. These changes could reflect shifts between the free amino acid compartments, but there were also some indications for muscle catabolism. The amino acid supply of sporting horses could be of specific significance for maintaining muscle integrity and for the improvement of post-exercise recovery of competition horses. PMID:19320929

  8. A Combined Metabolomic and Proteomic Analysis of Gestational Diabetes Mellitus.

    Science.gov (United States)

    Hajduk, Joanna; Klupczynska, Agnieszka; Dereziński, Paweł; Matysiak, Jan; Kokot, Piotr; Nowak, Dorota M; Gajęcka, Marzena; Nowak-Markwitz, Ewa; Kokot, Zenon J

    2015-12-16

    The aim of this pilot study was to apply a novel combined metabolomic and proteomic approach in analysis of gestational diabetes mellitus. The investigation was performed with plasma samples derived from pregnant women with diagnosed gestational diabetes mellitus (n = 18) and a matched control group (n = 13). The mass spectrometry-based analyses allowed to determine 42 free amino acids and low molecular-weight peptide profiles. Different expressions of several peptides and altered amino acid profiles were observed in the analyzed groups. The combination of proteomic and metabolomic data allowed obtaining the model with a high discriminatory power, where amino acids ethanolamine, L-citrulline, L-asparagine, and peptide ions with m/z 1488.59; 4111.89 and 2913.15 had the highest contribution to the model. The sensitivity (94.44%) and specificity (84.62%), as well as the total group membership classification value (90.32%) calculated from the post hoc classification matrix of a joint model were the highest when compared with a single analysis of either amino acid levels or peptide ion intensities. The obtained results indicated a high potential of integration of proteomic and metabolomics analysis regardless the sample size. This promising approach together with clinical evaluation of the subjects can also be used in the study of other diseases.

  9. Oxidative Stress Facilitates IFN-γ-Induced Mimic Extracellular Trap Cell Death in A549 Lung Epithelial Cancer Cells.

    Science.gov (United States)

    Lin, Chiou-Feng; Chen, Chia-Ling; Chien, Shun-Yi; Tseng, Po-Chun; Wang, Yu-Chih; Tsai, Tsung-Ting

    2016-01-01

    We previously demonstrated that IFN-γ induces an autophagy-regulated mimic extracellular trap cell death (ETosis) in A549 human lung cancer cells. Regarding reactive oxygen species (ROS) are involved in ETosis, this study investigated the role of oxidative stress. After IFN-γ stimulation, a necrosis-like cell death mimic ETosis occurred accompanied by the inhibition of cell growth, aberrant nuclear staining, and nucleosome release. ROS were generated in a time-dependent manner with an increase in NADPH oxidase component protein expression. STAT1-mediated IFN regulatory factor-1 activation was essential for upregulating ROS production. By genetically silencing p47phox, IFN-γ-induced ROS and mimic ETosis were significantly attenuated. This mechanistic study indicated that ROS may mediate DNA damage followed by histone H3 citrullination. Furthermore, ROS promoted IFN-γ-induced mimic ETosis in cooperation with autophagy. These findings further demonstrate that ROS regulates IFN-γ-induced mimic ETosis in lung epithelial malignancy. PMID:27575372

  10. Endothelial nitric oxide synthase gene polymorphisms (G894T in diabetes mellitus in Egypt

    Directory of Open Access Journals (Sweden)

    El-baz1 ; Farouk2; Tag Eldin2; Ezat2

    2010-06-01

    Full Text Available Objective: Diabetic nephropathy (DN is one of the major microvascular complications of diabetes. Genetic predisposition has been implicated in DN. The eNOS protein synthesizes nitric oxide constitutively via a reaction including the conversion of L-arginine to L-citrulline, which involves the transfer of five electrons provided by nicotinamide adenine dinucleotide phosphate The aim of this study is to evaluate the association of G894T polymorphisms of endothelial nitric oxide synthase(eNOS gene with the development of diabetic nephropathy (DN among Egyptian patients with type 1,2 diabetes mellitus in Egypt. Methods: Study subjects comprised 86 patients of type 2 diabetes with nephropathy,23 patients of type 1 diabetes with nephropathy and 46 patients of type 2 diabetes without nephropathy. G894T genotypes was determined by SSP- PCR analysis. Results: Mutant T allele, GT and TT genotypes of G894TSNP had no significant frequencies in type 1,2 diabetic patients with nephropathy compared to those without nephropathy.. Conclusion: These findings indicate that G894T polymorphism of eNOS gene may be not considered as genetic risk factors for DN among Egyptian type1, 2 diabetic patients. Abbreviations: T2DM: type 2 diabetes mellitus ­ DN: diabetic nephropathy eNOS : Endothelial nitric oxide synthase:­ SNP: single nucleotide polymorphism- SSP-PCR: sequence specific primer- polymerase chain reaction

  11. Pathomechanisms in rheumatoid arthritis--time for a string theory?

    Science.gov (United States)

    Weyand, Cornelia M; Goronzy, Jörg J

    2006-04-01

    RA is a quintessential autoimmune disease with a growing number of cells, mediators, and pathways implicated in this tissue-injurious inflammation. Now Kuhn and colleagues have provided convincing evidence that autoantibodies reacting with citrullinated proteins, known for their sensitivity and specificity as biomarkers in RA, enhance tissue damage in collagen-induced arthritis (see the related article beginning on page 961). This study adds yet another soldier to the growing army of autoaggressive mechanisms that underlie RA. With great success researchers have dismantled the pathogenic subunits of RA, adding gene to gene, molecule to molecule, and pathway to pathway in an ever more complex scheme of dysfunction. The complexity of the emerging disease model leaves us speechless. It seems that with this wealth of data available, we need to develop a new theory for this disease. We may want to seek guidance from our colleagues in physics and mathematics who have successfully integrated their knowledge of elementary particles and the complexity of their interacting forces by formulating the string theory.

  12. The effectiveness of a new domestic carbohydrate-protein product in the practice of training of high class

    Directory of Open Access Journals (Sweden)

    Vdovenko N.V.

    2013-08-01

    Full Text Available The effect of the use of the new course of the carbohydrate-protein product on the performance efficiency of skilled athletes (Greco-Roman. In the experiment involved 14 athletes aged 18-25 years who gave written consent to participate in the study. Developed and clinically tested a specialized carbohydrate-protein food product. The drink contains in its composition: glucose, sucrose, whey protein concentrate, creatine monohydrate, citrulline malate, mineral complex and ATP-lipid complex. The study was conducted in two micro-cycles (2 weeks. Athletes take a drink as follows: pre-workout (30-40 minutes - 250 ml of the drink, after a training session during the recovery period - 250 ml of the drink. It is established that the use of the drink has a positive effect on the performance of athletes performance during the execution of a sub-maximal anaerobic power in the area of energy supply. Argues that course application beverage reduces the severity of manifestations of lactate acidosis after exercise by improving the utilization of lactate. Found a significant decrease in the concentration of lactate in the blood of athletes in the 7th minute of recovery in relation to the original data.

  13. MSMA resistance studies.

    Science.gov (United States)

    Camper, N D; Keese, R J; Coker, P S

    2004-05-01

    Monosodium methanearsonate (MSMA)-resistant and -susceptible common cocklebur (Xanthium strumarium L.) and cotton (Gossypium hirsutum L.) were treated with MSMA. Plant parameters analyzed were: glutathione synthetase activity, selected amino acid (arginine, glutamic acid, alanine, citrulline, glutamine, and glutathione) content and arsenic content (MSMA, total arsenic, and arsonate). No reduction of arsenic from the parent pentavalent form present in MSMA to the trivalent form was detected. Arginine, glutamic acid, and glutamine content increased in tissue three days after MSMA treatment. Glutathione content decreased during the first three days after treatment; however, five days after treatment the resistant biotype of cocklebur and cotton had elevated glutathione levels (8-20 times greater, respectively). Glutathione Synthetase activity was higher in cotton than in either of the cocklebur biotypes; MSMA did not affect its activity in cotton or either cocklebur biotype. Resistant biotypes have a slightly higher activity than the susceptible biotype. Tolerance of cotton to MSMA may be related to glutathione synthetase activity and possibly to the presence of phytochelatins. Also, increased glutathione levels in the resistant biotype may implicate phytochelatin involvement in the resistance mechanism.

  14. Arginine metabolism in wounds

    International Nuclear Information System (INIS)

    Arginine metabolism in wounds was investigated in the rat in 1) lambda-carrageenan-wounded skeletal muscle, 2) Schilling chambers, and 3) subcutaneous polyvinyl alcohol sponges. All showed decreased arginine and elevated ornithine contents and high arginase activity. Arginase could be brought to the wound by macrophages, which were found to contain arginase activity. However, arginase was expressed by macrophages only after cell lysis and no arginase was released by viable macrophages in vitro. Thus the extracellular arginase of wounds may derive from dead macrophages within the injured tissue. Wound and peritoneal macrophages exhibited arginase deiminase activity as demonstrated by the conversion of [guanido-14C]arginine to radiolabeled citrulline during culture, the inhibition of this reaction by formamidinium acetate, and the lack of prokaryotic contamination of the cultures. These findings and the known metabolic fates of the products of arginase and arginine deiminase in the cellular populations of the wound suggest the possibility of cooperativity among cells for the production of substrates for collagen synthesis

  15. Clinical validation of surface-enhanced Raman scattering-based immunoassays in the early diagnosis of rheumatoid arthritis.

    Science.gov (United States)

    Chon, Hyangah; Wang, Rui; Lee, Sangyeop; Bang, So-Young; Lee, Hye-Soon; Bae, Sang-Cheol; Hong, Sung Hyun; Yoon, Young Ho; Lim, Dong Woo; deMello, Andrew J; Choo, Jaebum

    2015-11-01

    We assessed the clinical feasibility of conducting immunoassays based on surface-enhanced Raman scattering (SERS) in the early diagnosis of rheumatoid arthritis (RA). An autoantibody against citrullinated peptide (anti-CCP) was used as a biomarker, magnetic beads conjugated with CCP were used as substrates, and the SERS nanotags were comprised of anti-human IgG-conjugated hollow gold nanospheres (HGNs). We were able to determine the anti-CCP serum levels successfully by observing the distinctive Raman intensities corresponding to the SERS nanotags. At high concentrations of anti-CCP (>25 U/mL), the results obtained from the SERS assay confirmed those obtained via an ELISA-based assay. Nevertheless, quantitation via our SERS-based assay is significantly more accurate at low concentrations (25 U/mL) revealed a good correlation between the ELISA and SERS-based assays. However, in the anti-CCP-negative group (n = 43, <25 U/mL), the SERS-based assay was shown to be more reproducible. Accordingly, we suggest that SERS-based assays are novel and potentially useful tools in the early diagnosis of RA.

  16. Conservative Mechanisms of Extracellular Trap Formation by Annelida Eisenia andrei: Serine Protease Activity Requirement.

    Science.gov (United States)

    Homa, Joanna; Ortmann, Weronika; Kolaczkowska, Elzbieta

    2016-01-01

    Formation of extracellular traps (ETs) capturing and immobilizing pathogens is now a well-established defense mechanism added to the repertoire of vertebrate phagocytes. These ETs are composed of extracellular DNA (extDNA), histones and antimicrobial proteins. Formation of mouse and human ETs depends on enzymes (i) facilitating decondensation of chromatin by citrullination of histones, and (ii) serine proteases degrading histones. In invertebrates, initial reports revealed existence of ETs composed of extDNA and histones, and here we document for the first time that also coelomocytes, immunocompetent cells of an earthworm Eisenia andrei, cast ETs which successfully trap bacteria in a reactive oxygen species (ROS)-dependent and -independent manner. Importantly, the formation of ETs was observed not only when coelomocytes were studied ex vivo, but also in vivo, directly in the earthworm coelom. These ETs were composed of extDNA, heat shock proteins (HSP27) and H3 histones. Furthermore, the formation of E. andrei ETs depended on activity of serine proteases, including elastase-like activity. Moreover, ETs interconnected and hold together aggregating coelomocytes, a processes proceeding encapsulation. In conclusion, the study confirms ET formation by earthworms, and unravels mechanisms leading to ET formation and encapsulation in invertebrates. PMID:27416067

  17. Advances in Neonatal Pulmonary Hypertension.

    Science.gov (United States)

    Steinhorn, Robin H

    2016-01-01

    Persistent pulmonary hypertension of the newborn (PPHN) is a surprisingly common event in the neonatal intensive care unit, and affects both term and preterm infants. Recent studies have begun to elucidate the maternal, fetal and genetic risk factors that trigger PPHN. There have been numerous therapeutic advances over the last decade. It is now appreciated that oxygen supplementation, particularly for the goal of pulmonary vasodilation, needs to be approached as a therapy that has risks and benefits. Administration of surfactant or inhaled nitric oxide (iNO) therapy at a lower acuity of illness can decrease the risk of extracorporeal membrane oxygenation/death, progression of disease and duration of hospital stay. Milrinone may have specific benefits as an 'inodilator', as prolonged exposure to iNO plus oxygen may activate phosphodiesterase (PDE) 3A. Additionally, sildenafil and hydrocortisone may benefit infants exposed to hyperoxia and oxidative stress. Continued investigation is likely to reveal new therapies such as citrulline and cinaciguat that will enhance NO synthase and soluble guanylate cyclase function. Continued laboratory and clinical investigation will be needed to optimize treatment and improve outcomes. PMID:27251312

  18. Altered brain arginine metabolism in schizophrenia

    Science.gov (United States)

    Liu, P; Jing, Y; Collie, N D; Dean, B; Bilkey, D K; Zhang, H

    2016-01-01

    Previous research implicates altered metabolism of l-arginine, a versatile amino acid with a number of bioactive metabolites, in the pathogenesis of schizophrenia. The present study, for we believe the first time, systematically compared the metabolic profile of l-arginine in the frontal cortex (Brodmann's area 8) obtained post-mortem from schizophrenic individuals and age- and gender-matched non-psychiatric controls (n=20 per group). The enzyme assays revealed no change in total nitric oxide synthase (NOS) activity, but significantly increased arginase activity in the schizophrenia group. Western blot showed reduced endothelial NOS protein expression and increased arginase II protein level in the disease group. High-performance liquid chromatography and liquid chromatography/mass spectrometric assays confirmed significantly reduced levels of γ-aminobutyric acid (GABA), but increased agmatine concentration and glutamate/GABA ratio in the schizophrenia cases. Regression analysis indicated positive correlations between arginase activity and the age of disease onset and between l-ornithine level and the duration of illness. Moreover, cluster analyses revealed that l-arginine and its main metabolites l-citrulline, l-ornithine and agmatine formed distinct groups, which were altered in the schizophrenia group. The present study provides further evidence of altered brain arginine metabolism in schizophrenia, which enhances our understanding of the pathogenesis of schizophrenia and may lead to the future development of novel preventions and/or therapeutics for the disease. PMID:27529679

  19. The role of polyamines in protein-dependent hypoxic tolerance of Drosophila

    Directory of Open Access Journals (Sweden)

    Vigne Paul

    2008-12-01

    Full Text Available Abstract Background Chronic hypoxia is a major component of ischemic diseases such as stroke or myocardial infarction. Drosophila is more tolerant to hypoxia than most mammalian species. It is considered as a useful model organism to identify new mechanisms of hypoxic tolerance. The hypoxic tolerance of flies has previously been reported to be enhanced by low protein diets. This study analyses the mechanisms involved. Results Feeding adult Drosophila on a yeast diet dramatically reduced their longevities under chronic hypoxic conditions (5% O2. Mean and maximum longevities became close to the values observed for starving flies. The action of dietary yeast was mimicked by a whole casein hydrolysate and by anyone of the 20 natural amino acids that compose proteins. It was mimicked by amino acid intermediates of the urea cycle such as L-citrulline and L-ornithine, and by polyamines (putrescine, spermidine and spermine. α-difluoromethylornithine, a specific inhibitor of ornithine decarboxylase, partially protected hypoxic flies from amino acid toxicity but not from polyamine toxicity. N1-guanyl-1,7 diaminoheptane, a specific inhibitor of eIF5A hypusination, partially relieved the toxicities of both amino acids and polyamines. Conclusion Dietary amino acids reduced the longevity of chronically hypoxic flies fed on a sucrose diet. Pharmacological evidence suggests that the synthesis of polyamines and the hypusination of eIF5A contributed to the life-shortening effect of dietary amino acids.

  20. CD151 gene delivery increases eNOS activity and induces ECV304 migration, proliferation and tube formation

    Institute of Scientific and Technical Information of China (English)

    Zhen-zhong ZHENO; Zheng-xiang LIU

    2007-01-01

    Aim: To investigate the effects of CD151 on the activity of endothelial NO syn-thase (eNOS), and ECV304 migration, proliferation and tube formation. Methods:pAAV-CD151 and pAAV-anti-CD151 were constructed and used to transiently transfect ECV304 mediated with Lipofectamine 2000. After transfection, the ex-pression of CD151 was measured by Western blotting. Cell migration assay was performed using Boyden transwell; proliferation assay was evaluated using the 3-[4,5-dimethylthiazol-2-yl]-2,5, diphenyltetrazolium bromide (MTT) method, and tube formation test was examined on matrigel, eNOS activity was assayed by L-[3H]citrulline production from L-[3H]arginine. The involvement of eNOS was ex-plored using an eNOS inhibitor (L-NAME) and the effects in the process were observed. Results: CD151 promotes cell migration, proliferation and tube formation.In addition, CD151 increases eNOS activity. Moreover, cell migration, prolifera-tion and tube formation induced by CD151 are inhibited when L-NAME is used,which indicates that there is an involvement of eNOS in CD151-induced cell migration, cell proliferation and tube formation. Conclusion: CD151 promotes ECV304 migration, proliferation and tube formation. The mechanism is that CD151 increases eNOS activity. This result also suggests that eNOS is involved in the angiogenic effects of CD151.

  1. The role of arginine in infection and sepsis.

    Science.gov (United States)

    Luiking, Yvette C; Poeze, Martijn; Ramsay, Graham; Deutz, Nicolaas E P

    2005-01-01

    Sepsis is a systemic response to an infection, with high morbidity and mortality rates. Metabolic changes during infection and sepsis could be related to changes in metabolism of the amino acid L-arginine. In sepsis, protein breakdown is increased, which is a key process to maintain arginine delivery because both endogenous de novo arginine production from citrulline and food intake are reduced. Arginine catabolism, on the other hand, is markedly increased by enhanced use of arginine via the arginase and nitric oxide pathways. As a result, lowered plasma arginine levels are usually found. Arginine may therefore be considered as an essential amino acid in sepsis, and supplementation could be beneficial in sepsis by improving microcirculation and protein anabolism. L-Arginine supplementation in a hyperdynamic pig model of sepsis prohibits the increase in pulmonary arterial blood pressure, improves muscle and liver protein metabolism, and restores the intestinal motility pattern. Arguments raised against arginine supplementation are mainly pointed at stimulating nitric oxide (NO) production, with concerns about toxicity of increased NO and hemodynamic instability with refractory hypotension. NO synthase inhibition, however, increased mortality. Arginine supplementation in septic patients has transient effects on hemodynamics when supplied as a bolus but seems without hemodynamic side effects when supplied continuously. In conclusion, arginine could have an essential role in infection and sepsis.

  2. [Rheumatoid arthritis: diagnostics and therapy 2012].

    Science.gov (United States)

    Lorenz, H-M

    2012-07-01

    Rheumatoid Arthritis (RA) should be suspected if patients do not only complain of joint pain, but suffer from joint swelling, sensation of heat, hyperemia and warmth around the joints. An arthritic joint pain should be most prominent at night time or early in the morning and cause morning stiffness (> 30 min) of the joint, exercise will improve the symptoms. Diagnosis of RA will be even more likely if wrists, MCP- or PIP joints are affected. Serologic procedures will test for rheumatoid factor or anti-citrullinated antibodies (CCP Ab). One needs to keep in mind that positive results for rheumatoid factor or CCP Ab alone never proves the diagnosis of RA. After diagnosis therapy should be started immediately, recruiting physiotherapy, pain medication, corticosteroids and disease-modifying anti-rheumatic drugs (DMARDs), primarily methotrexate. At the latest after failure of two DMARDs biologics like TNF-α-blockers, an Interleukin-6-Receptor-antibody, a B-cell-specific antibody or a rather T-cell-specific biologic will be initiated. Aim of therapy is freedom of symptoms of an ongoing arthritis, low dosage of immunosuppressants (especially corticosteroids maximally 5 mg/day), stop of radiological progression and prevention of long term consequences of inflammation like myocardial infarction, stroke or lymphoma.

  3. Electrostatic transition state stabilization rather than reactant destabilization provides the chemical basis for efficient chorismate mutase catalysis.

    Science.gov (United States)

    Burschowsky, Daniel; van Eerde, André; Ökvist, Mats; Kienhöfer, Alexander; Kast, Peter; Hilvert, Donald; Krengel, Ute

    2014-12-01

    For more than half a century, transition state theory has provided a useful framework for understanding the origins of enzyme catalysis. As proposed by Pauling, enzymes accelerate chemical reactions by binding transition states tighter than substrates, thereby lowering the activation energy compared with that of the corresponding uncatalyzed process. This paradigm has been challenged for chorismate mutase (CM), a well-characterized metabolic enzyme that catalyzes the rearrangement of chorismate to prephenate. Calculations have predicted the decisive factor in CM catalysis to be ground state destabilization rather than transition state stabilization. Using X-ray crystallography, we show, in contrast, that a sluggish variant of Bacillus subtilis CM, in which a cationic active-site arginine was replaced by a neutral citrulline, is a poor catalyst even though it effectively preorganizes chorismate for the reaction. A series of high-resolution molecular snapshots of the reaction coordinate, including the apo enzyme, and complexes with substrate, transition state analog and product, demonstrate that an active site, which is only complementary in shape to a reactive substrate conformer, is insufficient for effective catalysis. Instead, as with other enzymes, electrostatic stabilization of the CM transition state appears to be crucial for achieving high reaction rates.

  4. The inflammatory role of phagocyte apoptotic pathways in rheumatic diseases.

    Science.gov (United States)

    Cuda, Carla M; Pope, Richard M; Perlman, Harris

    2016-08-23

    Rheumatoid arthritis affects nearly 1% of the world's population and is a debilitating autoimmune condition that can result in joint destruction. During the past decade, inflammatory functions have been described for signalling molecules classically involved in apoptotic and non-apoptotic death pathways, including, but not limited to, Toll-like receptor signalling, inflammasome activation, cytokine production, macrophage polarization and antigen citrullination. In light of these remarkable advances in the understanding of inflammatory mechanisms of the death machinery, this Review provides a snapshot of the available evidence implicating death pathways, especially within the phagocyte populations of the innate immune system, in the perpetuation of rheumatoid arthritis and other rheumatic diseases. Elevated levels of signalling mediators of both extrinsic and intrinsic apoptosis, as well as the autophagy, are observed in the joints of patients with rheumatoid arthritis. Furthermore, risk polymorphisms are present in signalling molecules of the extrinsic apoptotic and autophagy death pathways. Although research into the mechanisms underlying these pathways has made considerable progress, this Review highlights areas where further investigation is particularly needed. This exploration is critical, as new discoveries in this field could lead to the development of novel therapies for rheumatoid arthritis and other rheumatic diseases. PMID:27549026

  5. The HB22.7-vcMMAE antibody-drug conjugate has efficacy against non-Hodgkin lymphoma mouse xenografts with minimal systemic toxicity.

    Science.gov (United States)

    Abuhay, Mastewal; Kato, Jason; Tuscano, Emily; Barisone, Gustavo A; Sidhu, Ranjit S; O'Donnell, Robert T; Tuscano, Joseph M

    2016-10-01

    In this study, HB22.7, an anti-CD22 monoclonal antibody, was used for specific, targeted delivery of monomethyl auristatin E (MMAE) to non-Hodgkin lymphoma (NHL). MMAE was covalently coupled to HB22.7 through a valine-citrulline peptide linker (vc). Maleimide-functionalized vcMMAE (mal-vcMMAE) was reacted with thiols of the partially reduced mAb. Approximately 4 molecules of MMAE were conjugated to HB22.7 as determined by residual thiol measurement and hydrophobic interaction chromatography-HPLC (HIC-HPLC). HB22.7-vcMMAE antibody-drug conjugate (ADC) retained its binding to Ramos NHL cells and also exhibited potent and specific in vitro cytotoxicity on a panel of B cell NHL cell lines with IC50s of 20-284 ng/ml. HB22.7-vcMMAE also showed potent efficacy in vivo against established NHL xenografts using the DoHH2 and Granta 519 cell lines. One dose of the ADC induced complete and persistent response in all DoHH2 xenografts and 90 % of Granta xenografts. Minimal toxicity was observed. In summary, HB22.7-vcMMAE is an effective ADC that should be evaluated for clinical translation. PMID:27506529

  6. Role of Natural Antioxidants in the Modulation of Plasma Amino Acid Pattern in Rats Exposed to Hemic Hypoxia

    Directory of Open Access Journals (Sweden)

    Nouf Mohamed Al-Rasheed

    2015-10-01

    Full Text Available ABSTRACTThe aim of this work was to investigate whether the free radical scavengers, L-arginine (L-arg and/or carnosine, either alone, or in combination would modulate tissue injury induced by hypoxia by measuring Fischer's ratio [concentrations of branched chain amino acids (BCAAs/aromatic amino acids]. Decreased Fischer's ratios and increased malondialdehyde (MDA led to pathogeneses of many diseases. Rats were injected with sodium nitrite (60 mg/kg to establish hypoxia. They were treated with L-arg, (200 mg/ kg and/or carnosine (200 mg/ kg and their combination 24 and 1 h prior to sodium nitrite intoxication. The results revealed that hypoxia significantly decreased hemoglobin, arginine, citrulline and proline and increased sLDH, MDA , ammonia , urea, BCAAs (valine, leucine and isoleucine and aromatic amino acids (phenylalanine and tyrosine . The Fischer's ratio was decreased compared with the control; the administration of the aforementioned antioxidants ameliorated most of the previously altered parameters. It was concluded that Fischer's ratio was a valuable tool for understanding the pathology of hemic hypoxia, evaluating the degree of the modulatory effect of various natural antioxidants and the synergy between L-arg and carnosine in ameliorating the effect of sodium nitrite on amino acids pattern. Thus, it could be recommended to administer the combination of L-arg and carnosine in the areas of high altitudes to combat the hazard effect of hypoxia on hemoglobin concentration and MDA level.

  7. Nitric Oxide as a Mediator of Oxidant Lung Injury Due to Paraquat

    Science.gov (United States)

    Berisha, Hasan I.; Pakbaz, Hedayatollah; Absood, Afaf; Said, Sami I.

    1994-08-01

    At low concentrations, nitric oxide is a physiological transmitter, but in excessive concentrations it may cause cell and tissue injury. We report that in acute oxidant injury induced by the herbicide paraquat in isolated guinea pig lungs, nitric oxide synthesis was markedly stimulated, as evidenced by increased levels of cyclic GMP in lung perfusate and of nitrite and L-citrulline production in lung tissue. All signs of injury, including increased airway and perfusion pressures, pulmonary edema, and protein leakage into the airspaces, were dose-dependently attenuated or totally prevented by either N^G-nitro-L-arginine methyl ester or N^ω-nitro-L-arginine, selective and competitive inhibitors of nitric oxide synthase. Protection was reversed by excess L-arginine but not by its enantiomer D-arginine. When blood was added to the lung perfusate, the paraquat injury was moderated or delayed as it was when paraquat was given to anesthetized guinea pigs. The rapid onset of injury and its failure to occur in the absence of Ca2+ suggest that constitutive rather than inducible nitric oxide synthase was responsible for the stimulated nitric oxide synthesis. The findings indicate that nitric oxide plays a critical role in the production of lung tissue injury due to paraquat, and it may be a pathogenetic factor in other forms of oxidant tissue injury.

  8. P-selectin promotes neutrophil extracellular trap formation in mice.

    Science.gov (United States)

    Etulain, Julia; Martinod, Kimberly; Wong, Siu Ling; Cifuni, Stephen M; Schattner, Mirta; Wagner, Denisa D

    2015-07-01

    Neutrophil extracellular traps (NETs) can be released in the vasculature. In addition to trapping microbes, they promote inflammatory and thrombotic diseases. Considering that P-selectin induces prothrombotic and proinflammatory signaling, we studied the role of this selectin in NET formation. NET formation (NETosis) was induced by thrombin-activated platelets rosetting with neutrophils and was inhibited by anti-P-selectin aptamer or anti-P-selectin glycoprotein ligand-1 (PSGL-1) inhibitory antibody but was not induced by platelets from P-selectin(-/-) mice. Moreover, NETosis was also promoted by P-selectin-immunoglobulin fusion protein but not by control immunoglobulin. We isolated neutrophils from mice engineered to overproduce soluble P-selectin (P-selectin(ΔCT/ΔCT) mice). Although the levels of circulating DNA and nucleosomes (indicative of spontaneous NETosis) were normal in these mice, basal neutrophil histone citrullination and presence of P-selectin on circulating neutrophils were elevated. NET formation after stimulation with platelet activating factor, ionomycin, or phorbol 12-myristate 13-acetate was significantly enhanced, indicating that the P-selectin(ΔCT/ΔCT) neutrophils were primed for NETosis. In summary, P-selectin, cellular or soluble, through binding to PSGL-1, promotes NETosis, suggesting that this pathway is a potential therapeutic target for NET-related diseases.

  9. A rapid approach for characterization of thiol-conjugated antibody-drug conjugates and calculation of drug-antibody ratio by liquid chromatography mass spectrometry.

    Science.gov (United States)

    Firth, David; Bell, Leonard; Squires, Martin; Estdale, Sian; McKee, Colin

    2015-09-15

    We present the demonstration of a rapid "middle-up" liquid chromatography mass spectrometry (LC-MS)-based workflow for use in the characterization of thiol-conjugated maleimidocaproyl-monomethyl auristatin F (mcMMAF) and valine-citrulline-monomethyl auristatin E (vcMMAE) antibody-drug conjugates. Deconvoluted spectra were generated following a combination of deglycosylation, IdeS (immunoglobulin-degrading enzyme from Streptococcus pyogenes) digestion, and reduction steps that provide a visual representation of the product for rapid lot-to-lot comparison-a means to quickly assess the integrity of the antibody structure and the applied conjugation chemistry by mass. The relative abundance of the detected ions also offer information regarding differences in drug conjugation levels between samples, and the average drug-antibody ratio can be calculated. The approach requires little material (small-scale process development testing or as an early component of a complete characterization project facilitating informed decision making regarding which aspects of a molecule might need to be examined in more detail by orthogonal methodologies. PMID:26070852

  10. Variations in short term products of inorganic carbon fixation in exponential and stationary phase cultures of Aphanocapsa 6308.

    Science.gov (United States)

    Weathers, P J; Allen, M M

    1978-03-01

    Aphanocapsa 6308 metabolizes both NaHCO3 and Na2CO3. The short term incorporation (5-s) metabolic pattern and the patterns of incorporation of bicarbonate for exponential versus stationary phase cultures differ, however. Cells were equilibrated for 10 min in air and distilled water prior to injection of either NaH14CO3 at pH 8.0, or Na214CO3 at pH 11.0. Hot ethanol extracts were analyzed via paper chromatography and autoradiography for products of CO2 fixation. At 5 s, malate (51.5%) predominates slightly as a primary bicarbonate fixation product over 3-phosphoglycerate (40.3%); 3-phosphoglycerate is the primary product of carbonate fixation. At 60 s, the carbonate and bicarbonate labelling patterns are similar. Cells in stationary phase fix in 5 s a greater proportion of bicarbonate into malate (36% vs. 14% for 3-phosphoglycerate) than do cells in exponential growth. Likewise, 60 s incorporations show a large amount of bicarbonate fixed into aspartate (30.9%) in stationary phase cells over that of exponential phase (11.6%). These data suggest an operative C4 pathway for purposes not related to carbohydrate synthesis but rather as compensation for the incomplete tricarboxylic acid cycle in cyanobacteria. The enhancement of both aspartate fixation and CO2 fixation into citrulline in stationary phase correlates with an increase in cyanophycin granule production which requires both aspartate and arginine. PMID:417691

  11. Asymmetric Dimethylarginine in Chronic Obstructive Pulmonary Disease (ADMA in COPD

    Directory of Open Access Journals (Sweden)

    Jeremy A. Scott

    2014-04-01

    Full Text Available l-Arginine metabolism including the nitric oxide (NO synthase and arginase pathways is important in the maintenance of airways function. We have previously reported that accumulation of asymmetric dimethylarginine (ADMA in airways, resulting in changes in l-arginine metabolism, contributes to airways obstruction in asthma and cystic fibrosis. Herein, we assessed l-arginine metabolism in airways of patients with chronic obstructive pulmonary disease (COPD. Lung function testing, measurement of fractional exhaled NO (FeNO and sputum NO metabolites, as well as quantification of l-arginine metabolites (l-arginine, l-ornithine, l-citrulline, ADMA and symmetric dimethylarginine using liquid chromatography-mass spectrometry (LC-MS were performed. Concentrations of l-ornithine, the product of arginase activity, correlated directly with l-arginine and ADMA sputum concentrations. FeNO correlated directly with pre- and post-bronchodilator forced expiratory volume in one second (FEV1. Sputum arginase activity correlated inversely with total NO metabolite (NOx and nitrite concentrations in sputum, and with pre- and post-bronchodilator FEV1. These findings suggest that ADMA in COPD airways results in a functionally relevant shift of l-arginine breakdown by the NO synthases towards the arginase pathway, which contributes to airway obstruction in these patients.

  12. Rheumatoid arthritis onset in postmenopausal women: Does the ACPA seropositive subset result from genetic effects, estrogen deficiency, skewed profile of CD4(+) T-cells, and their interactions?

    Science.gov (United States)

    Sapir-Koren, Rony; Livshits, Gregory

    2016-08-15

    Rheumatoid arthritis (RA) incidence displays a differentiated age-dependent female-to-male ratio in which women outnumber men. Evidence that the peak incidence of RA in women coincides with menopause age, suggests a potential estrogenic role to disease etiology. Estrogens exert physiologically both stimulatory and inhibitory effects on the immune system. Epidemiologic and animal model studies with estrogen deprivation or supplementation suggested estrogens as to play, mainly, a protective role in RA immunopathology. In this review, we propose that some yet unidentified disturbances associated with estrogen circulating levels, differentiated by the menopausal status, play a major role in women's RA susceptibility. We focus on the interaction between estrogen deprivation and genetic risk alleles for anti-citrullinated protein antibodies (ACPA) seropositive RA, as a major driving force for increased immune reactivity and RA susceptibility, in postmenopausal women. This opens up new fields for research concerning the association among different irregular estrogenic conditions, the cytokine milieu, and age/menopausal status bias in RA. PMID:27178986

  13. Effect of fermentation and subsequent pasteurization processes on amino acids composition of orange juice.

    Science.gov (United States)

    Cerrillo, I; Fernández-Pachón, M S; Collado-González, J; Escudero-López, B; Berná, G; Herrero-Martín, G; Martín, F; Ferreres, F; Gil-Izquierdo, A

    2015-06-01

    The fermentation of fruit produces significant changes in their nutritional composition. An orange beverage has been obtained from the controlled alcoholic fermentation and thermal pasteurization of orange juice. A study was performed to determine the influence of both processes on its amino acid profile. UHPLC-QqQ-MS/MS was used for the first time for analysis of orange juice samples. Out of 29 amino acids and derivatives identified, eight (ethanolamine, ornithine, phosphoethanolamine, α-amino-n-butyric acid, hydroxyproline, methylhistidine, citrulline, and cystathionine) have not previously been detected in orange juice. The amino acid profile of the orange juice was not modified by its processing, but total amino acid content of the juice (8194 mg/L) was significantly increased at 9 days of fermentation (13,324 mg/L). Although the pasteurization process produced partial amino acid degradation, the total amino acid content was higher in the final product (9265 mg/L) than in the original juice, enhancing its nutritional value.

  14. Leptin Inhibits the Proliferation of Vascular Smooth Muscle Cells Induced by Angiotensin II through Nitric Oxide-Dependent Mechanisms

    Directory of Open Access Journals (Sweden)

    Amaia Rodríguez

    2010-01-01

    Full Text Available Objective. This study was designed to investigate whether leptin modifies angiotensin (Ang II-induced proliferation of aortic vascular smooth muscle cells (VSMCs from 10-week-old male Wistar and spontaneously hypertensive rats (SHR, and the possible role of nitric oxide (NO. Methods. NO and NO synthase (NOS activity were assessed by the Griess and 3H-arginine/citrulline conversion assays, respectively. Inducible NOS (iNOS and NADPH oxidase subutnit Nox2 expression was determined by Western-blot. The proliferative responses to Ang II were evaluated through enzymatic methods. Results. Leptin inhibited the Ang II-induced proliferative response of VSMCs from control rats. This inhibitory effect of leptin was abolished by NOS inhibitor, NMMA, and iNOS selective inhibitor, L-NIL, and was not observed in leptin receptor-deficient fa/fa rats. SHR showed increased serum leptin concentrations and lipid peroxidation. Despite a similar leptin-induced iNOS up-regulation, VSMCs from SHR showed an impaired NOS activity and NO production induced by leptin, and an increased basal Nox2 expression. The inhibitory effect of leptin on Ang II-induced VSMC proliferation was attenuated. Conclusion. Leptin blocks the proliferative response to Ang II through NO-dependent mechanisms. The attenuation of this inhibitory effect of leptin in spontaneous hypertension appears to be due to a reduced NO bioavailability in VSMCs.

  15. MSMA resistance studies.

    Science.gov (United States)

    Camper, N D; Keese, R J; Coker, P S

    2004-05-01

    Monosodium methanearsonate (MSMA)-resistant and -susceptible common cocklebur (Xanthium strumarium L.) and cotton (Gossypium hirsutum L.) were treated with MSMA. Plant parameters analyzed were: glutathione synthetase activity, selected amino acid (arginine, glutamic acid, alanine, citrulline, glutamine, and glutathione) content and arsenic content (MSMA, total arsenic, and arsonate). No reduction of arsenic from the parent pentavalent form present in MSMA to the trivalent form was detected. Arginine, glutamic acid, and glutamine content increased in tissue three days after MSMA treatment. Glutathione content decreased during the first three days after treatment; however, five days after treatment the resistant biotype of cocklebur and cotton had elevated glutathione levels (8-20 times greater, respectively). Glutathione Synthetase activity was higher in cotton than in either of the cocklebur biotypes; MSMA did not affect its activity in cotton or either cocklebur biotype. Resistant biotypes have a slightly higher activity than the susceptible biotype. Tolerance of cotton to MSMA may be related to glutathione synthetase activity and possibly to the presence of phytochelatins. Also, increased glutathione levels in the resistant biotype may implicate phytochelatin involvement in the resistance mechanism. PMID:15473645

  16. Nitric Oxide Synthesis Is Increased in Cybrid Cells with m.3243A>G Mutation

    Directory of Open Access Journals (Sweden)

    Juliana Gamba

    2012-12-01

    Full Text Available Nitric oxide (NO is a free radical and a signaling molecule in several pathways, produced by nitric oxide synthase (NOS from the conversion of L-arginine to citrulline. Supplementation of L-arginine has been used to treat MELAS (mitochondrial encephalopathy with lactic acidosis and stroke like syndrome, a mitochondrial disease caused by the m.3243A>G mutation. Low levels of serum arginine and endothelium dysfunction have been reported in MELAS and this treatment may increase NO in endothelial cells and promote vasodilation, decreasing cerebral ischemia and strokes. Although clinical benefits have been reported, little is known about NO synthesis in MELAS. In this study we found that osteosarcoma derived cybrid cells with high levels of m.3243A>G had increased nitrite, an NO metabolite, and increased intracellular NO, demonstrated by an NO fluorescent probe (DAF-FM. Muscle vessels from patients with the same mutation had increased staining in NADPH diaphorase, suggestive of increased NOS. These results indicate increased production of NO in cells harboring the m.3243A>G, however no nitrated protein was detected by Western blotting. Further studies are necessary to clarify the exact mechanisms of L-arginine effect to determine the appropriate clinical use of this drug therapy.

  17. Measurement of (15)N enrichment of glutamine and urea cycle amino acids derivatized with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate using liquid chromatography-tandem quadrupole mass spectrometry.

    Science.gov (United States)

    Nakamura, Hidehiro; Karakawa, Sachise; Watanabe, Akiko; Kawamata, Yasuko; Kuwahara, Tomomi; Shimbo, Kazutaka; Sakai, Ryosei

    2015-05-01

    6-Aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC) is an amino acid-specific derivatizing reagent that has been used for sensitive amino acid quantification by liquid chromatography-tandem quadrupole mass spectrometry (LC-MS/MS). In this study, we aimed to evaluate the ability of this method to measure the isotopic enrichment of amino acids and to determine the positional (15)N enrichment of urea cycle amino acids (i.e., arginine, ornithine, and citrulline) and glutamine. The distribution of the M and M+1 isotopomers of each natural AQC-amino acid was nearly identical to the theoretical distribution. The standard deviation of the (M+1)/M ratio for each amino acid in repeated measurements was approximately 0.1%, and the ratios were stable regardless of the injected amounts. Linearity in the measurements of (15)N enrichment was confirmed by measuring a series of (15)N-labeled arginine standards. The positional (15)N enrichment of urea cycle amino acids and glutamine was estimated from the isotopic distribution of unique fragment ions generated at different collision energies. This method was able to identify their positional (15)N enrichment in the plasma of rats fed (15)N-labeled glutamine. These results suggest the utility of LC-MS/MS detection of AQC-amino acids for the measurement of isotopic enrichment in (15)N-labeled amino acids and indicate that this method is useful for the study of nitrogen metabolism in living organisms.

  18. Triterpenoic Acids from Apple Pomace Enhance the Activity of the Endothelial Nitric Oxide Synthase (eNOS).

    Science.gov (United States)

    Waldbauer, Katharina; Seiringer, Günter; Nguyen, Dieu Linh; Winkler, Johannes; Blaschke, Michael; McKinnon, Ruxandra; Urban, Ernst; Ladurner, Angela; Dirsch, Verena M; Zehl, Martin; Kopp, Brigitte

    2016-01-13

    Pomace is an easy-accessible raw material for the isolation of fruit-derived compounds. Fruit consumption is associated with health-promoting effects, such as the prevention of cardiovascular disease. Increased vascular nitric oxide (NO) bioavailability, for example, due to an enhanced endothelial nitric oxide synthase (eNOS) activity, could be one molecular mechanism mediating this effect. To identify compounds from apple (Malus domestica Borkh.) pomace that have the potential to amplify NO bioavailability via eNOS activation, a bioassay-guided fractionation of the methanol/water (70:30) extract has been performed using the (14)C-L-arginine to (14)C-L-citrulline conversion assay (ACCA) in the human endothelium-derived cell line EA.hy926. Phytochemical characterization of the active fractions was performed using the spectrophotometric assessment of the total phenolic content, as well as TLC, HPLC-DAD-ELSD, and HPLC-MS analyses. Eleven triterpenoic acids, of which one is a newly discovered compound, were identified as the main constituents in the most active fraction, accompanied by only minor contents of phenolic compounds. When tested individually, none of the tested compounds exhibited significant eNOS activation. Nevertheless, cell stimulation with the reconstituted compound mixture restored eNOS activation, validating the potential of apple pomace as a source of bioactive components.

  19. Microparticles That Form Immune Complexes as Modulatory Structures in Autoimmune Responses

    Directory of Open Access Journals (Sweden)

    Catalina Burbano

    2015-01-01

    Full Text Available Microparticles (MPs are induced during apoptosis, cell activation, and even “spontaneous” release. Initially MPs were considered to be inert cellular products with no biological function. However, an extensive research and functional characterization have shown that the molecular composition and the effects of MPs depend upon the cellular background and the mechanism inducing them. They possess a wide spectrum of biological effects on intercellular communication by transferring different molecules able to modulate other cells. MPs interact with their target cells through different mechanisms: membrane fusion, macropinocytosis, and receptor-mediated endocytosis. However, when MPs remain in the extracellular milieu, they undergo modifications such as citrullination, glycosylation, and partial proteolysis, among others, becoming a source of neoantigens. In rheumatoid arthritis (RA and systemic lupus erythematosus (SLE, reports indicated elevated levels of MPs with different composition, content, and effects compared with those isolated from healthy individuals. MPs can also form immune complexes amplifying the proinflammatory response and tissue damage. Their early detection and characterization could facilitate an appropriate diagnosis optimizing the pharmacological strategies, in different diseases including cancer, infection, and autoimmunity. This review focuses on the current knowledge about MPs and their involvement in the immunopathogenesis of SLE and RA.

  20. Amino acid metabolism in the kidneys of genetic and nutritionally obese rats.

    Science.gov (United States)

    Herrero, M C; Remesar, X; Bladé, C; Arola, L

    1997-06-01

    The ability of the kidney to take up and/or release amino acids has been determined in two models of obesity in Zucker rats, one genetic and the other nutritional (diet-obese). There was a noticeable increase in gluconeogenic amino acids in the arterial blood of diet-obese animals whereas the genetically obese rats showed small variations in the levels of these amino acids. There were significant decreases in renal Gly and Ser, only in the genetically obese rats. Genetically obese animals showed an increase in Glutamine synthetase activity. The uptake and/or release of amino acids showed important variations between the groups. The diet-obese group exhibited greater variation, since this group took up Glu, Ala, Gy, Phe and Citrulline and released Gln, Ser, Arg and Tyr. Genetically obese rats took up Gln, His and Taurine and released Ser. These different patterns may be related to variations in the whole body metabolic rate, since the diet-obese group was more active than the genetically obese group.

  1. A novel mutation in the SLC25A15 gene in a Turkish patient with HHH syndrome: functional analysis of the mutant protein.

    Science.gov (United States)

    Ersoy Tunalı, Nagehan; Marobbio, Carlo M T; Tiryakioğlu, N Ozan; Punzi, Giuseppe; Saygılı, Seha K; Onal, Hasan; Palmieri, Ferdinando

    2014-05-01

    The hyperornithinemia-hyperammonemia-homocitrullinuria syndrome is a rare autosomal recessive disorder caused by the functional deficiency of the mitochondrial ornithine transporter 1 (ORC1). ORC1 is encoded by the SLC25A15 gene and catalyzes the transport of cytosolic ornithine into mitochondria in exchange for citrulline. Although the age of onset and the severity of the symptoms vary widely, the disease usually manifests in early infancy. The typical clinical features include protein intolerance, lethargy, episodic confusion, cerebellar ataxia, seizures and mental retardation. In this study, we identified a novel p.Ala15Val (c.44C>T) mutation by genomic DNA sequencing in a Turkish child presenting severe tantrum, confusion, gait disturbances and loss of speech abilities in addition to hyperornithinemia, hyperammonemia and homocitrullinuria. One hundred Turkish control chromosomes did not possess this variant. The functional effect of the novel mutation was assessed by both complementation of the yeast ORT1 null mutant and transport assays. Our study demonstrates that the A15V mutation dramatically interferes with the transport properties of ORC1 since it was shown to inhibit ornithine transport nearly completely.

  2. Hyperornithinemia-hyperammonemia-homocitrullinuria (HHH)-syndrome. Ultrastructural changes of mitochondria in cultured dermal fibroblasts of three patients.

    Science.gov (United States)

    Haust, M D; Dewar, R A; Gatfield, D P; Gordon, B A

    1996-03-01

    Mitochondria of fibroblasts cultured from the skin obtained at biopsy from three patients with the hyperornithinemia-hyperammonemia-homocitrullinuria (HHH)-syndrome, one of the autosomal recessive, heritable urea cycle disorders, were studied with appropriate controls ultrastructurally. The patients were two severely retarded 10- and 12-year-old boys, and a 22-year-old sister of the former whose mental status was at the low normal range; she never had motor impairments or seizures. The mitochondria, similar in all three patients, were increased in number, very long, branching and/or "looping," and tortuous. "Spurs" or "buddings" extended from their lateral surfaces and the terminal segments were often bulbous. Other unusual configurations were also present. In addition, giant forms with large diameter contained innumerable closely-packed and parallel cristae which traversed the entire width of these mitochondria; at times they assumed a "whirled" pattern. The mitochondrial matrix was usually of high electron density. These changes were not a feature of fibroblastic mitochondria of controls. Several changes resembled those of hepatic mitochondria in this disorder. All features are interpreted as an attempt at expanding the mitochondrial volume (via structural substratum) to compensate for the metabolic incompetence of these organelles (a block in transmembranous transfer of ornithine from hyaloplasm into mitochondria for conversion to citrulline).

  3. Participation of oleic acid in the formation of the aortic aneurysm in Marfan syndrome patients.

    Science.gov (United States)

    Soto, María Elena; Iturriaga Hernández, Alejandra Valeria; Guarner Lans, Verónica; Zuñiga-Muñoz, Alejandra; Aranda Fraustro, Alberto; Velázquez Espejel, Rodrigo; Pérez-Torres, Israel

    2016-03-01

    Marfan syndrome (MFS) is associated with progressive aortic dilatation and endothelial dysfunction that lead to early acute dissection and rupture of the aorta and sudden death. Alteration in fatty acid (FA) metabolism can stimulate nitric oxide (NO) overproduction which increases the activity of the inducible form of NO synthase (iNOS) that is involved in endothelial dysfunction. We evaluated the participation of FA in the formation of thoracic aneurysms in MFS and its relation to the iNOS. Oleic acid (OA), iNOS, citrulline, nitrates and nitrites, TGF-β1, TNF-α, monounsaturated FA and NO synthase activity were significantly increased (p<0.05) in tissue from the aortas of MFS. Saturated FA, eNOS and HDL were significantly decreased (p<0.05). Arachidonic acid, delta-9 desaturase tended to increase and histological examination showed an increase in cystic necrosis, elastic fibers and collagen in MFS. The increase in OA contributes to the altered pathway of iNOS, which favors endothelial dysfunction and formation of the aortic aneurysms in MFS. PMID:27163200

  4. Complete dissection of the Hb(64-76) determinant using T helper 1, T helper 2 clones, and T cell hybridomas

    DEFF Research Database (Denmark)

    Evavold, B D; Williams, S G; Hsu, B L;

    1992-01-01

    cells identifying more critical amino acids then did the Th2 cells. For the Th1 cells and also the T cell hybridomas, the phenylalanine at position 71 was critical for a T cell response. Analysis of peptide affinity for IEk molecules indicated that position 71 played a role in peptide binding to MHC...... to an immunogenic peptide. A panel of altered Hb peptides was made by replacing each amino acid in the Hb peptide (positions 64-76) with a conservative amino acid substitution or an alanine. Although none of the eleven T cell clones and hybridomas tested exhibited the same pattern of reactivity to the substituted....... Secondary T cell contact residues, which were important for many but not all of the T cells, were identified at positions 69, 70, and 76. Overall T cell responses were minimally affected by changes in the amino acid residues at positions 64-68, 74, and 75. We have also demonstrated that cloned Th1 cells, Th...

  5. Characterization of Protein and Peptide Binding to Nanogels Formed by Differently Charged Chitosan Derivatives

    Directory of Open Access Journals (Sweden)

    Anastasia Zubareva

    2013-07-01

    Full Text Available Chitosan (Chi is a natural biodegradable cationic polymer with remarkable potency as a vehicle for drug or vaccine delivery. Chi possesses multiple groups, which can be used both for Chi derivatization and for particle formation. The aim of this work was to produce stable nanosized range Chi gels (nanogels, NGs with different charge and to study the driving forces of complex formation between Chi NGs and proteins or peptides. Positively charged NGs of 150 nm in diameter were prepared from hexanoyl chitosan (HC by the ionotropic gelation method while negatively charged NGs of 190 nm were obtained from succinoyl Chi (SC by a Ca2+ coacervation approach. NGs were loaded with a panel of proteins or peptides with different weights and charges. We show that NGs preferentially formed complexes with oppositely charged molecules, especially peptides, as was demonstrated by gel-electrophoresis, confocal microscopy and HPLC. Complex formation was accompanied by a change in zeta-potential and decrease in size. We concluded that complex formation between Chi NGs and peptide/proteins is mediated mostly by electrostatic interactions.

  6. De novo sequencing of two novel peptides homologous to calcitonin-like peptides, from skin secretion of the Chinese Frog, Odorrana schmackeri

    Directory of Open Access Journals (Sweden)

    Geisa P.C. Evaristo

    2015-09-01

    Full Text Available An MS/MS based analytical strategy was followed to solve the complete sequence of two new peptides from frog (Odorrana schmackeri skin secretion. This involved reduction and alkylation with two different alkylating agents followed by high resolution tandem mass spectrometry. De novo sequencing was achieved by complementary CID and ETD fragmentations of full-length peptides and of selected tryptic fragments. Heavy and light isotope dimethyl labeling assisted with annotation of sequence ion series. The identified primary structures are GCD[I/L]STCATHN[I/L]VNE[I/L]NKFDKSKPSSGGVGPESP-NH2 and SCNLSTCATHNLVNELNKFDKSKPSSGGVGPESF-NH2, i.e. two carboxyamidated 34 residue peptides with an aminoterminal intramolecular ring structure formed by a disulfide bridge between Cys2 and Cys7. Edman degradation analysis of the second peptide positively confirmed the exact sequence, resolving I/L discriminations. Both peptide sequences are novel and share homology with calcitonin, calcitonin gene related peptide (CGRP and adrenomedullin from other vertebrates. Detailed sequence analysis as well as the 34 residue length of both O. schmackeri peptides, suggest they do not fully qualify as either calcitonins (32 residues or CGRPs (37 amino acids and may justify their classification in a novel peptide family within the calcitonin gene related peptide superfamily. Smooth muscle contractility assays with synthetic replicas of the S–S linked peptides on rat tail artery, uterus, bladder and ileum did not reveal myotropic activity.

  7. The structure of HLA-DR52c: Comparison to other HLA-DRB3 alleles

    Energy Technology Data Exchange (ETDEWEB)

    Dai, Shaodong; Crawford, Frances; Marrack, Philippa; Kappler, John W. (HHMI)

    2008-09-05

    Class II major histocompatibility complex (MHCII) molecules present antigens to CD4{sup +} T cells. In addition to the most commonly studied human MHCII isotype, HLA-DR, whose {beta} chain is encoded by the HLA-DRB1 locus, several other isotypes that use the same {alpha} chain but have {beta} chains encoded by other genes. These other DR molecules also are expressed in antigen-presenting cells and are known to participate in peptide presentation to T cells and to be recognized as alloantigens by other T cells. Like some of the HLA-DRB1 alleles, several of these alternate DR molecules have been associated with specific autoimmune diseases and T cell hypersensitivity. Here we present the structure of an HLA-DR molecule (DR52c) containing one of these alternate {beta} chains (HLA-DRB3*0301) bound to a self-peptide derived from the Tu elongation factor. The molecule shares structurally conserved elements with other MHC class II molecules but has some unique features in the peptide-binding groove. Comparison of the three major HLA-DBR3 alleles (DR52a, b, and c) suggests that they were derived from one another by recombination events that scrambled the four major peptide-binding pockets at peptide positions 1, 4, 6, and 9 but left virtually no polymorphisms elsewhere in the molecules.

  8. Further investigation of a peptide extraction method with mesoporous silica using high-performance liquid chromatography coupled with tandem mass spectrometry.

    Science.gov (United States)

    Du, Yan; Wu, Dapeng; Guan, Yafeng

    2016-06-01

    Mobil Composition of Matter No. 41 (MCM-41) was the most frequently used mesoporous silica material to extract peptides from complex biological samples. However, there were confusing extraction conditions and large extraction efficiency variance among related reports, which resulted from unclear understanding about the interaction between the material and peptides. In this study, the extraction mechanism was investigated with one set of tryptic peptides by using high-performance liquid chromatography coupled with triple quadrupole mass spectrometry. Generally, hydrophobic interaction and electrostatic attraction were two major driving forces for extraction of peptides, while electrostatic repulsion greatly weakened the interaction between the material and peptides with isoelectric points below the pH. With most peptides positively charged and MCM-41 slightly negatively charged, most efficient extraction was obtained at pH 3, and it was proved that electrostatic and hydrophobic interaction acted in synergy for extraction of all the peptides. A mixed solution of acetonitrile with buffers of high pH or ion strength was demonstrated to be favorable for elution, which performed much better than the commonly used eluate (mixture of acetonitrile with 0.1% trifluoroacetic acid). Finally, under optimum conditions, it was found that extraction efficiency of MCM-41 for protein digest and human serum was greatly improved. PMID:27059091

  9. A computational docking study on the pH dependence of peptide binding to HLA-B27 sub-types differentially associated with ankylosing spondylitis.

    Science.gov (United States)

    Serçinoğlu, Onur; Özcan, Gülin; Kabaş, Zeynep Kutlu; Ozbek, Pemra

    2016-07-01

    A single amino acid difference (Asp116His), having a key role in a pathogenesis pathway, distinguishes HLA-B*27:05 and HLA-B*27:09 sub-types as associated and non-associated with ankylosing spondylitis, respectively. In this study, molecular docking simulations were carried out with the aim of comprehending the differences in the binding behavior of both alleles at varying pH conditions. A library of modeled peptides was formed upon single point mutations aiming to address the effect of 20 naturally occurring amino acids at the binding core peptide positions. For both alleles, computational docking was applied using Autodock 4.2. Obtained free energies of binding (FEB) were compared within the peptide library and between the alleles at varying pH conditions. The amino acid preferences of each position were studied enlightening the role of each on binding. The preferred amino acids for each position of pVIPR were found to be harmonious with experimental studies. Our results indicate that, as the pH is lowered, the capacity of HLA-B*27:05 to bind peptides in the library is largely lost. Hydrogen bonding analysis suggests that the interaction between the main anchor positions of pVIPR and their respective binding pocket residues are affected from the pH the most, causing an overall shift in the FEB profiles. PMID:27506766

  10. Integrating solid-state NMR and computational modeling to investigate the structure and dynamics of membrane-associated ghrelin.

    Directory of Open Access Journals (Sweden)

    Gerrit Vortmeier

    Full Text Available The peptide hormone ghrelin activates the growth hormone secretagogue receptor 1a, also known as the ghrelin receptor. This 28-residue peptide is acylated at Ser3 and is the only peptide hormone in the human body that is lipid-modified by an octanoyl group. Little is known about the structure and dynamics of membrane-associated ghrelin. We carried out solid-state NMR studies of ghrelin in lipid vesicles, followed by computational modeling of the peptide using Rosetta. Isotropic chemical shift data of isotopically labeled ghrelin provide information about the peptide's secondary structure. Spin diffusion experiments indicate that ghrelin binds to membranes via its lipidated Ser3. Further, Phe4, as well as electrostatics involving the peptide's positively charged residues and lipid polar headgroups, contribute to the binding energy. Other than the lipid anchor, ghrelin is highly flexible and mobile at the membrane surface. This observation is supported by our predicted model ensemble, which is in good agreement with experimentally determined chemical shifts. In the final ensemble of models, residues 8-17 form an α-helix, while residues 21-23 and 26-27 often adopt a polyproline II helical conformation. These helices appear to assist the peptide in forming an amphipathic conformation so that it can bind to the membrane.

  11. The most common mutation of KRT9, c.C487T (p.R163W), in epidermolytic palmoplantar keratoderma in two large Chinese pedigrees.

    Science.gov (United States)

    Liu, Wen-Ting; Ke, Hai-Ping; Zhao, Yan; Chen, Xiao-Ling; Lu, Jia-Jun; Du, Zheng-Fang; Yu, Dan; Zhang, Xian-Ning

    2012-04-01

    Epidermolytic palmoplantar keratoderma (EPPK) is generally associated with dominant-negative mutations of the Keratin 9 gene (KRT9), and rarely with the Keratin 1 gene (KRT1). To date, a myriad of mutations has been reported with a high frequency of codon 163 mutations within the first exon of KRT9 in different populations. Notably, a distinct phenotypic heterogeneity, digital mutilation, was found recently in a 58-year-old female Japanese EPPK patient with p.R163W. Here, we report the most common mutation, c.C487T (p.R163W) of KRT9, in two large EPPK pedigrees from southeast China. The arginine residue in peptide position 163 remains almost constant in at least 47 intermediate filament proteins ranging from snail to human. A substitution in arginine alters both the charge and shape of the 1A rod domain and disrupts the function of the helix initiation motif of keratins, finally compromising the integrity of filaments and weakening their stability in the epidermis of palms and soles. We summarize the clinical symptoms of EPPK in Chinese and show that knuckle pads are associated with KRT9 mutations. We suggest that the frequency of p.R163W in Chinese EPPK patients (31.03%) is consistent with that in the general population (29.33%), and that codon 163 is truly a hotspot mutational site of KRT9.

  12. Down-Regulation of Porcine Heart Diaphorase Reactivity by Trimanganese Hexakis(3,5-Diisopropylsalicylate), Mn(3)(3,5-DIPS)6, and Down-Regulation of Nitric Oxide Synthase Reactivity by Mn(3)(3,5-DIPS)(6) and Cu(II)(2)(3,5-DIPS)(4).

    Science.gov (United States)

    Booth, B L; Pitters, E; Mayer, B; Sorenson, J R

    1999-01-01

    Purposes of this work were to examine the plausible down-regulation of porcine heart diaphorase (PHD) enzyme reactivity and nitric oxide synthase (NOS) enzyme reactivity by trimanganese hexakis(3,5-diisopropylsalicylate), [Mn(3)(3,5-DIPS)(6)] as well as dicopper tetrakis(3,5- diisopropylsalicylate, [Cu(II)(2)(3,5-DIPS)(4)] as a mechanistic accounting for their pharmacological activities.Porcine heart disease was found to oxidize 114 muM reduced nicotinamide-adenine- dinucleotide-'(3)-phosphate (NADPH) with a corresponding reduction of an equivalent concentration of 2,6-dichlorophenolindophenol (DCPIP). As reported for Cu(II)(2) (3,5-DIPS)(4), addition of Mn(3)(3,5-DIPS)(6) to this reaction mixture decreased the reduction of DCPIP without significantly affecting the oxidation of NADPH. The concentration of Mn(3)(3,5-DIPS)(6) that produced a 50% decrease in DCPIP reduction (IC(50)) was found to be 5muM. Mechanistically, this inhibition of DCPIP reduction with ongoing NADPH oxidation by PHD was found to be due to the ability of Mn(3)(3,5-DIPS)(6) to serve as a catalytic electron acceptor for reduced PHD as had been reported for Cu(II)(2)(3,5-DIPS)(4). This catalytic decrease in reduction of DCPIP by Mn(3)(3,5-DIPS)(6) was enhanced by the presence of a large concentration of DCPIP and decreased by the presence of a large concentration of NADPH, consistent with what had been observed for the activity of Cu(II)(2)(3,5-DIPS)(4)Oxidation of NADPH by PHD in the presence of Mn(3)(3,5-DIPS)(6) and the absence of DCPIP was linearly related to the concentration of added Mn(3)(3,5-DIPS)(6) through the concentration range of 2.4 muM to 38muM with a 50% recovery of NADPH oxidation by PHD at a concentration of 6 muM Mn(3)(3,5-DIPS)(6)Conversion of [(3)H] L-Arginine to [(3)H] L-Citrulline by purified rat brain nitric oxide synthase (NOS) was decreased in a concentrated related fashion with the addition of Mn(3)(3,5-DIPS)(6) as well as Cu(II)(2)(3,5-DIPS)(4) which is an extention of

  13. Ectopic lymphoid structures support ongoing production of class-switched autoantibodies in rheumatoid synovium.

    Directory of Open Access Journals (Sweden)

    Frances Humby

    2009-01-01

    Full Text Available BACKGROUND: Follicular structures resembling germinal centres (GCs that are characterized by follicular dendritic cell (FDC networks have long been recognized in chronically inflamed tissues in autoimmune diseases, including the synovium of rheumatoid arthritis (RA. However, it is debated whether these ectopic structures promote autoimmunity and chronic inflammation driving the production of pathogenic autoantibodies. Anti-citrullinated protein/peptide antibodies (ACPA are highly specific markers of RA, predict a poor prognosis, and have been suggested to be pathogenic. Therefore, the main study objectives were to determine whether ectopic lymphoid structures in RA synovium: (i express activation-induced cytidine deaminase (AID, the enzyme required for somatic hypermutation and class-switch recombination (CSR of Ig genes; (ii support ongoing CSR and ACPA production; and (iii remain functional in a RA/severe combined immunodeficiency (SCID chimera model devoid of new immune cell influx into the synovium. METHODS AND FINDINGS: Using immunohistochemistry (IHC and quantitative Taqman real-time PCR (QT-PCR in synovial tissue from 55 patients with RA, we demonstrated that FDC+ structures invariably expressed AID with a distribution resembling secondary lymphoid organs. Further, AID+/CD21+ follicular structures were surrounded by ACPA+/CD138+ plasma cells, as demonstrated by immune reactivity to citrullinated fibrinogen. Moreover, we identified a novel subset of synovial AID+/CD20+ B cells outside GCs resembling interfollicular large B cells. In order to gain direct functional evidence that AID+ structures support CSR and in situ manufacturing of class-switched ACPA, 34 SCID mice were transplanted with RA synovium and humanely killed at 4 wk for harvesting of transplants and sera. Persistent expression of AID and Igamma-Cmu circular transcripts (identifying ongoing IgM-IgG class-switching was observed in synovial grafts expressing FDCs/CD21L

  14. Correlations between immunogenicity, drug levels, and disease activity in an Italian cohort of rheumatoid arthritis patients treated with tocilizumab

    Science.gov (United States)

    Benucci, Maurizio; Meacci, Francesca; Grossi, Valentina; Infantino, Maria; Manfredi, Mariangela; Bellio, Emanuele; Bellio, Valerio; Li Gobbi, Francesca; Bazzichi, Laura; Moscato, Paolo; Caputo, Dario; Saviola, Gianantonio; Talotta, Rossella; Sarzi-Puttini, Piercarlo; Atzeni, Fabiola

    2016-01-01

    The aim of this study was to evaluate the real-life immunogenicity of anti-drug antibodies, drug levels, and disease activity in an Italian cohort of rheumatoid arthritis patients treated with tocilizumab (TCZ). We evaluated 126 TCZ-treated patients with rheumatoid arthritis (16 males and 110 females; mean age 59±12 years, range 26–83; mean disease duration 11±5 years) with inadequate 12-week response to any synthetic and biological disease-modifying anti-rheumatic drugs, in a retrospective analysis. One-hundred and seven patients were treated with methotrexate mean dose 12.6±1.3 mg/week in combination with TCZ, 13 received TCZ monotherapy, and six received leflunomide 20 mg/day plus TCZ; all patients were treated with prednisone mean dose 6.4±1.2 mg/day. They had a 28-joint Disease Activity Score (DAS28) of >3.2, an erythrocyte sedimentation rate (ESR) of >30 mm/hour, and CRP levels of >1.0 mg/dL. We evaluated at baseline and after 6 months of treatment: DAS28; rheumatoid factor (RF) IgM, IgA, and IgG; anti-citrullinated peptide antibody; ESR; CRP; TNF-α; and IL-6. TCZ and anti-TCZ antibodies were detected using LISA-TRACKER Duo TCZ. TCZ levels of 10 µg/mL high. After 6 months of treatment only one patient was positive for anti-TCZ antibodies. There were correlations between DAS28, ESR, and CRP and IL-6 levels in all patients. Comparison of the 84 patients with TCZ levels of 10 µg/mL showed the following differences: DAS28: 3.09±1.32 vs 2.78±1.32, P=0.0005; ESR: 27±14.8 vs 14±12 mm/hour, P=0.0001; CRP: 1.47±1.05 vs 0.65±0.80 mg/dL, P=0.0086; TNF-α: 10.2±1.2 vs 9.9±1.1 pg/mL, P=0.999; IL-6: 3.65±4.75 vs 3.62±4.41 pg/mL, P=0.97; anti-citrullinated peptide antibody: 85.2±93.7 vs 86.7±90.3 IU/mL, P=0.94; RF IgM: 72.4±62.7 vs 68.3±61.6 IU/mL, P=0.754; RF IgA: 41.7±36.4 vs 47.8±42.1 U/mL, P=0.449; and RF IgG: 46.4±46.1 vs 59.3±58.2 U/mL, P=0.212. These findings show that the occurrence of anti-drug antibodies against TCZ is very rare and that

  15. Urea cycle disorders in Thai infants: a report of 5 cases.

    Science.gov (United States)

    Wasant, Pornswan; Srisomsap, Chantragan; Liammongkolkul, Somporn; Svasti, Jisnuson

    2002-08-01

    Urea Cycle Disorders (UCD) is an inborn error of urea synthesis in which ammonium and other nitrogenous precursors of urea accumulate leading to episodic coma and a high mortality rate. Therapy with peritoneal dialysis, essential amino acids or their nitrogen-free analogues has increased survival. The authors report 5 cases of urea cycle disorders, all of whom developed and were rescued from hyperammonemic coma. However, the eventual outcome was quite variable. Argininosuccinate lyase deficiency (ALD) Case 1. A 2 month old male infant, a product of a consanguineous marriage (Suphanburi province); developed poor feeding on day 7, lethargy, convulsion, hepatomegaly and respiratory alkalosis leading to respiratory failure and coma. Hyperammonemia, elevation of glutamic acid and argininosuccinic acid and its anhydrides confirmed the diagnosis of ALD. He is now 9 years old and severely retarded. Case 2. A male infant with history of lethargy, poor feeding on day 3, treated as sepsis and required respiratory support for 6 days; subsequently readmitted at age 2 weeks with vomitting, lethargy, seizure activity and hyperammonemia, and was treated by a local pediatrician in Songkhla province. There was a history of parental consanguinity and he was referred to Siriraj Hospital on day 64 with severe essential amino acid deficiency and acrodermatitis enteropathica with markedly elevated plasma citrulline level. In spite of aggressive treatment; the patient developed sepsis and he expired on day 78. Ornithine transcarbamylase deficiency (OTC) Case 3. An eleven-month-old male infant, the product of a non-consanguineous marriage, developed neonatal onset of hyperammonemia on day 5 after poor feeding, lethargy, hypothermia, seizure, apnea and coma. He was rescued from neonatal hyperammonemic coma on day 9 after aggressive treatment, but expired at eleven months of age after overwhelming sepsis. Case 4. A male infant, sibling of case 3 was referred to Siriraj Hospital on day 8 with

  16. High IL-23 level is a marker of disease activity in rheumatoid arthritis.

    Science.gov (United States)

    Abu Al Fadl, Esam M; Fattouh, Mona; Allam, Ahmed A

    2013-01-01

    Rheumatoid arthritis (RA) is a chronic autoimmune systemic disorder characterized by inflammatory responses mainly affecting the synovial joints. Interleukin-23 (IL-23) is a heterodimeric pro-inflammatory cytokine secreted by activated dendritic cells and activated macrophages. IL-23 is the key cytokine controlling inflammation in peripheral tissues leading to the development of autoimmune diseases. The objective of our study was to determine the relationship between the IL-23 level and disease activity in RA patients. Sixty RA patients were included in the study with mean age of 40 years; they included 44 (73.3 %) females and 16 males (26.7 %). The clinical parameters of disease activity were determined, including the 28-joint disease activity score (DAS28), serum levels of C-reactive protein (CRP), Anti-citrullinated peptide antibody (ACPA), rheumatoid factor (RF), and TNF-alpha and the degree of bony erosions based on X-rays. Patients were subdivided into active disease group (n = 30) with DAS28 score higher than 5.1 (Group I); and remission group (n = 30) with DAS28 score less than 2.6 (Group II). Thirty healthy individuals in the same age group of RA patients including 22 (73.3%) females and 8 males (26.7%) were randomly selected as the control group (Group III). The levels of IL-23 were determined by enzyme-linked immunosorbent assay (ELISA) and the correlations between the serum levels of IL-23 and disease activity parameters of patients with RA were determined. Serum levels of IL-23 were significantly higher in RA patients during active stage of the disease in comparison to the patients in remission and the control group. There was a significant positive correlation between serum IL-23 levels in RA patients and individual disease activity parameters. It is concluded that elevated serum IL-23 level may be a useful marker to detect active RA and disease progression in patients with RA. PMID:24617049

  17. Reprogramming: A Preventive Strategy in Hypertension Focusing on the Kidney

    Directory of Open Access Journals (Sweden)

    You-Lin Tain

    2015-12-01

    Full Text Available Adulthood hypertension can be programmed in response to a suboptimal environment in early life. However, developmental plasticity also implies that one can prevent hypertension in adult life by administrating appropriate compounds during early development. We have termed this reprogramming. While the risk of hypertension has been assessed in many mother-child cohorts of human developmental programming, interventions necessary to prove causation and provide a reprogramming strategy are lacking. Since the developing kidney is particularly vulnerable to environmental insults and blood pressure is determined by kidney function, renal programming is considered key in developmental programming of hypertension. Common pathways, whereby both genetic and acquired developmental programming converge into the same phenotype, have been recognized. For instance, the same reprogramming interventions aimed at shifting nitric oxide (NO-reactive oxygen species (ROS balance, such as perinatal citrulline or melatonin supplements, can be protective in both genetic and developmentally programmed hypertension. Furthermore, a significantly increased expression of gene Ephx2 (soluble epoxide hydrolase was noted in both genetic and acquired animal models of hypertension. Since a suboptimal environment is often multifactorial, such common reprogramming pathways are a practical finding for translation to the clinic. This review provides an overview of potential clinical applications of reprogramming strategies to prevent programmed hypertension. We emphasize the kidney in the following areas: mechanistic insights from human studies and animal models to interpret programmed hypertension; identified risk factors of human programmed hypertension from mother-child cohorts; and the impact of reprogramming strategies on programmed hypertension from animal models. It is critical that the observed effects on developmental reprogramming in animal models are replicated in human

  18. Quantitative amino acid profiling and stable isotopically labeled amino acid tracer enrichment used for in vivo human systemic and tissue kinetics measurements.

    Science.gov (United States)

    Bornø, Andreas; van Hall, Gerrit

    2014-03-01

    An important area within clinical functional metabolomics is in vivo amino acid metabolism and protein turnover measurements for which accurate amino acid concentrations and stable isotopically labeled amino acid enrichments are mandatory not the least when tissue metabolomics is determined. The present study describes a new sensitive liquid chromatography tandem mass-spectrometry method quantifying 20 amino acids and their tracer(s) ([ring-(13)C6]/D5Phenylalanine) in human plasma and skeletal muscle specimens. Before analysis amino acids were extracted and purified via deprotonization/ion exchange, derivatized using a phenylisothiocyanate reagent and each amino acid was quantitated with its own stable isotopically labeled internal standard (uniformly labeled-(13)C/(15)N). The method was validated according to general recommendations for chromatographic analytical methods. The calibration curve correlations for amino acids were on average; r(2)=0.998. Interday accuracy for amino acids determined in spiked plasma was on average 97.3% and the coefficient of variation (CV) was 2.6%. The ([ring-(13)C6]/D5Phenylalanine) enrichment CV's for machine reproducibility in muscle tissue fluid and plasma were 4.4 and 0.8%, and the interday variability was 3.4% and the recovery was 90.5%, respectively. In conclusion, we have developed and validated a method for quantitative amino acid profiling that meets the requirements for systemic and tissue human in vivo amino acid and protein turnover kinetics measurements. Moreover, citrulline, ornithine, π-methyl-histidine, τ-methyl-l-histidine, hydroxy-proline and carnitine were analysed but when similar precision and accuray are required an additional stable istopically labeled internal standard for these meatablites should be be added.

  19. Autoantibodies in inflammatory arthritis.

    Science.gov (United States)

    Conigliaro, P; Chimenti, M S; Triggianese, P; Sunzini, F; Novelli, L; Perricone, C; Perricone, R

    2016-07-01

    Rheumatoid arthritis (RA) is a systemic chronic inflammatory disease characterized by extensive synovitis resulting in erosions of articular cartilage and marginal bone with joint destruction. The lack of immunological tolerance in RA represents the first step toward the development of autoimmunity. Susceptible individuals, under the influence of environmental factors, such as tobacco smoke, and silica exposure, develop autoimmune phenomena that result in the presence of autoantibodies. HLA and non-HLA haplotypes play a major role in determining the development of specific autoantibodies differentiating anti-citrullinated antibodies (ACPA)-positive and negative RA patients. Rheumatoid factor (RF) and ACPA are the serological markers for RA, and during the preclinical immunological phase, autoantibody titers increase with a progressive spread of ACPA antigens repertoire. The presence of ACPA represents an independent risk factor for developing RA in patients with undifferentiated arthritis or arthralgia. Moreover, anti-CarP antibodies have been identified in patients with RA as well as in individuals before the onset of clinical symptoms of RA. Several autoantibodies mainly targeting post-translational modified proteins have been investigated as possible biomarkers to improve the early diagnosis, prognosis and response to therapy in RA patients. Psoriatic arthritis (PsA) is distinguished from RA by infrequent positivity for RF and ACPA, together with other distinctive clinical features. Actually, specific autoantibodies have not been described. Recently, anti-CarP antibodies have been reported in sera from PsA patients with active disease. Further investigations on autoantibodies showing high specificity and sensibility as well as relevant correlation with disease severity, progression, and response to therapy are awaited in inflammatory arthritides. PMID:26970491

  20. HIPERAMONEMIA NEONATAL CAUSADA POR DEFECTOS DEL CICLO DE LA UREA Neonatal hyperammonemia in urea cycle disorders patients

    Directory of Open Access Journals (Sweden)

    Yolanda Cifuentes C

    2010-12-01

    Full Text Available Los defectos del ciclo de la úrea se deben a deficiencias de diferentes enzimas; las manifestaciones clínicas son similares y están relacionadas con la hiperamonemia. Se presentan las historias clínicas de tres neonatos a término, sin evidencia de alteración al nacimiento. Se les detectó hiperamonemia y se sospechó enfermedad metabólica. La cromatografía de aminoácidos sugirió defectos del ciclo de la úrea. El manejo incluyó dieta con restricción de proteínas, administración de benzoato de sodio, exsanguinotransfusión y diálisis peritoneal pese a lo cual fallecieron. Se revisan las causas de hiperamonemia en el neonato y se propone una secuencia para su diagnósticoThe urea cycle disorders result from deficiency of activity of enzymes N-acetyl glutamate synthetase, carbamyl phosphate synthase, ornithine transcarbamylase, argininosuccinic acid synthetase, argininosuccinic acid lyase and arginase. Except for the last one, the clinical features are similar and related with the hiperammonaemia. It reports three full term, newborn cases, they had encephalopathy and needed respiratory support after be well in neonatal period. They had hyperammonemia as inborn error. The thin layer amino acids chromatography showed alanine and glutamine, in the siblings appeared citruline, suggesting urea cycle disorders. Despite protein restriction diet, sodium benzoate administration, blood exchange and peritoneal dialysis,babies died. High argininosuccinic acid levels in the first case and high citrulline levels with argininosuccinic acid absence in the third case, which was diagnosed as argininosuccinic aciduria with citrullinemia. This report provide an overview of neonatal hyperammonemia causes and propose a secuency for diagnosis

  1. Orphan drugs in development for urea cycle disorders: current perspectives

    Directory of Open Access Journals (Sweden)

    Häberle J

    2014-09-01

    Full Text Available Johannes Häberle,1 Shawn E McCandless2 1Division of Metabolism and Children's Research Center, University Children's Hospital Zurich, Zurich, Switzerland; 2Center for Human Genetics, University Hospitals Case Medical Center, and Department of Genetics and Genome Sciences, Case Western Reserve University, Cleveland, OH, USA Abstract: The urea cycle disorders are caused by deficiency of one of the six hepatic enzymes or two transporters involved in detoxification of ammonia. The resulting hyperammonemia causes severe brain injury unless aggressive steps are taken to reduce the accumulation of ammonia, which is thought to be the most toxic metabolite. This review describes the current state of chronic management of urea cycle disorders, focusing on new and emerging therapies. Management strategies include the mainstay of treatment, namely dietary protein restriction and supplementation with l-arginine or l-citrulline. Several currently approved medications utilize and enhance alternative pathways of waste nitrogen excretion (sodium benzoate, sodium phenylacetate, sodium phenylbutyrate in several formulations, and glycerol phenylbutyrate, working through conjugation of the drug to either glycine (in the case of benzoate or glutamine, the products of which are excreted in the urine. Carglumic acid activates the first committed step of conversion of ammonia to urea, carbamoylphosphate synthetase, and thus effectively treats defective synthesis of the endogenous activator, N-acetylglutamate, whether due to genetic defects or biochemical inhibition of the N-acetylglutamate synthase enzyme. Approaches to neuroprotection during episodes of hyperammonemia are discussed, including the use of controlled hypothermia (brain cooling, as well as proposed, but as yet untested, pharmacologic therapies. Finally, cell-based therapies, including liver transplantation, infusion of fresh or cryopreserved hepatocytes, use of stem cells, and new approaches to gene

  2. Human neutrophils in auto-immunity.

    Science.gov (United States)

    Thieblemont, Nathalie; Wright, Helen L; Edwards, Steven W; Witko-Sarsat, Véronique

    2016-04-01

    Human neutrophils have great capacity to cause tissue damage in inflammatory diseases via their inappropriate activation to release reactive oxygen species (ROS), proteases and other tissue-damaging molecules. Furthermore, activated neutrophils can release a wide variety of cytokines and chemokines that can regulate almost every element of the immune system. In addition to these important immuno-regulatory processes, activated neutrophils can also release, expose or generate neoepitopes that have the potential to break immune tolerance and result in the generation of autoantibodies, that characterise a number of human auto-immune diseases. For example, in vasculitis, anti-neutrophil cytoplasmic antibodies (ANCA) that are directed against proteinase 3 or myeloperoxidase are neutrophil-derived autoantigens and activated neutrophils are the main effector cells of vascular damage. In other auto-immune diseases, these neutrophil-derived neoepitopes may arise from a number of processes that include release of granule enzymes and ROS, changes in the properties of components of their plasma membrane as a result of activation or apoptosis, and via the release of Neutrophil Extracellular Traps (NETs). NETs are extracellular structures that contain chromatin that is decorated with granule enzymes (including citrullinated proteins) that can act as neo-epitopes to generate auto-immunity. This review therefore describes the processes that can result in neutrophil-mediated auto-immunity, and the role of neutrophils in the molecular pathologies of auto-immune diseases such as vasculitis, rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE). We discuss the potential role of NETs in these processes and some of the debate in the literature regarding the role of this phenomenon in microbial killing, cell death and auto-immunity. PMID:27036091

  3. Potential biomarkers of fatigue identified by plasma metabolome analysis in rats.

    Science.gov (United States)

    Kume, Satoshi; Yamato, Masanori; Tamura, Yasuhisa; Jin, Guanghua; Nakano, Masayuki; Miyashige, Yukiharu; Eguchi, Asami; Ogata, Yoshiyuki; Goda, Nobuhito; Iwai, Kazuhiro; Yamano, Emi; Watanabe, Yasuyoshi; Soga, Tomoyoshi; Kataoka, Yosky

    2015-01-01

    In the present study, prior to the establishment of a method for the clinical diagnosis of chronic fatigue in humans, we validated the utility of plasma metabolomic analysis in a rat model of fatigue using capillary electrophoresis-mass spectrometry (CE-MS). In order to obtain a fatigued animal group, rats were placed in a cage filled with water to a height of 2.2 cm for 5 days. A food-restricted group, in which rats were limited to 10 g/d of food (around 50% of the control group), was also assessed. The food-restricted group exhibited weight reduction similar to that of the fatigued group. CE-MS measurements were performed to evaluate the profile of food intake-dependent metabolic changes, as well as the profile in fatigue loading, resulting in the identification of 48 metabolites in plasma. Multivariate analyses using hierarchical clustering and principal component analysis revealed that the plasma metabolome in the fatigued group showed clear differences from those in the control and food-restricted groups. In the fatigued group, we found distinctive changes in metabolites related to branched-chain amino acid metabolism, urea cycle, and proline metabolism. Specifically, the fatigued group exhibited significant increases in valine, leucine, isoleucine, and 2-oxoisopentanoate, and significant decreases in citrulline and hydroxyproline compared with the control and food-restricted groups. Plasma levels of total nitric oxide were increased in the fatigued group, indicating systemic oxidative stress. Further, plasma metabolites involved in the citrate cycle, such as cis-aconitate and isocitrate, were reduced in the fatigued group. The levels of ATP were significantly decreased in the liver and skeletal muscle, indicative of a deterioration in energy metabolism in these organs. Thus, this comprehensive metabolic analysis furthered our understanding of the pathophysiology of fatigue, and identified potential diagnostic biomarkers based on fatigue pathophysiology.

  4. Genetic Manipulation of Leishmania donovani to Explore the Involvement of Argininosuccinate Synthase in Oxidative Stress Management.

    Directory of Open Access Journals (Sweden)

    Abul Hasan Sardar

    2016-03-01

    Full Text Available Reactive oxygen and nitrogen species (ROS and RNS produced by the phagocytic cells are the most common arsenals used to kill the intracellular pathogens. However, Leishmania, an intracellular pathogen, has evolved mechanisms to survive by counterbalancing the toxic oxygen metabolites produced during infection. Polyamines, the major contributor in this anti-oxidant machinery, are largely dependent on the availability of L-arginine in the intracellular milieu. Argininosuccinate synthase (ASS plays an important role as the rate-limiting step required for converting L-citrulline to argininosuccinate to provide arginine for an assortment of metabolic processes. Leishmania produce an active ASS enzyme, yet it has an incomplete urea cycle as it lacks an argininosuccinate lyase (ASL. There is no evidence for endogenous synthesis of L-arginine in Leishmania, which suggests that these parasites salvage L-arginine from extracellular milieu and makes the biological function of ASS and the production of argininosuccinate in Leishmania unclear. Our previous quantitative proteomic analysis of Leishmania promastigotes treated with sub-lethal doses of ROS, RNS, or a combination of both, led to the identification of several differentially expressed proteins which included ASS. To assess the involvement of ASS in stress management, a mutant cell line with greatly reduced ASS activity was created by a double-targeted gene replacement strategy in L. donovani promastigote. Interestingly, LdASS is encoded by three copies of allele, but Western blot analysis showed the third allele did not appear to express ASS. The free thiol levels in the mutant LdASS-/-/+ cell line were decreased. Furthermore, the cell viability in L-arginine depleted medium was greatly attenuated on exposure to different stress environments and was adversely impacted in its ability to infect mice. These findings suggest that ASS is important for Leishmania donovani to counterbalance the stressed

  5. Pig kidney legumain: an asparaginyl endopeptidase with restricted specificity.

    Science.gov (United States)

    Dando, P M; Fortunato, M; Smith, L; Knight, C G; McKendrick, J E; Barrett, A J

    1999-01-01

    Legumain was recently discovered as a lysosomal endopeptidase in mammals [Chen, Dando, Rawlings, Brown, Young, Stevens, Hewitt, Watts and Barrett (1997) J. Biol. Chem. 272, 8090-8098], having been known previously only from plants and invertebrates. It has been shown to play a key role in processing of the C fragment of tetanus toxin for presentation by the MHC class-II system [Manoury, Hewitt, Morrice, Dando, Barrett and Watts (1998) Nature (London) 396, 695-699]. We examine here the specificity of the enzyme from pig kidney by use of protein, oligopeptide and synthetic arylamide substrates, all determinations being made at pH 5.8. In proteins, only about one in ten of the asparaginyl bonds were hydrolysed, and these were mostly predicted to be located at turns on the protein surface. Bonds that were not cleaved in tetanus toxin were cleaved when presented in oligopeptides, sometimes faster than an equivalent oligopeptide based on a bond that was cleaved in the protein. Legumain cleaved the bait region of rat alpha1-macroglobulin and was 'trapped' by the macroglobulin, as most other endopeptidases are, but did not interact with human alpha2-macroglobulin, which contains no asparagine residue in its bait region. Glycosylation of asparagine totally prevented hydrolysis by legumain. Specificity for arylamide substrates was evaluated with reference to benzyloxycarbonyl-Ala-Ala-Asn-aminomethylcoumarin, and the preference for the P3-position amino acid was Ala>Tyr(tertiary butyl)>Val>Pro>Phe=Tyr>Leu=Gly. There was no hydrolysis of substrate analogues containing mono- or di-N-methylasparagines, l-2-amino-3-ureidopropionic acid or citrulline in the P1 position. We conclude that mammalian legumain appears to be totally restricted to the hydrolysis of asparaginyl bonds in substrates of all kinds. There seem to be no strong preferences for particular amino acids in other subsites, and yet there are still unidentified factors that prevent hydrolysis of many asparaginyl bonds

  6. Metabolomics reveals amino acids contribute to variation in response to simvastatin treatment.

    Directory of Open Access Journals (Sweden)

    Miles Trupp

    Full Text Available UNLABELLED: Statins are widely prescribed for reducing LDL-cholesterol (C and risk for cardiovascular disease (CVD, but there is considerable variation in therapeutic response. We used a gas chromatography-time-of-flight mass-spectrometry-based metabolomics platform to evaluate global effects of simvastatin on intermediary metabolism. Analyses were conducted in 148 participants in the Cholesterol and Pharmacogenetics study who were profiled pre and six weeks post treatment with 40 mg/day simvastatin: 100 randomly selected from the full range of the LDL-C response distribution and 24 each from the top and bottom 10% of this distribution ("good" and "poor" responders, respectively. The metabolic signature of drug exposure in the full range of responders included essential amino acids, lauric acid (p<0.0055, q<0.055, and alpha-tocopherol (p<0.0003, q<0.017. Using the HumanCyc database and pathway enrichment analysis, we observed that the metabolites of drug exposure were enriched for the pathway class amino acid degradation (p<0.0032. Metabolites whose change correlated with LDL-C lowering response to simvastatin in the full range responders included cystine, urea cycle intermediates, and the dibasic amino acids ornithine, citrulline and lysine. These dibasic amino acids share plasma membrane transporters with arginine, the rate-limiting substrate for nitric oxide synthase (NOS, a critical mediator of cardiovascular health. Baseline metabolic profiles of the good and poor responders were analyzed by orthogonal partial least square discriminant analysis so as to determine the metabolites that best separated the two response groups and could be predictive of LDL-C response. Among these were xanthine, 2-hydroxyvaleric acid, succinic acid, stearic acid, and fructose. Together, the findings from this study indicate that clusters of metabolites involved in multiple pathways not directly connected with cholesterol metabolism may play a role in modulating

  7. Genetic Manipulation of Leishmania donovani to Explore the Involvement of Argininosuccinate Synthase in Oxidative Stress Management

    Science.gov (United States)

    Sardar, Abul Hasan; Jardim, Armando; Ghosh, Ayan Kumar; Mandal, Abhishek; Das, Sushmita; Saini, Savita; Abhishek, Kumar; Singh, Ruby; Verma, Sudha; Kumar, Ajay; Das, Pradeep

    2016-01-01

    Reactive oxygen and nitrogen species (ROS and RNS) produced by the phagocytic cells are the most common arsenals used to kill the intracellular pathogens. However, Leishmania, an intracellular pathogen, has evolved mechanisms to survive by counterbalancing the toxic oxygen metabolites produced during infection. Polyamines, the major contributor in this anti-oxidant machinery, are largely dependent on the availability of L-arginine in the intracellular milieu. Argininosuccinate synthase (ASS) plays an important role as the rate-limiting step required for converting L-citrulline to argininosuccinate to provide arginine for an assortment of metabolic processes. Leishmania produce an active ASS enzyme, yet it has an incomplete urea cycle as it lacks an argininosuccinate lyase (ASL). There is no evidence for endogenous synthesis of L-arginine in Leishmania, which suggests that these parasites salvage L-arginine from extracellular milieu and makes the biological function of ASS and the production of argininosuccinate in Leishmania unclear. Our previous quantitative proteomic analysis of Leishmania promastigotes treated with sub-lethal doses of ROS, RNS, or a combination of both, led to the identification of several differentially expressed proteins which included ASS. To assess the involvement of ASS in stress management, a mutant cell line with greatly reduced ASS activity was created by a double-targeted gene replacement strategy in L. donovani promastigote. Interestingly, LdASS is encoded by three copies of allele, but Western blot analysis showed the third allele did not appear to express ASS. The free thiol levels in the mutant LdASS-/-/+ cell line were decreased. Furthermore, the cell viability in L-arginine depleted medium was greatly attenuated on exposure to different stress environments and was adversely impacted in its ability to infect mice. These findings suggest that ASS is important for Leishmania donovani to counterbalance the stressed environments

  8. Treating sarcopenia in older and oldest old.

    Science.gov (United States)

    Martone, Anna Maria; Lattanzio, Fabrizia; Abbatecola, Angela Marie; Carpia, Domenico La; Tosato, Matteo; Marzetti, Emanuele; Calvani, Riccardo; Onder, Graziano; Landi, Francesco

    2015-01-01

    The presence of sarcopenia is not only rapidly rising in geriatric clinical practice and research, but is also becoming a significant concept in numerous medical specialties. This rapidly rising concept has encouraged the need to identify methods for treating sarcopenia. Physical activity measures using resistance training exercise, combined with nutritional interventions (protein and amino acid supplementation) have shown to significantly improve muscle mass and strength in older persons. Moreover, resistance training may improve muscle strength and mass by improving protein synthesis in skeletal muscle cells. Aerobic exercise has also shown to hold beneficial impacts on sarcopenia by improving insulin sensitivity. At the moment, the literature indicates that most significant improvement in sarcopenia is based on exercise programs. Thus, this type of intervention should be implemented in a persistent manner over time in elders, with or at risk of muscle loss. At the same time, physical training exercise should include correcting nutritional deficits with supplementation methods. For example, in older sarcopenic patients with adequate renal function, daily protein intake should be increased to >1. 0 grams of protein per kilogram of body weight. In particular, leucine, - hydroxy β-methylbutyrate (HMB), creatine and some milk-based proteins have been shown to improve skeletal muscle protein balance. In addition, it is also recommended for adjustment of for vitamin D deficiency, if present, considering the crucial role of vitamin D in the skeletal muscle. In this review, we provide evidence regarding the effects of different physical exercise protocols, specific nutritional intervention, and some new metabolic agents (HMB, citrulline malate, ornithine, and others) on clinical outcomes related to sarcopenia in older adults. PMID:25633117

  9. Argininosuccinate Synthetase Is a Functional Target for a Snake Venom Anti-hypertensive Peptide

    Science.gov (United States)

    Guerreiro, Juliano R.; Lameu, Claudiana; Oliveira, Eduardo F.; Klitzke, Clécio F.; Melo, Robson L.; Linares, Edlaine; Augusto, Ohara; Fox, Jay W.; Lebrun, Ivo; Serrano, Solange M. T.; Camargo, Antonio C. M.

    2009-01-01

    Bj-BPP-10c is a bioactive proline-rich decapeptide, part of the C-type natriuretic peptide precursor, expressed in the brain and in the venom gland of Bothrops jararaca. We recently showed that Bj-BPP-10c displays a strong, sustained anti-hypertensive effect in spontaneous hypertensive rats (SHR), without causing any effect in normotensive rats, by a pharmacological effect independent of angiotensin-converting enzyme inhibition. Therefore, we hypothesized that another mechanism should be involved in the peptide activity. Here we used affinity chromatography to search for kidney cytosolic proteins with affinity for Bj-BPP-10c and demonstrate that argininosuccinate synthetase (AsS) is the major protein binding to the peptide. More importantly, this interaction activates the catalytic activity of AsS in a dose-de pend ent manner. AsS is recognized as an important player of the citrulline-NO cycle that represents a potential limiting step in NO synthesis. Accordingly, the functional interaction of Bj-BPP-10c and AsS was evidenced by the following effects promoted by the peptide: (i) increase of NO metabolite production in human umbilical vein endothelial cell culture and of arginine in human embryonic kidney cells and (ii) increase of arginine plasma concentration in SHR. Moreover, α-methyl-dl-aspartic acid, a specific AsS inhibitor, significantly reduced the anti-hypertensive activity of Bj-BPP-10c in SHR. Taken together, these results suggest that AsS plays a role in the anti-hypertensive action of Bj-BPP-10c. Therefore, we propose the activation of AsS as a new mechanism for the anti-hypertensive effect of Bj-BPP-10c in SHR and AsS as a novel target for the therapy of hypertension-related diseases. PMID:19491403

  10. The subcellular compartmentalization of arginine metabolizing enzymes and their role in endothelial dysfunction

    Directory of Open Access Journals (Sweden)

    Feng eChen

    2013-07-01

    Full Text Available The endothelial production of nitric oxide (NO mediates endothelium-dependent vasorelaxation and restrains vascular inflammation, smooth muscle proliferation and platelet aggregation. Impaired production of NO is a hallmark of endothelial dysfunction and promotes the development of cardiovascular disease. In endothelial cells, NO is generated by endothelial nitric oxide synthase (eNOS through the conversion of its substrate, L-arginine to L-citrulline. Reduced access to L-arginine has been proposed as a major mechanism underlying reduced eNOS activity and NO production in cardiovascular disease. The arginases (Arg1 and Arg2 metabolize L-arginine to generate L-ornithine and urea and increased expression of arginase has been proposed as a mechanism of reduced eNOS activity secondary to the depletion of L-arginine. Indeed, supplemental L-arginine and suppression of arginase activity has been shown to improve endothelium-dependent relaxation and ameliorate cardiovascular disease. However, L-arginine concentrations in endothelial cells remain sufficiently high to support NO synthesis suggesting additional mechanisms. The compartmentalization of intracellular L-arginine into poorly interchangeable pools has been proposed to allow for the local depletion of L-arginine. Indeed the subcellular location of L-arginine metabolizing enzymes plays important functional roles. In endothelial cells, eNOS is found in discrete intracellular locations and the capacity to generate NO is heavily influenced by its localtion. Arg1 and Arg2 also reside in different subcellular environments and are thought to differentially influence endothelial function. The plasma membrane solute transporter, CAT-1 and the arginine recycling enzyme, ASL, co-localize with eNOS and facilitate NO release. This review highlights the importance of the subcellular location of eNOS and arginine transporting and metabolizing enzymes to NO release and cardiovascular disease.

  11. Liver transplantation versus conservative treatment for adult-onset type II citrullinemia: our experience and a review of the literature.

    Science.gov (United States)

    Kimura, N; Kubo, N; Narumi, S; Toyoki, Y; Ishido, K; Kudo, D; Umehara, M; Yakoshi, Y; Hakamada, K

    2013-11-01

    Adult-onset type II citrullinemia (CTLN2), an autosomal recessive disorder caused by a mutation in the SLC25A13 gene, is characterized by increased serum citrulline and ammonia levels. Patients with CTLN2 also display various neuropsychiatric symptoms. Many individuals with CTLN2 are fond of protein-rich and/or lipid-rich foods with an aversion to carbohydrate-rich foods. We herein report two cases of CTLN2 treated with living donor liver transplantation (LDLT) and provide a review of the pertinent literature. Case 1 was a 43-year-old man admitted to our hospital for repetitive episodes of consciousness disturbance. Case 2 was a 37-year-old man admitted to our hospital because of abnormal behavior associated with hyperammonemia. A definitive diagnosis of CTLN2 was accomplished by DNA analysis in both patients, who successfully underwent LDLT using liver segments from donor siblings with confirmed heterozygous gene expression. Case 2 also underwent conservative therapy with arginine and a high-fat, carbohydrate-restricted diet prior to LDLT. Postoperative recovery was uneventful and food was unrestricted in both patients. We also identified 77 cases of CTLN2 in the literature and reviewed them in terms of outcome of both liver transplantation and conservative therapy. The survival rate in patients treated by liver transplantation was 100%, whereas that in patients treated by conservative treatment showed improvement from 39.5% to 76.5% over the years. Liver transplantation is a practical treatment that fundamentally improves patient quality of life after transplantation. However, recent studies have suggested that arginine and sodium pyruvate administration combined with intensive nutritional support is also an effective therapy for CTLN2. Further development of conservative therapy may provide a safer, more affordable alternative to liver transplantation in the near future.

  12. Association of susceptible genetic markers and autoantibodies in rheumatoid arthritis

    Indian Academy of Sciences (India)

    Vasanth Konda Mohan; Nalini Ganesan; Rajasekhar Gopalakrishnan

    2014-08-01

    Rheumatoid arthritis (RA) is a chronic autoimmune disorder of unknown aetiology resulting in inflammation of the synovium, cartilage and bone. The disease has a heterogeneous character, consisting of clinical subsets of anti-citrullinated protein antibody (ACPA)-positive and APCA-negative disease. Although, the pathogenesis of RA is incompletely understood, genetic factors play a vital role in susceptibility to RA as the heritability of RA is between 50 and 60%, with the human leukocyte antigen (HLA) locus accounting for at least 30% of overall genetic risk. Non-HLA genes, i.e. tumour necrosis factor- (TNF-) within the MHC (major histocompatibility complex) have also been investigated for association with RA. Although, some contradictory results have originated from several studies on TNF- gene, the data published so far indicate the possible existence of TNF- gene promoter variants that act as markers for disease severity and response to treatment in RA. The correlation of HLA and non-HLA genes within MHC region is apparently interpreted. A considerable number of confirmed associations with RA and other autoimmune disease susceptibility loci including peptidylarginine deiminase type 4 (PADI4), protein tyrosine phosphatase non-receptor type 22 (PTPN22), signal transducer and activator of transcription (STAT4), cluster of differentiation 244 (CD244) and cytotoxic T lymphocyte-associated antigen 4 (CTLA4), located outside the MHC have been reported recently. In this review, we aim to give an update on recent progress in RA genetics, the importance of the combination of HLA-DRB1 alleles, non-HLA gene polymorphism, its detection and autoantibodies as susceptibility markers for early RA disease.

  13. Predicting the Risk of Rheumatoid Arthritis and Its Age of Onset through Modelling Genetic Risk Variants with Smoking

    Science.gov (United States)

    Scott, Ian C.; Seegobin, Seth D.; Steer, Sophia; Tan, Rachael; Forabosco, Paola; Hinks, Anne; Eyre, Stephen; Morgan, Ann W.; Wilson, Anthony G.; Hocking, Lynne J.; Wordsworth, Paul; Barton, Anne; Worthington, Jane; Cope, Andrew P.; Lewis, Cathryn M.

    2013-01-01

    The improved characterisation of risk factors for rheumatoid arthritis (RA) suggests they could be combined to identify individuals at increased disease risks in whom preventive strategies may be evaluated. We aimed to develop an RA prediction model capable of generating clinically relevant predictive data and to determine if it better predicted younger onset RA (YORA). Our novel modelling approach combined odds ratios for 15 four-digit/10 two-digit HLA-DRB1 alleles, 31 single nucleotide polymorphisms (SNPs) and ever-smoking status in males to determine risk using computer simulation and confidence interval based risk categorisation. Only males were evaluated in our models incorporating smoking as ever-smoking is a significant risk factor for RA in men but not women. We developed multiple models to evaluate each risk factor's impact on prediction. Each model's ability to discriminate anti-citrullinated protein antibody (ACPA)-positive RA from controls was evaluated in two cohorts: Wellcome Trust Case Control Consortium (WTCCC: 1,516 cases; 1,647 controls); UK RA Genetics Group Consortium (UKRAGG: 2,623 cases; 1,500 controls). HLA and smoking provided strongest prediction with good discrimination evidenced by an HLA-smoking model area under the curve (AUC) value of 0.813 in both WTCCC and UKRAGG. SNPs provided minimal prediction (AUC 0.660 WTCCC/0.617 UKRAGG). Whilst high individual risks were identified, with some cases having estimated lifetime risks of 86%, only a minority overall had substantially increased odds for RA. High risks from the HLA model were associated with YORA (P<0.0001); ever-smoking associated with older onset disease. This latter finding suggests smoking's impact on RA risk manifests later in life. Our modelling demonstrates that combining risk factors provides clinically informative RA prediction; additionally HLA and smoking status can be used to predict the risk of younger and older onset RA, respectively. PMID:24068971

  14. The role of rheumatoid arthritis genetic susceptibility markers in the prediction of erosive disease in patients with early inflammatory polyarthritis: results from the Norfolk Arthritis Register

    Science.gov (United States)

    Plant, Darren; Thomson, Wendy; Lunt, Mark; Flynn, Edward; Martin, Paul; Eyre, Steven; Farragher, Tracey; Bunn, Diane; Worthington, Jane; Symmons, Deborah

    2011-01-01

    Objectives. Recent whole-genome and candidate gene association studies in RA have identified a number of single nucleotide polymorphisms (SNPs) that predispose to disease with moderate risk. It remains poorly understood how recently identified genetic factors may contribute to RA severity. We therefore sought to investigate the role of recently identified RA susceptibility SNP markers in predicting erosive outcome in patients with recent-onset inflammatory polyarthritis (IP). Methods. DNA and X-ray data were available for 1049 patients who were registered between 1990 and 2003 with the Norfolk Arthritis Register (NOAR); a primary care-based inception cohort of patients with recent-onset IP. Demographic and clinical data were recorded at inclusion, and at yearly assessments thereafter. Patients were genotyped for 18 SNP markers. The presence of serum anti citrullinated peptide antibodies (ACPAs) was assessed in samples collected at inclusion to the NOAR. The association of serological and genetic markers with poor radiological (Larsen) score at Years 1 and 5, and erosions at Years 1 and 5 was investigated. Results. Baseline ACPA positivity was associated with erosive disease and higher radiological damage. SNP markers within the TRAF1/C5 locus were associated with erosive disease at Year 1 [rs2900180: odds ratio (OR) 1.53 (95% CI 1.14, 2.05)] and Year 5 [rs2900180: OR 1.47 (95% CI 1.07, 2.02)]. None of the SNP markers tested was associated with Larsen score. Conclusion. Our results are in keeping with a previous report and suggest that the TRAF1/C5 region is associated with risk of development of radiological erosions in IP/RA patients. The finding requires replication in other large data sets. PMID:20219786

  15. High-Density Genotyping of Immune Loci in Koreans and Europeans Identifies Eight New Rheumatoid Arthritis Risk Loci

    Science.gov (United States)

    Kim, Kwangwoo; Bang, So-Young; Lee, Hye-Soon; Cho, Soo-Kyung; Choi, Chan-Bum; Sung, Yoon-Kyoung; Kim, Tae-Hwan; Jun, Jae-Bum; Yoo, Dae Hyun; Kang, Young Mo; Kim, Seong-Kyu; Suh, Chang-Hee; Shim, Seung-Cheol; Lee, Shin-Seok; Lee, Jisoo; Chung, Won Tae; Choe, Jung-Yoon; Shin, Hyoung Doo; Lee, Jong-Young; Han, Bok-Ghee; Nath, Swapan K.; Eyre, Steve; Bowes, John; Pappas, Dimitrios A.; Kremer, Joel M.; Gonzalez-Gay, Miguel A; Rodriguez-Rodriguez, Luis; Ärlestig, Lisbeth; Okada, Yukinori; Diogo, Dorothée; Liao, Katherine P.; Karlson, Elizabeth W.; Raychaudhuri, Soumya; Rantapää-Dahlqvist, Solbritt; Martin, Javier; Klareskog, Lars; Padyukov, Leonid; Gregersen, Peter K.; Worthington, Jane; Greenberg, Jeffrey D.; Plenge, Robert M.; Bae, Sang-Cheol

    2015-01-01

    Objective A highly polygenic etiology and high degree of allele-sharing between ancestries have been well-elucidated in genetic studies of rheumatoid arthritis. Recently, the high-density genotyping array Immunochip for immune disease loci identified 14 new rheumatoid arthritis risk loci among individuals of European ancestry. Here, we aimed to identify new rheumatoid arthritis risk loci using Korean-specific Immunochip data. Methods We analyzed Korean rheumatoid arthritis case-control samples using the Immunochip and GWAS array to search for new risk alleles of rheumatoid arthritis with anti-citrullinated peptide antibodies. To increase power, we performed a meta-analysis of Korean data with previously published European Immunochip and GWAS data, for a total sample size of 9,299 Korean and 45,790 European case-control samples. Results We identified 8 new rheumatoid arthritis susceptibility loci (TNFSF4, LBH, EOMES, ETS1–FLI1, COG6, RAD51B, UBASH3A and SYNGR1) that passed a genome-wide significance threshold (p<5×10−8), with evidence for three independent risk alleles at 1q25/TNFSF4. The risk alleles from the 7 new loci except for the TNFSF4 locus (monomorphic in Koreans), together with risk alleles from previously established RA risk loci, exhibited a high correlation of effect sizes between ancestries. Further, we refined the number of SNPs that represent potentially causal variants through a trans-ethnic comparison of densely genotyped SNPs. Conclusion This study demonstrates the advantage of dense-mapping and trans-ancestral analysis for identification of potentially causal SNPs. In addition, our findings support the importance of T cells in the pathogenesis and the fact of frequent overlap of risk loci among diverse autoimmune diseases. PMID:24532676

  16. Predicting the risk of rheumatoid arthritis and its age of onset through modelling genetic risk variants with smoking.

    Directory of Open Access Journals (Sweden)

    Ian C Scott

    Full Text Available The improved characterisation of risk factors for rheumatoid arthritis (RA suggests they could be combined to identify individuals at increased disease risks in whom preventive strategies may be evaluated. We aimed to develop an RA prediction model capable of generating clinically relevant predictive data and to determine if it better predicted younger onset RA (YORA. Our novel modelling approach combined odds ratios for 15 four-digit/10 two-digit HLA-DRB1 alleles, 31 single nucleotide polymorphisms (SNPs and ever-smoking status in males to determine risk using computer simulation and confidence interval based risk categorisation. Only males were evaluated in our models incorporating smoking as ever-smoking is a significant risk factor for RA in men but not women. We developed multiple models to evaluate each risk factor's impact on prediction. Each model's ability to discriminate anti-citrullinated protein antibody (ACPA-positive RA from controls was evaluated in two cohorts: Wellcome Trust Case Control Consortium (WTCCC: 1,516 cases; 1,647 controls; UK RA Genetics Group Consortium (UKRAGG: 2,623 cases; 1,500 controls. HLA and smoking provided strongest prediction with good discrimination evidenced by an HLA-smoking model area under the curve (AUC value of 0.813 in both WTCCC and UKRAGG. SNPs provided minimal prediction (AUC 0.660 WTCCC/0.617 UKRAGG. Whilst high individual risks were identified, with some cases having estimated lifetime risks of 86%, only a minority overall had substantially increased odds for RA. High risks from the HLA model were associated with YORA (P<0.0001; ever-smoking associated with older onset disease. This latter finding suggests smoking's impact on RA risk manifests later in life. Our modelling demonstrates that combining risk factors provides clinically informative RA prediction; additionally HLA and smoking status can be used to predict the risk of younger and older onset RA, respectively.

  17. Influence of heme-thiolate in shaping the catalytic properties of a bacterial nitric-oxide synthase.

    Science.gov (United States)

    Hannibal, Luciana; Somasundaram, Ramasamy; Tejero, Jesús; Wilson, Adjele; Stuehr, Dennis J

    2011-11-11

    Nitric-oxide synthases (NOS) are heme-thiolate enzymes that generate nitric oxide (NO) from L-arginine. Mammalian and bacterial NOSs contain a conserved tryptophan (Trp) that hydrogen bonds with the heme-thiolate ligand. We mutated Trp(66) to His and Phe (W66H, W66F) in B. subtilis NOS to investigate how heme-thiolate electronic properties control enzyme catalysis. The mutations had opposite effects on heme midpoint potential (-302, -361, and -427 mV for W66H, wild-type (WT), and W66F, respectively). These changes were associated with rank order (W66H < WT < W66F) changes in the rates of oxygen activation and product formation in Arg hydroxylation and N-hydroxyarginine (NOHA) oxidation single turnover reactions, and in the O(2) reactivity of the ferrous heme-NO product complex. However, enzyme ferrous heme-O(2) autoxidation showed an opposite rank order. Tetrahydrofolate supported NO synthesis by WT and the mutant NOS. All three proteins showed similar extents of product formation (L-Arg → NOHA or NOHA → citrulline) in single turnover studies, but the W66F mutant showed a 2.5 times lower activity when the reactions were supported by flavoproteins and NADPH. We conclude that Trp(66) controls several catalytic parameters by tuning the electron density of the heme-thiolate bond. A greater electron density (as in W66F) improves oxygen activation and reactivity toward substrate, but decreases heme-dioxy stability and lowers the driving force for heme reduction. In the WT enzyme the Trp(66) residue balances these opposing effects for optimal catalysis.

  18. Dissecting structural and electronic effects in inducible nitric oxide synthase.

    Science.gov (United States)

    Hannibal, Luciana; Page, Richard C; Haque, Mohammad Mahfuzul; Bolisetty, Karthik; Yu, Zhihao; Misra, Saurav; Stuehr, Dennis J

    2015-04-01

    Nitric oxide synthases (NOSs) are haem-thiolate enzymes that catalyse the conversion of L-arginine (L-Arg) into NO and citrulline. Inducible NOS (iNOS) is responsible for delivery of NO in response to stressors during inflammation. The catalytic performance of iNOS is proposed to rely mainly on the haem midpoint potential and the ability of the substrate L-Arg to provide a hydrogen bond for oxygen activation (O-O scission). We present a study of native iNOS compared with iNOS-mesohaem, and investigate the formation of a low-spin ferric haem-aquo or -hydroxo species (P) in iNOS mutant W188H substituted with mesohaem. iNOS-mesohaem and W188H-mesohaem were stable and dimeric, and presented substrate-binding affinities comparable to those of their native counterparts. Single turnover reactions catalysed by iNOSoxy with L-Arg (first reaction step) or N-hydroxy-L-arginine (second reaction step) showed that mesohaem substitution triggered higher rates of Fe(II)O₂ conversion and altered other key kinetic parameters. We elucidated the first crystal structure of a NOS substituted with mesohaem and found essentially identical features compared with the structure of iNOS carrying native haem. This facilitated the dissection of structural and electronic effects. Mesohaem substitution substantially reduced the build-up of species P in W188H iNOS during catalysis, thus increasing its proficiency towards NO synthesis. The marked structural similarities of iNOSoxy containing native haem or mesohaem indicate that the kinetic behaviour observed in mesohaem-substituted iNOS is most heavily influenced by electronic effects rather than structural alterations.

  19. Individual variability in human blood metabolites identifies age-related differences

    Science.gov (United States)

    Murakami, Itsuo; Takada, Junko; Kondoh, Hiroshi; Yanagida, Mitsuhiro

    2016-01-01

    Metabolites present in human blood document individual physiological states influenced by genetic, epigenetic, and lifestyle factors. Using high-resolution liquid chromatography-mass spectrometry (LC-MS), we performed nontargeted, quantitative metabolomics analysis in blood of 15 young (29 ± 4 y of age) and 15 elderly (81 ± 7 y of age) individuals. Coefficients of variation (CV = SD/mean) were obtained for 126 blood metabolites of all 30 donors. Fifty-five RBC-enriched metabolites, for which metabolomics studies have been scarce, are highlighted here. We found 14 blood compounds that show remarkable age-related increases or decreases; they include 1,5-anhydroglucitol, dimethyl-guanosine, acetyl-carnosine, carnosine, ophthalmic acid, UDP-acetyl-glucosamine, N-acetyl-arginine, N6-acetyl-lysine, pantothenate, citrulline, leucine, isoleucine, NAD+, and NADP+. Six of them are RBC-enriched, suggesting that RBC metabolomics is highly valuable for human aging research. Age differences are partly explained by a decrease in antioxidant production or increasing inefficiency of urea metabolism among the elderly. Pearson’s coefficients demonstrated that some age-related compounds are correlated, suggesting that aging affects them concomitantly. Although our CV values are mostly consistent with those CVs previously published, we here report previously unidentified CVs of 51 blood compounds. Compounds having moderate to high CV values (0.4–2.5) are often modified. Compounds having low CV values, such as ATP and glutathione, may be related to various diseases because their concentrations are strictly controlled, and changes in them would compromise health. Thus, human blood is a rich source of information about individual metabolic differences. PMID:27036001

  20. NADPH Oxidase Activity in Cerebral Arterioles Is a Key Mediator of Cerebral Small Vessel Disease—Implications for Prevention

    Directory of Open Access Journals (Sweden)

    Mark F. McCarty

    2015-04-01

    Full Text Available Cerebral small vessel disease (SVD, a common feature of brain aging, is characterized by lacunar infarcts, microbleeds, leukoaraiosis, and a leaky blood-brain barrier. Functionally, it is associated with cognitive decline, dementia, depression, gait abnormalities, and increased risk for stroke. Cerebral arterioles in this syndrome tend to hypertrophy and lose their capacity for adaptive vasodilation. Rodent studies strongly suggest that activation of Nox2-dependent NADPH oxidase activity is a crucial driver of these structural and functional derangements of cerebral arterioles, in part owing to impairment of endothelial nitric oxide synthase (eNOS activity. This oxidative stress may also contribute to the breakdown of the blood-brain barrier seen in SVD. Hypertension, aging, metabolic syndrome, smoking, hyperglycemia, and elevated homocysteine may promote activation of NADPH oxidase in cerebral arterioles. Inhibition of NADPH oxidase with phycocyanobilin from spirulina, as well as high-dose statin therapy, may have potential for prevention and control of SVD, and high-potassium diets merit study in this regard. Measures which support effective eNOS activity in other ways—exercise training, supplemental citrulline, certain dietary flavonoids (as in cocoa and green tea, and capsaicin, may also improve the function of cerebral arterioles. Asian epidemiology suggests that increased protein intakes may decrease risk for SVD; conceivably, arginine and/or cysteine—which boosts tissue glutathione synthesis, and can be administered as N-acetylcysteine—mediate this benefit. Ameliorating the risk factors for SVD—including hypertension, metabolic syndrome, hyperglycemia, smoking, and elevated homocysteine—also may help to prevent and control this syndrome, although few clinical trials have addressed this issue to date.

  1. Steatogenesis in adult-onset type II citrullinemia is associated with down-regulation of PPARα.

    Science.gov (United States)

    Komatsu, Michiharu; Kimura, Takefumi; Yazaki, Masahide; Tanaka, Naoki; Yang, Yang; Nakajima, Takero; Horiuchi, Akira; Fang, Zhong-Ze; Joshita, Satoru; Matsumoto, Akihiro; Umemura, Takeji; Tanaka, Eiji; Gonzalez, Frank J; Ikeda, Shu-Ichi; Aoyama, Toshifumi

    2015-03-01

    SLC25A13 (citrin or aspartate-glutamate carrier 2) is located in the mitochondrial membrane in the liver and its genetic deficiency causes adult-onset type II citrullinemia (CTLN2). CTLN2 is one of the urea cycle disorders characterized by sudden-onset hyperammonemia due to reduced argininosuccinate synthase activity. This disorder is frequently accompanied with hepatosteatosis in the absence of obesity and ethanol consumption. However, the precise mechanism of steatogenesis remains unclear. The expression of genes associated with fatty acid (FA) and triglyceride (TG) metabolism was examined using liver samples obtained from 16 CTLN2 patients and compared with 7 healthy individuals. Although expression of hepatic genes associated with lipogenesis and TG hydrolysis was not changed, the mRNAs encoding enzymes/proteins involved in FA oxidation (carnitine palmitoyl-CoA transferase 1α, medium- and very-long-chain acyl-CoA dehydrogenases, and acyl-CoA oxidase 1), very-low-density lipoprotein secretion (microsomal TG transfer protein), and FA transport (CD36 and FA-binding protein 1), were markedly suppressed in CTLN2 patients. Serum concentrations of ketone bodies were also decreased in these patients, suggesting reduced mitochondrial β-oxidation activity. Consistent with these findings, the expression of peroxisome proliferator-activated receptor α (PPARα), a master regulator of hepatic lipid metabolism, was significantly down-regulated. Hepatic PPARα expression was inversely correlated with severity of steatosis and circulating ammonia and citrulline levels. Additionally, phosphorylation of c-Jun-N-terminal kinase was enhanced in CTLN2 livers, which was likely associated with lower hepatic PPARα. Collectively, down-regulation of PPARα is associated with steatogenesis in CTLN2 patients. These findings provide a novel link between urea cycle disorder, lipid metabolism, and PPARα.

  2. DAMPs-activated neutrophil extracellular trap exacerbates sterile inflammatory liver injury

    Science.gov (United States)

    Huang, Hai; Tohme, Samer; Al-Khafaji, Ahmed B; Tai, Sheng; Loughran, Patricia; Chen, Li; Wang, Shu; Kim, Jiyun; Billiar, Timothy; Wang, Yanming; Tsung, Allan

    2015-01-01

    Innate immunity plays a crucial role in the response to sterile inflammation such as liver ischemia/reperfusion (I/R) injury. The initiation of liver I/R injury results in the release of damage associated molecular patterns (DAMPs), which trigger innate immune and inflammatory cascade via pattern recognition receptors. Neutrophils are recruited to the liver after I/R and contribute to the organ damage, innate immune and inflammatory responses. Formation of neutrophil extracellular trap (NET) has been recently found in response to various stimuli. However, the role of NETs during liver I/R injury remains unknown. We show that NETs form in the sinusoids of ischemic liver lobes in vivo. This was associated with increased NET markers, serum level of myeloperoxidase (MPO)-DNA complexes and tissue level of citrullinated-histone H3 compared to control mice. Treatment with peptidyl-arginine-deiminase (PAD) 4 inhibitor or DNase I significantly protected hepatocytes and reduced inflammation after liver I/R as evidenced by inhibition of NET formation, indicating the pathophysiological role of NETs in liver I/R injury. In vitro, NETs increase hepatocyte death and induce Kupffer cells to release proinflammatory cytokines. DAMPs, such as HMGB1 and histones, released by injured hepatocytes stimulate NET formation through Toll-like receptor (TLR4)- and TLR9-MyD88 signaling pathways. After neutrophil depletion in mice, the adoptive transfer of TLR4 knockout (KO) or TLR9 KO neutrophils confers significant protection from liver I/R injury with significant decrease in NET formation. In addition, we found inhibition of NET formation by PAD4 inhibitor or DNase I reduces HMGB1 and histone-mediated liver I/R injury. Conclusion DAMPs released during liver I/R promotes NET formation through TLRs signaling pathway. Development of NETs subsequently exacerbates organ damage and initiates inflammatory responses during liver I/R. PMID:25855125

  3. Components of yeast (Sacchromyces cervisiae) extract as defined media additives that support the growth and productivity of CHO cells.

    Science.gov (United States)

    Spearman, Maureen; Chan, Sarah; Jung, Vince; Kowbel, Vanessa; Mendoza, Meg; Miranda, Vivian; Butler, Michael

    2016-09-10

    Yeast and plant hydrolysates are used as media supplements to support the growth and productivity of CHO cultures for biopharmaceutical production. Through fractionation of a yeast lysate and metabolic analysis of a fraction that had bioactivity equivalent to commercial yeast extract (YE), bioactive components were identified that promoted growth and productivity of two recombinant CHO cell lines (CHO-Luc and CHO-hFcEG2) equivalent to or greater than YE-supplemented media. Autolysis of the yeast lysate was not necessary for full activity, suggesting that the active components are present in untreated yeast cells. A bioactive fraction (3KF) of the yeast lysate was isolated from the permeate using a 3kDa molecular weight cut-off (MWCO) filter. Supplementation of this 3KF fraction into the base media supported growth of CHO-Luc cells over eight passages equivalent to YE-supplemented media. The 3KF fraction was fractionated further by a cation exchange spin column using a stepwise pH elution. Metabolomic analysis of a bioactive fraction isolated at high pH identified several arginine and lysine-containing peptides as well as two polyamines, spermine and spermidine, with 3.5× and 4.5× higher levels compared to a fraction showing no bioactivity. The addition of a mixture of polyamines and their precursors (putrescine, spermine, spermidine, ornithine and citrulline) as well as increasing the concentration of some of the components of the original base medium resulted in a chemically-defined (CD) formulation that produced an equivalent viable cell density (VCD) and productivity of the CHO-Luc cells as the YE-supplemented medium. The VCD of the CHO-hFcEG2 culture in the CD medium was 1.9× greater and with equivalent productivity to the YE-supplemented media.

  4. Rheumatoid arthritis-associated interstitial lung disease

    Directory of Open Access Journals (Sweden)

    Brown KK

    2012-03-01

    Full Text Available Joshua J Solomon, Kevin K BrownAutoimmune Lung Center and Interstitial Lung Disease Program, National Jewish Health, Denver, CO, USAAbstract: Rheumatoid arthritis (RA is a systemic inflammatory disorder affecting 1% of the US population. Patients can have extra-articular manifestations of their disease and the lungs are commonly involved. RA can affect any compartment of the respiratory system and high resolution computed tomography (HRCT of the lung is abnormal in over half of these patients. Interstitial lung disease is a dreaded complication of RA. It is more prevalent in smokers, males, and those with high antibody titers. The pathogenesis is unknown but data suggest an environmental insult in the setting of a genetic predisposition. Smoking may play a role in the pathogenesis of disease through citrullination of protein in the lung leading to the development of autoimmunity. Patients usually present in middle age with cough and dyspnea. Pulmonary function testing most commonly shows reduced diffusion capacity for carbon monoxide and HRCT reveals a combination of reticulation and ground glass abnormalities. The most common pattern on HRCT and histopathology is usual interstitial pneumonia (UIP, with nonspecific interstitial pneumonia seen less frequently. There are no large-scale well-controlled treatment trials. In severe or progressive cases, treatment usually consists of corticosteroids with or without a cytotoxic agent for 6 months or longer. RA interstitial lung disease is progressive; over half of patients show radiographic progression within 2 years. Patients with a UIP pattern on biopsy have a survival similar to idiopathic pulmonary fibrosis.Keywords: rheumatoid arthritis, interstitial lung disease, nonspecific interstitial pneumonia, usual interstitial pneumonia, anti-CCP

  5. Treatment of rheumatoid arthritis-associated interstitial lung disease: a perspective review.

    Science.gov (United States)

    Iqbal, Kundan; Kelly, Clive

    2015-12-01

    Rheumatoid arthritis (RA) is a systemic autoimmune disease affecting 0.5-1% of the worldwide population. Whilst predominantly causing chronic pain and inflammation in synovial joints, it is also associated with significant extra-articular manifestations in a large proportion of patients. Among the various pulmonary manifestations, interstitial lung disease (ILD), a progressive fibrotic disease of the lung parenchyma, is the commonest and most important, contributing significantly to increased morbidity and mortality. The most frequent patterns of RA-associated ILD (RA-ILD) are usual interstitial pneumonia and nonspecific interstitial pneumonia. New insights during the past several years have highlighted the epidemiological impact of RA-ILD and have begun to identify factors contributing to its pathogenesis. Risk factors include smoking, male sex, human leukocyte antigen haplotype, rheumatoid factor and anticyclic citrullinated protein antibodies (ACPAs). Combined with clinical information, chest examination and pulmonary function testing, high-resolution computed tomography of the chest forms the basis of investigation and allows assessment of subtype and disease extent. The management of RA-ILD is a challenge. Several therapeutic agents have been suggested in the literature but as yet no large randomized controlled trials have been undertaken to guide clinical management. Therapy is further complicated by commonly prescribed drugs of proven articular benefit such as methotrexate, leflunomide (LEF) and anti-tumour necrosis factor α agents having been implicated in both ex novo occurrence and acceleration of existing ILD. Agents that offer promise include immunomodulators such as mycophenolate and rituximab as well as newly studied antifibrotic agents. In this review, we discuss the current literature to evaluate recommendations for the management of RA-ILD and discuss key gaps in our knowledge of this important disease.

  6. Computed tomography of pulmonary changes in rheumatoid arthritis: carcinoembryonic antigen (CEA) as a marker of airway disease.

    Science.gov (United States)

    Koch, Milene Caroline; Pereira, Ivânio Alves; Nobre, Luiz Felipe Souza; Neves, Fabricio Souza

    2016-04-01

    Rheumatoid arthritis (RA) classically affects the joints, but can present extra-articular manifestations, including pulmonary disease. The present study aimed to identify possible risk factors or laboratory markers for lung involvement in RA, particularly the presence of rheumatoid factor (RF), anti-citrullinated peptide antibodies (ACPA), and tumor markers, by correlating them with changes observed on chest high-resolution computerized tomography (HRCT). This cross-sectional study involved RA patients who were examined and questioned by a specialist physician and later subjected to chest HRCT and blood collection for measurement of C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), rheumatoid factor (RF), ACPA (anti-vimentin and/or anti-CCP3), and the tumor markers carcinoembryonic antigen (CEA), CA 125, CA 15-3, and CA 19-9. A total of 96 patients underwent chest HRCT. The most frequent findings were bronchial thickening (27/28.1 %) and bronchiectasis (25/26 %). RF was present in 63.2 % of patients (55/87), and ACPA (anti-vimentin or anti-CCP3) was present in 72.7 % of patients (64/88). CEA levels were high in 14 non-smokers (37.8 %) and 23 smokers (62.2 %). CA-19-9 levels were high in 6 of 86 patients (7.0 %), CA 15-3 levels were high in 3 of 85 patients (3.5 %), and CA 125 levels were high in 4 of 75 patients (5.3 %). Multivariate analysis indicated a statistically significant association between high CEA levels and the presence of airway changes in patients with RA (p = 0.048). CEA can serve as a predictor of lung disease in RA and can help identify individuals who require more detailed examination for the presence of respiratory disorders.

  7. Anticholinesterase Toxicity and Oxidative Stress

    Directory of Open Access Journals (Sweden)

    Dejan Milatovic

    2006-01-01

    Full Text Available Anticholinesterase compounds, organophosphates (OPs and carbamates (CMs are commonly used for a variety of purposes in agriculture and in human and veterinary medicine. They exert their toxicity in mammalian system primarily by virtue of acetylcholinesterase (AChE inhibition at the synapses and neuromuscular junctions, leading into the signs of hypercholinergic preponderance. However, the mechanism(s involved in brain/muscle damage appear to be linked with alteration in antioxidant and the scavenging system leading to free radical-mediated injury. OPs and CMs cause excessive formation of F2-isoprostanes and F4-neuroprostanes, in vivo biomarkers of lipid peroxidation and generation of reactive oxygen species (ROS, and of citrulline, a marker of NO/NOS and reactive nitrogen species (RNS generation. In addition, during the course of these excitatory processes and inhibition of AChE, a high rate of ATP consumption, coupled with the inhibition of oxidative phosphorylation, compromise the cell's ability to maintain its energy levels and excessive amounts of ROS and RNS may be generated. Pretreatment with N-methyl D-aspartate (NMDA receptor antagonist memantine, in combination with atropine sulfate, provides significant protection against inhibition of AChE, increases of ROS/RNS, and depletion of high-energy phosphates induced by DFP/carbofuran. Similar antioxidative effects are observed with a spin trapping agent, phenyl-N-tert-butylnitrone (PBN or chain breaking antioxidant vitamin E. This review describes the mechanisms involved in anticholinesterase-induced oxidative/nitrosative injury in target organs of OPs/CMs, and protection by various agents.

  8. Distribution of nitric oxide synthase in normal and cirrhotic human liver

    Science.gov (United States)

    McNaughton, Lance; Puttagunta, Lakshmi; Martinez-Cuesta, Maria Angeles; Kneteman, Norm; Mayers, Irvin; Moqbel, Redwan; Hamid, Qutayba; Radomski, Marek W.

    2002-01-01

    Chronic liver disorders represent a serious health problem, considering that 300 million people worldwide are hepatitis B virus carriers, and 8,000–10,000 patients per year, in the U.S. alone, die as a result of liver failure caused by hepatitis C infection. Nitric oxide synthase (NOS) regulates hepatic vasculature; however, the patterns of expression and activity of NOS proteins in healthy and diseased human livers are unknown. Sections of diseased (n = 42) and control livers (n = 14) were collected during orthotopic liver transplants and partial hepatectomy. The diseased sections included alcoholic cirrhosis, viral hepatitis, cholestasis, acute necrosis, and uncommon pathologies including α1-anti-trypsin disorder. The endothelial NOS (eNOS), inducible NOS (iNOS), and neuronal NOS (nNOS) were studied by using the citrulline assay, Western immunoblot, immunohistochemistry, and in situ hybridization. The systemic generation of plasma NO metabolites was measured by HPLC. In control livers, Ca2+-dependent and –independent NOS activities were identified by Western analysis as eNOS and iNOS, respectively. The eNOS was uniformly distributed in the hepatocytes and also detected in the endothelium of hepatic arteries, terminal hepatic venules, sinusoids, and in biliary epithelium. The iNOS was detected in hepatocytes and localized mainly in the periportal zone of the liver acinus. This pattern of distribution of eNOS and iNOS in normal liver was confirmed by in situ hybridization. In diseased livers, there was a significant increase in Ca2+-independent NOS with the corresponding strong appearance of iNOS in the cirrhotic areas. The eNOS was translocated to hepatocyte nuclei. Thus, eNOS and iNOS proteins are differentially expressed in healthy human liver, and this expression is significantly altered in cirrhotic liver disorders. PMID:12482944

  9. Adenoviral gene transfer of endothelial nitric-oxide synthase (eNOS) partially restores normal pulmonary arterial pressure in eNOS-deficient mice

    Science.gov (United States)

    Champion, Hunter C.; Bivalacqua, Trinity J.; Greenberg, Stanley S.; Giles, Thomas D.; Hyman, Albert L.; Kadowitz, Philip J.

    2002-01-01

    It has been shown that mice deficient in the gene coding for endothelial nitric-oxide synthase (eNOS) have increased pulmonary arterial pressure and pulmonary vascular resistance. In the present study, the effect of transfer to the lung of an adenoviral vector encoding the eNOS gene (AdCMVeNOS) on pulmonary arterial pressure and pulmonary vascular resistance was investigated in eNOS-deficient mice. One day after intratracheal administration of AdCMVeNOS to eNOS−/− mice, there was an increase in eNOS protein, cGMP levels, and calcium-dependent conversion of l-arginine to l-citrulline in the lung. The increase in eNOS protein and activity in eNOS−/− mice was associated with a reduction in mean pulmonary arterial pressure and pulmonary vascular resistance when compared with values in eNOS-deficient mice treated with vehicle or a control adenoviral vector coding for β-galactosidase, AdCMVβgal. These data suggest that in vivo gene transfer of eNOS to the lung in eNOS−/− mice can increase eNOS staining, eNOS protein, calcium-dependent NOS activity, and cGMP levels and partially restore pulmonary arterial pressure and pulmonary vascular resistance to near levels measured in eNOS+/+ mice. Thus, the major finding in this study is that in vivo gene transfer of eNOS to the lung in large part corrects a genetic deficiency resulting from eNOS deletion and may be a useful therapeutic intervention for the treatment of pulmonary hypertensive disorders in which eNOS activity is reduced. PMID:12237402

  10. Characterization of NF-κB Reporter U937 Cells and Their Application for the Detection of Inflammatory Immune-Complexes.

    Directory of Open Access Journals (Sweden)

    Csilla Kecse-Nagy

    Full Text Available Our study tested the hypothesis that immunoglobulins differ in their ability to activate the nuclear factor-κB pathway mediated cellular responses. These responses are modulated by several properties of the immune complex, including the ratio of antibody isotypes binding to antigen. Immunoassays allow the measurement of antigen specific antibodies belonging to distinct immunoglobulin classes and subclasses but not the net biological effect of the combination of these antibodies. We set out to develop a biosensor that is suitable for the detection and characterization of antigen specific serum antibodies. We genetically modified the monocytoid U937 cell line carrying Fc receptors with a plasmid encoding NF-κB promoter-driven GFP. This clone, U937-NF-κB, was characterized with respect to FcR expression and response to solid-phase immunoglobulins. Human IgG3, IgG4 and IgG1 induced GFP production in a time- and dose-dependent manner, in this order of efficacy, while IgG2 triggered no activation at the concentrations tested. IgA elicited no response alone but showed significant synergism with IgG3 and IgG4. We confirmed the importance of activation via FcγRI by direct stimulation with monoclonal antibody and by competition assays. We used citrullinated peptides and serum from rheumatoid arthritis patients to generate immune complexes and to study the activation of U937-NF-κB, observing again a synergistic effect between IgG and IgA. Our results show that immunoglobulins have distinct pro-inflammatory potential, and that U937-NF-κB is suitable for the estimation of biological effects of immune-complexes, offering insight into monocyte activation and pathogenesis of antibody mediated diseases.

  11. Characterization of NF-κB Reporter U937 Cells and Their Application for the Detection of Inflammatory Immune-Complexes.

    Science.gov (United States)

    Kecse-Nagy, Csilla; Szittner, Zoltán; Papp, Krisztián; Hegyi, Zoltán; Rovero, Paolo; Migliorini, Paola; Lóránd, Veronika; Homolya, László; Prechl, József

    2016-01-01

    Our study tested the hypothesis that immunoglobulins differ in their ability to activate the nuclear factor-κB pathway mediated cellular responses. These responses are modulated by several properties of the immune complex, including the ratio of antibody isotypes binding to antigen. Immunoassays allow the measurement of antigen specific antibodies belonging to distinct immunoglobulin classes and subclasses but not the net biological effect of the combination of these antibodies. We set out to develop a biosensor that is suitable for the detection and characterization of antigen specific serum antibodies. We genetically modified the monocytoid U937 cell line carrying Fc receptors with a plasmid encoding NF-κB promoter-driven GFP. This clone, U937-NF-κB, was characterized with respect to FcR expression and response to solid-phase immunoglobulins. Human IgG3, IgG4 and IgG1 induced GFP production in a time- and dose-dependent manner, in this order of efficacy, while IgG2 triggered no activation at the concentrations tested. IgA elicited no response alone but showed significant synergism with IgG3 and IgG4. We confirmed the importance of activation via FcγRI by direct stimulation with monoclonal antibody and by competition assays. We used citrullinated peptides and serum from rheumatoid arthritis patients to generate immune complexes and to study the activation of U937-NF-κB, observing again a synergistic effect between IgG and IgA. Our results show that immunoglobulins have distinct pro-inflammatory potential, and that U937-NF-κB is suitable for the estimation of biological effects of immune-complexes, offering insight into monocyte activation and pathogenesis of antibody mediated diseases. PMID:27232500

  12. Autoantibodies in inflammatory arthritis.

    Science.gov (United States)

    Conigliaro, P; Chimenti, M S; Triggianese, P; Sunzini, F; Novelli, L; Perricone, C; Perricone, R

    2016-07-01

    Rheumatoid arthritis (RA) is a systemic chronic inflammatory disease characterized by extensive synovitis resulting in erosions of articular cartilage and marginal bone with joint destruction. The lack of immunological tolerance in RA represents the first step toward the development of autoimmunity. Susceptible individuals, under the influence of environmental factors, such as tobacco smoke, and silica exposure, develop autoimmune phenomena that result in the presence of autoantibodies. HLA and non-HLA haplotypes play a major role in determining the development of specific autoantibodies differentiating anti-citrullinated antibodies (ACPA)-positive and negative RA patients. Rheumatoid factor (RF) and ACPA are the serological markers for RA, and during the preclinical immunological phase, autoantibody titers increase with a progressive spread of ACPA antigens repertoire. The presence of ACPA represents an independent risk factor for developing RA in patients with undifferentiated arthritis or arthralgia. Moreover, anti-CarP antibodies have been identified in patients with RA as well as in individuals before the onset of clinical symptoms of RA. Several autoantibodies mainly targeting post-translational modified proteins have been investigated as possible biomarkers to improve the early diagnosis, prognosis and response to therapy in RA patients. Psoriatic arthritis (PsA) is distinguished from RA by infrequent positivity for RF and ACPA, together with other distinctive clinical features. Actually, specific autoantibodies have not been described. Recently, anti-CarP antibodies have been reported in sera from PsA patients with active disease. Further investigations on autoantibodies showing high specificity and sensibility as well as relevant correlation with disease severity, progression, and response to therapy are awaited in inflammatory arthritides.

  13. Interferon-Gamma-Induced Nitric Oxide Inhibits the Proliferation of Murine Renal Cell Carcinoma Cells

    Directory of Open Access Journals (Sweden)

    David J. Tate Jr., John R. Patterson, Cruz Velasco-Gonzalez, Emily N. Carroll, Janie Trinh, Daniel Edwards, Ashok Aiyar, Beatriz Finkel-Jimenez, Arnold H. Zea

    2012-01-01

    Full Text Available Renal cell carcinoma (RCC remains one of the most resistant tumors to systemic chemotherapy, radiotherapy, and immunotherapy. Despite great progress in understanding the basic biology of RCC, the rate of responses in animal models and clinical trials using interferons (IFNs has not improved significantly. It is likely that the lack of responses can be due to the tumor's ability to develop tumor escape strategies. Currently, the use of targeted therapies has improved the clinical outcomes of patients with RCC and is associated with an increase of Th1-cytokine responses (IFNγ, indicating the importance of IFNγ in inhibiting tumor proliferation. Thus, the present study was designed to investigate a new mechanism by which IFNγ mediates direct anti-proliferative effects against murine renal cell carcinoma cell lines. When cultured RCC cell lines were exposed to murine recombinant IFNγ, a dose dependent growth inhibition in CL-2 and CL-19 cells was observed; this effect was not observed in Renca cells. Growth inhibition in CL-2 and CL-19 cell lines was associated with the intracellular induction of nitric oxide synthase (iNOS protein, resulting in a sustained elevation of nitric oxide (NO and citrulline, and a decrease in arginase activity. The inhibition of cell proliferation appears to be due to an arrest in the cell cycle. The results indicate that in certain RCC cell lines, IFNγ modulates L-arginine metabolism by shifting from arginase to iNOS activity, thereby developing a potent inhibitory mechanism to encumber tumor cell proliferation and survival. Elucidating the cellular events triggered by IFNγ in murine RCC cell lines will permit anti-tumor effects to be exploited in the development of new combination therapies that interfere with L-arginine metabolism to effectively combat RCC in patients.

  14. [Immunological markers of rheumatoid arthritis].

    Science.gov (United States)

    Matuszewska, Agnieszka; Madej, Marta; Wiland, Piotr

    2016-03-25

    Rheumatoid arthritis (RA) is the most common connective tissue disease of autoimmune origin. The disease is characterized by chronic inflammation leading to bone erosions and organ involvement. RA is a progressive disease. It affects the quality of life, leading to disability and death mainly due to premature cardiovascular disease. Early diagnosis and appropriate treatment are essential for prognosis and quality of life improvement. In 2010 the American College of Rheumatology (ACR) and The European League Against Rheumatism (EULAR) established new RA classification criteria. Besides clinical symptoms it includes two immunologic criteria: rheumatoid factor (RF) and anti-citrullinated protein antibodies (anti-CCP antibodies). RF is the first well-known RA immunologic marker. It is observed in 80-85% of patients with RA. Elevated serum level of RF has been associated with increased disease activity, radiographic progression, and the presence of extraarticular manifestations. The sensitivity of RF is 50-90%, and specificity is 50-95%. Anti-CCP antibodies appear to be a more specific marker than RF. They are often present at the very beginning of the disease, or even years before the first symptoms. The prognostic value of anti-CCP antibodies is well established. High serum level of anti-CCP correlates with poor prognosis and early erosions of the joints. The sensitivity of anti-CCP2 is 48-80%, and specificity is 96-98%. New immunologic markers include anti-carbamylated protein antibodies (anti-CarP) and antibodies against heterogeneous nuclear ribonucleoproteins (anti-hnRNP A2/B1, RA33). Scientists aim to identify a highly sensitive and specific biomarker of the disease that not only has diagnostic and prognostic value but also may predict the response to treatment.

  15. LEVELS OF ANGIOGENESIS-REGULATORY CHEMOKINES IN THE SYNOVIAL FLUID OF PATIENTS WITH RHEUMATOID ARTHRITIS

    Directory of Open Access Journals (Sweden)

    D. A. Zhebrun

    2015-01-01

    Full Text Available The role of chemokines in the immunopathogenesis of rheumatoid arthritis (RA has been actively investigated in recent years. Angiogenic and angiostatic chemokines are important mediators of angiogenesis in the development and extent of pannus. Peripheral blood and synovial fluid (SF is a major biomaterial in clinical and immunological studies. At the same time, it is the SF test that may yield the most informative results since that gives an idea of the processes that occur locally within a joint. Objective: to perform a comparative analysis of the levels of a number of CXC, CC, and CX3C chemokines in the SF of patients with RA, osteoarthritis (OA, and joint injuries. Subjects and methods. The multiplex analysis using xMAP technology (Luminex, USA was used to analyze levels of CXC, CC, and CX3C chemokines in SF and serum of patients with RA (n = 20, OA (n = 9 and controls (n = 9. Results and discussion. The SF levels of CCL24/eotaxin-2, as well as those of the angiostatic chemokines CXCL9/MIG, CXCL10/IP10, CXCL11/ITAC, and CXCL13/BCA-1 were higher in the RA group than in the control and OA groups. There was a direct correlation between SF levels of CCL5/RANTES and DAS28, as well as patient global disease activity assessment on visual analogue scale, and that between the level of CCL2/MCP-1 in the SF and that of anticyclic citrullinated peptide (anti-CCP antibodies in the serum. The SF concentrations of CXCL5/ENA78 and CXCL7/NAP-2 were shown to depend on the presence of serum anti-CCP. Serum CXCL13/BCA-1 levels were higher in RA than those in OA, as that of CXCL7/NAP-2 than in the control group.

  16. Individual variability in human blood metabolites identifies age-related differences.

    Science.gov (United States)

    Chaleckis, Romanas; Murakami, Itsuo; Takada, Junko; Kondoh, Hiroshi; Yanagida, Mitsuhiro

    2016-04-19

    Metabolites present in human blood document individual physiological states influenced by genetic, epigenetic, and lifestyle factors. Using high-resolution liquid chromatography-mass spectrometry (LC-MS), we performed nontargeted, quantitative metabolomics analysis in blood of 15 young (29 ± 4 y of age) and 15 elderly (81 ± 7 y of age) individuals. Coefficients of variation (CV = SD/mean) were obtained for 126 blood metabolites of all 30 donors. Fifty-five RBC-enriched metabolites, for which metabolomics studies have been scarce, are highlighted here. We found 14 blood compounds that show remarkable age-related increases or decreases; they include 1,5-anhydroglucitol, dimethyl-guanosine, acetyl-carnosine, carnosine, ophthalmic acid, UDP-acetyl-glucosamine,N-acetyl-arginine,N6-acetyl-lysine, pantothenate, citrulline, leucine, isoleucine, NAD(+), and NADP(+) Six of them are RBC-enriched, suggesting that RBC metabolomics is highly valuable for human aging research. Age differences are partly explained by a decrease in antioxidant production or increasing inefficiency of urea metabolism among the elderly. Pearson's coefficients demonstrated that some age-related compounds are correlated, suggesting that aging affects them concomitantly. Although our CV values are mostly consistent with those CVs previously published, we here report previously unidentified CVs of 51 blood compounds. Compounds having moderate to high CV values (0.4-2.5) are often modified. Compounds having low CV values, such as ATP and glutathione, may be related to various diseases because their concentrations are strictly controlled, and changes in them would compromise health. Thus, human blood is a rich source of information about individual metabolic differences. PMID:27036001

  17. NMR-based metabonomic study of the sub-acute toxicity of titanium dioxide nanoparticles in rats after oral administration

    International Nuclear Information System (INIS)

    As titanium dioxide nanoparticles (TiO2 NPs) are widely used commercially, their potential toxicity on human health has attracted particular attention. In the present study, the oral toxicological effects of TiO2 NPs (dosed at 0.16, 0.4 and 1 g kg-1, respectively) were investigated using conventional approaches and metabonomic analysis in Wistar rats. Serum chemistry, hematology and histopathology examinations were performed. The urine and serum were investigated by 1H nuclear magnetic resonance (NMR) using principal components and partial least squares discriminant analysis. The metabolic signature of urinalysis in TiO2 NP-treated rats showed increases in the levels of taurine, citrate, hippurate, histidine, trimethylamine-N-oxide (TMAO), citrulline, α-ketoglutarate, phenylacetylglycine (PAG) and acetate; moreover, decreases in the levels of lactate, betaine, methionine, threonine, pyruvate, 3-D-hydroxybutyrate (3-D-HB), choline and leucine were observed. The metabonomics analysis of serum showed increases in TMAO, choline, creatine, phosphocholine and 3-D-HB as well as decreases in glutamine, pyruvate, glutamate, acetoacetate, glutathione and methionine after TiO2 NP treatment. Aspartate aminotransferase (AST), creatine kinase (CK) and lactate dehydrogenase (LDH) were elevated and mitochondrial swelling in heart tissue was observed in TiO2 NP-treated rats. These findings indicate that disturbances in energy and amino acid metabolism and the gut microflora environment may be attributable to the slight injury to the liver and heart caused by TiO2 NPs. Moreover, the NMR-based metabolomic approach is a reliable and sensitive method to study the biochemical effects of nanomaterials.

  18. Interaction analysis between HLA-DRB1 shared epitope alleles and MHC class II transactivator CIITA gene with regard to risk of rheumatoid arthritis.

    Directory of Open Access Journals (Sweden)

    Marcus Ronninger

    Full Text Available HLA-DRB1 shared epitope (SE alleles are the strongest genetic determinants for autoantibody positive rheumatoid arthritis (RA. One of the key regulators in expression of HLA class II receptors is MHC class II transactivator (CIITA. A variant of the CIITA gene has been found to associate with inflammatory diseases.We wanted to explore whether the risk variant rs3087456 in the CIITA gene interacts with the HLA-DRB1 SE alleles regarding the risk of developing RA. We tested this hypothesis in a case-control study with 11767 individuals from four European Caucasian populations (6649 RA cases and 5118 controls.We found no significant additive interaction for risk alleles among Swedish Caucasians with RA (n = 3869, attributable proportion due to interaction (AP = 0.2, 95%CI: -0.2-0.5 or when stratifying for anti-citrullinated protein antibodies (ACPA presence (ACPA positive disease: n = 2945, AP = 0.3, 95%CI: -0.05-0.6, ACPA negative: n = 2268, AP = -0.2, 95%CI: -1.0-0.6. We further found no significant interaction between the main subgroups of SE alleles (DRB1*01, DRB1*04 or DRB1*10 and CIITA. Similar analysis of three independent RA cohorts from British, Dutch and Norwegian populations also indicated an absence of significant interaction between genetic variants in CIITA and SE alleles with regard to RA risk.Our data suggest that risk from the CIITA locus is independent of the major risk for RA from HLA-DRB1 SE alleles, given that no significant interaction between rs3087456 and SE alleles was observed. Since a biological link between products of these genes is evident, the genetic contribution from CIITA and class II antigens in the autoimmune process may involve additional unidentified factors.

  19. Identification of novel autoantigen in the synovial fluid of rheumatoid arthritis patients using an immunoproteomics approach.

    Directory of Open Access Journals (Sweden)

    Sagarika Biswas

    Full Text Available Rheumatoid arthritis (RA is a chronic, autoimmune and inflammatory joint disease with a poorly understood etiology. Despite widespread diagnostic use of anti-citrullinated protein antibodies and rheumatoid factor proteins there is a strong demand for novel serological biomarkers to improve the diagnosis this disease. The present study was aimed to identify novel autoantigens involved in rheumatoid arthritis (RA pathogenesis through immune-proteomic strategy. Synovial fluid samples from clinically diagnosed RA patients were separated on two-dimensional gel electrophoresis (2-DE. Samples from patients with non-RA rheumatisms (osteoarthritis and trauma were used as controls. Immunoreactive proteins were spotted by Western blotting followed by identification through Q-TOF mass spectrometer analysis. Forty Western blots were generated using plasma from ten individual RA patients and 33 reactive spots were identified, 20 from the high molecular weight (HMW gel and 13 from the low molecular weight (LMW gel. Among the 33 common immunogenic spots, 18 distinct autoantigens were identified, out of which 14 are novel proteins in this context. Expression analysis of five important proteins, vimentin, gelsolin, alpha 2 HS glycoprotein (AHSG, glial fibrillary acidic protein (GFAP, and α1B-glycoprotein (A1BG by Western blot analysis using their specific antibodies revealed their higher expression in RA synovial fluid as compared to non-RA samples. Recombinantly expressed GFAP and A1BG protein were used to develop an in-house ELISA to quantify the amount of autoantibodies in the RA patients. RA patients revealed an increase in the expression of GFAP and A1BG in the plasma as compared to osteoarthritis patients. Therefore, GFAP and A1BG can be proposed as potential new autoantigens of diagnostic importance for RA subjects. Further characterization of these proteins in rheumatoid arthritis will be helpful in understanding the role of these proteins in the disease

  20. Platelet hyperaggregability in obesity: is there a role for nitric oxide impairment and oxidative stress?

    Science.gov (United States)

    Leite, Natália Rodrigues Pereira; Siqueira de Medeiros, Mariana; Mury, Wanda Vianna; Matsuura, Cristiane; Perszel, Monique Bandeira Moss; Noronha Filho, Gerson; Brunini, Tatiana Mc; Mendes-Ribeiro, Antônio Claúdio

    2016-08-01

    Epidemiological evidence has shown that platelet activation markers are consistently elevated in obesity, contributing to its prothrombotic state. In order to improve the understanding of the regulation of platelet function in obesity, the aim of this study was to investigate the l-arginine-nitric oxide (NO) pathway in obese adults without other cardiovascular risk factor. Seventeen obese (body mass index [BMI] 35.9±1.0 kg/m(2) ) and eighteen age-matched normal weight subjects (BMI 22.0±0.6 kg/m(2) ) were included in this study. l-arginine influx was measured with incubation of l-[(3) H]-arginine. NO synthase (NOS) and arginase activities were determined by the citrulline assay and the conversion of l-[(14) C]-arginine to [(14) C]-urea, respectively. Cyclic guanosine monophosphate (cGMP) content was evaluated by enzyme-linked immunosorbent assay. In addition, the study analyzed: platelet aggregation; intraplatelet antioxidant enzymes, via superoxide dismutase (SOD) and catalase activities; and systemic levels of l-arginine, fibrinogen, and C-reactive protein (CRP). Obese patients presented a significant decrease of platelet l-arginine influx, NOS activity, and cGMP levels, along with platelet hyperaggregability. On the presence of NO donor, platelet aggregation was similar between the groups. The fibrinogen and CRP systemic levels were significantly higher and SOD activity was reduced in obesity. No significant differences were observed in plasma levels of l-arginine and intraplatelet arginase and catalase activities between groups. The diminished NO bioavailability associated with inflammatory status and impaired enzymatic antioxidant defence may contribute to future cardiovascular complications in obesity. PMID:27145241

  1. Peptidylarginine deiminases: novel drug targets for prevention of neuronal damage following hypoxic ischemic insult (HI) in neonates.

    Science.gov (United States)

    Lange, Sigrun; Rocha-Ferreira, Eridan; Thei, Laura; Mawjee, Priyanka; Bennett, Kate; Thompson, Paul R; Subramanian, Venkataraman; Nicholas, Anthony P; Peebles, Donald; Hristova, Mariya; Raivich, Gennadij

    2014-08-01

    Neonatal hypoxic ischaemic (HI) injury frequently causes neural impairment in surviving infants. Our knowledge of the underlying molecular mechanisms is still limited. Protein deimination is a post-translational modification caused by Ca(+2) -regulated peptidylarginine deiminases (PADs), a group of five isozymes that display tissue-specific expression and different preference for target proteins. Protein deimination results in altered protein conformation and function of target proteins, and is associated with neurodegenerative diseases, gene regulation and autoimmunity. In this study, we used the neonatal HI and HI/infection [lipopolysaccharide (LPS) stimulation] murine models to investigate changes in protein deimination. Brains showed increases in deiminated proteins, cell death, activated microglia and neuronal loss in affected brain areas at 48 h after hypoxic ischaemic insult. Upon treatment with the pan-PAD inhibitor Cl-amidine, a significant reduction was seen in microglial activation, cell death and infarct size compared with control saline or LPS-treated animals. Deimination of histone 3, a target protein of the PAD4 isozyme, was increased in hippocampus and cortex specifically upon LPS stimulation and markedly reduced following Cl-amidine treatment. Here, we demonstrate a novel role for PAD enzymes in neural impairment in neonatal HI Encephalopathy, highlighting their role as promising new candidates for drug-directed intervention in neurotrauma. Hypoxic Ischaemic Insult (HI) results in activation of peptidylarginine deiminases (PADs) because of calcium dysregulation. Target proteins undergo irreversible changes of protein bound arginine to citrulline, resulting in protein misfolding. Infection in synergy with HI causes up-regulation of TNFα, nuclear translocation of PAD4 and change in gene regulation as a result of histone deimination. Pharmacological PAD inhibition significantly reduced HI brain damage.

  2. Prevalence and clinical significance of nonorgan specific antibodies in patients with autoimmune thyroiditis as predictor markers for rheumatic diseases.

    Science.gov (United States)

    Elnady, Basant M; Kamal, Naglaa M; Shaker, Raneyah H M; Soliman, Amal F; Hasan, Waleed A; Alghamdi, Hamed A; Algethami, Mohammed M; Jajah, Mohamed Bilal

    2016-09-01

    Autoimmune diseases are considered the 3rd leading cause of morbidity and mortality in the industrialized countries. Autoimmune thyroid diseases (ATDs) are associated with high prevalence of nonorgan-specific autoantibodies, such as antinuclear antibodies (ANA), antidouble-stranded deoxyribonucleic acid (anti-dsDNA), antiextractable-nuclear antigens (anti-ENAs), rheumatoid factor (RF), and anticyclic-citrullinated peptides (anti-CCP) whose clinical significance is unknown.We aimed to assess the prevalence of various nonorgan-specific autoantibodies in patients with ATD, and to investigate the possible association between these autoantibodies and occurrence of rheumatic diseases and, if these autoantibodies could be considered as predictor markers for autoimmune rheumatic diseases in the future.This study had 2 phases: phase 1; in which 61 ATD patients free from rheumatic manifestations were assessed for the presence of these nonorgan-specific autoantibodies against healthy 61 control group, followed by 2nd phase longitudinal clinical follow-up in which cases are monitored systematically to establish occurrence and progression of any rheumatic disease in association to these autoantibodies with its influences and prognosis.Regarding ATD patients, ANA, anti-dsDNA, Anti-ENA, and RF were present in a percentage of (50.8%), (18%), (21.3%), and (34.4%), respectively, with statistically significance difference (P controls. Nearly one third of the studied group (32.8%) developed the rheumatic diseases, over 2 years follow-up. It was obvious that those with positive anti-dsDNA had higher risk (2.45 times) to develop rheumatic diseases than those without. There was a statistically significant positive linear relationship between occurrence of disease in months and (age, anti-dsDNA, anti-CCP, RF, and duration of thyroiditis). Anti-dsDNA and RF are the most significant predictors (P management of the disease, as early disease control will allow better quality of life. PMID

  3. Metabolic changes in summer active and anuric hibernating free-ranging brown bears (Ursus arctos.

    Directory of Open Access Journals (Sweden)

    Peter Stenvinkel

    Full Text Available The brown bear (Ursus arctos hibernates for 5 to 6 months each winter and during this time ingests no food or water and remains anuric and inactive. Despite these extreme conditions, bears do not develop azotemia and preserve their muscle and bone strength. To date most renal studies have been limited to small numbers of bears, often in captive environments. Sixteen free-ranging bears were darted and had blood drawn both during hibernation in winter and summer. Samples were collected for measurement of creatinine and urea, markers of inflammation, the calcium-phosphate axis, and nutritional parameters including amino acids. In winter the bear serum creatinine increased 2.5 fold despite a 2-fold decrease in urea, indicating a remarkable ability to recycle urea nitrogen during hibernation. During hibernation serum calcium remained constant despite a decrease in serum phosphate and a rise in FGF23 levels. Despite prolonged inactivity and reduced renal function, inflammation does not ensue and bears seem to have enhanced antioxidant defense mechanisms during hibernation. Nutrition parameters showed high fat stores, preserved amino acids and mild hyperglycemia during hibernation. While total, essential, non-essential and branched chain amino acids concentrations do not change during hibernation anorexia, changes in individual amino acids ornithine, citrulline and arginine indicate an active, although reduced urea cycle and nitrogen recycling to proteins. Serum uric acid and serum fructose levels were elevated in summer and changes between seasons were positively correlated. Further studies to understand how bears can prevent the development of uremia despite minimal renal function during hibernation could provide new therapeutic avenues for the treatment of human kidney disease.

  4. Potential biomarkers of fatigue identified by plasma metabolome analysis in rats.

    Directory of Open Access Journals (Sweden)

    Satoshi Kume

    Full Text Available In the present study, prior to the establishment of a method for the clinical diagnosis of chronic fatigue in humans, we validated the utility of plasma metabolomic analysis in a rat model of fatigue using capillary electrophoresis-mass spectrometry (CE-MS. In order to obtain a fatigued animal group, rats were placed in a cage filled with water to a height of 2.2 cm for 5 days. A food-restricted group, in which rats were limited to 10 g/d of food (around 50% of the control group, was also assessed. The food-restricted group exhibited weight reduction similar to that of the fatigued group. CE-MS measurements were performed to evaluate the profile of food intake-dependent metabolic changes, as well as the profile in fatigue loading, resulting in the identification of 48 metabolites in plasma. Multivariate analyses using hierarchical clustering and principal component analysis revealed that the plasma metabolome in the fatigued group showed clear differences from those in the control and food-restricted groups. In the fatigued group, we found distinctive changes in metabolites related to branched-chain amino acid metabolism, urea cycle, and proline metabolism. Specifically, the fatigued group exhibited significant increases in valine, leucine, isoleucine, and 2-oxoisopentanoate, and significant decreases in citrulline and hydroxyproline compared with the control and food-restricted groups. Plasma levels of total nitric oxide were increased in the fatigued group, indicating systemic oxidative stress. Further, plasma metabolites involved in the citrate cycle, such as cis-aconitate and isocitrate, were reduced in the fatigued group. The levels of ATP were significantly decreased in the liver and skeletal muscle, indicative of a deterioration in energy metabolism in these organs. Thus, this comprehensive metabolic analysis furthered our understanding of the pathophysiology of fatigue, and identified potential diagnostic biomarkers based on fatigue

  5. Immunological markers of rheumatoid arthritis

    Directory of Open Access Journals (Sweden)

    Agnieszka Matuszewska

    2016-03-01

    Full Text Available Rheumatoid arthritis (RA is the most common connective tissue disease of autoimmune origin. The disease is characterized by chronic inflammation leading to bone erosions and organ involvement. RA is a progressive disease. It affects the quality of life, leading to disability and death mainly due to premature cardiovascular disease. Early diagnosis and appropriate treatment are essential for prognosis and quality of life improvement. In 2010 the American College of Rheumatology (ACR and The European League Against Rheumatism (EULAR established new RA classification criteria. Besides clinical symptoms it includes two immunologic criteria: rheumatoid factor (RF and anti-citrullinated protein antibodies (anti-CCP antibodies. RF is the first well-known RA immunologic marker. It is observed in 80-85% of patients with RA. Elevated serum level of RF has been associated with increased disease activity, radiographic progression, and the presence of extraarticular manifestations. The sensitivity of RF is 50-90%, and specificity is 50-95%. Anti-CCP antibodies appear to be a more specific marker than RF. They are often present at the very beginning of the disease, or even years before the first symptoms. The prognostic value of anti-CCP antibodies is well established. High serum level of anti-CCP correlates with poor prognosis and early erosions of the joints. The sensitivity of anti-CCP2 is 48-80%, and specificity is 96-98%. New immunologic markers include anti-carbamylated protein antibodies (anti-CarP and antibodies against heterogeneous nuclear ribonucleoproteins (anti-hnRNP A2/B1, RA33. Scientists aim to identify a highly sensitive and specific biomarker of the disease that not only has diagnostic and prognostic value but also may predict the response to treatment.

  6. Activities of asymmetric dimethylarginine-related enzymes in white adipose tissue are associated with circulating lipid biomarkers

    Directory of Open Access Journals (Sweden)

    Iwasaki Hiroaki

    2012-04-01

    Full Text Available Abstract Background Asymmetric NG,NG-dimethylarginine (ADMA, an endogenous inhibitor of nitric oxide synthase, is regulated by the enzymatic participants of synthetic and metabolic processes, i.e., type I protein N-arginine methyltransferase (PRMT and dimethylarginine dimethylaminohydrolase (DDAH. Previous reports have demonstrated that circulating ADMA levels can vary in patients with type 1 and type 2 diabetes mellitus (T2DM. White adipose tissue expresses the full enzymatic machinery necessary for ADMA production and metabolism; however, modulation of the activities of adipose ADMA-related enzymes in T2DM remains to be determined. Methods A rodent model of T2DM using 11- and 20-week old Goto-Kakizaki (GK rats was used. The expression and catalytic activity of PRMT1 and DDAH1 and 2 in the white adipose tissues (periepididymal, visceral and subcutaneous fats and femur skeletal muscle tissue were determined by immunoblotting, in vitro methyltransferase and in vitro citrulline assays. Results Non-obese diabetic GK rats showed low expression and activity of adipose PRMT1 compared to age-matched Wistar controls. Adipose tissues from the periepididymal, visceral and subcutaneous fats of GK rats had high DDAH1 expression and total DDAH activity, whereas the DDAH2 expression was lowered below the control value. This dynamic of ADMA-related enzymes in white adipose tissues was distinct from that of skeletal muscle tissue. GK rats had lower levels of serum non-esterified fatty acids (NEFA and triglycerides (TG than the control rats. In all subjects the adipose PRMT1 and DDAH activities were statistically correlated with the levels of serum NEFA and TG. Conclusion Activities of PRMT1 and DDAH in white adipose tissues were altered in diabetic GK rats in an organ-specific manner, which was reflected in the serum levels of NEFA and TG. Changes in adipose ADMA-related enzymes might play a part in the function of white adipose tissue.

  7. Serum Vaspin Levels Are Associated with the Development of Clinically Manifest Arthritis in Autoantibody-Positive Individuals.

    Directory of Open Access Journals (Sweden)

    Karen I Maijer

    Full Text Available We have previously shown that overweight may increase the risk of developing rheumatoid arthritis (RA in autoantibody positive individuals. Adipose tissue could contribute to the development of RA by production of various bioactive peptides. Therefore, we examined levels of adipokines in serum and synovial tissue of subjects at risk of RA.Fifty-one individuals positive for immunoglobulin M rheumatoid factor (IgM-RF and/or anti-citrullinated protein antibodies (ACPA, without arthritis, were included in this prospective study. Levels of adiponectin, vaspin, resistin, leptin, chemerin and omentin were determined in baseline fasting serum samples (n = 27. Synovial tissue was obtained by arthroscopy at baseline and we examined the expression of adiponectin, resistin and visfatin by immunohistochemistry.The development of clinically manifest arthritis after follow-up was associated with baseline serum vaspin levels (HR1.5 (95% CI 1.1 to 2.2; p = 0.020, also after adjustment for overweight (HR1.7 (95% CI 1.1 to 2.5; p = 0.016. This association was not seen for other adipokines. Various serum adipokine levels correlated with BMI (adiponectin r = -0.538, leptin r = 0.664; chemerin r = 0.529 and systemic markers of inflammation such as CRP levels at baseline (adiponectin r = -0.449, omentin r = -0.557, leptin r = 0.635, chemerin r = 0.619, resistin r = 0.520 and ESR (leptin r = 0.512, chemerin r = 0.708, p-value<0.05. Synovial expression of adiponectin, resistin and visfatin was not associated with development of clinically manifest arthritis.In this exploratory study, serum adipokines were associated with an increased inflammatory sta