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Asthma is a chronic eosinophilic inflammatory disease with an increasing prevalence worldwide. Endocannabinoids are known to have immunomodulatory biological effects. However, the contribution of oleoylethanolamide (OEA) to airway inflammation remains to be elucidated. To investigate the effect of OEA, the expression of proinflammatory cytokines was measured by RT-qPCR and ELISA in airway epithelial (A549) cells. The numbers of airway inflammatory cells and cytokine levels in bronchoalveolar lavage fluid, airway hyperresponsiveness, and type 2 innate lymphoid cells (ILC2s) were examined in BALB/c mice after 4 days of OEA treatment. Furthermore, eosinophil activation after OEA treatment was evaluated by measuring cellular CD69 levels in eosinophils from human peripheral eosinophils using flow cytometry. OEA induced type 2 inflammatory responses in vitro and in vivo. OEA increased the levels of proinflammatory cytokines, such as IL-6, IL-8, and IL-33, in A549 cells. In addition, it also induced eosinophilic inflammation, the production of IL-4, IL-5, IL-13, and IL-33 in bronchoalveolar lavage fluid, and airway hyperresponsiveness. OEA increased the numbers of IL-5- or IL-13-producing ILC2s in a mouse model. Finally, we confirmed that OEA increased CD69 expression (an eosinophil activation marker) on purified eosinophils from patients with asthma compared to those from healthy controls. OEA may play a role in the pathogenesis of asthma by activating ILC2s and eosinophils., Asthma: A physiological cannabinoid promotes inflammation Oleoylethanolamide (OEA), an endocannabinoid similar to those found in cannabis, occurs naturally in the body, and has now been implicated in promoting symptoms of asthma. OEA activates white blood cells called eosinophils and ILC2 cells, which are both parts of our immune response. Researchers in South Korea, led by Ga-Young Ban at Kangdong Sacred Heart Hospital, Hallym University College of Medicine, Seoul, studied the effect of OEA on respiratory system cells from people with asthma and in mice used to model the condition in humans. OEA-induced inflammatory responses associated with asthma by stimulating the release of signaling molecules called cytokines. However, some other endocannabinoids have anti-inflammatory effects. Studying the effects of all endocannabinoids could yield new insights into asthma, with this study opening new possibilities for treating asthma by interfering with OEA-induced signaling responses.

Background Dysregulation of the arachidonic acid metabolic pathway is the most widely known pathomechanism of AERD. We performed integrative analysis of transcriptomic and epigenomic profiling with network analysis to determine the novel pathogenic features of AERD. Methods Ten patients with asthma including 5 patients with AERD and another 5 patients with aspirin tolerant asthma (ATA) were enrolled. Nasal scraping was performed and nasal mucosa was used in omics profiling. Peripheral eosinophil counts, sputum eosinophil counts, FeNO levels, and pulmonary function test results were evaluated. Differentially expressed genes (DEGs), differentially methylated probes (DMPs) and differentially correlated genes (DCGs) between patients with AERD and those with ATA were analyzed. Network analysis using Ingenuity Pathway Analysis (IPA) was performed to determine the gene connection network and signaling pathways. Results In total, 1,736 DEGs, 1,401 DMPs, and 19 pairs for DCGs were identified. Among DCGs, genes related to vesicle transport (e.g. RAB3B and STX2) and sphingolipid dysregulation (e.g. SMPD3) were found to be hypo-methylated and up-regulated in AERD. Using the canonical pathway analysis of IPA with 78 asthma-related DEGs, signaling pathways of T helper cell differentiation/activation and Fcε receptor I were generated. Up-regulation of RORγt and FCER1A were noted in AERD. Gene expression levels of RAB3B, SYNE1, STX2, SMPD3 and RORγt significantly affected sputum eosinophil counts. Quantitative real-time PCR was performed and mRNA expression levels of STX2, SMPD3, and RORγt were significantly higher in AERD compared to ATA. Conclusions Distinct pathogenic features were identified by using integrative multi-omics data analysis in patients with AERD.

OBJECTIVES To evaluate the effectiveness of a three-dimensional (3D) printed transparent kidney model as a surgical navigator for robot-assisted partial nephrectomy (RPN) in patients with complex renal tumours, defined by a R.E.N.A.L. (Radius, Exophytic/Endophytic, Nearness, Anterior/Posterior, Location) nephrometry score of ≥7. PATIENTS AND METHODS A total of 80 patients who underwent RPN were included in the present prospective case-matched study (case group [n = 40, application of 3D-printed transparent kidney model during RPN] vs matching group [n = 40, routine protocol]). The RPNs were performed by a single experienced surgeon. The RPN procedure consisted of six steps: (i) preparation of the renal hilar vessel for clamping, (ii) tumour detection and dissection, (iii) robotic ultrasonography, (iv) tumour resection, (v) calyx repair and haemostasis, and (vi) renorrhaphy. The time for each step, console time, and warm ischaemia time were compared between the two groups as a surrogate marker for surgical effectiveness. RESULTS Both groups were well-balanced for all baseline characteristics. The use of the model reduced the console time by ~20% compared to the matched group (64.6 vs 78.5 min, P = 0.001). On multivariate logistic regression analysis, tumour radius (P

Triterpenes, consisting of six isoprene units, are one of the largest classes of natural compounds in plants. The genus Taraxacum is in the family Asteraceae and is widely distributed in the Northern Hemisphere. Various triterpenes, especially taraxerol and taraxasterol, are present in Taraxacum plants. Triterpene biosynthesis occurs through the action of oxidosqualene cyclase (OSC), which generates various types of triterpenes from 2,3-oxidosqualene after the rearrangement of the triterpene skeleton. However, no functional characterization of the OSC genes involved in triterpene biosynthesis, except for a lupeol synthase in Taraxacum officinale, has been performed. Taraxacum coreanum, or Korean dandelion, grows in Korea and China. Putative OSC genes in T. coreanum plants were isolated by transcriptome analysis, and four of these (TcOSC1, TcOSC2, TcOSC3 and TcOSC4) were functionally characterized by heterologous expression in yeast. Both TcOSC1 and TcOSC2 were closely related to dammarenediol-II synthases. TcOSC3 and TcOSC4 were strongly grouped with β-amyrin synthases. Functional analysis revealed that TcOSC1 produced several triterpenes, including taraxasterol; Ψ-taraxasterol; α-, β- and δ-amyrin; and dammarenediol-II. TcOSC2 catalyzed the production of bauerenol and another unknown triterpene, TcOSC3 catalyzed the production of β-amyrin. TcOSC4 catalyzed the production of taraxerol. Moreover, we identified taraxasterol, ψ-taraxasterol, taraxerol, lupeol, δ-amyrin, α-amyrin, β-amyrin and bauerenol in the roots and leaves of T. coreanum. Our results suggest that TcOSC1, TcOSC2, TcOSC3 and TcOSC4 are key triterpene biosynthetic enzymes in T. coreanum. These enzymes are novel triterpene synthases involved in the production of taraxasterol, bauerenol and taraxerol.

Long-term exposure to ethanol simultaneously causes gastrointestinal inflammation, liver injury, and steatosis. In the present study, we investigated the effects of Bifidobacterium longum LC67, Lactobacillus plantarum LC27, and their mixture (LM) against ethanol-induced steatosis in mice. Exposure to ethanol caused liver damage: it increased ALT, AST, TG, TC, and lipopolysaccharide levels in the blood and induced NF-κB activation in the liver. Oral administration of LC27, LC67, or LM in mice reduced ethanol-induced ALT, AST, TG, and TC levels in the blood and liver. These also suppressed ethanol-induced NF-κB activation and α-smooth muscle actin expression in the liver and increased ethanol-suppressed AMPK activation. Treatment with LC27, LC67, or LM increased ethanol-suppressed alcohol dehydrogenase and acetaldehyde dehydrogenase activities in the liver, as well as tight junction protein expression in the liver and colon. Moreover, treatment with LC27, LC67, or LM restored the ethanol-disturbed gut microbiota composition, such as the increased population of Proteobacteria, and inhibited fecal and blood lipopolysaccharide levels. These inhibited NF-κB activation and increased tight junction protein expression in ethanol- or lipopolysaccharide-stimulated Caco-2 cells. These findings suggest that LC27, LC67, and LM can alleviate alcoholic steatosis by inhibiting LPS-mediated NF-κB activation through restoration of the disturbed gut microbiota.

Assessing the model fit quality of statistical models for network data is an ongoing and under-examined topic in statistical network analysis. Traditional metrics for evaluating model fit on tabular data such as the Bayesian Information Criterion are not suitable for models specialized for network data. We propose a novel self-developed goodness of fit (GOF) measure, the `stratified-sampling cross-validation' (SCV) metric, that uses a procedure similar to traditional cross-validation via stratified-sampling to select dyads in the network's adjacency matrix to be removed. SCV is capable of intuitively expressing different models' ability to predict on missing dyads. Using SCV on real-world social networks, we identify the appropriate statistical models for different network structures and generalize such patterns. In particular, we focus on conditionally independent dyad (CID) models such as the Erdos Renyi model, the stochastic block model, the sender-receiver model, and the latent space model.

Type I interferons (IFNs) play an important role in antiviral immunity as well as immunopathogenesis of diverse chronic viral infections. However, the precise mechanisms regulating the multifaceted effects of type I IFNs on the immune system and pathological inflammation still remain unclear. In order to assess the immunological dynamics associated with rapid viral clearance in chronic hepatitis C patients during the acute phase of type I IFN therapy, we analyzed multiple parameters of virological and immunological responses in a cohort of 59 Korean hepatitis C patients who received pegylated IFN-alpha and ribavirin (IFN/RBV). Most of the Korean patients had favorable alleles in the IFN-lambda loci for responsiveness to IFN/RBV (i.e., C/C in rs12979860, T/T in rs8099917, and TT/TT in rs368234815). Rapid virological response (RVR) was determined mainly by the hepatitis C virus genotype. Among the cytokines analyzed, higher plasma levels of IL-17A and FGF were observed in non-RVR patients infected with viral genotype 1 and IP-10 was consistently elevated in RVR group infected with genotype 2 during the early phase of antiviral therapy. In addition, these three cytokines were correlated each other, suggesting a functional linkage of the cytokines in antiviral responses during IFN/RBV therapy. A low baseline frequencies of regulatory T cells and.d T cells, but high level of group 2 innate lymphoid cells, in peripheral bloods were also significantly associated with the RVR group, implicating a potential role of the cellular immunity during the early phase of IFN/RBV therapy. Therefore, the immunological programs established by chronic hepatitis C and rapid disruption of the delicate balance by exogenous type I IFN might be associated with the subsequent virological outcomes in chronic hepatitis C patients.

We demonstrate a novel route to synthesize Fe3O4–CdSe/ZnS multifunctional nanoclusters (MNCs) with excellent optical and magnetic properties and biocompatibility. The successful fabrication of highly fluorescent and magnetic MNCs is achieved via a coupling process based on a partial ligand exchange reaction at the aqueous–organic solution interface. In addition, we show that dendritic cells (DCs), the sentinel of the immune system, can uptake the MNCs without significant change in cell viability. The MNCs uptaken by the DCs can be used for imaging, tracking, and separating the DCs. Furthermore, the MNCs can be loaded with a pathogen-associated molecular pattern, lipid A, via a hydrophobic–hydrophobic interaction. Ex vivo labeling of DCs with the MNC–lipid A complex enhances the DC migration to draining lymph nodes and tumor antigen-specific T cell responses in vivo. Our work may contribute to the development of synthetic routes to various multifunctional nanoclusters and DC-based cancer immunotherapies.

Members of the herpesviral family use multiple strategies to hijack infected host cells and exploit cellular signaling for their pathogenesis and latent infection. Among the most intriguing weapons in the arsenal of pathogenic herpesviruses are the constitutively active virally-encoded G protein-coupled receptors (vGPCRs). Even though vGPCRs contribute to viral pathogenesis such as immune evasion and proliferative disorders, the molecular details of how vGPCRs continuously activate cellular signaling are largely unknown. Here, we report that the vGPCR of Herpesvirus saimiri (HVS), an oncogenic γ2-herpesvirus, constitutively activates T cells via a heteromeric interaction with cellular CXCR4. Constitutive T cell activation also occurs with expression of the vGPCR of Kaposi's sarcoma-associated herpesvirus (KSHV), but not the vGPCR of Epstein-Barr virus. Expression of HVS vGPCR down-regulated the surface expression of CXCR4 but did not induce the degradation of the chemokine receptor, suggesting that vGPCR/CXCR4 signaling continues in cytosolic compartments. The physical association of vGPCR with CXCR4 was demonstrated by proximity ligation assay as well as immunoprecipitation. Interestingly, the constitutive activation of T cells by HVS vGPCR is independent of proximal T cell receptor (TCR) signaling molecules, such as TCRβ, Lck, and ZAP70, whereas CXCR4 silencing by shRNA abolished T cell activation by vGPCRs of HVS and KSHV. Furthermore, previously identified inactive vGPCR mutants failed to interact with CXCR4. These findings on the positive cooperativity of vGPCR with cellular CXCR4 in T cell activation extend our current understanding of the molecular mechanisms of vGPCR function and highlight the importance of heteromerization for GPCR activity.

Mucopolysaccharidosis type VI (MPS VI; Maroteaux-Lamy syndrome, OMIM #253200) is a rare disorder involving multiple organs and manifested particularly by severe skeletal abnormalities. Bone marrow transplantation (BMT) improves cardiopulmonary function and facial features, but has limited success in ameliorating skeletal abnormalities and short stature. Here, we report the outcome of enzyme replacement therapy (ERT) with recombinant human arylsulfatase-B (ASB, Naglazyme, BioMarin, Novato, CA) in an MPS VI patient who received BMT 10 years prior to ERT induction. Administration of weekly Naglazyme for 18 months was effective in improving range of motion in several joints [shoulders (improvement of flexion (Right/Left): 40°/55°; improvement of extension 30°/40°; improvement of abduction 10°/10°), elbows (improvement of flexion 25°/25°; improvement of extension 10°/15°), hips (improvement of flexion 25°/10°), and knees (improvement of flexion 45°/40°; improvement of extension 50°/60°)]. Improvement in the outcome of the 12-min walk test (70% increase) and 3-min stair-climbing test (29% increase) was also noted after ERT. Because ERT improved clinical features in an MPS VI patient who had undergone prior BMT, the role of ERT post successful BMT in MPS VI needs further investigation.

Context: Prader-Willi syndrome (PWS), a genetic disorder characterized by obesity in early childhood, is reported to have elevated levels of adiponectin. The effects of adiponectin are mediated by adiponectin receptors (ADIPORs) that include ADIPOR1 and ADIPOR2. There is evidence that several cytokines, including adiponectin, TNF-α, and IL-6, are involved in insulin sensitivity. Objective and Methods: We measured the relative expression of adiponectin, ADIPORs, several proinflammatory cytokines including TNF-α, and IL-6 expression in peripheral blood mononuclear cells (PBMCs) of children with PWS and obese comparators using real-time PCR. Their correlation with homeostasis model assessment insulin resistance index (HOMA-IR) was analyzed. Patients: Thirty children with PWS (median age 7.1 yr, 18 males, 12 females) that were being treated with GH and 32 obese children not receiving GH treatment (median age 9.1 yr, 15 males, 17 females) for comparison were enrolled. Results: The PWS children had increased expression of ADIPOR2 (P = 0.02) and decreased expression of IL-6 (P = 0.03) compared with the comparison group. Moreover, there was a significant positive correlation between the ADIPORs and TNF-α (ADIPOR1 vs. TNF-α: r = 0.66, P < 0.001 in PWS, r = 0.80, P < 0.001 in comparison group; ADIPOR2 vs. TNF-α: r = 0.69, P < 0.001 in comparison group). The ADIPORs in the comparison group showed significant negative correlation with HOMA-IR (ADIPOR1 vs. HOMA-IR; ρ = −0.41, P = 0.02, ADIPOR2 vs. HOMA-IR; ρ = −0.46, P < 0.01). Conclusion: The results of this study showed that inflammatory cytokine expression was closely associated with the expression of the ADIPORs in the PBMCs of both the children with PWS and the comparison group. Moreover, ADIPOR2 expression was highly expressed in the PBMCs of the children with PWS. A further study on the mechanism of increased expression of ADIPOR2 and its correlation with the expression of TNF-α in the PBMCs using the non-GH-treated PWS and obese control will be warranted because this study compared GH-treated PWS with an obese comparator group.

Rosa damascena has been manufactured as various food products, including tea, in Korea. A new flavonoid glycoside, kaempferol-3-O-beta-D-glucopyranosyl(1-->4)-beta-D-xylopyranoside, named roxyloside A was isolated from the buds of this plant, along with four known compounds, isoquercitrin, afzelin, cyanidin-3-O-beta-glucoside, and quercetin gentiobioside. The chemical structures of these compounds were determined by spectroscopic analyses, including FAB-MS, UV, IR, (1)H and (13)C NMR, DEPT, and 2D NMR (COSY, HSQC, and HMBC). All the isolated compounds except cyanidin-3-O-beta-glucoside exhibited high levels of inhibitory activity against 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase with IC(50) values ranging from 47.1 to 80.6 microM. Cyanidin-3-O-beta-glucoside significantly suppressed angiotensin I-converting enzyme (ACE) activity, with an IC(50) value of 138.8 microM, while the other four compounds were ineffective. These results indicate that R. damascena and its flavonoids may be effective to improve the cardiovascular system.

Objective: To evaluate the effects of recombinant human epidermal growth factor (rhEGF) on proliferation and radiation survival of normal fibroblast and cancer cell lines both in vitro and in vivo. Methods: Using normal fibroblast and cancer cell lines, we evaluated the expression of EGFR, and determined their pro- liferation and survival with rhEGF alone or in combination with radiation. For the combination treatments, we applied 10 nM rhEGF and delivered single radiation doses of 0, 2, 5, and 10 Gy. In the animal study, we introduced EMT-6 cells into BALB/c mice to assay for tumor growth delay. We applied single radiation dose of 10 or 20 Gy, with or without 1.0 mg/Kg of rhEGF, three times a day, for 7 days. Results: In a dose-dependent manner, rhEGF stimulated proliferation of the normal fibroblast, but not the cancer cell lines with low or intermediate expression of EGFR. rhEGF inhibited proliferation of the cancer cell line with the highest EGFR expression. Administration of rhEGF in combination with radiation attenuated the cell killing effect of radiation in normal fibroblast, but it had no effect or even augmented the radiation effect in cancer cell lines. In the animal study, we observed no difference in tumor growth rates when rhEGF was combined with radiation compared to radiation alone. Conclusions: Our results suggested that rhEGF might be useful in preventing and/or treating radiation-induced injury without stimulating tumor growth. Mini-Abstract: rhEGF effectively stimulated proliferation and radiation recovery in normal fibroblasts without promoting the growth of tumor cells. Therefore, it might be useful in preventing and/or treating radiation-induced tissue injury.

Pine mushroom (Tricholoma matsutake Sing.) is an expensive and highly prized delicacy in Korean and Japanese cuisines with its unique flavor and functional properties. The biological activities of pine mushroom juice (soluble solid contents 4.3 o Brix) were evaluated using different tests; DPPH radical scavenging assay for its antioxidant activity, fibrin plate method for fibrinolysis and angiotensin Ⅰ converting enzyme (ACE) inhibitory activity for anti-hypertensive effect. Free radical scavenging activity of the pine mushroom juice was 48.3±2.2% at the concentration of 1.0 mg/mL. The fibrinolytic activity of pine mushroom was about 2 times greater than that of plasmin used as positive control and the activity increased dose-dependently. The pine mushroom juice inhibited ACE activities dose-dependently and IC50 value of ACE activity was 1.03 o Brix. These results suggest that pine mushroom is a healthy delicacy.

The antifatigue properties of six Korean medicinal herb extracts were studied by evaluating forced swimming capacity and biochemical parameters in ICR mice. The treatment groups were orally administered 30% ethanolic extracts (500 mg/kg/day) of Rubus coreanus Miquel, Cyperus rotundus Linn., Acanthopanax sessiliflorus, Saururus chinensis Baili, Epimedium koreanumNakai, or Houttuynia cordata Thunb. for 4 weeks. Swimming time to exhaustion was found to be longer for the group fed R. coreanus than for the control group (P < .05). No significant differences were found in the plasma levels of either glucose or lactate between the control group and the group fed R. coreanus, which swam longer than the control. The plasma ammonia levels were significantly lower in the groups fed R. coreanus and A. sessiliflorus, when compared to the control group (P < .05). No significant differences were found in gastrocnemius muscle or liver glycogen content between the control group and any treatment group. These results suggest that R. coreanus extract, and none of the other herbs, has antifatigue effects in mice, as demonstrated by the increased forced swimming capacity and decreased plasma ammonia accumulation.

The plasma ghrelin has been reported to be elevated in Prader-Willi syndrome (PWS) and modulated by insulin. It was hypothesized that insulin might have a more pronounced effect on reducing plasma ghrelin in PWS patients, which would influence appetite. This study investigated the degree of ghrelin suppression using an euglycemic hyperinsulinemic clamp in children with PWS (n=6) and normal children (n=6). After a 90-min infusion of insulin, the plasma ghrelin level decreased from a basal value of 0.86+/-0.15 to 0.58+/-0.12 ng/mL in the controls, and from 2.38+/-0.76 to 1.12+/-0.29 ng/mL in children with PWS (p=0.011). The area under the curve below the baseline level over the 90 min insulin infusion was larger in children with PWS than in controls (-92.82+/-44.4 vs. -10.41+/-2.87 ng/mL/90 min) (p=0.011). The insulin sensitivity measured as the glucose infusion rate at steady state was similar in the two groups (p=0.088). The decrease in the ghrelin levels in response to insulin was more pronounced in the children with PWS than in the controls. However, the level of ghrelin was always higher in the children with PWS during the clamp study. This suggests that even though insulin sensitivity to ghrelin is well maintained, an increase in the baseline ghrelin levels is characteristic of PWS.

This paper analyzed the effects of group bibliotherapy in junior high school assigned by Daegu Metropolitan Office or Education as a model school. Seventy seven students haying problems were selected and divided into the two different programs which developed for internalizing problems and externalizing problems respectively. Each bibliotherapy program has been carried out 15 times. The effects of two programs were proved positive and statistically significant. The effect of each program were differ from the other, owing to the purpose and reading materials. The study skewed bibliotherapy are more effective for students having internalizing Problems and female students. Bibliotherapy also affected student`s reading attitude. In order to reach self bibliotherapy, developmental bibliotherapy should be connected to the reading guidance.

This study investigated the influences of reciprocal peer tutoring strategy and field independencedependence in the instruction enhancing student questions by using weekly reports. Seventh graders (N=152) from a middle school were assigned to WR (weekly reports) and WR-RPT (weekly reports-reciprocal peer tutoring) groups. Students were taught about ‘three states of matter', ‘motion of molecules', and ‘change of states and thermal energy for eighteen class hours and wrote weekly reports six times for the period. The students in the WR-RPT group also conducted reciprocal peer tutoring with the questions of weekly reports which they wrote. The results revealed that the scores of the WR-RPT group were significantly higher than those of the WR group in a conception test regardless of students' field independence-dependence. The field dependent students in the WR-RPT group performed better in an achievement test than those in WR group, while there was no significant difference for the field independent stu dents between the two groups. Additionally, field independent students in each group scored significantly higher than field dependent students in the two tests. Many students, especially having more field independence in the WR-RPT group, perceived WR-RPT positively.

Decreased fasting ghrelin levels and decreased ghrelin suppression in overweight children have been reported to be associated with insulin resistance. However, Prader-Willi syndrome (PWS) is associated with increased total ghrelin levels and relative hypoinsulinemia.The objective of the study was to analyze changes in acylated ghrelin (AG) and des-acylated ghrelin (DAG) levels after glucose loading and characterize correlations between insulin sensitivity and ghrelin suppression.Plasma glucose, insulin, AG, and DAG levels were measured in PWS children (n = 11) and normal obese controls (n = 10) during oral glucose tolerance testing.All subjects were admitted to the Samsung Medical Center.Oral glucose tolerance testing was performed in all subjects after an overnight fast.Plasma levels of the hormones AG, DAG, and insulin, and those of glucose at 0, 30, 60, 90, and 120 min after glucose challenge were measured, and whole-body insulin sensitivity index (WBISI) values were calculated.AG levels fell markedly more from fasting levels in PWS children than normal healthy obese controls at 30, 60, and 90 min after glucose challenge, but no significant differences in DAG levels were observed at any time between PWS patients and controls. Fasting AG and DAG levels were found to correlate with WBISI in PWS, and absolute suppressions (Delta from baseline) in AG at 30 min after glucose challenge (nadir) were also correlated with WBISI in PWS (r = 0.64, P = 0.035).Our results suggest that AG is sensitively suppressed by insulin and that this suppression correlated with insulin sensitivity in PWS children.

Lipid rafts are membrane microdomains that are proposed to function as platforms for both receptor signaling and trafficking. Our previous studies have demonstrated that Tip of herpesvirus saimiri (HVS), which is a T-lymphotropic tumor virus, is constitutively targeted to lipid rafts and interacts with cellular Lck tyrosine kinase and p80 WD repeat-containing endosomal protein. Through the interactions with Lck and p80, HVS Tip modulates diverse T-cell functions, which leads to the downregulation of T-cell receptor (TCR) and CD4 coreceptor surface expression, the inhibition of TCR signal transduction, and the activation of STAT3 transcription factor. In this study, we investigated the functional significance of Tip association with lipid rafts. We found that Tip expression remarkably increased lipid raft fractions in human T cells by enhancing the recruitment of lipid raft-resident proteins. Genetic analysis showed that the carboxyl-terminal transmembrane, but not p80 and Lck interaction, of Tip was required for the lipid raft localization and that lipid raft localization of Tip was necessary for the efficient downregulation of TCR and CD4 surface expression. Correlated with this, treatment with Filipin III, a lipid raft-disrupting agent, effectively reversed the downregulation of CD3 and CD4 surface expression induced by Tip. On the other hand, Tip mutants that were no longer present in lipid rafts were still capable of inhibiting TCR signaling and activating STAT3 transcription factor activity as efficiently as wild-type (wt) Tip. These results indicate that the association of Tip with lipid rafts is essential for the downregulation of TCR and CD4 surface expression but not for the inhibition of TCR signal transduction and the activation of STAT3 transcription factor. These results also suggest that the signaling and targeting activities of HVS Tip rely on functionally and genetically separable mechanisms, which may independently modulate T-cell function for viral persistence or pathogenesis.

Reactive oxygen species (ROS) play an important role in cellular signaling as second messengers. However, studying the role of ROS in physiological redox signaling has been hampered by technical difficulties in controlling their generation within cells. Here, we utilize two inert components, a photosensitizer and light, to finely manipulate the generation of intracellular ROS and examine their specific role in activating dendritic cells (DCs). Photoswitchable generation of intracellular ROS rapidly induced cytosolic mobilization of Ca(2+), differential activation of mitogen-activated protein kinases, and nuclear translocation of NF-κB. Moreover, a transient intracellular ROS surge could activate immature DCs to mature and potently enhance migration in vitro and in vivo. Finally, we observed that intracellular ROS-stimulated DCs enhanced antigen specific T-cell responses in vitro and in vivo, which led to delayed tumor growth and prolonged survival of tumor-bearing mice when immunized with a specific tumor antigen. Therefore, a transient intracellular ROS surge alone, if properly manipulated, can cause immature DCs to differentiate into a motile state and mature forms that are sufficient to initiate adaptive T cell responses in vivo.

The levels of ghrelin, an orexigenic hormone secreted by oxyntic cells in the digestive tract, are elevated in Prader-Willi syndrome (PWS) and GH deficiency (GHD) patients. In this study, we hypothesized that the hyperghrelinemia observed in PWS is related to IGF-I or GH/IGF axis deficiency.We investigated the densities of ghrelin-expressing cells (GECs), the amounts of ghrelin in gastric tissues, and ghrelin levels in plasma in 16 PWS patients and compared these results with those of 13 GHD patients and comparison groups (19 normal lean and 10 normal obese subjects).In the gastric body and fundus, 2- to 3-fold increases in the numbers of GECs (P0.001) and in the amounts of ghrelin (P0.018) were noted in PWS patients vs. comparison groups, whereas GEC numbers in GHD patients were similar to those of the comparison group despite elevated fasting plasma ghrelin levels. In addition, IGF-I sd scores in PWS were not found to be correlated with GEC densities, the amounts of ghrelin expressed in gastric tissues, or plasma ghrelin levels.Our results suggest that IGF-I or GH/IGF axis deficiency appears to be unrelated to observed GEC increases in the stomach of patients with PWS.

Prader-Willi syndrome (PWS) is the most common form of syndromic obesity associated with hyperphagia. Because ghrelin stimulates gastric motility in rodents, and PWS patients have 3- to 4-fold higher fasting plasma ghrelin concentrations than normal subjects, we hypothesized that hyperphagia associated with PWS may be partly explained by rapid gastric emptying due to the increased gastric motility caused by ghrelin. We determined gastric emptying times (GETs) and measured ghrelin levels in 11 PWS children and 11 age-, sex-, and body mass index-matched controls using a standard meal containing [ 99m Tc]diaminetriaminepentacetate. Median plasma ghrelin levels before (precibum) and after the GET study were higher in PWSpatientsthanincontrols(P0.004andP0.001,respectively). Median percent gastric retentions at 90 min after the standard meal were 57.1% (range, 34.0–83.2%) in PWS patients and 40.2% (range, 27.2–60.2%) in controls (P 0.03). In particular, precibum ghrelin concentrations were not significantly correlated with the rate of gastric emptying in PWS patients (P 0.153; r 0.461) or controls (P 0.911; r 0.048). Our results show that gastric emptying in PWS is reduced despite higher ghrelin levels, and that the voracious appetite associated with PWS is related to another mechanism. (J Clin Endocrinol Metab 90: 3367–3370, 2005)

This paper investigates the Bunko, the center of Japanese children`s reading movement, and its impact on the development of Japanese school library. Paper is consisted by two parts , one subject is the programs and the law made by government which is related to either the school library or children`s reading, the other is Bunko movements initiated by citizen mostly mothers who wish to improve their children`s reading environment. The Bunko movements started in 1960`s and motivate the establishment of public libraries in 1970`s, and it has triggered activating the school library after early 1990`s. By analyzing the relationships of the two subjects it can be concluded that they are tightly connected each other by emphasizing the function of school library not as a teaching-learning center but as a reading facility. It makes difficult for the Japanese school library to establish the most important function of supporting curriculum.

Ghrelin is a GH-releasing acylated peptide found in the stomach and a centrally acting food intake stimulator. Prader-Willi syndrome (PWS) is a genetic disorder characterized by a voracious appetite and increased fasting ghrelin levels. In this report we describe 24-h ghrelin profiles in PWS children (n = 5) and compare these with age, sex, and body mass index (BMI)-matched controls (n = 5). A 3- to 4-fold increase in ghrelin levels was found in PWS over a 24-h period, compared with controls (P < 0.001). Interestingly, there was a greater tendency for the up-regulation of ghrelin level in lean PWS than in obese PWS. To confirm this finding, we measured fasting ghrelin levels in 39 patients with PWS. Inverse correlations were found between plasma ghrelin levels and the following: age (r = -0.408, P = 0.005), BMI (r = -0.341, P = 0.017), percentage of the ideal weight for age (r = -0.382, P = 0.008), and BMI percentile (r = -0.311, P = 0.027). Our data show that there may be a suppressive (or up-regulating) controlling mechanism of ghrelin secretion in obese (or lean) PWS children. We hope that our data may further explain the mechanisms underlying the insatiable appetite and obesity characteristic of PWS.

We demonstrate a novel route to synthesize Fe3O4-CdSe/ZnS multifunctional nanoclusters (MNCs) with excellent optical and magnetic properties and biocompatibility. The successful fabrication of highly fluorescent and magnetic MNCs is achieved via a coupling process based on a partial ligand exchange reaction at the aqueous-organic solution interface. In addition, we show that dendritic cells (DCs), the sentinel of the immune system, can uptake the MNCs without significant change in cell viability. The MNCs uptaken by the DCs can be used for imaging, tracking, and separating the DCs. Furthermore, the MNCs can be loaded with a pathogen-associated molecular pattern, lipid A, via a hydrophobic-hydrophobic interaction. Ex vivo labeling of DCs with the MNC-lipid A complex enhances the DC migration to draining lymph nodes and tumor antigen-specific T cell responses in vivo. Our work may contribute to the development of synthetic routes to various multifunctional nanoclusters and DC-based cancer immunotherapies.

Background: Orientia tsutsugamushi, the causative agent of scrub typhus, is an obligate intracellular bacterium. Previously, a large number of genes that encode proteins containing eukaryotic protein-protein interaction motifs such as ankyrin-repeat (Ank) domains were identified in the O. tsutsugamushi genome. However, little is known about the Ank protein function in O. tsutsugamushi. Methodology/Principal Findings: To characterize the function of Ank proteins, we investigated a group of Ank proteins containing an F-box-like domain in the C-terminus in addition to the Ank domains. All nine selected ank genes were expressed at the transcriptional level in host cells infected with O. tsutsugamushi, and specific antibody responses against three Ank proteins were detected in the serum from human patients, indicating an active expression of the bacterial Ank proteins post infection. When ectopically expressed in HeLa cells, the Ank proteins of O. tsutsugamushi were consistently found in the nucleus and/or cytoplasm. In GST pull-down assays, multiple Ank proteins specifically interacted with Cullin1 and Skp1, core components of the SCF1 ubiquitin ligase complex, as well as the eukaryotic elongation factor 1 alpha (EF1 alpha). Moreover, one Ank protein co-localized with the identified host targets and induced downregulation of EF1 alpha potentially via enhanced ubiquitination. The downregulation of EF1 alpha was observed consistently in diverse host cell types infected with O. tsutsugamushi. Conclusion/Significance: These results suggest that conserved targeting and subsequent degradation of EF1 alpha by multiple O. tsutsugamushi Ank proteins could be a novel bacterial strategy for replication and/or pathogenesis during mammalian host infection.