Total proteins from Mtb subcellular fractions (10 or 50 μg per lane) were subjected to SDS-PAGE (Invitrogen, USA) and transferred to a nitrocellulose membrane. The membrane was blocked with 5% non-fat milk in tris-buffered saline containing 0.1% Tween RG7420 cell line 20 (TBS-T) and probed with the monoclonal supernatant (Clone 276.B7/IgG1κ) or anti-19 KD lipoprotein mAb (clone IT-19; kindly provided by Dr. Antônio Rothfuchs, NIH/NIAID-TVTRM Contract) at 1:1000 dilution followed by incubation with HRP-conjugated secondary Ab (1:2000). Detection was

performed by ECL analysis (Pierce, USA). Thirty-four patients with active pulmonary TB in the Division of Respiratory Diseases of the Central Public Health Clinic of Juiz de Fora, Minas Gerais State and 11 active-diseased patients from Hospital Octávio Mangabeira, Bahia, Brazil were selected. Only those patients with detectable AFB in the sputum bacilloscopy or culture-confirmed disease and who had selleck undergone clinical and chest X-ray examinations, as prescribed by the Brazilian Ministry of Health, were included in the study. AIDS, diabetes, hepatitis, hypertension, pregnancy, and alcoholism were exclusion

criteria. All patients included in the study have been confirmed to present negative bacilloscopy following treatment. Thirty-eight healthy BCG-vaccinated, which constituted the endemic control (EC) group formed by medical students and staff from UFJF, five foreign PPD-negative non-BCG-vaccinated subjects (the non-endemic group) and six PPD-negative BCG-vaccinated individuals were included in the control groups without prior history of Mtb infection. All patients and control

subjects have been informed of the study and have given consent for blood sampling. The UFJF Medical Ethics as well as the Oswaldo Cruz Foundation Committees have approved the study protocols (UFJF-1495.186.2008; CPqGM-219 (CAAE) 2221.0.000.225-06). Histological sections from pleural TB patients or control leprosy patients were deparaffinized in xylene, rehydrated in alcohol and water. Quenching of endogenous peroxidase was performed with a 1.5% hydrogen peroxide-methanol solution for 20 min. Sections were incubated Megestrol Acetate with normal goat serum (30 min 37°C) and then exposed to monoclonal anti-sMTL-13 supernatant (Clone 276.B7). Incubations with biotinylated goat anti-mouse Ab with streptavidin−HRP complex (Vectastain Elite ABC reagent, Vector Laboratories, CA, USA) were performed for 30 min at 37°C. Positive reactions was detected with 3,3′-diaminobenzidine (Dako Cytomation, CA, USA), followed by Harris’s hematoxylin counterstaining. Sections were examined microscopically and images were acquired using a Sight DS-5M-L1 digital camera (Nikon, Melville, NY, USA) connected to an Eclipse 50i light microscope (Nikon). Maxsorb plates (Nunc, Denmark) were coated with rec-sMTL-13 in carbonate buffer overnight at 4°C. Plates were washed with PBS/0.05% Tween-20.

We acknowledge the Wellcome Trust, NIHR Biomedical Research Centre Programme

(Oxford) and the MRC. None. “
“Inflammatory DCM (iDCM) may be related to autoimmune processes. An immunoadsorption (IA) has been reported to improve cardiac hemodynamics. The benefit of IA is probably related to the removal of autoantibodies. A recent study suggests additional effects of IA on the T cell–mediated immune reactions, especially on regulatory T cells (Tregs). In this prospective study, the correlation between the level of Tregs and improvement of myocardial contractility in response to IA in patients with iDCM was investigated. Patients (n = 18) with iDCM, reduced left ventricular (LV) ejection fraction (<35%), were enrolled for IA. Before and 6 months KPT-330 order after IA, LV systolic function was assessed by echocardiography, and blood levels of Tregs were quantified by FACS analysis. Patients (n = 12) with chronic ischaemic heart failure and comparable reduced LV-EF served as controls. IA improved see more LV-EF in 12 of 18 patients at 6-month follow-up. These patients were classified as ‘IA responder’. In 6 patients, LV-EF remained unchanged. At baseline, IA responder and non-responder subgroups showed similar values for C-reactive protein,

white blood cells, lymphocytes and T helper cells, but they differ for the number of circulating Tregs (responder: 2.32 ± 1.38% versus non-responder: 4.86 ± 0.28%; P < 0.01). Tregs increased significantly in the IA responders, but remained unchanged in the IA non-responders. In patients with ischaemic

cardiomyopathy, none of these values changed over Tau-protein kinase time. A low level of Tregs in patients with chronic iDCM may characterize a subset of patients who do best respond to IA therapy. Dilated cardiomyopathy (DCM) is defined by an impairment of myocardial contractile function and ventricular dilation. In a subset of patients, the etiopathophysiology of DCM is linked to autoimmune reactions, characterized by the appearance of cardiotoxic autoantibodies in the blood and signs of myocardial inflammation. In about 2/3 of patients with autoantibodies, viral or bacterial RNA or DNA can be detected in myocardial biopsies, suggesting that these immunological features are initiated by an infectious process [1-3]. A (non-ischaemic) DCM with an autoimmune- or immune-mediated infectious background has been termed as inflammatory DCM (iDCM). A variety of autoantibodies against cardiac cell proteins have been identified in patients with iDCM [3]. Of note, many of these autoantibodies (e.g. targeting ß1-adrenergic receptor, muscarinic M2-acetylcholine receptor, myosin, Na-K-ATPase, troponin I) belong to the IgG subclass 3 that has the highest antibody-dependent potency for cellular toxicity [4]. Wallukat et al.

The gata3 siRNA is a mixture of three kinds of double-stranded Tanespimycin solubility dmso RNA. The sequences gata3 siRNA are as follow. gata3-1 (sense): 5′-GACGGAAGAGGUGGACGUA(dTdT)-3′; gata3-1 (anti-sense): 5′-UACGUCCACCUCUUCCGUC(dTdT)-3′; gata3-2 (sense): 5′-UCGUACAUGGAAGCUCAGU(dTdT)-3′; gata3-2 (anti-sense): 5′-ACUGAGCUUCCAUGUACGA(dTdT)-3′; gata3-3 (sense): 5′-GAUUUCAGAUCUGGGC-AAU(dTdT)-3′; gata3-3 (anti-sense): 5′-AUUGCCCAGAUCUGAAAUC(dTdT)-3′. The sequences of control siRNA are as follows. Control (sense): 5′-CCUACGCCACCAAUUUCGU(dTdT)-3′; control (anti-sense): 5′-ACGAAAUUGGUGGCGUAGG(dTdT)-3′. Results were expressed as mean ± standard

deviation (SD). Differences between groups were determined Buparlisib order by a Student’s t-test. To investigate the molecular mechanism of GATA-3 in the regulation of Th2 cytokine and ifng loci, we searched for GATA-3-interacting proteins. We overexpressed HA-tagged GATA-3 in 293T cells. Cell extracts from these cells were passed through an HA-affinity column. Then, Th2 cell extracts were passed through this column. After washing and elution, GATA-3-interacting proteins were

analysed by MS/MS spectrometry. As the profile of GATA-3-interacting proteins is huge, we narrowed down the list to transcription factors and chromatin-remodelling factors (Table 2). Among the GATA-3-interacting proteins, we were particularly interested in MTA-2 and selected it for subsequent study, because MTA-2 has been shown to be involved in il4 transcription and chromatin regulation.22 We confirmed the binding of GATA-3 with MTA-2 by co-immunoprecipitation. We made cell extracts from in vitro-stimulated Th2 cells from C57/BL6 mice, and

immunoprecipitated with either the anti-GATA-3 selleck inhibitor or anti-MTA-2 antibody, then immunoblotted the anti-MTA-2 or anti-GATA-3 antibody, respectively. GATA-3 and MTA-2 co-immunoprecipitated with either the anti-GATA-3 or anti-MTA-2 antibody (Fig. 1a,b), indicating that these proteins interact with each other, which validated our affinity purification and MS/MS data. We next examined the relative amount of MTA-2 between Th1 and Th2 cells. We prepared cell extracts from Th1 and Th2 cells and measured the relative amount of MTA-2 protein by immunoblotting. The amount of MTA-2 protein was comparable between Th1 and Th2 cells (Fig. 1c). Acetylation of GATA-3 at the lysine residues has been shown to affect the function of GATA-3, in particular, in T-cell survival and homing to secondary lymphoid tissues.23 As the NuRD complex has deacetylase activity,18 we examined whether the acetylation status of GATA-3 can affect the binding with MTA-2. We found that an acetylated protein the same size as GATA-3 was co-immunoprecipitated with MTA-2, suggesting indirectly that acetylated GATA-3 may bind to MTA-2 (Fig. S1).

T cells were purified with negative magnetic bead selection using the “Pan T cell isolation Kit” (Miltenyi Biotech, Bergisch Gladbach, Germany). Antibodies that were used in this study were specific for following markers: CD3ε,

LFA-1, CD2 (BD-Bioscience, Heidelberg, Germany), calmodulin (Zymed, Munich, Germany) and LPL 17. W7 was from Calbiochem (Darmstadt, Germany), Hoechst 33342 from Invitrogen (Karlsruhe, Germany), and BPB and Cytochalasin D from ACP-196 molecular weight Sigma-Aldrich (Taufkirchen, Germany). For cDNA transfection into T cells, the “Human T Cell Nucleofector™ Kit” (Amaxa Biosystems, Cologne, Germany) was used. For the siRNA approaches, cells were electroporated with LPL-specific or control siRNA (Dharmacon, Lafayette, IN, USA). Thereafter, cells were stimulated with 2 μg/mL PHA for 16 h. The PHA was removed and the cells were transferred in medium containing 25 U/mL IL-2. After 2 days incubation cells were electroporated again and incubated for another 2 days at 37°C. The IL-2 was www.selleckchem.com/products/INCB18424.html removed and the cells were incubated in medium without IL-2 for 24 h prior to further experiments. Conjugates were formed between T cells and superantigen-loaded Raji B cells as described 17. Subcellular localization of proteins was determined by immunofluorescence and subsequent analysis with confocal LSM, TLV 17 or MIFC. Cells were stimulated and stained with

fluorescently labeled antibodies and nuclear dyes (Hoechst) as indicated. Data acquisition was performed with an ImageStream (IS100) and data were analyzed with IDEAS 3.0 (Amnis, Seattle, WA, USA). To find the contact zone between T cells and APC, a Hoechst-dependent

valley mask was defined between T-cell/APC couples and combined with a T-cell mask (Supporting Information Fig. 1). Thereafter, protein accumulation was calculated as ratio between the fluorescence intensities Dehydratase of the respective protein in the IS- and T-cell mask. The data were controlled by manually evaluation of 100 cells per sample. The size of the IS was calculated with the major axis feature and the size of T cells with the diameter feature on the T-cell mask. Both algorithms return the results in microns. The F-actin content in the cells was calculated as mean fluorescence intensity of the phalloidin staining within the T-cell mask. The plasmid pEGFP containing the wt-LPL cDNA was generated in our own laboratory 17. The plasmid was used to create mutants of LPL as follows: the two EF-hand calcium-binding domains of LPL at positions 22–27 (ΔEF1-LPL) and 62–73 (ΔEF2-LPL) or both calcium-binding domains (ΔEF1/2-LPL) 36, 37 were deleted using the QuickChange site-directed Mutagenesis XL Kit (Stratagene, La Jolla, CA, USA) according to the manufacturer’s instructions. The actin-binding domains at position 120–627 were removed by PCR amplification of the first 120 aa, which were subsequently introduced in pEGFP via EcoRI and XhoI.

This complex is called the death-inducing signaling complex [5], at which procaspase-8 is activated and cleaved. Since cellular FLICE-inhibitory

(c-FLIP) proteins contain death effector domains as well, these proteins compete with caspase-8 for FADD binding [6]. Recruitment of c-FLIP results in altered death-inducing signaling complex composition and apoptosis inhibition. The Cflar gene encodes different c-FLIP protein isoforms, which can have opposing functions [7, 8]. For instance, c-FLIPL contains a caspase-like domain lacking proteolytic activity and acts in a pro-apoptotic manner when expressed in low amounts but in an anti-apoptotic manner when highly expressed

[7]. In addition, Selleckchem RG 7204 Doxorubicin humans generate two solely anti-apoptotic acting short isoforms called c-FLIPS and c-FLIPR [6, 9], whose expression is regulated by a single nucleotide polymorphism [10]. Strikingly, c-FLIPR expression in humans is associated with an increased risk of follicular lymphoma [10]. The role of c-FLIPS in the human immune response has been extensively analyzed. For instance, c-FLIPS is highly induced in recently activated human T cells and contributes to resistance to CD95-induced apoptosis during the early phase of an immune response [11-14]. In contrast, the function of the c-FLIPR isoform remains enigmatic. Mouse models established so far for analysis of the physiological functions of short c-FLIP isoforms express human c-FLIPS in a T-cell-specific manner [15, 16]. Both studies reported decreased CD95-mediated apoptosis in transgenic T cells, normal lymphocyte

Amoxicillin cellularity, and decreased proliferation of T cells upon activation [15, 16]. However, since the murine Cflar gene (encoding c-FLIP) allows expression of c-FLIPR as the short isoform next to c-FLIPL but not c-FLIPS expression [17], the murine system seems to be a more suitable model for gaining a better understanding of the function of c-FLIPR in vivo. Of note, it is currently not known whether murine c-FLIPR is expressed endogenously at the protein level and whether it modulates the immune response during infection. To address these questions, we analyzed endogenous c-FLIP protein expression and show that murine c-FLIPR is induced during T-cell activation in a similar way as we previously reported for c-FLIPS in the human system [11]. Moreover, we generated vavFLIPR mice expressing a c-FLIPR transgene under the control of the vav-promoter leading to expression in all hemato-poietic cells [18]. vavFLIPR mice had normal numbers and frequencies of immune cells in the steady state. Upon challenge with Listeria monocytogenes, vavFLIPR mice exhibited less liver necrosis and a higher frequency of CD8+ T cells.

With respect to the other cytokines analysed [6], in NALT and NP it was observed that the frequency of IL-2, INF-γ and TNF-α-producing T cells was very low, compared to those producing IL-4, IL-5 and IL-10. They also were increased in immunized mice in relation to control mice (excepting TNF-α-producing cells in NP which did not change). Therefore, although the percentage of T cells that produce IL-2 and IFN-γ represent the lowest values in both groups and in both tissues, these data suggest https://www.selleckchem.com/products/Y-27632.html at least a small number of T cells in NALT and NP produce Th1

cytokines, and their frequency is slightly increased by the effects of Cry1Ac. In previous studies, we have reported that Cry1Ac is highly immunogenic and confers mucosal and systemic adjuvant selleck chemical effects when is administered to mice by systemic or mucosal routes [9–12]. In addition, we have observed that Cry1Ac increases protective immunity against experimental N. fowleri meningoencephalitis in mice [14]. Considering that the immune response elicited, following intranasal immunization with Cry1Ac protoxin, had not yet been analysed in the nasal tract, in this work we evaluated specific antibody cell responses, as well as the activation

and cytokine production, in the lymphocyte populations residing at the nasal compartments of the NALT and NP. On the basis of our previous results, and considering the additional advantages that Cry1Ac has over other mucosal adjuvants [10, 17], in that it is non-toxic to vertebrates, and its production costs are low, we had suggested that this protein may be an attractive candidate to improve the efficacy of vaccination against pathogens invading the nasal mucosa. While the outcomes

of present study contribute to explaining the potent immunogenicity of Cry1Ac via i.n. route. Because in both NALT Baricitinib and NP lymphocytes from immunized mice we found that: (1) significant specific IgA and IgG cell responses were induced, especially in NP, (2) the proportion of activated lymphocytes was increased and (3) the proportion of T cells spontaneously producing cytokines, especially a Th2 profile of cytokines, was increased. In mice immunized with Cry1Ac, the response found in nasal lymphocytes was as good, or even higher to that attained with CT, which was used as a reference of the most potent mucosal immunogen known. Nevertheless, the immunization scheme used may be not the optimal to achieve the maximal anti-CT responses. Besides, given the different conditions used for each protein, the immune responses achieved are not suitable to compare, because a higher dose of Cry1Ac was used, and because higher doses than 10 μg of CT can not be assayed in mice because of its toxicity.

A number of studies suggested that Treg exert their suppressive function on effector T cells indirectly by modifying the function of antigen-presenting

dendritic cells. Interestingly, a recent in vitro study showed that LFA-1 is important for the formation of dendritic cell/Treg aggregates, because LFA-1−/− Treg were no longer able to inhibit the maturation of cocultured dendritic cells 20. Similar effects were also observed in a mixed human/mouse suppression system 21. We CHIR-99021 chemical structure show here that LFA-1 deficiency results in a reduced Treg/effector cell ratio in the inflamed CNS. The reduction in Treg was already established in the spleen and thymi of unimmunized LFA-1−/− mice. Hence, besides a possible functional impairment of LY294002 price Treg lacking LFA-1, these results indicate a more fundamental role for LFA-1 in generation of FoxP3+ Treg in the thymus. ICAM-1, a ligand of LFA-1, is expressed on thymic stromal cells 22. Therefore, LFA-1 potentially increases the

physical contact between thymocytes and stromal cells, resulting in enhanced T-cell receptor triggering. Increased T-cell receptor signaling during thymocyte selection favors the generation of naturally occurring Treg 23, which would explain the contribution of LFA-1 to the generation of naturally occurring Treg. So far, LFA-1 has been mainly recognized as a molecule regulating the migration of lymphocytes. Generally, the migration of LFA-1-deficient T cells to the peripheral lymph nodes is impaired, resulting in significantly smaller lymph nodes 10, 14. However, upon immunization with MOG-peptide, we observed that these differences in cellularity in lymph nodes between WT and LFA-1 KO mice are more or less levelled out (data not shown). In the context of EAE and transendothelial migration, Laschinger et al. 11 demonstrated that encephalitogenic ID-8 T cells do not use LFA-1 for the initial adhesion to the endothelium of the blood/brain barrier. Instead, LFA-1 was involved in the later phases of migration into the CNS parenchyma. However, it should be noted that these results were obtained for the healthy spinal cord and that the role of LFA-1 for migration

could be different during later stages of an EAE disease, in which other integrin interactions may compensate for the lack of LFA-1. In our study, we did not directly address the question of lymphocyte migration via the blood/brain barrier. However, the observation that the frequency of MOG reactive CD4+ T cells in LFA-1−/− mice is already higher outside the CNS suggests an impaired suppression of effector T cells by Treg rather than an altered migration as cause for the higher ratio of effector versus Treg in the inflamed CNS in LFA-1−/−. Overall, the exacerbated EAE in the absence of LFA-1 seems to be due to the impaired suppression of autoantigen-specific effector T cells by Treg, which in LFA-1−/− mice show a more extensive expansion in secondary lymphoid organs upon immunization with the MOG-peptide.

The benefits and effects of mTORi were assessed in our centre’s cohort. Methods: We analysed graft function, rejection rates, tolerability and discontinuation rates in a retrospective cohort analysis of 44 adult kidney transplant recipients (29 male and 15 female) treated

with mTORi between 2006 to 2012. Results: All patients switched from CNI to mTORi, the reasons for conversion were skin cancers (37%), CNI toxicity/ intolerance (25%), CT99021 datasheet planned reduction in immunosuppression (14%), study trials (7%), BK nephropathy (5%) and others (12%). mTORi had to be discontinued in 15 (34%) patients within 24 months and in 7 (16%) after 24 months because of either rejection, severe see more proteinuria, oedema, muco-cutaneous

effects, leukopenia, pneumonitis, or cerebral venous thrombosis. The eGFR pre-conversion was 56 ± 22 mL/min/1.73 m2 and 63 ± 24 mL/min/1.73 m2 (P < 0.01) at 1 month, but did not differ from pre-conversion at 3, 6, 12 and 24 months. Fourteen (32%) patients experienced biopsy proven rejection (n = 9 cellular, 2 mixed and 3 borderline changes) without association to HLA mismatches, or time of conversion after transplantation. Conclusions: In this retrospective analysis of a small subset of patients, mTORi treatment is associated with early adverse effects

or acute rejection leading to discontinuation of mTORi in up to 50% of patients. mTOR inhibitors are a reasonable therapeutic alternative to CNIs for a only a subset of renal transplantation recipients. 265 HIGH-SENSITIVITY TROPONIN T AS A PREDICTOR OF CARDIOVASCULAR MORBIDITY IN RENAL TRANSPLANT RECIPIENTS Digestive enzyme K FERNANDEZ, C MUNRO, M SURANYI, A MAKRIS, J WONG, H HASSAN Renal Unit Liverpool Hospital, Australia Aim: Determine if any significant change in High-sensitivity troponin T (hsTnT) occurs following renal transplantation. Background: hsTnT is a biomarker for detecting myocardial injury. Its use as a predictor of cardiac events in stable dialysis patients has previously been investigated. It remains uncertain if pre-transplant hsTnT levels offer any predictive value in determining cardiac events post-transplant. Methods: We designed a prospective cohort study in South West Sydney in a non-transplant centre. Serum hsTnT was analysed from 30 dialysis patients pre-transplant and post-transplant. Patients were then classified and analysed according to their pre-transplant hsTnT levels: normal (Group 1 – levels < 14 ng/L) and those with elevated hsTnT (Group 2).

The few HD transplanted cases that have undergone autopsy [22,42–46] offer a unique window into the events that take place around and within grafted tissue when placed in a pathological context. The information derived from each post-mortem analysis is invaluable and critical to the implementation of significant improvements of transplantation strategies. Bachoud-Lévi et al., Lancet 2000 [48]

(1 year) Gaura et al., Brain 2004 [49] (2 years) Bachoud-Lévi et al., Lancet Neurol 2006 [50] (6 years) Krystkowiak et al., PLoS ONE 2007 [51] (n = 13) Rosser et al., J Neurol Neurosurg Psychiatry 2002 [19] (6 months) Barker et al., J Neurol Neurosurg buy RAD001 Psychiatry 2013 [41] (3–10 years) Gallina et al., Exp Neurol 2008 [52] (15 months) Gallina et al., Exp Neurol 2010 [21] (18 to 34 months) 1–2/7–9 weeks 25–43 mm Keene et al., Neurology 2007 [46] (6–7 years) Keene et al., Acta Neuropathol 2009 [45] (10 years) Napabucasin in vitro Freeman et al., Proc Natl Acad Sci USA 2000 [42] (18 months) Cicchetti et al., Proc Natl Acad Sci USA 2009 [43] (9, 9.5 and 10 years) Cisbani et al., Brain 2013 [44] (9 and 12 years) In the last decade, our group has undertaken a series of unique studies on the post-mortem analysis of brains obtained from HD patients who have taken part in a clinical trial initiated by the University of South Florida (Table 1)

[17,42–44]. A few additional cases from American and European cohorts have been investigated post-mortem (Table 2). Capetian et al. have recently described one case from the University of Freiburg trial who died 6 months following the transplant procedure [22]. The group of Keene and collaborators who leads the California trial, have published the post-mortem analyses

of three of their cases who have come to autopsy 6, 7 [46] and 10 years [45] after transplantation. In total, the post-mortem analyses of nine cases originating from three distinct clinical trials have been reported (Tables 2 and 3) [22,42–46]. Despite this limited number of cases, each of them has yielded critical and unique information on how grafted foetal tissue behaves in a severely diseased brain and how this may account for the suboptimal clinical outcomes reached. Notwithstanding Dynein discrepancies in the methodologies used in each of the three trials, these post-mortem studies further lead one to hypothesize about how long-term graft survival may be affected by factors such as tissue dissection, cell preparation methods and patient selection. Finally, this review discusses the possible factors influencing graft survival, with a particular emphasis on the post-mortem data. 8/10 (9 years) 9/11 (9.5 years) 1/16 (10 years) None Cysts and mass lesions 8/10 (9 years) 11/11 (12 years) In all clinical trials of cell transplantation in HD patients, postoperative magnetic resonance imaging (MRI) has been used to confirm graft placement (Table 1).

Patients’ outcomes were reviewed for 2 years from the admission of acute coronary syndrome. Primary outcomes of the study included re-hospitalization for acute coronary syndrome and all- cause mortality. Results: Thienopyridines users experienced significantly more re-hospitalization for acute coronary syndrome than aspirin users (26.64% vs. 17.48%, P < 0.001), whereas adjusted hazard buy MI-503 ratio [HR] was 1.56 (95% confidence interval [CI]: 1.30 to 1.88)

and all cause of mortality adjusted HR was 1.15 (95% CI: 0.99 to 1.34). Conclusion: In this retrospective analysis, aspirin treatment appeared more effective than thienopyridines for secondary prevention of acute coronary syndrome and showed a non-significant trend towards lower all-cause mortality. LIN CHIH-CHING1,2, YANG WU-CHANG1,2 1Taipei Veterans General Hospital; 2National Yang Ming University Introduction: Elevated

plasma asymmetric dimethylarginine (ADMA) has been reported to be associated with restenosis after percutaneous transluminal angioplasty (PTA) of AVF in hemodialysis (HD) patients. Dimethylarginine dimethylaminohydrolase 1 (DDAH1) is the major enzyme eliminating ADMA, but the effect of genetic variations in DDAH1 on the outcome of vascular access after PTA in HD patients remained unknown. Methods: We assessed the association between polymorphisms in DDAH1 and vascular access outcome in 473 maintenance HD patients, who were prospectively followed up for one Tigecycline cost year after PTA for vascular access dysfunction. Eleven single nucleotide polymorphisms (SNPs) in endothelial function related genes were analyzed and plasma ADMA levels were determined at baseline. Results: After adjustment of demographic,

access, and risk factors, individuals with high baseline plasma ADMA (>0.9 μM) levels had higher rates of re-intervention at 6 months after PTA (74% vs. 53%, p = 0.05). DDAH1 rs233112 was significantly associated with increased levels of plasma ADMA levels. Compared with individuals with rs233112 AA genotypes, individuals with rs233112 GA or GG genotypes had higher risks for re-intervention (58% vs. 45%, p = 0.003) after PTA at 6 months. Phosphoglycerate kinase In the same multivariate- adjusted model, the clinical factors predicting higher risk of re-intervention at 6 months include current smoker, graft access, and rs233112 GG+GA genotypes of DDAH1 gene (HR 2.302, 95% CI 1.557–3.407). Conclusion: Our study demonstrate that rs233112 GG+GA genotypes of DDAH1 gene predict early and frequent restenosis of vascular accesses after PTA in HD patients. SEONG LIM PAIK, CHUNG JENG YA, YING WU MING Tungs’ Taichung Metroharbour Hospital Introduction: Chronic inflammation in dialysis patients may cause malnutrition and progressive atherosclerotic CVD and available data suggest that pro-inflammatory cytokines play a central role.