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Unlike conventional servers housed in a centralized and secured indoor environment (e.g., data centers), Internet-of-Things (IoT) devices such as sensor/actuator are geographically distributed and may be closely located to the physical systems where IoT devices are utilized. However, the resource-constrained nature of IoT devices limits their capacity to deploy sophisticated security solutions. The proposed approach assumes that a device can be compromised and hence, the need to be able to automatically isolate the compromised device(s). In order to enforce security policies even when devices are compromised, we propose using blockchain in the monitoring framework. Unlike existing centralized or distributed security solutions (which do not consider the possibility that the solutions themselves can be compromised), the proposed blockchain-based framework can enforce the security policies as long as a majority of the devices are not compromised. By employing the permissioned blockchain (Hyperledger Fabric) and add-on hardware modules, the proposed framework offers significantly lower latency and overhead compared to permissionless blockchain frameworks (e.g., Ethereum) and allows existing IoT devices to join the framework without modification.

Patients with aspirin-exacerbated respiratory disease (AERD) regularly exhibit severe nasal polyposis. Studies suggest that chronic rhinosinusitis with nasal polyps (CRSwNP) is characterized by excessive fibrin deposition associated with a profound decrease in epithelial tissue plasminogen activator (tPA). Retinoids, including vitamin A and its active metabolite retinoic acid (RA), are necessary for maintaining epithelial function and well-known inducers of tPA in endothelial cells.This study sought to determine whether endogenous retinoids are involved in NP pathophysiology and disease severity in patients with CRSwNP and AERD.NP tissue was collected from patients with AERD or CRSwNP, and concentrations of retinoids and fibrinolysis markers were measured using ELISA. Normal human bronchial epithelial cells were stimulated alone or in combination with RA and IL-13 for 24 hours.This study observed lower retinoid levels in nasal polyps of patients with AERD than those with CRSwNP or healthy controls (P lt; .01). Levels of the fibrin-breakdown product d-dimer were the lowest in AERD polyps (P lt; .01), which is consistent with lower tPA expression (P lt; .01). In vitro, all-trans RA upregulated tPA levels in normal human bronchial epithelial cells by 15-fold and reversed the IL-13-induced attenuation of tPA expression in cultured cells (P lt; .01).RA, a potent inducer of epithelial tPA in vitro, is reduced in tissue from patients with AERD, a finding that may potentially contribute to decreased levels of tPA and fibrinolysis in AERD. RA can induce tPA in epithelial cells and can reverse IL-13-induced tPA suppression in vitro, suggesting the potential utility of RA in treating patients with CRSwNP and/or AERD.

This paper describes work in progress to analyze the cyber-physical resiliency of a microgrid when operated in islanded mode, as would be the case during an extreme event. The analysis must include each of the four components of a microgrid: sources or generation, energy storage, load, and power electronics interfaces. Two networks with disparate time constants must be accurately modeled. The electrical (and mechanical if there are generators or wind turbines) network is simulated with the power flow equations, a challenging control problem that requires accurate state estimation for its solution. In addition, there is the sensor-communication network consisting of legacy devices such as MV90 meters and SCADA hardware as well as IoT devices such as PMUs and grid management equipment.

We report that S100 proteins were reduced in patients with chronic rhinosinusitis (CRS). S100A8/9, which is important in epithelial barrier function, was particularly decreased in elderly patients with CRS. Epithelial expression of S100A8/9 is partly regulated by the IL-6 trans-signaling pathway. The goal of this study was to investigate whether or not age-related reduction of S100A8/9 in CRS is associated with blunting of IL-6 trans-signaling. The levels of IL-6, soluble IL-6 receptor (sIL-6R), soluble gp130 (sgp130), and S100A8/9 from control subjects (n = 10), and patients with CRS without nasal polyps (n = 13) and those with CRS with nasal polyps (CRSwNP) (n = 14), were measured by ELISA. Age-related differences in the level of each protein were investigated. Normal human bronchial epithelial cells were cultured in air-liquid interface and stimulated with IL-6/sIL-6R and tumor necrosis factor (TNF)-α with or without the addition of sgp130, a natural inhibitor of IL-6 trans-signaling. There was a significant age-related decline in S100A8/9 and an increase in sgp130 in nasal tissue samples from patients with CRSwNP, although there was no age-related difference in IL-6/sIL-6R production. Additionally, expression of the S100A8/9 gene and protein was increased significantly by IL-6/sIL-6R plus TNF-α in normal human bronchial epithelial cells. This increase was blocked by sgp130. These results suggest that increased sgp130 in older patients may inhibit IL-6 trans-signaling, impair barrier function, and decrease S1008/9 production in elderly patients with CRSwNP. Restoration of barrier function by targeting sgp130 may be a novel treatment strategy.

Objectives/Hypothesis Chronic Rhinosinusitis (CRS) is accompanied by evidence of a vigorous adaptive immune response, and emerging studies demonstrate that some nasal polyps manifest a polyclonal autoantibody response. We previously found that antibodies against BP180, a component of the hemidesmosome complex and the dominant epitope in autoimmune bullous pemphigoid, were found at elevated levels in nasal polyp tissue. Given the critical role of hemidesmosomes in maintaining epithelial integrity, we sought to investigate the distribution of BP180 in nasal tissue and evaluate for evidence of systemic autoimmunity against this antigen in CRS. Study Design Case-control experimental study. Methods The expression and distribution of BP180 in cultured nasal epithelial cells and normal nasal tissue were confirmed using real-time polymerase chain reaction (PCR), Western immunoblotting, immunofluorescence and immunohistochemistry. Sera were collected from three groups: control, CRSsNP, and CRSwNP. A commercially available ELISA was utilized to compare anti-BP180 autoantibody levels in sera. Results BP180 is expressed in nasal epithelium, but is not confined to the basement membrane as it is in human skin. In cultured nasal epithelial cells, confocal immunofluorescence showed a punctate distribution of BP180 along the basal surface, consistent with its distribution in epithelial keratinocytes. There are significantly higher levels of circulating nonpathologic anti-BP180 autoantibodies in CRS patients compared with normal controls (P

Interferon-γ (IFN-γ) production by natural killer (NK) cells and cytotoxic lymphocytes is a key component of innate and adaptive immune responses. Because inhibitor of κB-ζ (IκBζ), a Toll-like receptor (TLR)/interleukin-1 receptor (IL-1R) inducible transcription factor, regulates IFN-γ production in KG-1 cells, we tested IκBζ's role in the classic lymphocyte pathway of IL-12/IL-18–induced IFN-γ. Upon stimulation with IL-12/IL-18, monocyte-depleted human peripheral blood lymphocytes expressed the 79-kDa form of IκBζ and released IFN-γ. CD56+ NK cells were shown to be the IκBζ-producing lymphocyte subpopulation, which also released abundant IFN-γ in response to IL-12/IL-18. Importantly, IκBζ was undetectable in CD56− lymphocytes where IFN-γ release was 10-fold lower. In addition, small interfering RNA knockdown of IκBζ suppressed IFN-γ expression in CD56+ cells. The association of IκBζ with the IFN-γ promoter was documented by chromatin immunoprecipitation. IFN-γ promoter activity from IκBζ overexpression was confirmed by luciferase reporter assay. Finally, IκBζ coprecipitated with p65 and p50 NF-κB in NK cells in response to IL-12/IL-18, suggesting that IκBζ's effects on IFN-γ promoter activity are coregulated by NF-κB. These results suggest that IκBζ functions as an important regulator of IFN-γ in human NK cells, further expanding the class of IκBζ-modulated genes.

Glucocorticoids (GCs) are commonly prescribed to patients with a variety of inflammatory disorders, including inflammatory bowel disease (IBD). IBD is the manifestation of a dysregulated mucosal immune response, usually against commensal bacteria, involving many different immune cells. GCs mediate their immunomodulatory effects through many different mechanisms and target multiple signaling pathways. The GC dexamethasone down-modulates both innate and adaptive immune cell activation. Group 3 innate lymphocytes (ILC3s) have critical roles in mucosal inflammation. ILC3s secrete high levels of the cytokine IL-22, promoting epithelial proliferation, antimicrobial peptides and mucins. The effects of dexamethasone on T cell function are well described, however, it is not known if dexamethasone modulates the critical function of ILC3s or if dexamethasone regulates the cytokine IL-22. In this study, we examined the effects of dexamethasone on IL-22 production by ILC3s. We found that dexamethasone suppressed IL-23-mediated IL-22 production in homeostatic and activated human and mouse ILC3s. Dexamethasone did not modulate the IL-23 canonical signaling pathway of JAK2/STAT3, but did suppress phosphorylation of IkBa, an important regulator in NF-kB activation. These findings implicate NF-kB as a regulator of IL-22 production in ILC3s and have likely repercussions on GC treatment of IBD patients.

IL-6 is a pleiotropic cytokine implicated in the pathogenesis of disorders such as sepsis and cancer. We noted that human monocytes are excellent producers of IL-6 as compared with monocyte-derived macrophages. Because macrophages from molecule containing ankyrin repeats induced by LPS (MAIL) knockout animals have suppressed IL-6 production, we hypothesized that regulation of MAIL is key to IL-6 production in humans and may explain the differences between human monocytes and macrophages. To test this hypothesis fresh human monocytes and monocyte-derived macrophages were compared for MAIL expression in response to LPS. LPS-induced monocyte MAIL expression was highly inducible and transient. Importantly for our hypothesis MAIL protein expression was suppressed during differentiation of monocytes to macrophages. Of note, the human MAIL protein detected was the 80 kDa MAIL-L form and human MAIL showed nuclear localization. Human MAIL-L bound to p50 subunit of the NF-κB and increased IL-6 luciferase promoter activity in a cEBPβ, NF-κB, and AP-1-dependent fashion. Like the differences in MAIL induction, monocytes produced 6-fold more IL-6 compared with macrophages (81.7 ± 29.7 vs 12.6 ± 6.8 ng/ml). Furthermore, suppression of MAIL by small interfering RNA decreased the production of IL-6 significantly in both THP-1 cells and in primary monocytes. Costimulation of monocytes with LPS and muramyl dipeptide induced an enhanced IL-6 response that was suppressed by siMAIL. Our data suggests that MAIL is a key regulator of IL-6 production in human monocytes and plays an important role in both TLR and NOD-like receptor ligand induced inflammation.

Relative to monocytes, human macrophages are deficient in their ability to process and release IL-1β. In an effort to explain this difference, we used a model of IL-1β processing and release that is dependent upon bacterial escape into the cytosol. Fresh human blood monocytes were compared with monocyte-derived macrophages (MDM) for their IL-1β release in response to challenge with Francisella novicida. Although both cell types produced similar levels of IL-1β mRNA and intracellular pro-IL-1β, only monocytes readily released processed mature IL-1β. Baseline mRNA expression profiling of candidate genes revealed a remarkable deficiency in the pyrin gene, MEFV, expression in MDM compared with monocytes. Immunoblots confirmed a corresponding deficit in MDM pyrin protein. To determine whether pyrin levels were responsible for the monocyte/MDM difference in mature IL-1β release, pyrin expression was knocked down by nucleofecting small interfering RNA against pyrin into monocytes or stably transducing small interfering RNA against pyrin into the monocyte cell line, THP-1. Pyrin knockdown was associated with a significant drop in IL-1β release in both cell types. Importantly, M-CSF treatment of MDM restored pyrin levels and IL-1β release. Similarly, the stable expression of pyrin in PMA-stimulated THP-1-derived macrophages induces caspase-1 activation, associated with increased IL-1β release after infection with F. novicida. In summary, intracellular pyrin levels positively regulate MDM IL-1β responsiveness to Francisella challenge.

Bacillus anthracis, the causative agent of anthrax, secretes lethal toxin that down-regulates immune functions. Translocation of B. anthracis across mucosal epithelia is key for its dissemination and pathogenesis. Group 3 innate lymphocytes (ILC3s) are important in mucosal barrier maintenance due to their expression of the cytokine IL-22, a critical regulator of tissue responses and repair during homeostasis and inflammation. We found that B. anthracis lethal toxin perturbed ILC3 function in vitro and in vivo, revealing an unknown IL-23-mediated MAPK signaling pathway. Lethal toxin had no effects on the canonical STAT3-mediated IL-23 signaling pathway. Rather lethal toxin triggered the loss of several MAP2K kinases, which correlated with reduced activation of downstream ERK1/2 and p38, respectively. Inhibition studies showed the importance of MAPK signaling in IL-23-mediated production of IL-22. Our finding that MAPK signaling is required for optimal IL-22 production in ILC3s may lead to new approaches for targeting IL-22 biology., Author summary Bacillus anthracis, the bacterium that causes the deadly disease anthrax, is commonly known for its use in bioterrorism. We have used B. anthracis to study the effect of MAPK signaling pathways in group 3 innate immune lymphocytes (ILC3s). B. anthracis enters the host through the lungs or gastrointestinal (GI) tract. To cause disease, the pathogen requires translocation across barrier tissue layers to disseminate. During this infection process, the bacterium produces many virulence factors, including a lethal toxin, that allow it to subvert the host immune response. In many immune cells, lethal toxin turns off signaling events through degradation of intermediate signaling components. In this study, we show that lethal toxin interferes in a specific signaling pathway, MAPK, which is significant because it is not usually associated with IL-23-mediated signaling. We specifically studied the interference of MAPK in ILC3s, rare immune cells found in the lungs and GI tract. These cells secrete high levels of interleukin-22 (IL-22), a key cytokine for maintaining barrier tissues during inflammation. Lethal toxin significantly reduced in vitro IL-23-mediated IL-22 production in mouse and human ILC3s. Furthermore, in vivo administration of lethal toxin reduced IL-22 production in ILC3s. This negative effect on IL-22 implicated MAPK signaling as one of the required pathways in ILC3s for IL-22 production, which means that B. anthracis may inhibit ILC3 function during infection, potentially aiding its bacterial dissemination. These findings increase our understanding of the molecular factors that control IL-22 regulation in ILC3s and aid in development of immune therapeutics and preventive measures.

Chronic Rhinosinusitis (CRS) is accompanied by evidence of a vigorous adaptive immune response, and emerging studies demonstrate that some nasal polyps manifest a polyclonal autoantibody response. We previously found that antibodies against BP180, a component of the hemidesmosome complex and the dominant epitope in autoimmune bullous pemphigoid, were found at elevated levels in nasal polyp tissue. Given the critical role of hemidesmosomes in maintaining epithelial integrity, we sought to investigate the distribution of BP180 in nasal tissue and evaluate for evidence of systemic autoimmunity against this antigen in CRS.Case-control experimental study.The expression and distribution of BP180 in cultured nasal epithelial cells and normal nasal tissue were confirmed using real-time polymerase chain reaction (PCR), Western immunoblotting, immunofluorescence and immunohistochemistry. Sera were collected from three groups: control, CRSsNP, and CRSwNP. A commercially available ELISA was utilized to compare anti-BP180 autoantibody levels in sera.BP180 is expressed in nasal epithelium, but is not confined to the basement membrane as it is in human skin. In cultured nasal epithelial cells, confocal immunofluorescence showed a punctate distribution of BP180 along the basal surface, consistent with its distribution in epithelial keratinocytes. There are significantly higher levels of circulating nonpathologic anti-BP180 autoantibodies in CRS patients compared with normal controls (P0.05).BP180 is more widely expressed in nasal epithelium versus skin, although it appears to play a similar role in the formation of hemidesmosomes along the basement membrane. Further investigations are ongoing to characterize the pathogenicity of the anti-epithelial antibody response in CRS.

IkappaBzeta is a novel member of the IkappaB family of NFkappaB regulators, which modulates NFkappaB activity in the nucleus, rather than controlling its nuclear translocation. IkappaBzeta is specifically induced by IL-1beta and several TLR ligands and positively regulates NFkappaB-mediated transcription of genes such as IL-6 and NGAL as an NFkappaB binding co-factor. We recently reported that the IL-1 family cytokines, IL-1beta and IL-18, strongly synergize with TNFalpha for IFNgamma production in KG-1 cells, whereas the same cytokines alone have minimal effects on IFNgamma production. Given the striking similarities between the IL-1R and IL-18R signaling pathways we hypothesized that a common signaling event or gene product downstream of these receptors is responsible for the observed synergy. We investigated IkappaBzeta protein expression in KG-1 cells upon stimulation with IL-1beta, IL-18 and TNFalpha. Our results demonstrated that IL-18, as well as IL-1beta, induced moderate IkappaBzeta expression in KG-1 cells. However, TNFalpha synergized with IL-1beta and IL-18, whereas by itself it had a minimal effect on IkappaBzeta expression. NFkappaB inhibition resulted in decreased IL-1beta/IL-18/TNFalpha-stimulated IFNgamma release. Moreover, silencing of IkappaBzeta expression led to a specific decrease in IFNgamma production. Overall, our data suggests that IkappaBzeta positively regulates NFkappaB-mediated IFNgamma production in KG-1 cells.

Macrophages and their precursors, monocytes, are key cells involved in the innate immune response. Although both monocytes and macrophages produce caspase-1, the key enzyme responsible for pro-IL-1β processing; macrophages are limited in their ability to activate the enzyme and release functional IL-1β. In this context, because mutations in the pyrin gene (MEFV) cause the inflammatory disorder familial Mediterranean fever, pyrin is believed to regulate IL-1β processing. To determine whether variations in pyrin expression explain the difference between monocytes and macrophages in IL-1β processing and release, pyrin was studied in human monocytes and monocyte-derived macrophages. Although monocytes express pyrin mRNA and protein, which is readily inducible by endotoxin, monocyte-derived macrophages express significantly less pyrin mRNA and protein. Pyrin levels directly correlated with IL-1β processing in monocytes and macrophages; therefore, we asked whether pyrin might promote IL-1β processing and release. HEK293 cells were transfected with pyrin, caspase-1, apoptotic speck protein with a caspase recruitment domain, and IL-1β. Pyrin induced IL-1β processing and release in a dose-dependent manner. Conversely, pyrin small interference RNA suppressed pro-IL-1β processing in both THP-1 cells and fresh human monocytes. In summary, both pyrin expression and IL-1β processing and release are diminished upon the maturation of monocytes to macrophages. When pyrin is ectopically expressed or silenced, IL-1β processing and release parallels the level of pyrin. In conclusion, in the context of endotoxin-induced activation of mononuclear phagocytes, pyrin augments IL-1β processing and release.

Epstein-Barr virus-induced G-protein coupled receptor 2 (EBI2) is important for the migration of B cells and induction of extrafollicular plasma cells within secondary lymphoid tissues. Recent studies from our group have demonstrated that many molecules associated with B cell survival and plasma cell differentiation (e.g. BAFF and immunoglobulins) were elevated in nasal polyps from patients with chronic rhinosinusitis (CRS). To investigate whether B cells or EBI2 played a role in CRS pathogenesis, expression of B cell- and EBI2-related genes in nasal tissue from normal controls and CRS patients with (CRSwNP) or without (CRSsNP) nasal polyps was assessed. EBI2 mRNA was quantitated by qRT-PCR, EBI2 protein was measured by western blot, and B cell subsets were characterized by flow cytometry. Expression of EBI2 mRNA was significantly elevated in polyp tissue (p

S U N D A Y 531 Elevated Expression of CC Chemokine Ligand 18 in Chronic Rhinosinusitis A. Kato, S. Peterson, J. A. Poposki, D. R. Nagarkar, R. T. Chustz, A. T. Peters, L. A. Suh, R. Carter, J. Norton, K. E. Harris, L. C. Grammer, B. Tan, R. K. Chandra, D. B. Conley, R. C. Kern, R. P. Schleimer; Division of Allergy-Immunology, Northwestern University Feinberg School of Medicine, Chicago, IL, Department of Otolaryngology, Northwestern University Feinberg School of Medicine, Chicago, IL. RATIONALE: Chronic rhinosinusitis with nasal polyps (CRSwNP) is associated with Th2-dominant inflammation including eosinophilia, in contrast to non-polypoid CRS (CRSsNP). CCL18/PARC is a CC chemokine known to recruit na€ive T cells, B cells, and immature dendritic cells, and thought to be involved in Th2-related inflammatory diseases including asthma and atopic dermatitis. However, expression of CCL18 in CRS has not been extensively studied. METHODS:Using nasal polyp tissue (NP) and uncinate tissue (UT) from controls and patients with CRS, we examined the expression of CCL18 mRNA by real-time PCR and assayed CCL18 protein by ELISA, immunohistochemistry and western blot. RESULTS: Compared to control subjects (n59), CCL18 mRNAwas significantly increased in NP (432-fold, p

Macrophages and their precursors, monocytes, are key cells involved in the innate immune response. Although both monocytes and macrophages produce caspase-1, the key enzyme responsible for the processing of proIL-1β; macrophages are limited in their ability to process and release functional IL-1β. In this context, since mutations in the pyrin gene (MEFV) cause the inflammatory disorder familial Mediterranean fever, the protein pyrin is believed to regulate IL-1β. To determine whether variations in pyrin levels explain the difference between monocyte and macrophages in IL-1β processing and release, pyrin was studied in fresh human monocytes and monocyte derived macrophages. While fresh monocytes express pyrin protein and MEFV mRNA, monocyte derived macrophages have significantly less pyrin mRNA and protein. Since pyrin levels correlated with IL-1β processing, we asked whether pyrin may promote IL-1β processing and release. Pyrin induced IL-1β processing and release in a dose dependent fashion in HEK293 cells and pyrin overexpression in THP-1 cells confirmed the effect. Conversely, pyrin siRNA suppressed proIL-1β processing in both THP-1 cells and fresh human monocytes. In summary, pyrin expression and IL-1β processing and release is diminished upon the maturation of monocytes to macrophages. In the setting of endotoxin induced activation of mononuclear phagocytes, pyrin appears to augment IL-1β processing and release. Supported by grant NIH-NHLBI-HL40871 to MDW