bluuebunny/crossref_metadata_2025 huggingface dataset:
Critical Role of IL-1 Receptor-Associated Kinase-M in Regulating Chemokine-Dependent Deleterious Inflammation in Murine Influenza Pneumonia
Authors: Seki Masafumi , Kohno Shigeru , Newstead Michael W. , Zeng Xianying , Bhan Urvashi , Lukacs Nicholas W. , Kunkel Steven L. , Standiford Theodore J.
Published: December 2009

Influenza virus is a common cause of respiratory infection and morbidity, which is often due to deleterious host immune responses directed against the pathogen. We investigated the role of IL-1 receptor-associated kinase-M (IRAK-M), an inhibitor of MyD88-dependent TLR signaling, in modulating the innate inflammatory response during influenza pneumonia using a murine model. The intranasal administration of influenza resulted in the upregulation of IRAK-M mRNA and protein levels in the lungs within 2 d after infectious challenge. Pulmonary influenza infection in mice deficient in IRAK-M (IRAK-M−/−) resulted in substantially increased mortality compared with similarly treated wild-type animals. Increased mortality in IRAK-M−/− mice was associated with enhanced early influx of neutrophils, high permeability edema, apoptosis of lung epithelial cells, markedly increased expression of inflammatory cytokines/chemokines, and release of neutrophil-derived enzymes, including myeloperoxidase and neutrophil elastase. Early viral clearance was not different in mutant mice, whereas viral titers in lungs and blood were significantly higher in IRAK-M−/−mice compared with wild-type animals. Increased lethality observed in IRAK-M−/− mice after influenza challenge was abrogated by Ab-mediated blockade of CXCR2. Collectively, our findings indicate that IRAK-M is critical to preventing deleterious neutrophil-dependent lung injury during influenza infection of the respiratory tract.
LL-37 Promotes Rapid Sensing of CpG Oligodeoxynucleotides by B Lymphocytes and Plasmacytoid Dendritic Cells
Authors: Hurtado Plinio , Au Peh Chen
Published: December 2009

LL-37 is a cationic antimicrobial peptide derived from neutrophils and keratinocytes. It plays an important role in protection against bacterial infection in the skin and mucosal surfaces. However, its role within the blood compartment remains unclear given that serum inhibits its bactericidal property. In this study, we show that LL-37 promotes very rapid and highly efficient sensing of CpG motifs in bacterial DNA by human B lymphocytes and plasmacytoid dendritic cells (pDCs) in serum-containing media and in whole blood. LL-37 allowed detection of CpG oligodeoxynucleotide (ODN) within minutes of exposure. Without LL-37, 20–30 times more CpG was required to produce the same effect. The promotion of CpG detection by LL-37 was independent of the backbone of the ODN, as the effect was observed not only in ODNs with modified phosphorothioate backbone, but also in ODNs with natural phosphodiester backbone, as found in genomic DNA. Unmethylated CpG motifs within the phosphodiester ODN and LL-37–mediated delivery are required for pDCs to respond. In keeping with the above, cells responded to CpG-rich bacterial DNA and LL-37, but not to human DNA and LL-37. The ability of LL-37 to enhance delivery of CpG to stimulate immune cells is independent of its amphipathic structure and its bactericidal property. LL-37 aids the delivery of CpG to B cells and pDCs, but not T cells. These findings are pertinent to rapid recognition of microbial DNA and are highly relevant to contemporary studies of CpG/TLR9 agonists in vaccines and cancer therapy.
IL-17 Regulates CXCL1 mRNA Stability via an AUUUA/Tristetraprolin-Independent Sequence
Authors: Datta Shyamasree , Novotny Michael , Pavicic Paul G. , Zhao Chenyang , Herjan Tomasz , Hartupee Justin , Hamilton Thomas
Published: December 2009

IL-17 contributes to inflammatory response in part by promoting enhanced expression of chemokines, such as CXCL1, by prolonging the t1/2 of this constitutively unstable mRNA. Although IL-17 is a weak stimulus for transcription of the CXCL1 gene, it strongly potentiates message accumulation via stabilization when the mRNA is transcribed in cells stimulated with TNF. In myeloid cells, LPS-induced CXCL1 mRNA stabilization is dependent on AUUUA-containing sequence motifs that are recognized by the RNA binding protein tristetraprolin (TTP). Using deletion and site-specific mutagenesis, we report that IL-17–mediated stabilization of CXCL1 mRNA in nonmyeloid cells depends on a sequence that does not contain the AUUUA motif. Furthermore, a specific two-nucleotide mutation within this region markedly abrogates sensitivity for IL-17–mediated stabilization. Consistent with this finding, the IL-17–sensitive sequence does not exhibit increased instability in the presence of TTP, and CXCL1 mRNA remains unstable and can be stabilized in response to treatment with IL-17 in embryo fibroblasts from mice in which the TTP gene has been deleted. Whereas the RNA binding protein KSRP has been shown to participate in regulating the instability of human CXCL8 mRNA, inhibitory RNA-based reduction in KSRP does not effect the instability mediated by the IL-17–sensitive sequence motif. These findings suggest that IL-17–mediated chemokine mRNA stabilization in nonmyeloid cells uses a mechanism that is distinct from that operating to control AU-rich mRNA stability in myeloid cells.
Regulation of Lentivirus Neurovirulence by Lipopolysaccharide Conditioning: Suppression of CXCL10 in the Brain by IL-10
Authors: Maingat Ferdinand , Viappiani Serena , Zhu Yu , Vivithanaporn Pornpun , Ellestad Kristofor K. , Holden Janet , Silva Claudia , Power Christopher
Published: December 2009

Lentivirus infections including HIV and feline immunodeficiency virus (FIV) cause neurovirulence, which is largely mediated by innate immunity. To investigate the interactions between neurovirulence and repeated conditioning by innate immune activation, models of lentivirus infection were exposed to LPS. Gene expression in HIV-infected (HIV+) and control (HIV−) patient brains was compared by real time RT-PCR and immunocytochemistry. Supernatants from mock and HIV-infected monocyte-derived macrophages exposed to LPS were applied to human neurons. FIV-infected (FIV+) and control (FIV−) animals were exposed repeatedly to LPS postinfection together with concurrent neurobehavioral testing, viral load, and host gene analyses. Brains from HIV+ individuals exhibited induction of CD3ε, CXCL10, and granzyme A expression (p < 0.05). Supernatants from HIV+ monocyte-derived macrophages induced CXCL10 expression in neurons, which was diminished by IL-10 treatment (p < 0.05). LPS-exposed FIV+ animals demonstrated lower plasma and brain viral loads (p < 0.05). Neuronal CXCL10 expression was increased in FIV+ animals but was suppressed by LPS exposure, together with reduced brain CD3ε and granzyme A expression (p < 0.05). In conjunction with preserved NeuN-positive neuronal counts in parietal cortex (p < 0.05), FIV+ animals exposed to LPS also showed less severe neurobehavioral deficits (p < 0.05). Repeated LPS exposures suppressed CXCL10 in the brain and ensuing T cell infiltration with a concomitant reduction in neurovirulence. Thus, innate immune chronic conditioning exerted beneficial effects on neurovirulence through suppression of a specific chemotactic factor, CXCL10, mediated by IL-10, leading to reduced leukocyte infiltration and release of neurotoxic factors.
Cell Type-Specific Recognition of Human Metapneumoviruses (HMPVs) by Retinoic Acid-Inducible Gene I (RIG-I) and TLR7 and Viral Interference of RIG-I Ligand Recognition by HMPV-B1 Phosphoprotein
Authors: Goutagny Nadege , Jiang Zhaozhao , Tian Jane , Parroche Peggy , Schickli Jeanne , Monks Brian G. , Ulbrandt Nancy , Ji Hong , Kiener Peter A. , Coyle Anthony J. , Fitzgerald Katherine A.
Published: December 2009

Human metapneumoviruses (HMPVs) are recently identified Paramyxoviridae that contribute to respiratory tract infections in children. No effective treatments or vaccines are available. Successful defense against virus infection relies on early detection by germ line-encoded pattern recognition receptors and activation of cytokine and type I IFN genes. Recently, the RNA helicase retinoic acid-inducible gene I (RIG-I) has been shown to sense HMPV. In this study, we investigated the abilities of two prototype strains of HMPV (A1 [NL\1\00] and B1 [NL\1\99]) to activate RIG-I and induce type I IFNs. Despite the abilities of both HMPV-A1 and HMPV-B1 to infect and replicate in cell lines and primary cells, only the HMPV-A1 strain triggered RIG-I to induce IFNA/B gene transcription. The failure of the HMPV-B1 strain to elicit type I IFN production was dependent on the B1 phosphoprotein, which specifically prevented RIG-I–mediated sensing of HMPV viral 5′ triphosphate RNA. In contrast to most cell types, plasmacytoid dendritic cells displayed a unique ability to sense both HMPV-A1 and HMPV-B1 and in this case sensing was via TLR7 rather than RIG-I. Collectively, these data reveal differential mechanisms of sensing for two closely related viruses, which operate in cell type‑specific manners.
Mycolactone Suppresses T Cell Responsiveness by Altering Both Early Signaling and Posttranslational Events
Authors: Boulkroun Sheerazed , Guenin-Macé Laure , Thoulouze Maria-Isabel , Monot Marc , Merckx Anaïs , Langsley Gordon , Bismuth Georges , Di Bartolo Vincenzo , Demangel Caroline
Published: December 2009

Mycolactone is a diffusible lipid toxin produced by Mycobacterium ulcerans, the causative agent of a necrotizing skin disease referred to as Buruli ulcer. Intriguingly, patients with progressive lesions display a systemic suppression of Th1 responses that resolves on surgical excision of infected tissues. In this study, we examined the effects of mycolactone on the functional biology of T cells and identified two mechanisms by which mycolactone suppresses cell responsiveness to antigenic stimulation. At noncytotoxic concentrations, mycolactone blocked the activation-induced production of cytokines by a posttranscriptional, mammalian target of rapamycin, and cellular stress-independent mechanism. In addition, mycolactone triggered the lipid-raft association and activation of the Src-family kinase, Lck. Mycolactone-mediated hyperactivation of Lck resulted in the depletion of intracellular calcium stores and downregulation of the TCR, leading to impaired T cell responsiveness to stimulation. These biochemical alterations were not observed when T cells were exposed to other bacterial lipids, or to structurally related immunosuppressors. Mycolactone thus constitutes a novel type of T cell immunosuppressive agent, the potent activity of which may explain the defective cellular responses in Buruli ulcer patients.
Peripheral Blood CCR4+CCR6+ and CXCR3+CCR6+ CD4+ T Cells Are Highly Permissive to HIV-1 Infection
Authors: Gosselin Annie , Monteiro Patricia , Chomont Nicolas , Diaz-Griffero Felipe , Said Elias A. , Fonseca Simone , Wacleche Vanessa , El-Far Mohamed , Boulassel Mohamed-Rachid , Routy Jean-Pierre , Sekaly Rafick-Pierre , Ancuta Petronela
Published: December 2009

There is limited knowledge on the identity of primary CD4+ T cell subsets selectively targeted by HIV-1 in vivo. In this study, we established a link between HIV permissiveness, phenotype/homing potential, and lineage commitment in primary CD4+ T cells. CCR4+CCR6+, CCR4+CCR6−, CXCR3+CCR6+, and CXCR3+CCR6− T cells expressed cytokines and transcription factors specific for Th17, Th2, Th1Th17, and Th1 lineages, respectively. CCR4+CCR6+ and CXCR3+CCR6+ T cells expressed the HIV coreceptors CCR5 and CXCR4 and were permissive to R5 and X4 HIV replication. CCR4+CCR6− T cells expressed CXCR4 but not CCR5 and were permissive to X4 HIV only. CXCR3+CCR6− T cells expressed CCR5 and CXCR4 but were relatively resistant to R5 and X4 HIV in vitro. Total CCR6+ T cells compared with CCR6− T cells harbored higher levels of integrated HIV DNA in treatment-naive HIV-infected subjects. The frequency of total CCR6+ T cells and those of CCR4+CCR6+ and CXCR3+CCR6+ T cells were diminished in chronically infected HIV-positive subjects, despite viral-suppressive therapy. A high-throughput analysis of cytokine profiles identified CXCR3+CCR6+ T cells as a major source of TNF-α and CCL20 and demonstrated a decreased TNF-α/IL-10 ratio in CXCR3+CCR6− T cells. Finally, CCR4+CCR6+ and CXCR3+CCR6+ T cells exhibited gut- and lymph node-homing potential. Thus, we identified CCR4+CCR6+ and CXCR3+CCR6+ T cells as highly permissive to HIV replication, with potential to infiltrate and recruit more CCR6+ T cells into anatomic sites of viral replication. It is necessary that new therapeutic strategies against HIV interfere with viral replication/persistence in discrete CCR6+ T cell subsets.
IL-33 Induces IL-13–Dependent Cutaneous Fibrosis
Authors: Rankin Andrew L. , Mumm John B. , Murphy Erin , Turner Scott , Yu Ni , McClanahan Terrill K. , Bourne Patricia A. , Pierce Robert H. , Kastelein Rob , Pflanz Stefan
Published: December 2009

IL-33 is constitutively expressed in epithelial barrier tissues, such as skin. Although increased expression of IL-33/IL-33R has been correlated with fibrotic disorders, such as scleroderma and progressive systemic sclerosis, the direct consequences of IL-33 release in skin has not been reported. To determine the effects of dysregulated IL-33 signaling in skin, we administered IL-33 s.c. and monitored its effects at the injection site. Administration of IL-33 resulted in IL-33R–dependent accumulation of eosinophils, CD3+ lymphocytes, F4/80+ mononuclear cells, increased expression of IL-13 mRNA, and the development of cutaneous fibrosis. Consistent with extensive cutaneous tissue remodeling, IL-33 resulted in significant modulation of a number of extracellular matrix-associated genes, including collagen VI, collagen III, and tissue inhibitor of metalloproteases-1. We establish that IL-33–induced fibrosis requires IL-13 using IL-13 knockout mice and eosinophils using ΔdblGATA mice. We show that bone marrow-derived eosinophils secrete IL-13 in response to IL-33 stimulation, suggesting that eosinophil-derived IL-13 may promote IL-33–induced cutaneous fibrosis. Collectively, our results identify IL-33 as a previously unrecognized profibrotic mediator in skin and highlight the cellular and molecular pathways by which this pathology develops.
Thiophene–Benzothiadiazole Co‐Oligomers: Synthesis, Optoelectronic Properties, Electrical Characterization, and Thin‐Film Patterning
Authors: Melucci Manuela , Favaretto Laura , Zanelli Alberto , Cavallini Massimiliano , Bongini Alessandro , Maccagnani Piera , Ostoja Paolo , Derue Gwennaelle , Lazzaroni Roberto , Barbarella Giovanna
Published: January 2010

Newly synthesized thiophene (T) and benzothiadiazole (B) co‐oligomers of different size, alternation motifs, and alkyl substitution types are reported. Combined spectroscopic data, electrochemical analysis, and theoretical calculations show that the insertion of a single electron‐deficient B unit into the aromatic backbone strongly affects the LUMO energy level. The insertion of additional B units has only a minor effect on the electronic properties. Cast films of oligomers with two alternated B rings (B–T–B inner core) display crystalline order. Bottom‐contact FETs based on films cast on bare SiO2 show hole‐charge mobilities of 1 × 10−3–5 × 10−3 cm2 V−1s−1 and Ion/Ioff ratios of 105–106. Solution‐cast films of cyclohexyl‐substituted compounds are amorphous and do not show FET behavior. However, the lack of order observed in these films can be overcome by nanorubbing and unconventional wet lithography, which allow for fine control of structural order in thin deposits.
Usefulness of the discrete choice methodology for marketing decision‐making in new product development: an example from the European functional foods market
Authors: Krystallis Athanasios , Linardakis Michalis , Mamalis Spyridon
Published: January 2010

Understanding consumer needs should be a high‐priority strategic objective in new product development (NPD). Despite their importance, however, NPD processes do not take full advantage of the consumer survey methodologies available. The authors investigate+ parents” preferences for three new functional children's snacks marketed in Greece. Using a very simple discrete choice experimental design, the authors aim (a) to provide an example of consumer research implementation in food‐related NPD and to assess its marketing value, and (b) to evaluate the quality of the acquired consumer‐related information for marketing decision‐making in food consumer‐led NPD processes against a number of criteria set in the relevant literature. “Functionality” is found to be a statistically significant attribute for all three children's snacks of the research design compared to other attributes such as flavor and price. Parents seem to perceive the functional snack alternative as offering greater utility as compared to its conventional (i.e., nonfunctional) counterpart. Overall, the authors conclude that, despite some inherent weaknesses, the advantage of the discrete choice methodology in connection with consumer‐led food NPD lies in its ability to describe current consumer needs and desires realistically and to contribute to the optimization of existing products. [EconLit Classification: M310, C920, C930, D100]. © 2010 Wiley Periodicals, Inc.
Surface modification effects of core–shell rubber particles on the toughening of poly(butylene terephthalate)
Authors: Yang Hoichang , Cho Kilwon
Published: January 2010

We toughened poly(butylene terephthalate) (PBT) by loading core–shell rubber (CSR) type impact modifiers, consisting of a rubbery poly(n‐butyl acrylate) core and a rigid poly(methyl methacrylate) shell. To optimize the dispersion of CSR particles into the PBT matrix during melt compounding, the shell surface was modified with different grafting ratios of glycidyl methacrylate (GMA) reactive with PBT chain ends. In PBT blends with a 20 wt % CSR loading, the dispersed rubbery phases showed discernible shapes depending on the grafted GMA content, from predetermined spheres with 0.25 ± 0.05 μm diameters to their aggregates in the 2–3 μm diameter range. As a result, the interparticle spacing (τ) could be controlled from 0.25 to 4.0 μm in the PBT blends containing the fixed rubber loading. The Izod impact strengths of these samples increased significantly below τ = 0.4 μm. Additional thermal and morphological analyses strongly supported the hypothesis that the marked increase in toughness of the blends was related to less ordered lamellar formation of the PBT matrix under the confined geometry. © 2010 Wiley Periodicals, Inc. J Appl Polym Sci, 2010
Layered double hydroxide/NaSb(OH)6–poly(vinyl chloride) nanocomposites: Preparation, characterization, and thermal stability
Authors: Chen Xue‐Gang , Wu Dai‐Dai , Lv Shuang‐Shuang , Zhang Lu , Ye Ying , Cheng Ji‐Peng
Published: January 2010

A novel layered double hydroxide/NaSb(OH)6‐based nanocomposite (Sb‐LDH) has been prepared via intercalation of thio‐antimonite (SbS33−) and reconstruction of LDH using Mg‐Al LDH as precursors. It is composed of LDH nanolayers with thickness of 25 nm and NaSb(OH)6 nanoparticles with diameter of 3–25 nm. The presence of NaSb(OH)6 will decrease the decomposition intensity and hinder the decomposition of Mg‐Al LDH because of the potential synergetic effect. When applied to poly(vinyl chloride) (PVC) composites, both Mg‐Al LDH and Sb‐LDH can enhance the thermal stability and increase the decomposition temperature of PVC. Compared with Mg‐Al LDH, Sb‐LDH results in higher decomposition temperatures and whiteness and higher initial and long‐term stabilities due to the presence of NaSb(OH)6, which can react with HCl and coordinate with Cl in the PVC chains. Because Mg‐Al LDH will accelerate the dehydrochlorination of PVC driving by the Lewis acid such as AlCl3, the thermal stability of PVC decreases with increasing nanofiller loading. When 1 wt % Sb‐LDH was added, the color change time and Congo red time of PVC composites are 140 min and 154 min, respectively. With enhanced thermal stabilization, this novel LDH nanocomposite could gain promising application in thermal stabilizer for PVC resins. © 2010 Wiley Periodicals, Inc. J Appl Polym Sci, 2010
Influence of temperature and penetration depth on photopolymerization of n‐butyl acrylate using a narrow channel reactor
Authors: Nekkanti V. , Jachuck R. J. J.
Published: January 2010

The influence of temperature and reactor size on photopolymerization kinetics of n‐butyl acrylate (n‐BA) using narrow channel reactors has been investigated. Experimental results have shown that increase in temperature and decrease in size of the reactor increases the monomer conversion. The effect of temperature was studied by immersing the reactor in a water bath maintained at different temperatures. A narrow channel glass reactor, 900 mm long, with diameters 1.5 mm, 1.0 mm, and 0.5 mm respectively were used to study the effect of light penetration depth on the rate of polymerization and molecular weights of the polymer produced. The degree of branching in the polymer produced was also estimated and compared with published data. © 2010 Wiley Periodicals, Inc. J Appl Polym Sci, 2010
Synthesis and properties of copolymer epoxy resins prepared from copolymerization of bisphenol A, epichlorohydrin, and liquefied Dendrocalamus latiflorus
Authors: Wu Chiou‐Chang , Lee Wen‐Jau
Published: January 2010

Dendrocalamus latiflorus Munro (ma bamboo) was liquefied in phenol and polyhydric alcohol (polyethylene glycol/glycerol cosolvent) with H2SO4 as catalyst. Liquefied bamboos reacted with bisphenol A and epichlorohydrin were then employed to prepare copolymer epoxy resins. The curing property and thermal property of copolymer epoxy resins were investigated. The results showed that copolymer epoxy resins could cure at room temperature after the hardener was added, and its curing process was an exothermic reaction. Comparison showed that copolymer epoxy resins prepared with phenol‐liquefied bamboo as raw material had higher heat released than those prepared with polyhydric alcohol‐liquefied bamboo during curing. The DSC analysis showed that heat treatment could enhance the crosslinking of copolymer epoxy resins cured at room temperature. However, resins prepared with polyhydric alcohol‐liquefied bamboo had a lower glass transition temperature. The TGA analysis showed that resins prepared with phenol‐liquefied bamboo had better thermal stability. © 2010 Wiley Periodicals, Inc. J Appl Polym Sci, 2010
Rheological, morphological, and heat seal properties of linear low density polyethylene and cyclo olefine copolymer (LLDPE/COC) blends
Authors: Lamnawar K. , Vion‐Loisel F. , Maazouz A.
Published: January 2010

The rheological and morphological properties of linear low‐density polyethylene/cyclo olefin copolymer (LLDPE/COC) blends were investigated, as were their peel seal characteristics when heat sealed to films of either polyethylene (PE) or polyethylene terephtalate (PET). Three blend systems, with COC volume percentages of 5, 10, and 15%, were melt blended in a twin‐screw extruder. A partial phase miscibility/compatibility was initially suggested by Cole–Cole and equivalent plots of the dynamic rheological properties and subsequently confirmed by scanning electron microscopy. The investigation of the systems' heat sealing properties pointed out an interesting industrial potential for PET sheets covered with a fine layer of a LLDPE/COC blend before heat sealing to a PE film for packaging applications. © 2010 Wiley Periodicals, Inc. J Appl Polym Sci, 2010
Investigating the micellization behavior of cetyltrimethylammonium bromide: As affected by the contents of poly(ethylene oxide) and temperature of the system
Authors: Baloch Musa Kaleem , Rauf Abdur , Ahmad Farzana , Durani Gulrez Fatima , Hameed Gulzar
Published: January 2010

The micellization behavior of cetyltrimethylammonium bromide (CTAB) is investigated in the absence and presence of poly(ethylene oxide) employing surface tension, light scattering, and viscosity and conductance measurement. It has been concluded that the critical micellization concentration (CMC) of CTAB in the presence of polymer increases with the increase in polymer concentration. The three ranges of interaction among the polymer and CTAB with respect to concentration of CTAB are identified at surface tension plot and noted to depend upon polymer concentration. The effect of temperature over these interactions and CMC of CTAB has been investigated. It is observed that the CMC, ΔH, and flow activation energy of the system increases with the increase in polymer concentration whereas CMC of CTAB decreases with the temperature. RH and Rg/RH values determined by light scattering also indicate such type of behavior. The CMC values have also been obtained by viscosity and conductivity measurement. The results and conclusions drawn through all the techniques were consistent. New methodology has also been introduced to interpret the data obtained by surface tension, viscosity, and conductance measurement, and get information about the point of interaction and saturation of polymer with surfactant, which was noted to be very useful and informative. © 2010 Wiley Periodicals, Inc. J Appl Polym Sci, 2010
Up‐regulation of cytokines and chemokines predates the onset of rheumatoid arthritis
Authors: Kokkonen Heidi , Söderström Ingegerd , Rocklöv Joacim , Hallmans Göran , Lejon Kristina , Rantapää Dahlqvist Solbritt
Published: January 2010

Objective

To identify whether cytokines, cytokine‐related factors, and chemokines are up‐regulated prior to the development of rheumatoid arthritis (RA).

Methods

A nested case–control study was performed in 86 individuals who had donated blood samples before experiencing any symptoms of disease (pre‐patients) and 256 matched control subjects (1:3 ratio). In 69 of the pre‐patients, blood samples were also obtained at the time of the diagnosis of RA. The plasma levels of 30 cytokines, related factors, and chemokines were measured using a multiplex system.

Results

The levels of several of the cytokines, cytokine receptors, and chemokines were significantly increased in individuals before disease onset compared with the levels in control subjects; i.e., those representing signs of general immune activation (interleukin‐1β [IL‐1β], IL‐2, IL‐6, IL‐1 receptor antagonist, and tumor necrosis factor), activation of Th1 cells (interferon‐γ, IL‐12), Th2 cells (IL‐4, eotaxin), Treg cells (IL‐10), bone marrow–derived factors (IL‐7, granulocyte–macrophage colony‐stimulating factor, and granulocyte colony‐stimulating factor), as well as chemokines (monocyte chemotactic protein 1 and macrophage inflammatory protein 1α). The levels were particularly increased in anti–cyclic citrullinated peptide antibody– and rheumatoid factor–positive individuals, and the concentration of most of these increased further after disease onset. The concentration of IL‐17 in individuals before disease onset was significantly higher than that in patients after disease onset. Individuals in whom RA subsequently developed were discriminated from control subjects mainly by the presence of Th1 cells, Th2 cells, and Treg cell–related cytokines, while chemokines, stromal cell–derived cytokines, and angiogenic‐related markers separated patients after the development of RA from individuals before the onset of RA.

Conclusion

Individuals in whom RA later developed had significantly increased levels of several cytokines, cytokine‐related factors, and chemokines representing the adaptive immune system (Th1, Th2, and Treg cell–related factors); after disease onset, the involvement and activation of the immune system was more general and widespread.
The association of a nonsynonymous single‐nucleotide polymorphism in TNFAIP3 with systemic lupus erythematosus and rheumatoid arthritis in the Japanese population
Authors: Shimane Kenichi , Kochi Yuta , Horita Tetsuya , Ikari Katsunori , Amano Hirofumi , Hirakata Michito , Okamoto Akiko , Yamada Ryo , Myouzen Keiko , Suzuki Akari , Kubo Michiaki , Atsumi Tatsuya , Koike Takao , Takasaki Yoshinari , Momohara Shigeki , Yamanaka Hisashi , Nakamura Yusuke , Yamamoto Kazuhiko
Published: January 2010

Objective

Genome‐wide association (GWA) studies in systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA) in Caucasian populations have independently identified risk variants in and near the tumor necrosis factor α (TNFα)–induced protein 3 gene (TNFAIP3), which is crucial for the regulation of TNF‐mediated signaling and Toll‐like receptor signaling. The aim of this study was to assess the role of TNFAIP3 in the development of SLE and RA in Japanese subjects.

Methods

We selected 2 single‐nucleotide polymorphisms (SNPs) from previous GWA studies. Rs2230926 is a nonsynonymous SNP in TNFAIP3 and is associated with SLE, while rs10499194 is an intergenic SNP associated with RA. We then performed 2 independent sets of SLE case–control comparisons (717 patients and 1,362 control subjects) and 3 sets of RA case–control comparisons (3,446 patients and 2,344 control subjects) using Japanese subjects. We genotyped SNPs using TaqMan assays.

Results

We observed a significant association between rs2230926 and an increased risk of SLE and RA in the Japanese population (for SLE, odds ratio [OR] 1.92, 95% confidence interval [95% CI] 1.53–2.41, P = 1.9 × 10−8; for RA, OR 1.35, 95% CI 1.18–1.56, P = 2.6 × 10−5). The intergenic SNP rs10499194 was also associated with SLE and RA, while the risk allele for RA in Caucasians was protective against the diseases in our population.

Conclusion

We demonstrated a significant association between the nonsynonymous variant in TNFAIP3 and the risk for SLE and RA in the Japanese population. TNFAIP3, similar to STAT4 and IRF5, may be a common genetic risk factor for SLE and RA that is shared between the Caucasian and Japanese populations.
Proliferative remodeling of the spatial organization of human superficial chondrocytes distant from focal early osteoarthritis
Authors: Rolauffs Bernd , Williams James M. , Aurich Matthias , Grodzinsky Alan J. , Kuettner Klaus E. , Cole Ada A.
Published: January 2010

Objective

Human superficial chondrocytes show distinct spatial organizations, and they commonly aggregate near osteoarthritic (OA) fissures. The aim of this study was to determine whether remodeling or destruction of the spatial chondrocyte organization might occur at a distance from focal (early) lesions in patients with OA.

Methods

Samples of intact cartilage (condyles, patellofemoral groove, and proximal tibia) lying distant from focal lesions of OA in grade 2 joints were compared with location‐matched nondegenerative (grade 0–1) cartilage samples. Chondrocyte nuclei were stained with propidium iodide, examined by fluorescence microscopy, and the findings were recorded in a top‐down view. Chondrocyte arrangements were tested for randomness or significant grouping via point pattern analyses (Clark and Evans Aggregation Index) and were correlated with the OA grade and the surface cell densities.

Results

In grade 2 cartilage samples, superficial chondrocytes were situated in horizontal patterns, such as strings, clusters, pairs, and singles, comparable to the patterns in nondegenerative cartilage. In intact cartilage samples from grade 2 joints, the spatial organization included a novel pattern, consisting of chondrocytes that were aligned in 2 parallel lines, building double strings. These double strings correlated significantly with an increased number of chondrocytes per group and an increased corresponding superficial zone cell density. They were observed in all grade 2 condyles and some grade 2 tibiae, but never in grade 0–1 cartilage.

Conclusion

This study is the first to identify a distinct spatial reorganization of human superficial chondrocytes in response to distant early OA lesions, suggesting that proliferation had occurred distant from focal early OA lesions. This spatial reorganization may serve to recruit metabolically active units as an attempt to repair focal damage.
Blocking ERK‐1/2 reduces tumor necrosis factor α–induced interleukin‐18 bioactivity in rheumatoid arthritis synovial fibroblasts by induction of interleukin‐18 binding protein A
Authors: Marotte Hubert , Ahmed Salahuddin , Ruth Jeffrey H. , Koch Alisa E.
Published: January 2010

Objective

To examine the mechanism of regulation of interleukin‐18 (IL‐18) bioactivity by IL‐18 binding protein (IL‐18BP) induction.

Methods

Levels of IL‐18 and IL‐18BPa in synovial fluid samples from patients with osteoarthritis (OA) or rheumatoid arthritis (RA) were determined by enzyme‐linked immunosorbent assays (ELISAs), followed by calculation of free IL‐18. IL‐18 and IL‐18BPa synthesis in RA synovial fibroblasts that had been treated with proinflammatory and antiinflammatory cytokines were assessed by quantitative real‐time polymerase chain reaction and ELISA, respectively, followed by IL‐18 bioactivity determination using KG‐1 cells. Chemical signaling inhibitors were used for determination of the signal transduction pathways involved in IL‐18BPa/IL‐18 regulation. Tumor necrosis factor α (TNFα)–induced caspase 1 activity was determined by a colorimetric assay.

Results

IL‐18BPa was lower in RA synovial fluid than in OA synovial fluid (P < 0.05; n = 8), and free IL‐18 was higher in RA synovial fluid than in OA synovial fluid. TNFα induced RA synovial fibroblast IL‐18BPa and IL‐18 in a time‐dependent manner (P < 0.05). Evaluation of signaling pathways suggested that TNFα induced IL‐18 production through the ERK‐1/2, protein kinase Cδ (PKCδ), and Src pathways, whereas IL‐18BPa synthesis was mediated through the NFκB, PKC, Src, and JNK pathways. Furthermore, addition of exogenous IL‐18BPa‐Fc reduced the RA synovial fibroblast phosphorylation of ERK‐1/2 induced by TNFα.

Conclusion

These results suggest that IL‐18BPa reduces IL‐18 bioactivity induced by TNFα, by regulating the ERK‐1/2 pathway in RA synovial fibroblasts. Targeting IL‐18 bioactivity by induction or addition of IL‐18BPa may provide another therapeutic option in the management of RA.
Smoking increases rheumatoid arthritis susceptibility in individuals carrying the HLA–DRB1 shared epitope, regardless of rheumatoid factor or anti–cyclic citrullinated peptide antibody status
Authors: Bang So‐Young , Lee Kyoung‐Ho , Cho Soo‐Kyung , Lee Hye‐Soon , Lee Kyung Wha , Bae Sang‐Cheol
Published: January 2010

Objective

Smoking is associated with rheumatoid arthritis (RA) in individuals with the HLA–DRB1 shared epitope (SE). SE alleles have been shown to be predominantly associated with anti–cyclic citrullinated peptide (anti‐CCP)–positive RA. These risk factors have not been identified for anti‐CCP–negative RA. The aim of this study was to investigate whether SE‐containing HLA–DRB1 alleles, smoking, or the combination of these factors contributes to the development of RA, depending on the presence or absence of serologic markers, in a Korean population.

Methods

All of the patients with RA (n =1,482) and all of the control subjects (n = 1,119) were Korean. Four‐digit HLA–DRB1 typing was performed by a conventional polymerase chain reaction–sequence‐based typing method. Information about smoking history was obtained through a questionnaire. The patients with RA were tested for anti‐CCP antibodies and rheumatoid factor (RF).

Results

The SE alleles had significant effects on anti‐CCP antibody and RF formation. The DRB1*0901 allele was associated with the presence of anti‐CCP antibodies (odds ratio [OR] 2.49) and RF (OR 2.09). SE alleles and smoking were associated with both anti‐CCP–positive and anti‐CCP–negative RA. The combination of smoking and double copies of the SE allele increased the risk of anti‐CCP–positive RA 36.11‐fold and increased the risk of anti‐CCP–negative RA 12.29‐fold, compared with the risk among nonsmokers not carrying SE alleles. Interactions between SE alleles and smoking were observed for both anti‐CCP–positive and RF‐positive RA, although the associations of RF‐positive RA could be consequences of the underlying anti‐CCP antibody status.

Conclusion

We demonstrated that the combination of SE alleles and smoking is associated with RA susceptibility regardless of anti‐CCP antibody or RF status, but that the combination shows stronger effects in anti‐CCP–positive/RF‐positive patients with RA than in anti‐CCP–negative/RF‐negative patients with RA. The SE–smoking interactions were present in anti‐CCP–positive and RF‐positive RA.
Differential expression of protease‐activated receptors in monocytes from patients with primary antiphospholipid syndrome
Authors: López‐Pedrera Chary , Aguirre Maria Ángeles , Buendía Paula , Barbarroja Nuria , Ruiz‐Limón Patricia , Collantes‐Estevez Eduardo , Velasco Francisco , Khamashta Munther , Cuadrado Maria José
Published: January 2010

Objective

To investigate the expression of protease‐activated receptors (PARs), their potential regulation by anticardiolipin antibodies (aCL), and their association with the expression of other molecules relevant to thrombosis in monocytes obtained from 62 patients with primary antiphospholipid syndrome (APS).

Methods

Monocytes were isolated from peripheral blood mononuclear cells by magnetic depletion of nonmonocytes. Expression of tissue factor (TF) and PARs 1–4 genes was measured by quantitative real‐time reverse transcription–polymerase chain reaction. Cell surface TF and PARs 1–4 expression was analyzed by flow cytometry. For in vitro studies, purified normal monocytes were incubated with purified APS patient IgG, normal human serum IgG, or lipopolysaccharide, in the presence or absence of specific monoclonal antibodies anti–PAR‐1 (ATAP2) or anti–PAR‐2 (SAM11) to test the effect of blocking the active site of PAR‐1 or PAR‐2.

Results

Analysis of both mRNA and protein for the 4 PARs revealed significantly increased expression of PAR‐2 as compared with the control groups. PAR‐1 was significantly overexpressed in APS patients with thrombosis and in the control patients with thrombosis but without APS. PAR‐3 expression was not significantly altered. PAR‐4 expression was absent in all groups analyzed. In addition, we demonstrated a correlation between the levels of PAR‐2 and the titers of IgG aCL, as well as parallel behavior of TF and PAR‐2 expression. In vitro, IgG from APS patients significantly increased monocyte expression of PAR‐1 and PAR‐2. Inhibition studies suggested that there was direct cross‐talk between TF and PAR‐2, such that inhibition of PAR‐2 prevented the aCL‐induced expression of TF.

Conclusion

These results provide the first demonstration of increased expression of PARs in monocytes from patients with APS. Thus, PAR antagonists might have therapeutic potential as antithrombotic agents in APS.
Antimalarial treatment may have a time‐dependent effect on lupus survival: Data from a multinational Latin American inception cohort
Authors: Shinjo Samuel K. , Bonfá Eloísa , Wojdyla Daniel , Borba Eduardo F. , Ramirez Luis A. , Scherbarth Hugo R. , Brenol João C. Tavares , Chacón‐Diaz Rosa , Neira Oscar J. , Berbotto Guillermo A. , de la Torre Ignacio Garcia , Acevedo‐Vázquez Eduardo M. , Massardo Loreto , Barile‐Fabris Leonor A. , Caeiro Francisco , Silveira Luis H. , Sato Emilia I. , Buliubasich Sandra , Alarcón Graciela S. , Pons‐Estel Bernardo A.
Published: January 2010

Objective

To evaluate the beneficial effect of antimalarial treatment on lupus survival in a large, multiethnic, international longitudinal inception cohort.

Methods

Socioeconomic and demographic characteristics, clinical manifestations, classification criteria, laboratory findings, and treatment variables were examined in patients with systemic lupus erythematosus (SLE) from the Grupo Latino Americano de Estudio del Lupus Eritematoso (GLADEL) cohort. The diagnosis of SLE, according to the American College of Rheumatology criteria, was assessed within 2 years of cohort entry. Cause of death was classified as active disease, infection, cardiovascular complications, thrombosis, malignancy, or other cause. Patients were subdivided by antimalarial use, grouped according to those who had received antimalarial drugs for at least 6 consecutive months (user) and those who had received antimalarial drugs for <6 consecutive months or who had never received antimalarial drugs (nonuser).

Results

Of the 1,480 patients included in the GLADEL cohort, 1,141 (77%) were considered antimalarial users, with a mean duration of drug exposure of 48.5 months (range 6–98 months). Death occurred in 89 patients (6.0%). A lower mortality rate was observed in antimalarial users compared with nonusers (4.4% versus 11.5%; P< 0.001). Seventy patients (6.1%) had received antimalarial drugs for 6–11 months, 146 (12.8%) for 1–2 years, and 925 (81.1%) for >2 years. Mortality rates among users by duration of antimalarial treatment (per 1,000 person‐months of followup) were 3.85 (95% confidence interval [95% CI] 1.41–8.37), 2.7 (95% CI 1.41–4.76), and 0.54 (95% CI 0.37–0.77), respectively, while for nonusers, the mortality rate was 3.07 (95% CI 2.18–4.20) (P for trend < 0.001). After adjustment for potential confounders in a Cox regression model, antimalarial use was associated with a 38% reduction in the mortality rate (hazard ratio 0.62, 95% CI 0.39–0.99).

Conclusion

Antimalarial drugs were shown to have a protective effect, possibly in a time‐dependent manner, on SLE survival. These results suggest that the use of antimalarial treatment should be recommended for patients with lupus.
Association of the FAM167A–BLK region with systemic sclerosis
Authors: Ito Ikue , Kawaguchi Yasushi , Kawasaki Aya , Hasegawa Minoru , Ohashi Jun , Kawamoto Manabu , Fujimoto Manabu , Takehara Kazuhiko , Sato Shinichi , Hara Masako , Tsuchiya Naoyuki
Published: January 2010

Objective

An association of single‐nucleotide polymorphisms (SNPs) in the FAM167A (previously referred to as C8orf13)–BLK region with systemic lupus erythematosus (SLE) has been demonstrated in Caucasians and in Asians. Recent studies have shown that many genes, including IRF5, STAT4, and PTPN22, are shared susceptibility genes in multiple autoimmune diseases. We undertook the current study to examine whether the FAM167A–BLK region is also associated with susceptibility to systemic sclerosis (SSc).

Methods

Japanese patients with SSc (n = 309) and healthy controls (n = 769) were enrolled in a 2‐tiered case–control association study. In tier 1, 124 patients and 412 controls were tested to determine association of 16 tag SNPs encompassing the FAM167A–BLK region with SSc. In tier 2, an additional 185 patients and 357 controls were analyzed for SNP rs13277113.

Results

Two haplotype blocks that correspond approximately to FAM167A and BLK were observed. In tier 1 of the study, the rs13277113A allele in the BLK block exhibited the most significant association with SSc after correction for multiple testing (permutated P = 0.024). Two SNP haplotypes formed by rs13277113 and the most significant SNP in the FAM167A block did not exhibit stronger association. When samples from tier 1 and tier 2 were combined, the rs13277113A allele was significantly associated with SSc (odds ratio 1.45 [95% confidence interval 1.17–1.79], P = 6.1 × 10−4). Association or a tendency toward association of rs13277113A with SSc was observed regardless of a patient's autoantibody profile or whether a patient had diffuse cutaneous or limited cutaneous SSc.

Conclusion

Our findings indicate that the rs13277113A allele is associated not only with SLE but also with SSc and that the FAM167A–BLK region is a common genetic risk factor for both SLE and SSc.
Evaluating the effects of reactivity to supervisor presence on staff response to tactile prompts and self‐monitoring in a group home setting
Authors: Mowery Judy M. , Miltenberger Raymond G. , Weil Timothy M.
Published: January 2010

Because staff management research in group home settings typically involves direct observation of staff performance, reactivity to being observed is a potential limitation in these studies. The current study evaluated the use of a tactile prompt, self‐monitoring, and feedback to increase positive interaction in a group home setting. Reactivity was assessed by having a confederate observe staff positive interaction when the supervisor was present and when the supervisor was absent. The effect of supervisor presence was evaluated using an alternating treatment within a multiple baseline across participants' research design. Results showed that two of four participants increased positive interactions only when a supervisor was present and the two other participants increased positive interaction only when a supervisor was present and provided feedback. Copyright © 2010 John Wiley & Sons, Ltd.
The Role of Flexibility in the Rational Design of Modularly Assembled Ligands Targeting the RNAs that Cause the Myotonic Dystrophies
Authors: Disney Matthew D. , Lee Melissa M. , Pushechnikov Alexei , Childs‐Disney Jessica L.
Published: January 2010

Modularly assembled ligands were designed to target the RNAs that cause two currently untreatable neuromuscular disorders, myotonic dystrophy types 1 (DM1) and 2 (DM2). DM1 is caused by an expanded repeating sequence of CUG, and DM2 is caused by expanded CCUG repeats. Both are present in noncoding regions and fold into hairpins with either repeating 1×1 nucleotide UU (DM1) or 2×2 nucleotide 5′‐CU/3′‐UC (DM2) internal loops separated by two GC pairs. The repeats are toxic because they sequester the RNA splicing regulator muscleblind‐like 1 protein (MBNL1). Rational design of ligands targeting these RNAs was enabled by a database of RNA motif–ligand partners compiled by using two‐dimensional combinatorial screening (2DCS). One 2DCS study found that the 6′′‐azido‐kanamycin A module binds internal loops similar to those found in DM1 and DM2. In order to further enhance affinity and specificity, the ligand was assembled on a peptoid backbone to precisely control valency and the distance between ligand modules. Designed compounds are more potent and specific binders to the toxic RNAs than MBNL1 and inhibit the formation of the RNA–protein complexes with nanomolar IC50 values. This study shows that three important factors govern potent inhibition: 1) the surface area sequestered by the assembled ligands; 2) the spacing between ligand modules since a longer distance is required to target DM2 RNAs than DM1 RNAs; and 3) flexibility in the modular assembly scaffold used to display the RNA‐binding module. These results have impacts on the general design of assembled ligands targeting RNAs present in genomic sequence.
Copper‐Catalysed Ring Opening of Polycyclic meso‐Hydrazines with Trialkylaluminium Reagents and SimplePhos Ligands
Authors: Palais Laëtitia , Bournaud Chloée , Micouin Laurent , Alexakis Alexandre
Published: January 2010

meso‐Hydrazines could be desymmetrised by ring‐opening reactions by using various metal catalysts to form substituted cyclopentenes, which have a high synthetic potential. Herein, we report the asymmetric copper‐catalysed ring opening of a range of polycyclic hydrazines with trialkylaluminium reagents and SimplePhos ligands with both high isolated yields and enantioselectivities of up to 95 %.