In the latter case, the secretory mechanism involves

intragranular compartments organized as tubular vesicles or tubular networks, which bud from donor granules and relocate specific granule products in response to stimulation 24. Consequently, PMD would accomplish discharge of secretory constituents from storage granules without granule-to-granule and granule-to-plasma membrane fusion events and without direct granule opening to the cell exterior, as we have observed in our experiment. PMD has been demonstrated to occur in case of cytokine secretion 23, 24, but the molecular mechanisms underlying PMD are largely unknown. In particular, very little is known about what governs the cell decision to opt for either release of entire granules or PMD, and the precise molecular mechanisms that regulate mobilization of vesicle-associated secretory Akt inhibitor aliquots in a PMD manner. In light of these results, it can be speculated that the lowered availability of cytosolic Ca2+ in activated MCs interacting with Tregs could be responsible for unsuccessful exocytosis but could be enough for promoting PMD. This could explain the selective inhibitory effect of Tregs on the secretion of pre-stored and usually early released mediators and the delay of TNF-α release observed at early time point. In conclusion, this study describes the dynamic and functional profile

of MC–Treg interactions. This cross-talk is not restricted to BMMCs but is a common feature of mature MCs and human MCs. Importantly, Ku-0059436 supplier Acetophenone we found that this cross-talk is regulated on a single-cell level also providing the first morphological evidence for a role of the OX40–OX40L axis in Treg inhibition of MC function. However, the dynamics of Treg–MC conjugates reflects a complex synaptic structure and a more detailed analysis is necessary to understand the molecular composition of this interaction. Moreover, the evidence of PMD in MCs interacting with Tregs underlines the necessity to understand all events and mechanisms governing differential sorting, packing and

secretion of granule-stored mediators. Our findings pave the road to identify selective secretory pathways that are still partially unknown and might regulate MC degranulation without modifying their innate immune functions. C57BL/6 mice were purchased from Harlan (Harlan Italy), C57BL/6 OX40-deficient mice were kindly provided by M. Colombo in Milan, Italy. CD4+CD25+ cells were purified using the CD25+ T cell isolation kit (Miltenyi Biotec, Bergisch Gladbach, Germany) according to the manufacturer’s instructions. By flow cytometry analysis, cells were more than 90% Foxp3+. BMMCs were obtained by in vitro differentiation of BM cells taken from mouse femur as described 4. After 5 wk, BMMCs were monitored for c-kit and FcεRI expression by flow cytometry. Purity was usually more than 97%.

From these observations, they concluded that the cloned lipase was lipase/phospholipase A1. We purified the lipase and identified that it possesses the ability to degrade tributyrin and to cleave pNp-fatty acyl esters. Because our purified lipase resembled lipase/phospholipase A1, which had been reported by Merino et al. (11), we considered that it is lipase/phospholipase A1 of A. sobria. We measured the phospholipase A1 activity of our sample using an EnzCheck phospholipase A1 assay kit (Invitrogen; Carlsbad, CA, USA), and found that the purified lipase has significant activity (data not shown). Thus, this website we confirmed that this lipase possesses phospholipase A1 activity. Subsequently we tried

to detect the ability of purified lipase to hydrolyze PC to LPC or LPC to GPC. We measured check details the activity by the hydroxamate method using commercially available egg-yolk lecithin as a substrate (29). In this method, the amount of fatty acid ester residues of the phospholipids is measured. Contrary

to our expectation, the purified lipase did not hydrolyze the substrate (egg-yolk lecithin) under the conditions used (data not shown). We postulate that the lipase did not hydrolyze the egg-yolk lecithin because the sensitivity of the lipase to egg-yolk lecithin is very low. Actually, the lipase does hydrolyze pNp-fatty acyl esters, however, its RNA Synthesis inhibitor efficacy in cleaving esters containing long-chain fatty acids is low (Fig. 4). It is possible that the lipase hydrolyzes lipids containing short-chain fatty acids, such as the substrate in the assay kit, but has low activity when it comes to hydrolyzing lipids containing long-chain fatty acids. Further studies on the reactions of the lipase with various substrates are needed to clarify its characteristics. It has been reported that the hemolytic, cytotoxic, and enterotoxic activities of A. hydrophila AH-3 are not reduced by destruction of the gene for lipase/phospholipase A1, suggesting that it is not involved in these pathogenic activities (11).

We also examined the purified lipase for cytotoxic effects on cultured cells (HeLa cells) and found that it had none (data not shown), supporting that the lipase is not involved in A. sobria’s cytotoxicity under the conditions used. Generally, phospholipase A produced by bacteria does not show severe cytotoxicity. However, in the presence of phospholipids, some lipase/phospholipase A1s such as the phospholipase A of Serratia marcescens show cytotoxicity (30). It has been considered that the lysophospholipids produced by these lipases affect cell membranes, resulting in cell lysis. Therefore, the lipase might show cytotoxic effects in the presence of phospholipids. That is, lipase produced in vivo might immediately react with phospholipids in the milieu of the bacteria and produce lysophospholipids.

Controls were 115 voluntary healthy bone marrow donors recruited at the blood bank of the Service of Immunology at the Hospital de Clínicas de Porto Alegre, most of them resident in the urban area of Porto Alegre/RS (83 women and 32 men; 86·1% European descendents and 13·9% African descendents). Individuals presenting chronic or acute diseases were excluded from the sample, as well as those presenting family history of genetic diseases (X-linked, autosomal or chromosomal abnormalities). 3-MA in vitro Amerindians and subjects

with Asiatic origin were not included. All patients were interviewed and examined according to an extensive questionnaire directed to the evaluation of end-organ damage [14]. Disease subtype was classified as follows: diffuse cutaneous SSc (truncal and acral skin tautness), limited cutaneous SSc (skin tautness restricted to extremities

and/or face) and limited SSc (sine scleroderma) [13,15]. Clinical characteristics of the disease were observed and recorded as described previously [14]. Blood samples were collected for serology [anti-nuclear antibodies (ANA), anti-centromere and anti-topoisomerase I antibodies] and DNA extraction. Pulmonary high-resolution computed tomography (HRCT) was performed in most patients. Doppler echocardiography was used to estimate the pulmonary systolic arterial pressure (PSAP), and patients with PSAP ≥ 40 mmHg were considered to have pulmonary arterial hypertension. This study was approved by the Research Ethics Board of Hospital de Clínicas de Porto Alegre (IRB0000921). www.selleckchem.com/products/OSI-906.html Farnesyltransferase All patients and controls signed a written informed consent before participating in this study. DNA was extracted from blood buffy coat using a modified salting-out technique, as described by Miller SA et al.[16]. Fifteen KIR genes (2DS1, 2DS2, 2DS3, 2DS4, 2DS5, 3DS1, 2DL1, 2DL2,

2DL3, 2DL4, 2DL5, 3DL1, 3DL2, 3DL3 and 2DP1) were typed in patients and controls using a polymerase chain reaction with sequence specific primers (PCR–SSP) method, as described by Gomez-Lozano et al.[17]. For the PCR, 10 ng DNA, 50 mM MgCl2, 1 µl PCR buffer, 25 mM deoxyribonucleoside triphosphates (dNTPs), 500 nM primers, 100 nm internal control and 2·5 units of Taq polymerase were mixed in a total volume of 10 µl [internal control primers amplify a 796 base pairs (bp) fragment of the third intron of human leucocyte antigen (HLA) DRB1]. PCR products were amplified by the GeneAmp PCR system 9600 (Perkin-Elmer, Norwalk, CA, USA), with denaturation for 3 min at 94°C, followed by four cycles of 15 s at 94°C, 15 s at 65°C, 15 s at 72°C; 21 cycles of 15 s at 94°C, 15 s at 60°C, 30 s at 72°C; five cycles of 15 s at 94°C, 1 min at 55°C, 2 min at 72°C; and a final elongation step at 72°C for 7 min. The PCR products were analysed on 1% agarose gel after electrophoresis.

casei showed a similar pattern of these Th1 cytokines and would have an influence on the results when associated with the vaccine. IL-2 would exert a strong influence

on the proliferative capacity and maintenance of memory T cells [40], which would be a desirable characteristic in the selection of an efficacious long-term vaccine. Some lactobacilli used as adjuvants in vaccination protocols increased systemic protection through an increase in the Th1 response [19]. In addition, an immune response based on the Th1 population participates actively in the resolution of S. pneumoniae infection in humans [41]. Considering our results, the probiotic strain would exert an immunostimulatory effect on the Th1 cells and on the release of their cytokines in the lung. On the other hand, regulation of the inflammatory response is most important in infectious diseases. In this sense, the probiotic administered by the oral and nasal routes was able to increase Epacadostat in vivo the regulatory Th2 Z-VAD-FMK clinical trial IL-10 cytokine. This would be of great importance to ensure a balanced immune response that would enable resolution of the infectious process, limiting a possible exacerbated inflammatory response and avoiding damage to the host’s tissues. The greatest IL-10 production was obtained on day 42 in the

groups that received the live and inactivated vaccine associated with orally administered L. casei. In contrast, the nasal administration of Lc and D-LL + Lc induced an IFN-γ/IL-10 ratio > 1, which could have negative implications for the host after infection if the Th1 response was exacerbated. However, other factors must be considered. Thus, recent works have associated IL-17 with stimulation in the production of chemokines capable of recruiting IFN-γ-producing CD4+ T cells [42,43]. In addition, IL-17 and IL-22 produced by Th17 induce the attraction of neutrophils and macrophages into the parenchymal tissue, favouring pathogen clearance [44]. It was also demonstrated that this cytokine, being a key factor in the adaptive

immunity against the above pathogen, would mediate the death of pneumococci in the presence or absence of specific antibodies [45]. Moreover, using knock-out Thiamine-diphosphate kinase mice, IL-17 was shown to be of fundamental importance to reduce nasal colonization by S. pneumoniae. Oral and nasal administration of L. casei in association with LL vaccination induced the highest IL-17 levels. It also increased IL-2 and IFN-γ cytokine levels and afforded full protection against pneumoccocal challenge. In contrast, the dead vaccine failed to prevent pneumococcal colonization by both serotypes 3 and 14 of the pathogen, although it induced high IL-17 and Th1 cytokine levels, indicating the complexity of the protective response. On the other hand, it should be pointed out that too-high levels of IL-17 could be associated with autoimmunity [44], so that a balanced response is desirable after vaccination.

Conversely, an increase in Bim could have interesting consequences. Activation of Bim-mediated lymphocyte killing upon pro-apoptotic BH3-mimetics could adjust the balance between activated and regulatory lymphocyte populations and ameliorate colitis. Inducing apoptosis of autoreactive lymphocytes could be a new promising therapeutic

strategy for CD patients. This work was supported by the Swiss National Foundation (M.H., 31003A_127247) and the Broad Medical Research Program (M.H., IBD-0324R). We thank the microscopy centre at the University of Zurich (ZMB) for technical assistance. K.L., M.K., M.F. and M.H. have no conflicts Panobinostat mw of interest to disclose. G.R. discloses grant support from Abbot, Ardeypharm, Essex, FALK, Flamentera, Novartis, Roche, Tillots, UCB and Zeller.

“
“The adenosine A2A receptor (A2AR) is the major cellular adenosine receptor commonly associated with immunosuppression. Here, we investigated whether A2AR activation holds the potential for impacting the severity of experimental autoimmune myasthenia gravis (EAMG) induced following immunization of Lewis rats with the acetylcholine receptor (AChR) R97–116 peptide. This Kinase Inhibitor Library report demonstrates reduced A2AR expression by both T cells and B cells residing in spleen and lymph nodes following EAMG induction. A2AR stimulation inhibited anti-AChR antibody production and proliferation of AChR-specific lymphocytes in vitro. Inhibition was blocked with the A2AR antagonists or protein kinase A inhibitor. We also determined that the development of EAMG was accompanied by a T-helper cell imbalance that could be restored following A2AR stimulation that resulted in increased Treg cell levels and a reduction in Th1-, Th2-, and Th17-cell subtypes. An EAMG-preventive treatment regimen was established that consisted of (2-(p-(2-carbonylethyl)phenylethylamino)-5-N-ethylcarboxamidoadenosine) (CGS21680; A2AR agonist) administration 1 day prior to EAMG induction. Administration

of CGS21680 3-oxoacyl-(acyl-carrier-protein) reductase 29 days post EAMG induction (therapeutic treatment) also ameliorated disease severity. We conclude that A2AR agonists may represent a new class of compounds that can be developed for use in the treatment of myasthenia gravis or other T-cell- and B-cell-mediated autoimmune diseases. Myasthenia gravis (MG) is a B-cell-mediated, T-cell-dependent autoimmune disease characterized by excessive muscle weakness and fatigue [[1]]. The development of an autoimmune response to the neural acetylcholine receptor (nAChR) present at neuromuscular junctions leads to the production of function-blocking anti-nAChR antibodies and this results in symptoms characteristic to MG [[2, 3]].

Part of the work presented here was supported by a grant from the University of Saarland Medical Faculty (Homfor, to I.J.) and from the German Federal Ministry for Education and Science (BMBF) (grant #01 KI 07103 Skin Staph to M.H.). “
“Chaperone production is an essential step for proper folding of certain proteins. Accumulation of misfolded/unfolded proteins within the endoplasmic reticulum (ER) lumen triggers a signalling pathway named unfolded protein response (UPR). Upon activation,

the UPR pathway Dasatinib cost augments transcription of ER chaperones increasing protein folding, decreases protein translation to ameliorate the ER overload, increases protein degradation, and activates the apoptotic programme if all previous measures fail. In this review, we will cover the chaperones involved in folding of proteins related to the immune response, followed by an overview of the UPR pathway. Lastly, we will discuss data from this last decade that demonstrate how the improper 3-deazaneplanocin A activation of the UPR pathway has been uncovered as a mechanism responsible for failure to mount a proper immune response, both innate

and adaptive. The accumulation of unfolded/misfolded proteins within the endoplasmic reticulum (ER), followed by inability of the cell to cope with this excessive protein load defines the ER stress. The unfolded protein response (UPR) corresponds to the signalling pathway that cells have evolved in order to trigger those mechanisms that aim at properly folding and exporting intra-luminal protein load. This aim is achieved by means of (1) induction of molecular chaperones to increase the rate of protein folding; (2) attenuation of protein translation; (3) increased degradation of misfolded proteins; and ultimately, when all previous fail; (4) activation of apoptotic pathways for cell termination. In this review, we will cover some aspects of immunity-related proteins folding, followed by an overview of the activation and regulation steps of the UPR pathway. Lastly, we will describe the scenarios known so far in which improper protein folding was uncovered as a mechanism responsible for failure of a proper immune response. Pyruvate dehydrogenase This review will focus specifically in immune responses from the innate

system and B cells, so far characterized as being most sensitive to failure of activation of the UPR pathway. Chaperone production is an essential step for proper folding of certain proteins. ER chaperones bind to unfolded polypeptide chains while they are being synthesized, preventing them from aggregating and becoming non-functional. Some chaperones are important for proper assembly of macromolecular structures, as immunoglobulins, for example. Chaperones assist the folding and assembly of several macromolecules but are not components of the final structure. Chaperones are divided into three groups: chaperones of the heat-shock family, chaperone lectins, and substrate-specific chaperones (for an extensive review on chaperone classification see [1]).

The emerging literature in several areas points to complex interactions between exposure to microbial pathogens and susceptibility to the induction and expression of allergic diseases exemplified by atopic asthma. Earlier notions that infections protect against allergy by enhancing Th1-associated immunity have been supplanted by a broader understanding of the relevance of issues, such as timing, type and intensity of infections, to the underlying disease process. Many issues Bafilomycin A1 remain to be resolved, and this will remain a ‘hot topic’ in the allergy field for some time

to come. The authors have no conflicts of interest to declare. “
“Immunoglobulin (Ig) replacement therapy has substantially changed the life of patients with primary antibody deficiency (PAD). In the majority of cases, patients with common variable immunodeficiency (CVID) or X-linked agammaglobulinaemia (XLA) now live to lead a near-normal life. Modern Smoothened Agonist cell line production facilities, a series of safety measures and a choice of several ways of

administration make Ig replacement a safe and relatively easy therapy to use. The well-known presentations of PAD, such as pneumonia, septicaemia and other invasive bacterial infections [1], would continue to occur in PAD patients without regular replacement therapy. In this paper, we comment on the success and limitations of our present Ig replacement therapy in PAD. We also speculate how further improvement can be achieved in the treatment of complications from which PAD patients continue to suffer. IgG replacement effectively prevents pneumonia and invasive bacterial infections, as shown in several large cohorts.

For instance, in a large Italian cohort of CVID patients, the prevalence of pneumonia was reduced from 49·0 to 20·5% upon initiation of Ig therapy [2]. Prevention of pneumonia by Ig replacement therapy appears to be possible in a dose-dependent fashion. In a meta-analysis on IgG trough levels of 676 patients, the risk of pneumonia declined by 27% with each 0·1 g/kg body weight increment in the monthly IgG dose [3], although other factors, such as individual IgA levels, may determine the risk of pneumonia even more (-)-p-Bromotetramisole Oxalate strongly [4]. However, the effect of IgG replacement therapy on bacterial bronchitis and sinusitis in PAD patients is less clear. In the Italian CVID cohort, prevalence of chronic bacterial airway infections rose markedly from time at diagnosis through an observation period of a mean of 11 years of performed IgG replacement therapy. Frequency of both chronic bronchitis and sinusitis increased from 33·9 to 46·4% and from 36·6 to 54·0%, respectively [2]. The increase of these conditions during Ig therapy was described similarly in XLA patients [1, 4]. Chronic bronchitis and sinusitis in PAD is due almost exclusively to chronic bacterial infection.

, 1999) but may also be suspended in host material as seen in many chronic infections (Burmølle et al., 2010). Microbiologists have up until the last few decades focused and emphasized the planktonic state over the biofilm state. However, the importance of the biofilm mode of growth is becoming increasingly

recognized as improved methods to study sessile bacteria have become available, and hence the subsequent accumulation of evidence for its widespread presence. It has been suggested that bacteria are predominantly growing as sessile communities rather than as single cells (Costerton et al., 1987; Davey & O’Toole, 2000). Sessile growing bacteria are defined as an assemblage of cells embedded ‘in a self-produced polymeric matrix’. This matrix is Sunitinib mouse very important for the properties of the biofilm, because it offers structural stability and increased tolerance to antimicrobials and immune cells (Stoodley Sorafenib cost et al., 2002; Anderson & O’Toole, 2008; Mulcahy et al., 2008; Ma et al., 2009). To gain further information on this phenomenon, one has to investigate how a biofilm is established and propagated. The most

common method is the continuous-culture once-through flow system using the model organism Pseudomonas aeruginosa. In this system, media are slowly passed over the biofilm-growing bacteria, which have attached to a cover slip on a flow cell. This in vitro process of P. aeruginosa biofilm formation can be divided into at least five stages: in the first stage, planktonic cells reversibly attach to a vacant surface. Irreversible binding follows this attachment and then multiplication into microcolonies. The microcolonies produce an extracellular polymeric matrix, which in turn envelopes the colonies. After a couple of days, the microcolonies Interleukin-3 receptor attain tower- or mushroom-like structures measuring up to 50 μm in the flow cell (Costerton et al., 1995; Davey & O’Toole, 2000;

Stoodley et al., 2002). The extracellular matrix contains a mixture of polysaccharides, proteins, and DNA (Wingender et al., 2001; Whitchurch et al., 2002; Costerton et al., 2003). When the biofilm grows to a size not beneficial for bacterial survival and growth (e.g., owing to nutrient limitations), focal areas of the biofilm are sloughed off. It is hypothesized this enables the otherwise sessile biofilm bacteria to spread and colonize new surfaces and biofilms to spread. Hence, it seems that the biofilm lifecycle by P. aeruginosa is a dynamic process capable of renewing itself (Costerton et al., 1995; Davey & O’Toole, 2000; Stoodley et al., 2002). The biofilm lifecycle and the matrix components have preferably been investigated by means of confocal laser scanning microscopy (CLSM). This method has provided valuable insight into the biofilm development; however, the information on the detailed ultrastructure of the biofilm is difficult to image by light microscopes.

Supernatants of NK cells (1×106/mL) incubated in the presence of 1 μg of HPV16-VLPs or with positive and negative controls were stored at −80°C and were analyzed for TNF-α and IFN-γ production in a specific capture ELISA according to the manufacturer’s instructions (BioSource, Merelbeke, Belgium). NK cells (2×105/200 μL) were incubated with 10 μg of CFSE-VLPs or LYNX-VLPs in complete RPMI for 1 h at 4°C (binding step). After washing, cells were placed at 37°C for different incubation times. For the experiments investigating the entry pathway,

different reagents (Sigma) were added 1 h before the DMXAA ic50 incubation with labeled VLPs: 2 μM of cytochalasin D, 25 μg/mL of chlorpromazine for the clathrin-dependent pathway, 25 μg/mL of nystatin for the caveolin-dependent pathway, or 1 U/mL heparinase Selleckchem GDC-0068 II. For blocking experiments with anti-CD16 mAb (BD Biosciences, 1 μg/mL), cells were incubated with this antibody for 40 min before the addition of labeled VLPs. After incubation with fluorescent VLPs, 0.5×106 cells were incubated for 1 h with Hoechst 33342 DNA stain (10 μM, Acros Organics, Geel, Belgium). After washing, cells were deposited on a polylysine-coated coverslip, fixed with PAF (4%) and cell membranes were stained with phalloidin 633 (Invitrogen) for 45 min at room temperature in the dark. Then, coverslips were fixed with 20 μL of Mowiol (Hoechst GmbH, Frankfurt, Germany). Images were acquired using an

Olympus Fluoview FV1000 confocal system (Olympus, Aartselaar, Belgium) equipped with an Olympus IX81 inverted microscope (objective UPLSAPO 60X/NA 1.35). To check VLP conformation, 10 μL of each VLP pool were deposited on copper grids coated with a carbon film (EMS, Abiraterone datasheet Brussels, Belgium). Grids were stained twice with 2% uranyl acetate (Fluka, Bornem, Belgium) for 1 min and washed with filtered demineralized water. To analyze VLP entry, NK cells were incubated with VLPs as described above for 10 min to 18 h. Cells were then centrifuged, fixed at room temperature in 4% glutaraldehyde (Laborimpex, Brussels,

Belgium) and post-fixed in 1% osmium tetroxide (Laborimpex) for 1 h at 4°C. Pellets were dehydrated with ethanol solutions (VWR International, Leuven, Belgium) and embedded in Epon (Serva, Breda, The Netherlands) and propylene oxide (Laborimpex) at 60°C. Ultrathin sections were stained with uranyl acetate (Fluka) and lead citrate (Leica, Groot Bijgaarden, Belgium). Grids were examined using a transmission electron microscope EM Jeol 100 CX II (Jeol, Zaventem, Belgium). A fluid-uptake assay with FITC-dextran was performed on cells (2×105/200 μL) incubated in the presence of HPV16–VLPs (10μg/mL) or positive and negative controls in complete RPMI containing FITC-dextran (1 mg/mL, average mol wt. 20 000, Sigma) at 37°C. After incubation, cells were washed three times and fixed (PAF 1%). Cells treated with 2 μM of cytochalasin D (Sigma) for 30 min before the incubation of VLPs were also used as controls.

Results were entered on a computerised database and discussed at a multi-disciplinary meeting on a fortnightly basis. Methods: This was an observational retrospective cohort study of patients aged 18 years and above, who had been on haemodialysis for at least 1.5 years before September, 2010. Targets monitored included Haemoglobin, Ferritin, Transferrin saturation, Calcium, Phosphate, Calcium Phosphate product, PTH, kt/V and Urea Reduction Ratio (URR). Values achieved for each parameter, before and after commencement of this periodic review system were compared for each patient. Results: More values were within the

targeted range for Transferrin saturation, Ferritin, Phosphate, Calcium Phosphate product, kt/V and URR although statistical significance was observed only with Transferrin saturation and Phosphate. Values for Haemoglobin, Calcium and

PTH were less likely to be within the target range however this was check details not statistically significant. Conclusions: A systematic periodical review system of haematological and biochemical results is helpful in attaining targets in patients on haemodialysis as opposed to standard review of results on routine clinical visits. 233 VARIABILITY IN THE MANAGEMENT OF LITHIUM POISONING DM ROBERTS1,2, S GOSSELIN3,4 1Addenbrooke’s Hospital, Cambridge, UK; 2University of Queensland, Brisbane, Australia; 3McGill University Health Centre, Montreal; 4Centre Antipoison du Quebec, Quebec City, Canada Aim: To assess decision-making by clinical toxicologists, including the role of Lenvatinib manufacturer extracorporeal treatment, in the treatment of lithium poisoning. Background: Three patterns of lithium poisoning are recognized: acute, acute-on-chronic, and chronic. Intravenous fluids with or without an extracorporeal treatment are the mainstay of treatment and their respective roles may differ depending on the mode of poisoning being treated. Existing

tuclazepam recommendations for treatment are based on a small observational studies and their uptake by clinicians is not known. Methods: Four case presentations of lithium poisoning were presented in a stepwise manner to experts in clinical toxicology who were attending a workshop at a meeting in Europe. Opinions on the treatment of these cases were determined anonymously using a hand-held audience response system, and a frequency evaluation was performed. Results: 163 health professionals, mostly physicians and poison information specialists, from 33 countries participated. Variability in treatment decisions was evident, in addition to discordance with published recommendations. Participants did not consistently indicate that haemodialysis was the first-line treatment, instead opting for a conservative approach. Continuous modalities were considered favourably, being selected in approximately 30% of cases where an extracorporeal therapy was recommended.