To assess the number of intracellular bacteria, plates were washed

and then incubated for another 60 min in a fresh medium. Then, extracellular bacteria were killed by incubation with a medium containing gentamicin IGF-1R inhibitor (100 μg mL−1) for 30 min. After washes with warm PBS, the cells were lysed and lysates were plated as above. Bacterial recovery was determined after an overnight incubation. The invasion rate was determined as the relation of intracellular bacteria to the total count from the same experiment. To determine the possible influence of ARA290 on cell proliferation and viability, the XTT assay was used (Sigma-Aldrich, St. Louis, MO). Cells were grown in 96-well plates (Costar) until reaching confluence and stimulated for 24 h as described above. Cells incubated in medium alone served as controls. Triplicates were

analyzed for each condition. After 24 h, cells were washed three times in PBS and incubated for 4 h with 250 μL freshly prepared XTT–menadione solution (1 mg mL−1 and 12.5 μM, respectively) at 37 °C. The formazan concentration was then measured at 490 nm. For immunoprecipitation, cells were seeded in six-well plates (Costar). After reaching confluence, the cells were stimulated and infected as described for cell infection assays. After centrifugation at 300 g for 5 min, cells were incubated for further 5, 15 or 25 min at 37 °C or collected directly. Cells were washed with ice-cold PBS, lysed with lysis buffer [137 nM NaCl, 1% IGEPAL CA-630, 20 mM Tris IWR 1 (pH 8.0), 200 μM phenylmethylsulfonyl fluoride, 10% glycerol, complete protease inhibitor (1 : 100, Sigma-Aldrich), phosphatase inhibitor cocktail (1 : 100, Sigma-Aldrich)] and cleared by

centrifugation for 20 min at 10 000 g and 4 °C. The protein concentration in the lysates was measured using BCA Protein Assay reagent (Pierce, Thermo Scientific, Rockford, IL) and samples were adjusted to equal protein concentrations. Lysates were then incubated for 1 h at room temperature with Protein G-coated Sclareol beads (Dynabeads Protein G; Dynal, Oslo, Norway) to remove unspecifically bound proteins. Cleared lysate was incubated with goat anti-focal adhesion kinase (anti-FAK) antibody A-17 (Santa Cruz Biotechnology, Santa Cruz, CA) overnight at 4 °C. The FAK–antibody complex was then precipitated with Protein G-coated beads for 1 h at room temperature. After three washes with PBS, collected proteins were eluted from the beads by heating the samples in sodium dodecyl sulfate (SDS) sample buffer (Bio-Rad Laboratories, Hercules, CA) supplemented with 0.5%β-mercaptoethanol at 95 °C for 5 min. Proteins were subjected to SDS-polyacrylamide gel electrophoresis on a 10% polyacrylamide gel (Tris-HCl Ready Gel Precast Gel, Bio-Rad Laboratories) and transferred to a polyvinylidene fluoride membrane (Invitrogen, Carlsbad, CA). The membrane was blocked with 5% milk in 0.

All the Fabs kept their peptide-specific, MHC-restricted binding to the MOG-35-55 loaded empty RTL302-5D (Fig. 3B), excluding any binding dependence to non-native sequences of RTL1000. Additionally, we tested Fab binding to RTL1000 in different buffer conditions and found the Fabs to be conformationally sensitive, losing their ability to react with denatured RTL1000 (Supporting Information Fig. 1). Taken together, these data indicate selective Fab binding to the α1β1 DR2–MOG-35-55 native sequence of the folded RTL1000. We next tested the ability of the anti-RTL1000 Fabs to Carfilzomib bind the native full-length four-domain form of MHC-II complexes as expressed on APCs. L-cell DR*1501 transfectants

(L466.1 cells) were loaded with MOG-35-55 or control peptide. The loaded cells were incubated with the purified Fabs following anti-Fab-FITC incubation. As shown in Fig. 4A, no specific binding of Fabs was observed for MOG-35-55 loaded cells. MOG-35-55 and control-peptide loaded cells produced the same fluorescence

intensity as background. MHC expression on the APC surface was confirmed by anti-DR mAb (L243). A portion of the loaded cells that were used for the FACS analysis was incubated with the H2-1 T-cell hybridoma specific for the DR2–MOG-35-55 complex. Following 72 h incubation, cell supernatants were transferred to IL-2-dependent CTLL cells for detection of IL-2 levels secreted from the H2-1 hybridoma (Fig. 4B). H2-1 cells were activated Osimertinib cell line only by the MOG-35-55 pulsed cells, secreting eightfold higher levels of IL-2 compared to non-pulsed or control peptide-pulsed APCs. Peptide-specific H2-1 activation confirmed a successful loading of MOG-35-55 peptide to the native MHC on the APCs used for the FACS analysis. Despite the presence of a biologically active determinant in the form of DR2–MOG-35-55

molecules presented by the APCs, no staining of such a complex was obtained by any of our anti RTL1000 Fabs. Considering the high affinity of the selected Fabs and the permissive conditions used for this experiment, we conclude that the Fabs do not bind the native DR2–MOG-35-55 complex presented by APCs. Further support for this finding came from blocking experiments which tested the Fabs ability to inhibit peptide-specific activation of the H2-1 hybridoma by DR2 filipin APCs pulsed with MOG-35-55 peptide (Fig. 4C). None of our selected Fabs were able to block this peptide-specific, MHC-restricted activation, as compared to a control TCRL Fab (D2) specific for RTL2010 (DR4–GAD-555-567) that also failed to block H2-1 activation. In contrast, complete blocking was achieved by the control anti-MHC-II mAb (TU39). The failure of the Fabs to interfere with MHC presentation to TCR implies an inability to bind native four domain DR2–MOG-35-55 complexes. This was indeed the case, as demonstrated by ELISA (Fig. 4D).

Methods: From 1997 to 2010, a total of 1605 women with bothersome LUTS received video-urodynamic study in our unit. We reviewed the charts of 212 women diagnosed with BOO based on video-urodynamic criteria and 264 women without abnormal findings. LUTS and urodynamic parameters were compared Panobinostat mw between obstructed and unobstructed cases and among the BOO subgroups. Results: The mean ages of the BOO (58.2 years) and control groups (58.8 years) were

similar. The mean values of detrusor pressure at maximum urinary flow rate (PdetQmax)/maximum flow rate (Qmax) of the BOO and control groups were 51.83 cm H2O/10.22 mL/s versus 18.81 cm H2O/20.52 mL/s. In the BOO group, cinefluoroscopy revealed dysfunctional voiding in 168 patients (79.2%), urethral stricture in 17 (8.0%), and bladder neck dysfunction in 27 (12.7%). Patients with dysfunctional voiding had significantly lower urethral resistance compared with the other two BOO subgroups. Combined lower urinary tract symptoms were present most often in all BOO patients (69.3%), followed by isolated storage symptoms (30.2%) and isolated voiding symptoms (0.5%). Seventy-seven patients (37.3%) had Kinase Inhibitor Library dysuria and 79 patients (36.3%) had frequency as their main symptom. Conclusion: Women with BOO usually

have nonspecific LUTS. Dysfunctional voiding was the most common form among women with clinically unsuspected BOO, but the degree of obstruction was less severe than with primary bladder neck obstruction and urethral stricture. “
“Objectives: We evaluated the association of lower urinary tract symptoms (LUTS) and sleep disorders (SD) in patients with benign prostatic hyperplasia (BPH). We also examined improvement 3-oxoacyl-(acyl-carrier-protein) reductase of SD following the α1-blocker

therapy for LUTS. Methods: Sixty-eight male patients were enrolled in the study, consisting of 38 cases with LUTS and BPH (BPH group), and 30 men without significant LUTS or BPH (non-BPH group). The degree of LUTS and SD was evaluated by the International Prostate Symptom Score and the Pittsburg Sleep Quality Index (PSQI), respectively. The patients of BPH group then were treated with α1-blocker for 4 weeks, and were re-examined by all the questionnaires to evaluate the therapeutic efficacies. Results: The correlation analyses showed a significant association of LUTS with SD in BPH group (r = 0.4995, P = 0.0068). Twenty cases (52.6%) in BPH group showed 5.5 or more PSQI scores. Following 4 weeks of α1-blocker administration, the average PSQI decreased significantly from 6.3 to 4.8 points (P < 0.001). Significant improvement was observed in domains of “sleep quality” and “sleep disturbances” among PSQI (P = 0.0215 and 0.0391, respectively). Moreover, significant association between α1-blocker induced improvements of nocturia and SD was identified in patients with 5.5 or more PSQI score at baseline (r = 0.445, P = 0.0334).

gondii infection. Therefore, this https://www.selleckchem.com/products/LBH-589.html disparity led to an increased Tact cell elimination by the mAb in B6 mice (67%), whereas in BALB/c animals, the same treatment led to the elimination of 45.3% of Tact. Because CD25 expression is not restricted to Tregs or Tact, we analyzed CD8+, CD19+ and natural killer

(NK) cells, which are also activated during T. gondii infection and could be eliminated after depletion. As can be observed in Fig. 3, in uninfected animals from both strains, the proportion of these activated subsets was very low (<3.6%), and after depletion, a slight nonsignificant reduction was detected. At the time point of infection analyzed, the proportion of these activated populations was dramatically increased in B6, but not in BALB/c mice (Fig. 3), a pattern similar to that observed in the CD4+ subset (Fig. 1). Despite the slight increase of activated CD8+, CD19+ and NK cells in BALB/c mice after infection, treatment with PC61 before infection did not modify these proportions significantly (Fig. 3). However, INCB024360 depleted/infected B6 mice showed

a significantly reduced proportion of activated CD19+ and NK cells. Therefore, PC61 treatment before infection eliminates other activated cells, and the different pattern of depletion observed between strains is a consequence of the contrasting expansion and activation of effector cells. A summary of the effect of depletion on all cell types analyzed is shown in Table 1. Because of the potent immune response generated in B6 mice, the injection of PC61 mAb eliminates a very high proportion of most activated cell subtypes (up to 69%), but only low levels of Tregs (38.1%). Hence, it is impossible to analyze the role of Tregs in T. gondii-infected B6 animals using classical CD25 depletion experiments, and any interpretation drawn from this model, including mortality rates, could be more related to a role of activated cells than to the role of Tregs. Our results

agree with a previous report (Couper et al., 2009) next and extend the current knowledge on the effect of depletion in other cell types using an infectious model. Our results were obtained using a single low dose of mAb (200 μg); therefore, it is clear that repeated injections of the mAb or the use of higher concentrations are unnecessary and would lead to the complete elimination of all subtypes expressing CD25. Even though other activated cell subtypes are also eliminated in BALB/c mice using the same treatment, Tregs are the largest eliminated cell subtype in this strain. Thus, the results obtained by Tregs depletion with anti-CD25 mAbs could provide an insight into the role of Tregs during T. gondii infection only in the BALB/c strain. As a consequence of the contrasting immune response against the same pathogen generated by two mice strains of different haplotype, the depleted cell subtypes differ.

“
“Aim:  The aim of this study is to assess the characteristics of urinary system diseases and the role of the ultrasound screening and urinalysis screening for chronic kidney disease (CKD) in asymptomatic children in China. Methods: 

Between September 2008 and November 2008, 14 256 children excluding those with obvious symptoms and signs were enrolled in our study. All the subjects accepted ultrasound and urinary screening. A case–control study was performed to evaluate the relative risk of having stones in those children exposed to melamine formula. Results:  Of the enrolled children, 6.10% (869 of 14 256) showed abnormalities, of which 409 (2.87%) were established by ultrasound, 572 (4.01%) by urinalysis and 112 (0.79%) Pexidartinib ic50 by both ultrasound screening and urinalysis. The abnormalities included congenital anomalies of kidney and urinary tract, urinary stones and/or hydronephrosis, leucocyturia Alisertib purchase and haematuria and/or proteinuria. Children exposed to melamine formula were 5.17 times as likely to have kidney stones as children exposed to no-melamine formula (95% confidence interval, 3.28–8.14; P < 0.001); the probability of kidney stones in melamine-fed infants were 6.28 times

as likely as those no melamine-fed (95% confidence interval, 3.71–10.65; P < 0.001). Conclusion:  Ultrasonography and urinalysis could complement each other and play important roles in the early diagnosis of anomalies of the urinary system, but urinalysis is a more cost-effective screening tool for CKD in children in China. Exposure to melamine-contaminated formula associated with urinary stones, especially in infants, was significantly higher than the control group. "
“Aim:  The ankle brachial index (ABI) is a marker for peripheral artery disease and can predict mortality in advanced chronic kidney disease (CKD) and haemodialysis patients, respectively. However, it is CHIR-99021 chemical structure seldom studied in Taiwan, an area with high prevalence of CKD and end-stage renal disease. The aim of this study was to investigate the predictors for mortality by using ABI value in patients with CKD and undergoing haemodialysis in Taiwan. Methods:  One hundred and sixty-nine

patients with CKD stage 3–5 and 231 haemodialysis patients were enrolled in one regional hospital. The mean follow-up period was 23.3 ± 3.3 months. Patients were stratified into three groups according to ABI value (<0.9, ≥0.9 to <1.3, and ≥1.3). The relative mortality risk was analyzed by Cox-regression methods. Results:  In multivariate analysis, an ABI of 1.3 or more (hazard ratio, 3.846; P = 0.043) and coronary artery disease (P = 0.012) were positively associated with overall mortality, and serum low-density lipoprotein cholesterol level (P = 0.042) was negatively associated with overall mortality. In addition, an ABI of less than 0.9 (P = 0.049), an ABI of 1.3 or more (P = 0.033), coronary artery disease (P = 0.024) and haemodialysis treatment (P = 0.

The finding that VCAM-1+ stroma express 4–1BBL, CCL19, CXCL12, and IL-7 and that adoptively transferred CD8+ memory T cells are often found in

proximity to VCAM-1+CD45− cells in the BM demonstrates the plausibility of the VCAM-1+ stromal cell as Selleck Venetoclax the radioresistant cell that provides 4–1BBL to memory CD8+ T cells in the BM. These data support a model in which a radioresistant VCAM-1+ stromal cell attracts the VLA-4+ CD8+ memory T cells via CCL19, where they can receive 4–1BB-4–1BBL induced survival signals. As the VCAM-1-positive stromal population is very abundant in the BM, there may be heterogeneity in the VCAM-1+ stroma with respect to 4–1BBL, cytokines, and chemokines that contribute to CD8+ T-cell memory maintenance. Further analysis will be required to definitively identify the 4–1BBL-expressing radioresistant cell that contributes to CD8+ T-cell memory. C57BL/6 WT mice were obtained from Charles River Laboratories (St. Constant, QC, Canada).

4–1BB−/− mice [47] extensively backcrossed to the C57BL/6 (n = 10) background were bred in our facility. These mice were previously provided to us by Dr. Byoung S. Kwon (National Cancer Center, Ilsan, Korea). 4–1BBL-deficient (4–1BBL−/−) mice were originally obtained under a materials transfer agreement from Immunex (Amgen, Thousand Oaks, CA, USA) and further backcrossed to the C57BL/6 background in our facility (total n = 9). OT-I

and CD45.1 congenic mice were obtained from Jackson Laboratories (Bar Harbor, ME, USA) and crossed to buy AUY-922 generate CD45.1+/+ or CD45.1+/− OT-I mice. TCRα−/– mice were kindly provided by Dr. Cynthia Guidos (Hospital for Sick Children, Toronto). FoxP3gfp knock-in mice on the C57BL/6 background were kindly provided by Dr. Mohamed Oukka (Harvard Medical School) [48]. ACTB-DsRed transgenic mice expressing DsRed protein under control of the β-actin promoter and backcrossed to B6 mice for five generations (B6.Cg-Tg (ACTB-DsRed*MST) 1Nagy/J) were obtained from the Jackson laboratories and crossed with OT-I mice to obtain OT-I ACTB-DsRed mice (OT-I-DsRed). Mice were maintained under specific pathogen-free conditions in sterile microisolators at the University of Toronto. All mouse experiments were approved Sucrase by the University of Toronto animal care committee in accordance with the regulations of the Canadian Council on animal care (University of Toronto approved protocol #20007828). CD8+ T cells with a central memory phenotype were generated by culture with Ag followed by IL-15 using a variation of a previous protocol [7, 29]. In brief, OT-I splenocytes were stimulated with 0.1 μg/mL SIINFEKL peptide and 1 μg/mL of LPS for 1 day, and then the nonadherent cells were rested for 2 days in fresh media (RPMI-1640 with 10% heat-inactivated FCS, 0.03% L-glutamine, antibiotics, and 2-mercaptoethanol).

4).[3] Also, Weisholzer et al. in his study of 430 haemodialysis patients showed stroke rate was not statistically different in patients with and without atrial fibrillation when on no anti-thrombotic therapy (P = 0.22).[28] In this study, antithrombotic therapy with warfarin or salicylates was associated with a higher incidence of stroke (8.3/100 patient-years vs 2.6/100 patient-years; P = 0.0002).[28] An observational study on Dialysis Outcomes and Practice Patterns Study (DOPPS) data showed that use of warfarin was MK-1775 order associated with higher risk of stroke in patients with AF.[1]

This observation was perhaps due to confounding variables or inherent higher risk in these warfarin users or cause due to haemorrhagic stroke.[1]

Chan et al. study also showed that compared with non-use, warfarin use (44.7% of AF cohort) associated with a significantly increased risk for new stroke (hazard ratio (HR) 1.93; 95% confidence interval (CI) 1.29–2.90).[23] However, there XL765 in vitro were several limitations in this retrospective study, which makes it difficult to draw any firm conclusions. Most importantly, international normalization ratio (INR) monitoring was perhaps suboptimal in these studies that may lead to wrong interpretation. Platelet abnormalities including subnormal dense granule content Reduction in intracellular ADP and serotonin Impaired released of the platelet alpha granule protein and beta thromboglobulin Enhanced intracellular cAMP and abnormal mobilization of platelet calcium Abnormal platelet arachidonic acid metabolism Defective cyclo-oxygenase activity Abnormality of the activation-dependent binding activity of GPIIb/IIIa Increased formation of vascular (PG)12 Altered von Willebrand factor Indirectly Presence pentoxifylline of uraemic toxins, especially parathyroid hormone Anaemia/altered blood rheology Erythropoietin deficiency Specific drug treatment (e.g. non-steroidal anti-inflammatory drugs) Atherosclerosis and diffuse endothelia damage Dysfunctional activated

protein C metabolism Both elevated plasminogen activator inhibitor-1 to tissue type plasminogen activator ratios and inhibition of plasmin by increased levels of lipoprotein (a) Defects in the expression of glycoprotein GPIb (the receptor for von Willebrand factor) To the contrary, a recent large observational study showed that warfarin treatment in dialysis population was associated with a significantly decreased risk of stroke or systemic thromboembolism (HR 0.44; 95% CI 0.26–0.74; P = 0.002) but not with aspirin (HR 0.88; 95% CI 0.59–1.32; P = 0.54).[11] Studies in Table 5 were observational and heterogeneous so that the absolute risk of stroke could not be precisely determined.[1, 3, 7, 10, 20, 23, 28] As epidemiological analysis can identify only an association, causal relationships need to be shown by clinical trials. Hence, the results of epidemiological data analysis should be interpreted with caution.

Our analyses revealed five major

findings: (1) HII and CON show similar behavioral indices of memory as indexed by VPC novelty preference across three delays, (2) PSW responses were greatest over left scalp regions, (3) over temporal electrode sites HII infants show differential patterns of Nc responses to the three faces as compared to CON, (4) at temporal electrode sites, the PSW showed largest responses to the recent familiar face condition, and (5) in examining the relation between the VPC and ERP measures, CON showed a significant positive correlation between VPC novelty preference after a 24-h delay and PSW mean amplitude. The first two findings mentioned demonstrate click here the similarities found between infants who have experienced HII and typically developing infants in the present study. With https://www.selleckchem.com/products/byl719.html regard to the VPC task, both groups exhibit a VPC novelty preference only when tested immediately after familiarization but not after a 2-min or 24-h delay. This result is similar to the findings of Morgan and Hayne (2011), who used 3D pictures of cartoon-like faces, and also showed that 1-year-olds exhibited a VPC novelty preference immediately after familiarization but not after 24-h delay. Furthermore, they

found it was not until age 2 years when their participants exhibited novelty preference after 24-h delay; their study did not evaluate a 2-min delay. In contrast to our findings, studies on younger infants using slightly different testing methods than our own found novelty preference after varying time Inositol oxygenase delays. One study, which similarly used pictures of female faces but differed in their familiarization methods, found that 6-month-olds exhibited a novelty preference

after both a 2-min and 24-h delay (Pascalis et al., 1998). Another study, which used pictures of black-and-white sunburst and diamond patterns, found that 4-month-olds exhibited a novelty preference after a short delay lasting approximately the length of a feeding (Geva et al., 1999). It is difficult to compare these studies, as their VPC testing methods were slightly different from one another and from our own, but based on our study and that of Morgan and Hayne (2011), 12-months-old infants appear to demonstrate visual recognition memory retention on behavioral testing of less than 2 min. A second finding that showed no group differences was greater PSW mean amplitude over the left region. For the temporal electrode sites, this meant greater PSW over the left as compared to the right region, and for the frontocentral electrode sites, greater PSW over left as compared to right and middle regions. The regionalization of PSW to the left or right hemisphere has been under debate in prior studies.

Screening the diabetes population for DKD and intervening with ACE inhibitors and ARB as indicated, Pictilisib order together with appropriate glycaemic control and management of lifestyle-related risk factors, is a priority in responding to the health burden of diabetes

in Australia. The first priority in screening for DKD should be the detection of microalbuminuria Since the vast majority of DKD is associated with the presence of albuminuria, testing for microalbuminuria is key to screening strategies for the detection of DKD. Numerous studies have evaluated the cost-effectiveness of screening for albuminuria in the diabetes population, concluding that screening in diabetics based on dipstick urinalysis and/or measurement of urinary albumin to creatinine

ratio, followed by intervention with an ACE inhibitor or ARB, is cost-effective across all age groups.[33-35] Screening the diabetes population for DKD on the basis of eGFR has also been shown to be cost-effective,[36] although is most favourable above 50–60 years of age;[37] thus, these two markers potentially have complementary roles in screening different age groups.[38] The underlying burden of DKD will increase as long as diabetes prevalence is increasing, and this challenge must be met with lifestyle change The underlying burden of DKD in Australia is rising and will continue to do so as an inevitable AZD0530 result of increasing diabetes prevalence, driven by rates of obesity second and population aging. Therefore, averting the burden of DKD in Australia requires engagement with lifestyle change and healthy aging. A 2012 review from the American Heart Association of interventions to promote healthy lifestyles concluded

that, whereas interventions oriented around the individual were unlikely to have significant impact, population-based multicomponent interventions involving government mandated economic incentives and changes to the physical environment were able to effect change in lifestyle behaviours and health outcomes.[39] Nephrologists should consider themselves stakeholders in these types of population interventions for the primary prevention of diabetes and DKD. Health services planning requires accurate projections of the future burden of DKD and ESKD There is an urgent need to gather Australian data on longitudinal trends in the incidence and prevalence of diabetes and DKD, and more accurate information regarding attributable costs. Predicting future rates of DM-ESKD for the purposes of health services planning is complex and requires data on the current and future population at risk, longitudinal data on disease incidence trends and rates of progression, mortality data indicating trends in competing risks, and information on changing demographics of the diabetes population.

[Eur. J. Immunol. 2013. 43, 2126–2137] show that the NLRP3 inflammasome contributes to oxidative DNA damage. In addition, activation of the NLRP3 inflammasome modulates a number of pathways involved in DNA damage repair, cell cycle, and apoptosis, suggesting a novel role for the NLRP3 inflammasome in DNA damage responses following cellular stress. From microbes to radiation and other carcinogens, the environment in which we live can seem like a veritable minefield. Fortunately, the cells and molecules of the innate immune system have evolved, along with cell-intrinsic processes, to respond swiftly in defense of our cellular and genomic integrity. These multilayered and redundant mechanisms combat

the potentially deleterious effects of diverse environmental stresses by promoting either resolution or cell buy Selumetinib death in an attempt to return to homeostasis. An important component of the innate immune system is the NLRP3 inflammasome. Following detection of cellular damage,

the cytoplasmic nucleotide-binding domain leucine-rich repeat containing (NLR) molecule NLRP3 forms a multiprotein complex, along with the adaptor molecule ASC and the cysteine protease caspase-1 [1]. This process culminates in the activation of caspase-1 and the subsequent maturation and secretion of the proinflammatory cytokines, IL-1β and IL-18 [2-5]. Interestingly, oligomerization and activation of the NLRP3 inflammasome can be induced by a heterogeneous collection of pathogen- and damage-associated molecular patterns (PAMPs and DAMPs, respectively), although the means KPT-330 purchase by which this occurs is unclear. It has been proposed that these inflammasome activating signals actually

work indirectly via a common downstream ligand, such as reactive oxygen species (ROS) [6, 7] generated following mitochondrial damage [8, 9]. Cellular cation fluxes, including a potassium efflux and a calcium influx, have also been shown to be critical DNA ligase for activation of the NLRP3 inflammasome [10, 11]. In addition to its role in immune surveillance, dysregulation of the NLRP3 inflammasome has been reported to contribute to the pathogenesis of a number of human diseases that have an underlying component of chronic inflammation, such as type 2 diabetes mellitus, atherosclerosis, and inflammatory bowel disease [12]. As well, mutations within the gene encoding NLRP3 have been associated with the autoinflammatory cryopyrin-associated periodic syndromes [13]. Such widespread effects underscore the complexity of pathways through which the well-studied NLRP3 inflammasome functions, and emerging literature on the subject indicates there is much left to learn. In this issue of the European Journal of Immunology, Licandro et al. [14] explore noncanonical roles for the NLRP3 inflammasome, i.e. proinflammatory cytokine-independent effects under conditions of cellular stress.