Preconception exposure to EtOH through the paternal route may affect neurobehavioral and developmental features of offspring. This study investigates the effects of paternal exposure to EtOH before conception on the hyperactivity, inattention, and impulsivity behavior of male offspring in mice. Sire mice were treated with EtOH in a concentration range approximating human binge drinking (0-4 g/kg/day EtOH) for 7 weeks and mated with untreated females mice to produce offspring. EtOH exposure to sire mice induced attention deficit hyperactivity disorder (ADHD)-like hyperactive, inattentive, and impulsive behaviors in offspring. As a mechanistic link, both protein and mRNA expression of dopamine transporter (DAT), a key determinant of ADHD-like phenotypes in experimental animals and humans, were significantly decreased by paternal EtOH exposure in cerebral cortex and striatum of offspring mice along with increased methylation of a CpG region of the DAT gene promoter. The increase in methylation of DAT gene promoter was also observed in the sperm of sire mice, suggesting germline changes in the epigenetic methylation signature of DAT gene by EtOH exposure. In addition, the expression of two key regulators of methylation-dependent epigenetic regulation of functional gene expression, namely, MeCP2 and DNMT1, was markedly decreased in offspring cortex and striatum sired by EtOH-exposed mice. These results suggest that preconceptional exposure to EtOH through the paternal route induces behavioral changes in offspring, possibly via epigenetic changes in gene expression, which is essential for the regulation of ADHD-like behaviors.

We verified our findings using a mouse model, primary human hepatocytes and human liver tissues. Our data elucidate a mechanism by which HBV evades the host innate immune system.

Recent studies indicate that Toll-like receptors (TLRs), originally identified as infectious agent receptors, also mediate sterile inflammatory responses during tissue damage. In this study, we investigated the role of TLR2 in excitotoxic hippocampal cell death using TLR2 knock-out (KO) mice. TLR2 expression was up-regulated in microglia in the ipsilateral hippocampus of kainic acid (KA)-injected mice. KA-mediated hippocampal cell death was significantly reduced in TLR2 KO mice compared with wild-type (WT) mice. Similarly, KA-induced glial activation and proinflammatory gene expression in the hippocampus were compromised in TLR2 KO mice. In addition, neurons in organotypic hippocampal slice cultures (OHSCs) from TLR2 KO mouse brains were less susceptible to KA excitotoxicity than WT OHSCs. This protection is partly attributed to decreased expression of proinflammatory genes, such as TNF-␣ and IL-1␤ in TLR2 KO mice OHSCs. These data demonstrate conclusively that TLR2 signaling in microglia contributes to KA-mediated innate immune responses and hippocampal excitotoxicity.Toll-like receptors (TLRs) 3 are a group of transmembrane proteins that play a central role in innate immune responses.To date, more than 10 different TLR members have been identified that each recognizes a specific set of pathogen-associated molecular patterns (PAMPs) expressed by microorganisms (1, 2). Interestingly, emerging data indicate that TLRs function as receptors not only for PAMPs, but also for endogenous molecules released from damaged tissue or cells. For example, TLR2 and -4 recognize various endogenous molecules including heat shock proteins, hyaluronan, and high mobility group box-1 (HMGB-1) (3-6). In addition, TLR3 binds mRNA released from necrotic cells (7). It is possible that TLR recognition of these endogenous molecules is involved in the inflammatory response during "sterile" tissue damage.In the central nervous system (CNS), TLRs including TLR2, -3, and -4 are expressed in microglia and astrocytes, suggesting a role as innate immune cells in the CNS (8). Based on their function as receptors for "danger signals" (9), TLR expression in glial cells is implicated in various "sterile" neurological disorders including mouse cerebral ischemia/reperfusion injury (10, 11), spinal cord injury (12), and axonal transection (13). We have also reported that TLR2 plays a critical role in nerve injury-induced spinal cord glial activation and subsequent pain hypersensitivity (14), and traumatic brain injury (15). In mouse epileptic seizure model, TLR2 transcripts are up-regulated in hippocampal microglia/ macrophages upon pilocarpine injection (16) implicating TLR2 in hippocampal excitotoxicity, although this possibility has not yet been explored.Excitotoxicity is an underlying mechanism of various neurological disorders including traumatic brain injury and stroke, and is also implicated in chronic neurodegenerative diseases such as amyotrophic lateral sclerosis and epileptic seizure (17). NMDA and AMPA/kainate glutamate receptor overstimu...

Objective. Interleukin-32 (IL-32) is a recently discovered cytokine that appears to play a critical role in human rheumatoid arthritis (RA). It is highly expressed in synovium and fibroblast-like synoviocytes (FLS) from RA patients, but not in patients with osteoarthritis (OA). This study was undertaken to assess IL-32 levels in RA synovial fluid (SF) and to investigate the secretion and regulation of IL-32 in RA FLS.Methods. FLS and SF were obtained from the joints of RA patients. The secretion and expression of IL-32 and activation of signaling molecules were examined by enzyme-linked immunosorbent assay, immunoblotting, immunoprecipitation, reverse transcriptasepolymerase chain reaction, and small interfering RNA (siRNA) transfection.Results. IL-32 levels were high in RA SF compared with OA SF. Furthermore, RA FLS expressed and secreted IL-32 when stimulated with tumor necrosis factor ␣ (TNF␣). TNF␣-induced expression of IL-32 was significantly suppressed, in a dose-dependent manner, by inhibitors of Syk, protein kinase C␦ (PKC␦), and JNK and by knockdown of these kinases and c-Jun with siRNA. We also observed that PKC␦ mediated the activation of JNK and c-Jun, and experiments using specific inhibitors and siRNA demonstrated that Syk was the upstream kinase for the activation of PKC␦.Conclusion. The present findings suggest that IL-32 may be a newly identified prognostic biomarker in RA, thereby adding valuable knowledge to the understanding of this disease. The results also demonstrate that the production of IL-32 in RA FLS is regulated by Syk/PKC␦-mediated signaling events.

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