Plasmacytoid dendritic cells (pDCs) are innate sensors that produce IFN-α in response to viral infections. Determining how aging alters the cellular and molecular function of these cells may provide an explanation of increased susceptibility of older people to viral infections. Hence, we examined whether aging critically impairs pDC function during infection with HSV-2, a viral pathogen that activates TLR9. We found that impaired IFN-α production by aged murine pDCs led to impaired viral clearance with aging. Upon TLR9 activation, aged pDCs displayed defective up-regulation of IFN-regulatory factor 7, a key adaptor in the type I IFN pathway, as compared with younger counterparts. Aged pDCs had more oxidative stress, and reducing oxidative stress in aged pDCs partly recovered the age-induced IFN-α defect during TLR9 activation. In sum, aging impairs the type I IFN pathway in pDCs, and this alteration may contribute to the increased susceptibility of older people to certain viral infections.

Recent evidence supports a role of Toll-like receptor (TLR) signaling in the development of atherosclerotic lesions. In this study, we tested whether TLR4 signaling promotes a proinflammatory phenotype in human and mouse arterial smooth muscle cells (SMC), characterized by increased cytokine and chemokine synthesis and increased TLR expression. Human arterial SMC were found to express mRNA encoding TLR4 and the TLR4-associated molecules MD-2 and CD14 but not TLR2 mRNA. Mouse aortic SMC, on the other hand, expressed both TLR2 and TLR4 mRNA constitutively. Human SMC derived from the coronary artery, but not those from the pulmonary artery, were found to express cell surface-associated CD14. Low concentrations (ng/ml) of Escherichia coli LPS, the prototypical TLR4 agonist, markedly stimulated extracellular regulated kinase 1/2 (ERK1/2) activity, induced release of monocyte-chemoattractant protein-1 (MCP-1) and interleukin (IL)-6, and stimulated IL-1alpha expression in human aortic SMC, and exogenous CD14 enhanced these effects. Expression of a dominant negative form of TLR4 in human SMC attenuated LPS-induced ERK1/2 and MCP-1 release. LPS was a potent inducer of NF-kappaB activity, ERK1/2 phosphorylation, MCP-1 release, and TLR2 mRNA expression in wild-type mice but not in TLR4-signaling deficient mouse aortic SMC. These studies show that TLR4 signaling promotes a proinflammatory phenotype in vascular smooth muscle cells (VSMC) and suggest that VSMC may potentially play an active role in vascular inflammation via the release of chemokines, proinflammatory cytokines, and increased expression of TLR2.

Pathogen associated molecular patterns (PAMPs) are signals detected by plants that activate basal defenses. One of these PAMPs is chitin, a carbohydrate present in the cell walls of fungi and in insect exoskeletons. Previous work has shown that chitin treatment of Arabidopsis thaliana induced defense-related genes in the absence of a pathogen and that the response was independent of the salicylic acid (SA), jasmonic acid (JA) and ethylene (ET) signaling pathways. One of these genes is ATL9 ( = ATL2G), which encodes a RING zinc-finger like protein. In the current work we demonstrate that ATL9 has E3 ubiquitin ligase activity and is localized to the endoplasmic reticulum. The expression pattern of ATL9 is positively correlated with basal defense responses against Golovinomyces cichoracearum, a biotrophic fungal pathogen. The basal levels of expression and the induction of ATL9 by chitin, in wild type plants, depends on the activity of NADPH oxidases suggesting that chitin-mediated defense response is NADPH oxidase dependent. Although ATL9 expression is not induced by treatment with known defense hormones (SA, JA or ET), full expression in response to chitin is compromised slightly in mutants where ET- or SA-dependent signaling is suppressed. Microarray analysis of the atl9 mutant revealed candidate genes that appear to act downstream of ATL9 in chitin-mediated defenses. These results hint at the complexity of chitin-mediated signaling and the potential interplay between elicitor-mediated signaling, signaling via known defense pathways and the oxidative burst.

ObjectivesTo determine whether the associations with key risk factors in patients with diagnosed and undiagnosed type 2 diabetes mellitus (T2DM) are different using data from the National Health and Nutrition Examination Survey (NHANES) from 1999 to 2010.MethodsThe study analysed the prevalence and association with risk factors of undiagnosed and diagnosed T2DM using a regression model and a multinomial logistic regression model. Data from the NHANES 1999–2010 were used for the analyses.ResultsThe study analysed data from 10 570 individuals. The overall prevalence of diagnosed and undiagnosed T2DM increased significantly from 1999 to 2010. The prevalence of undiagnosed T2DM was significantly higher in non-Hispanic whites, in individuals <30 years old and in those with near optimal (130–159 mg/dl) or very high (≥220 mg/dl) non-high-density lipoprotein cholesterol levels compared with diagnosed T2DM. Body mass index, low economic status or low educational level had no effect on T2DM diagnosis rates. Though diagnosed T2DM was associated with favourable diet/carbohydrate intake behavioural changes, it had no effect on physical activity levels.ConclusionThe overall T2DM prevalence increased between 1999 and 2010, particularly for undiagnosed T2DM in patients that were formerly classified as low risk.

Objective-The intracellular bacterium Chlamydia pneumoniae is present in many atherosclerotic lesions, where it could promote inflammation. This study determined whether monocyte chemoattractant protein 1 (MCP-1) release is stimulated in vascular smooth muscle cells (VSMCs) that are exposed to or infected by C pneumoniae and whether toll-like receptor 2 (TLR2) or TLR4 mediate these effects. Methods and Results-TLR2 mRNA was expressed constitutively and was upregulated by C pneumoniae exposure in mouse aortic SMC and was inducible by C pneumoniae and TLR3 and TLR4 agonists in human coronary artery SMCs. Exposure to inactivated or viable extracellular C pneumoniae evoked a robust increase in MCP-1 release and activated nuclear factor-B and extracellular signal-regulated kinase 1/2 in wild-type and TLR4 signaling-deficient mouse aortic SMCs but not in TLR2-deficient SMCs, probably because of TLR2-mediated recognition of a chlamydial antigen. Brief exposure to viable C pneumoniae led to active infection of VSMCs, shown by chlamydial protein synthesis, and caused a persistent (Ͼ48-hour) MCP-1 release that was also TLR2 dependent. Conclusions-The results show that VSMCs express functional TLR2 and that TLR2 mediates both a persistent activation of chemokine release in C pneumoniae-infected VSMCs and its acute stimulation by extracellular C pneumoniae. Therefore, TLR2 expressed in VSMCs may promote inflammation within the arterial wall. Key Words: monocyte chemoattractant protein-1 Ⅲ nuclear factor-B Ⅲ extracellular signal-regulated kinase 1/2 Ⅲ heat shock protein 60 Ⅲ lipopolysaccharide I nflammation is a key component of atherosclerosis, the earliest stages of which are characterized by invasion of the intima by mononuclear phagocytes. 1 Growing evidence indicates that infectious agents, including Chlamydia pneumoniae, a Gram-negative obligate intracellular bacterium, may promote arterial inflammation. C pneumoniae is frequently found in atherosclerotic lesions residing within vascular smooth muscle cells (VSMCs) and endothelial cells as well as in macrophages, but it is rare or absent in normal arteries. [2][3][4][5][6] In animal models of respiratory infection, the organism localizes to the arterial wall and promotes atherogenesis by acting in concert with traditional risk factors such as hyperlipidemia, 7-13 raising the possibility that C pneumoniae may affect vascular cell phenotype and function directly. In vitro studies support the hypothesis that C pneumoniae can promote cellular changes similar to those occurring during atherogenesis, including increased lipid accumulation in macrophages 14 and proliferation of human VSMCs. 15 Despite substantial evidence supporting a pathogenic role of C pneumoniae in atherosclerosis, little is known about the molecular mechanisms by which it may alter vascular cell function.Effective defense against invading microbes requires a sensitive pathogen recognition system. The toll-like receptor (TLR) family includes at least 10 transmembrane pathogenrecognition receptors, which a...

Inflammation plays a key role in atherogenesis, perhaps promoted by bacterial and viral products present within the artery wall. Vascular smooth muscle cells (VSMC) can express certain bacterially responsive Toll-like receptors (TLR), which promote a proinflammatory and proliferative VSMC phenotype when activated, but it is unknown whether virally activated TLR can regulate VSMC phenotype. Here we tested the role in VSMC of TLR3, which is activated by double-stranded (dsRNA), a molecular signature of viruses. VSMC from multiple vessel types, including human coronary artery (HCoASMC) and mouse aorta (MAoSMC), expressed TLR3 constitutively, and HCoASMC were exquisitely sensitive to dsRNA-stimulated release of monocyte chemoattractant protein-1 (MCP-1) and interleukin-6. dsRNA-induced MCP-1 release was abolished by small interfering RNA-mediated TLR3 knockdown in HCoASMC and was absent in TLR3-/- MAoSMC but was unimpaired in TLR2-/- and in TLR4 signaling-deficient MAoSMC. Exposure to dsRNA also activated ERK1/2 and NF-kappaB in both human and murine SMC, but these effects were absent in SMC from TLR3-deficient mice, demonstrating a crucial role of TLR3 signaling. dsRNA also stimulated proliferation of HCoASMC, indicated by increased DNA synthesis, and induced persistent elevations in the intracellular levels of growth-promoting mediators, including interleukin-1alpha and phospho-ERK1/2. We conclude that exposure of HCoASMC to dsRNA elicits dramatic TLR3-mediated proinflammatory and proproliferative phenotypic changes, responses that could potentially be triggered by viral infection of cells within the arterial wall.

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Background We examined the effects of integrated cognitive-behavioral therapy for depression and insomnia (CBT-D + CBT-I) delivered via videoconferening in rural, middle aged and older adults with depressive and insomnia symptoms. Method Forty patients with depressive and insomnia symptoms were randomized to receive either 10 sessions of CBT-D + CBT-I or usual care (UC). Patients in the integrated CBT condition were engaged in telehealth treatment through Skype at their primary care clinic. Assessments were conducted at baseline, post-treatment, and 3-month follow-up. Results CBT-D +CBT-I participants had significantly greater improvements in sleep at post-treatment and 3-month follow-up as compared to the UC participants. The time by group interaction for depression was not significant; both the CBT-D + CBT-I and UC conditions had a decrease in depressive symptoms over time. Conclusion While integrated CBT benefits both depression and insomnia symptoms, its effects on depression are more equivocal. Further research should consider expanding the depression treatment component of integrated CBT to enhance effectiveness.