Modulation of innate immunity by nucleotide binding: biochemical and functional characterization of a CATERPILLER/NLR protein, Monarch-1/NLRP12

The recently discovered Nucleotide Binding Domain-Leucine Rich Repeat (NLR) gene family is conserved from plants to mammals and several members are associated with human autoinflammatory or immunodeficiency disorders. This family is defined by a central nucleotide binding domain that contains the highly conserved Walker A and Walker B motifs. Although the nucleotide binding domain is a defining feature of this family, it has not been extensively studied in its purified form. In this thesis, we show that purified Monarch-1/NLRP12, an NLR protein that negatively regulates NF-[kappa]B signaling, specifically binds ATP and exhibits ATP hydrolysis activity. Intact Walker A/B motifs are required for this activity. These motifs are also required for Monarch-1 to undergo self-oligomerization, TLR- or CD40L- activated association with NIK and IRAK-1, degradation of NIK, and inhibition of IRAK-1 phosphorylation. Stable expression of a Walker A/B mutant in THP-1 monocytes results in increased production of proinflammatory cytokines and chemokines to an extent comparable to cells in which Monarch-1 is silenced via shRNA. In addition, the functional role of conserved motifs in Monarch-1 NBD domain is examined. The results of this study are consistent with a model wherein ATP binding regulates the anti-inflammatory activity of Monarch-1

NLRP3 Plays a Critical Role in the Development of Experimental Autoimmune Encephalomyelitis by Mediating Th1 and Th17 Responses

The interplay between innate and adaptive immunity is important in multiple sclerosis (MS). The inflammasome complex, which activates caspase-1 to process pro–IL-1β and pro–IL-18, is rapidly emerging as a pivotal regulator of innate immunity, with nucleotide-binding domain, leucine-rich repeat containing protein family, pyrin domain containing 3 (NLRP3) (cryopyrin or NALP3) as a prominent player. Although the role of NLRP3 in host response to pathogen associated molecular patterns and danger associated molecular patterns is well documented, its role in autoimmune diseases is less well studied. To investigate the role of NLRP3 protein in MS, we used a mouse model of MS, experimental autoimmune encephalomyelitis (EAE). Nlrp3 expression was elevated in the spinal cords during EAE, and Nlrp3−/− mice had a dramatically delayed course and reduced severity of disease. This was accompanied by a significant reduction of the inflammatory infiltrate including macrophages, dendritic cells, CD4, and CD8+ T cells in the spinal cords of the Nlrp3−/− mice, whereas microglial accumulation remained the same. Nlrp3−/− mice also displayed improved histology in the spinal cords with reduced destruction of myelin and astrogliosis. Nlrp3−/− mice with EAE produced less IL-18, and the disease course was similar to Il18−/− mice. Furthermore, Nlrp3−/− and Il18−/− mice had similarly reduced IFN-γ and IL-17 production. Thus, NLRP3 plays a critical role in the induction of the EAE, likely through effects on capase-1–dependent cytokines which then influence Th1 and Th17

Cutting Edge: NLRP12 Controls Dendritic and Myeloid Cell Migration To Affect Contact Hypersensitivity

Nucleotide-binding domain leucine-rich repeat (NLR) proteins are regulators of inflammation and immunity. Although first described 8 y ago, a physiologic role for NLRP12 has remained elusive until now. We find that murine Nlrp12, an NLR linked to atopic dermatitis and hereditary periodic fever in humans, is prominently expressed in dendritic cells (DCs) and neutrophils. Nlrp12-deficient mice exhibit attenuated inflammatory responses in two models of contact hypersensitivity that exhibit features of allergic dermatitis. This cannot be attributed to defective Ag processing/presentation, inflammasome activation, or measurable changes in other inflammatory cytokines. Rather, Nlrp12(-/-) DCs display a significantly reduced capacity to migrate to draining lymph nodes. Both DCs and neutrophils fail to respond to chemokines in vitro. These findings indicate that NLRP12 is important in maintaining neutrophils and peripheral DCs in a migration-competent state

The innate immune sensor NLRC3 attenuates Toll-like receptor signaling via modification of the signaling adaptor TRAF6 and transcription factor NF-κB

Several members of the NLR family of sensors activate innate immunity. In contrast, we found here that NLRC3 inhibited Toll-like receptor (TLR)-dependent activation of the transcription factor NF-κB by interacting with the TLR signaling adaptor TRAF6 to attenuate Lys63 (K63)-linked ubiquitination of TRAF6 and activation of NF-κB. We used bioinformatics to predict interactions between NLR and TRAF proteins, including interactions of TRAF with NLRC3. In vivo, macrophage expression of Nlrc3 mRNA was diminished by the administration of lipopolysaccharide (LPS) but was restored when cellular activation subsided. To assess biologic relevance, we generated Nlrc3−/− mice. LPS-treated Nlrc3−/− macrophages had more K63-ubiquitinated TRAF6, nuclear NF-κB and proinflammatory cytokines. Finally, LPS-treated Nlrc3−/− mice had more signs of inflammation. Thus, signaling via NLRC3 and TLR constitutes a negative feedback loop. Furthermore, prevalent NLR-TRAF interactions suggest the formation of a ‘TRAFasome’ complex

Identifying SYNE1 Ataxia With Novel Mutations in a Chinese Population

Objective: Variants in SYNE1 have been widely reported in ataxia patients in Europe, with highly variable clinical phenotype. Until now, no mutation of SYNE1 ataxia has been reported among the Chinese population. Our aim was to screen for SYNE1 ataxia patients in China and extend the clinicogenetic spectrum.Methods: Variants in SYNE1 were detected by high-throughput sequencing on a cohort of 126 unrelated index patients with unexplained autosomal recessive or sporadic ataxia. Pathogenicity assessments of SYNE1 variants were interpreted according to the ACMG guidelines. Potential pathogenic variants were confirmed by Sanger sequencing. Clinical assessments were conducted by two experienced neurologists.Results: Two Chinese families with variable ataxia syndrome were identified (accounting for 1.6%; 2/126), separately caused by the novel homozygous SYNE1 mutation (NM_033071.3: c.21568C>T, p.Arg7190Ter), and compound heterozygous SYNE1 mutation (NM_033071.3: c.18684G>A, p.Trp6228Ter; c.17944C>T, p.Arg5982Ter), characterized by motor neuron impairment, mental retardation and arthrogryposis.Conclusions:SYNE1 ataxia exists in the Chinese population, as a rare form of autosomal recessive ataxia, with a complex phenotype. Our findings expanded the ethnic, phenotypic and genetic diversity of SYNE1 ataxia

Determination of relevance between surface free energy and adsorption capacity of cement particles

ABSTRACT The compatibility between superplasticizer and cement was influenced by the adsorption capacity of cement particles. This study investigated the relevance between the adsorption capability and surface free energy. Adsorption capacity and surface free energy of both sulphoaluminate cement and portland cement were measured. The adsorption capacity of cement particles was measured by ultraviolet spectrophotometry. The test showed that particles of sulphoaluminate cement adsorbed more molecules of superplasticizer than portland cement particles. The weight of superplasticizer adsorbed by 2g of sulphoaluminate cement and portland cement were 0.28mg and 0.159mg respectively. Surface free energy of cement particles was calculated by contact angle and the contact angles were determined by the thin-layer wicking technique and washburn equation which is theoretical basis of thin-layer wiching technique presented by Chibowski E. The sulphoaluminate cement, portland cement's surface free energy were 51.46 mJ·m-2 and 49.36 mJ·m-2 respectively. The results showed that the higher adsorption capacity of particles was usual accompanied by higher surface free energy. The fluidity of cement paste was influenced by the adsorption capacity of cement particles because the more molecules of superplasticizer was adsorbed by cement particles there were lacking superplasticizer in the paste. The macro-behaviour of higher adsorption capacity is that the cement paste need more superplasticizer to reach the needed fluidity

Design, fabrication and characterization of quantum dot infrared photodetectors

textElectrical and Computer Engineerin

Security Authentication of Dual Chaotic Image Watermarking in Spatial Domain with Spatial and Frequency Domain Characteristics Analysis

This article presents an advanced dual chaotic watermarking scheme to improve information security. To ensure confidentiality in digital image transmission, a secure dual watermarking scheme is proposed by applying the chaotic logistic system and hyper-chaotic dynamical system. Chaotic watermarking was conducted in the spatial domain, where suboptimal secure hashing with a variable length was selected in preprocessing stages. The secret key was generated by the chaotic sequence for pixel shuffling using a chaotic logistic map, so that a controlled amount of distortion was inserted into the host digital image. Watermarking was proceeded after the chaotic watermark had been embedded into the shuffled image. To strengthen the security, the hyper-chaotic system was used to generate chaotic blocks for block scrambling in order to achieve dual chaotic watermarking. Characteristics analysis was conducted for multiple examples in both spatial and frequency domains. Potential effects induced by the chaotic driving parameter on processing time and integrity authentication of chaotic dual watermarking were also analyzed in detail