Search

The purpose of this study is to educate foreign university students on the practice of task representation and to examine its effects on their level of writing. The writing of foreign university students is greatly influenced by task representation. Therefore, providing more education on task representation will improve the quality of the texts submitted by students. First, the writing scores of foreign university students were evaluated. Group A was provided corrective feedback on writing without being given any training for task representation, while groups B and C were educated in this area. However, group B did not receive corrective feedback, but group C did. As a result of the experiment, there was a significant improvement found in the texts for groups B and C, but not in group A. In addition, the proportion of texts suitable for academic writing was higher in the former two groups. Therefore, we can conclude that education regarding task representation has a positive effect on improving the writing score of foreign university students.

Mycobacterium abscessus(MAB) is one of the rapidly growing, multidrug-resistant non-tuberculous mycobacteria (NTM) causing various diseases including pulmonary disorder. Although it has been known that type I interferons (IFNs) contribute to host defense against bacterial infections, the role of type I IFNs against MAB infection is still unclear. In the present study, we show that rIFN-β treatment reduced the intracellular growth of MAB in macrophages. Deficiency of IFN-α/β receptor (IFNAR) led to the reduction of nitric oxide (NO) production in MAB-infected macrophages. Consistently, rIFN-β treatment enhanced the expression of iNOS gene and protein, and NO production in response to MAB. We also found that NO is essential for the intracellular growth control of MAB within macrophages in an inhibitor assay using iNOS-deficient cells. In addition, pretreatment of rIFN-β before MAB infection in mice increased production of NO in the lungs at day 1 after infection and promoted the bacterial clearance at day 5. However, when alveolar macrophages were depleted by treatment of clodronate liposome, rIFN-β did not promote the bacterial clearance in the lungs. Moreover, we found that a cytosolic receptor nucleotide-binding oligomerization domain 2 (NOD2) is required for MAB-induced TANK binding kinase 1 (TBK1) phosphorylation and IFN-β gene expression in macrophages. Finally, increase in the bacterial loads caused by reduction of NO levels was reversed by rIFN-β treatment in the lungs of NOD2-deficient mice. Collectively, our findings suggest that type I IFNs act as an intermediator of NOD2-induced NO production in macrophages and thus contribute to host defense against MAB infection.

Influenza viruses cause significant morbidity and mortality worldwide. Long-term or frequent use of approved anti-influenza agents has resulted in drug-resistant strains, thereby necessitating the discovery of new drugs. In this study, we found aprotinin, a serine protease inhibitor, as an anti-influenza candidate through screening of compound libraries. Aprotinin has been previously reported to show inhibitory effects on a few influenza A virus (IAV) subtypes (e.g., seasonal H1N1 and H3N2). However, because there were no reports of its inhibitory effects on the other types of influenza viruses, we investigated the inhibitory effects of aprotinin in vitro on a wide range of influenza viruses, including avian and oseltamivir-resistant influenza virus strains. Our cell-based assay showed that aprotinin had inhibitory effects on seasonal human IAVs (H1N1 and H3N2 subtypes), avian IAVs (H5N2, H6N5, and H9N2 subtypes), an oseltamivir-resistant IAV, and a currently circulating influenza B virus. We have also confirmed its activity in mice infected with a lethal dose of influenza virus, showing a significant increase in survival rate. Our findings suggest that aprotinin has the capacity to inhibit a wide range of influenza virus subtypes and should be considered for development as a therapeutic agent against influenza.

Problem Chorioamnionitis is caused by a bacterial infection that ascends from the vagina and can cause adverse pregnancy outcomes (APOs). Fusobacterium nucleatum (F. nucleatum) is a periodontal pathogen associated with the occurrence of APOs. In this study, we evaluated whether receptor-interacting protein kinase 2 (Ripk2), an adaptor protein of the cytosolic receptors nucleotide-binding oligomerization domain (NOD)1 and NOD2, in macrophages and human decidual stromal cells (hDSCs) contributes to immune responses against F. nucleatum. Method of study Bone marrow-derived macrophages (BMDMs) isolated from wild-type (WT) and Ripk2-deficient mice and hDSCs were cultured with F. nucleatum (MOI 1, 10, 100). BMDMs and hDSCs were assessed using enzyme-linked immunosorbent assay, Western blot analysis, real-time PCR, and nitrite assay. Results Fusobacterium nucleatum-induced production of IL-6, but not of TNF-α and IL-10, was lower in Ripk2-deficient BMDMs than in WT cells. Western blotting revealed a decrease in F. nucleatum-induced p65 phosphorylation in Ripk2-deficient macrophages, whereas mitogen-activated protein kinases activation was comparable between WT and Ripk2-deficient cells. The production of nitric oxide (NO) in response to F. nucleatum and the gene and protein expression of inducible NO synthase was impaired in Ripk2-deficient BMDMs. In hDSCs, F. nucleatum upregulated the gene and protein expression of NOD1, NOD2, and Ripk2 in a time-dependent manner. F. nucleatum also increased the production of IL-6, CXCL8, and CCL2, whereas this production was decreased by the Ripk2 inhibitors SB203580 and PP2. Conclusions In conclusion, Ripk2 signaling appears to contribute to the F. nucleatum-induced immune response and can be a preventive and therapeutic target against APOs.

Previously, we found that the water extract of Artermisia scoparia Waldst.Kit suppressed the cytokine production of lipopolysaccharide (LPS)-stimulated macrophages and alleviated carrageenan-induced acute inflammation in mice. Artemisia contains various sesquiterpene lactones and most of them exert immunomodulatory activity.In the present study, we investigated the immunomodulatory effect of estafiatin (EST), a sesquiterpene lactone derived from A. scoparia, on LPS-induced inflammation in macrophages and mouse sepsis model.Murine bone marrow-derived macrophages (BMDMs) and THP-1 cells, a human monocytic leukemia cell line, were pretreated with different doses of EST for 2 h, followed by LPS treatment. The gene and protein expression of pro-inflammatory cytokines interleukin (IL)-6, tumor necrosis factor (TNF)-α, and inducible nitric oxide synthase (iNOS) were measured by quantitative real-time polymerase chain reaction (qPCR) and Western blot analysis. The activation of nuclear factor kappa B (NF-κB) and mitogen-activated protein kinases (MAPKs) was also evaluated at the level of phosphorylation. The effect of EST on inflammatory cytokine production, lung histopathology, and survival rate was assessed in an LPS-induced mice model of septic shock. The effect of EST on the production of cytokines in LPS-stimulated peritoneal macrophages was evaluated by in vitro and ex vivo experiments and protective effect of EST on cecal ligation and puncture (CLP) mice was also assessed.The LPS-induced expression of IL-6, TNF-α, and iNOS was suppressed at the mRNA and protein levels in BMDMs and THP-1 cells, respectively, by pretreatment with EST. The half-maximal inhibitory concentration (ICThese results demonstrated that EST exerts anti-inflammatory effects on LPS-stimulated macrophages and protects mice from sepsis. Our study suggests that EST could be developed as a new therapeutic agent for sepsis and various inflammatory diseases.

Background: Long-term or frequent use of currently approved anti-influenza agents has resulted in the emergence of drug-resistant viruses, necessitating the discovery of new drugs. In this study, we found aprotinin, a serine protease inhibitor, as an anti-influenza candidate through screening of compound libraries. Aprotinin has been previously reported to show inhibitory effects on a few subtypes (e.g., seasonal H1N1 and H3N2) of influenza A virus (IAV). However, there were no reports of its inhibitory effects on the other types of influenza virus. Therefore, in this study, we investigated the inhibitory effects of aprotinin against a wide range of influenza viruses in vitro and in vivo. Methods: We tested the antiviral activity of aprotinin in Madine-Darby canine kidney (MDCK) cells against seasonal human IAVs, avian influenza viruses with zoonotic potential, oseltamivir-resistant IAVs, and influenza B virus. We also tested the antiviral activity of aprotinin against A/PR/8/34 (H1N1) virus in a mouse model. Results: Our cell-based assay showed that aprotinin had inhibitory effects on seasonal human IAVs (H1N1 and H3N2 subtypes), avian IAVs (H5N2, H6N5, and H9N2 subtypes), an oseltamivir-resistant IAV, and a currently circulating influenza B virus. We have also confirmed its activity in mice infected with a lethal dose of influenza virus, showing a significant increase in survival rate. Conclusions: Our findings suggest that aprotinin has the capacity to inhibit a wide range of influenza virus subtypes and should be considered for development as a therapeutic agent against influenza.

The effectiveness of current influenza vaccines is considered suboptimal, and 1 way to improve the vaccines is using adjuvants. However, the current pool of adjuvants used in influenza vaccination is limited due to safety concerns. Aloe vera, or aloe, has been shown to have immunomodulatory functions and to be safe for oral intake. In this study, we explored the potential of orally administered processed Aloe vera gel (PAG) as an adjuvant for influenza vaccines in C57BL/6 mice. We first evaluated its adjuvanticity with a split-type pandemic H1N1 (pH1N1) Ag by subjecting the mice to lethal homologous influenza challenge. Oral PAG administration with the pH1N1 Ag increased survival rates in mice to levels similar to those of alum and MF59, which are currently used as adjuvants in influenza vaccine formulations. Similarly, oral PAG administration improved the survival of mice immunized with a commercial trivalent influenza vaccine against lethal homologous and heterologous virus challenge. PAG also increased hemagglutination inhibition and virus neutralization Ab titers against homologous and heterologous influenza strains following immunization with the split-type pH1N1 Ag or the commercial trivalent vaccine. Therefore, this study demonstrates that PAG may potentially be used as an adjuvant for influenza vaccines.

Acinetobacter baumannii is known for its multidrug antibiotic resistance. New approaches to treating drug-resistant bacterial infections are urgently required. Cathelicidin-related antimicrobial peptide (CRAMP) is a murine antimicrobial peptide that exerts diverse immune functions, including both direct bacterial cell killing and immunomodulatory effects. In this study, we sought to identify the role of CRAMP in the host immune response to multidrug-resistant Acinetobacter baumannii. Wild-type (WT) and CRAMP knockout mice were infected intranasally with the bacteria. CRAMP−/− mice exhibited increased bacterial colony-forming units (CFUs) in bronchoalveolar lavage (BAL) fluid after A. baumannii infection compared to WT mice. The loss of CRAMP expression resulted in a significant decrease in the recruitment of immune cells, primarily neutrophils. The levels of IL-6 and CXCL1 were lower, whereas the levels of IL-10 were significantly higher in the BAL fluid of CRAMP−/− mice compared to WT mice 1 day after infection. In an in vitro assay using thioglycollate-induced peritoneal neutrophils, the ability of bacterial phagocytosis and killing was impaired in CRAMP−/− neutrophils compared to the WT cells. CRAMP was also essential for the production of cytokines and chemokines in response to A. baumannii in neutrophils. In addition, the A. baumannii-induced inhibitor of κB-α degradation and phosphorylation of p38 MAPK were impaired in CRAMP−/− neutrophils, whereas ERK and JNK phosphorylation was upregulated. Our results indicate that CRAMP plays an important role in the host defense against pulmonary infection with A. baumannii by promoting the antibacterial activity of neutrophils and regulating the innate immune responses.

Acinetobacter baumannii is known for its multidrug antibiotic resistance. New approaches to treating drug-resistant bacterial infections are urgently required. Cathelicidin-related antimicrobial peptide (CRAMP) is a murine antimicrobial peptide that exerts diverse immune functions, including both direct bacterial cell killing and immunomodulatory effects. In this study, we sought to identify the role of CRAMP in the host immune response to multidrug-resistant Acinetobacter baumannii. Wild-type (WT) and CRAMP knockout mice were infected intranasally with the bacteria. CRAMP-/- mice exhibited increased bacterial colony-forming units (CFUs) in bronchoalveolar lavage (BAL) fluid after A. baumannii infection compared to WT mice. The loss of CRAMP expression resulted in a significant decrease in the recruitment of immune cells, primarily neutrophils. The levels of IL-6 and CXCL1 were lower, whereas the levels of IL-10 were significantly higher in the BAL fluid of CRAMP-/- mice compared to WT mice 1 day after infection. In an in vitro assay using thioglycollate-induced peritoneal neutrophils, the ability of bacterial phagocytosis and killing was impaired in CRAMP-/- neutrophils compared to the WT cells. CRAMP was also essential for the production of cytokines and chemokines in response to A. baumannii in neutrophils. In addition, the A. baumannii-induced inhibitor of κB-α degradation and phosphorylation of p38 MAPK were impaired in CRAMP-/- neutrophils, whereas ERK and JNK phosphorylation was upregulated. Our results indicate that CRAMP plays an important role in the host defense against pulmonary infection with A. baumannii by promoting the antibacterial activity of neutrophils and regulating the innate immune responses.

The effectiveness of current influenza vaccines is considered suboptimal, and 1 way to improve the vaccines is using adjuvants. However, the current pool of adjuvants used in influenza vaccination is limited due to safety concerns. Aloe vera, or aloe, has been shown to have immunomodulatory functions and to be safe for oral intake. In this study, we explored the potential of orally administered processed Aloe vera gel (PAG) as an adjuvant for influenza vaccines in C57BL/6 mice. We first evaluated its adjuvanticity with a split-type pandemic H1N1 (pH1N1) Ag by subjecting the mice to lethal homologous influenza challenge. Oral PAG administration with the pH1N1 Ag increased survival rates in mice to levels similar to those of alum and MF59, which are currently used as adjuvants in influenza vaccine formulations. Similarly, oral PAG administration improved the survival of mice immunized with a commercial trivalent influenza vaccine against lethal homologous and heterologous virus challenge. PAG also increased hemagglutination inhibition and virus neutralization Ab titers against homologous and heterologous influenza strains following immunization with the split-type pH1N1 Ag or the commercial trivalent vaccine. Therefore, this study demonstrates that PAG may potentially be used as an adjuvant for influenza vaccines.

Influenza B virus remains a major cause of respiratory diseases worldwide. Because of limited epidemiological and genetic data, the local and global transmission patterns of influenza B virus are not fully understood. Here we report the molecular and phylogenetic characterization of 163 influenza B virus isolates from pediatric inpatients with influenza-like illness in the winter of 2011-2012 in South Korea. Analysis of haemagglutinin and neuraminidase genes of the influenza B isolates revealed that both B/Victoria (62 %) and B/Yamagata lineages (38 %) co-circulated during that influenza season, and a considerable number of the isolates carried several amino acid substitutions in the four major antigenic epitopes of their haemagglutinin protein.

Background Aloe vera is a functional food with various pharmacological functions, including an immune-modulating effect. Until now, A. vera has never been studied as an adjuvant in influenza vaccine, and its effects on upper respiratory tract infection (URI) are unknown. Purpose The objective of our study was to investigate the effect of processed A. vera gel (PAG) on immunogenicity of quadrivalent inactivated influenza vaccine and URI in healthy adults. Study design A randomized, double-blind, placebo-controlled clinical trial was performed. Methods This study was conducted in 100 healthy adults at a single center from September 2017 to May 2018. Subjects were randomly divided into a PAG group (n = 50) and a placebo group (n = 50). The enrolled subjects were instructed to ingest the study drug for 8 weeks. The participants received a single dose of quadrivalent inactivated influenza vaccine after taking the study drug for the first 4 weeks of the study. The primary endpoint was seroprotection rate against at least one viral strain at 4 weeks post-vaccination. Other outcomes were seroprotection rate at 24 weeks post-vaccination, seroconversion rate, geometric mean fold increase (GMFI) at 4 and 24 weeks post-vaccination, seroprotection rate ratio and geometric mean titer ratio (GMTR) at 4 weeks post-vaccination between PAG and placebo groups, and incidence, severity, and duration of URI. Results The European Committee for proprietary medicinal products (CPMP) evaluation criteria were met at least one in the PAG and placebo groups for all strains. However, there was no significant difference in the seroprotection rate at 4 weeks post-vaccination against all strains in both PAG and placebo groups. Among secondary endpoints, the GMFI at 4 weeks post-vaccination for the A/H3N2 was significantly higher in the PAG than in placebo group. The GMTR as adjuvant effect was 1.382 (95% CI, 1.014-1.1883). Kaplan–Meier curve analysis showed a reduction in incidence of URI (p = 0.035), and a generalized estimating equation model identified a decrease in repeated URI events (odds ratio 0.57; 95% CI, 0.39-0.83; p = 0.003) in the PAG group. Conclusions Oral intake of PAG did not show a significant increase in seroprotection rate from an immunogenicity perspective. However, it reduced the number of URI episodes. A well-designed further study is needed on the effect of PAG's antibody response against A/H3N2 in the future.

Zika virus (ZIKV) is a mosquito-borne member of the Flaviviridae family. ZIKV infection has been associated with neurological complications such as microcephaly in newborns and Guillain-Barré syndrome in adults; thus, therapeutic agents are urgently needed. Statins are clinically approved for lowering cholesterol levels to prevent cardiovascular disease but have shown potential as antiviral drugs. In this study, we explored the possibility of utilizing statins as anti-ZIKV drugs. We found that, generally, lipophilic statins (atorvastatin, cerivastatin, fluvastatin, lovastatin, mevastatin, and simvastatin) could reduce ZIKV production in vitro and result in smaller foci of infection. Time-of-drug-addition assay revealed that early treatment with statins is more beneficial than late treatment; however, statins could not completely inhibit the entry stage of ZIKV infection. Furthermore, individual lipophilic statins differed in anti-ZIKV capacity, with fluvastatin being the most efficient at low concentrations. Taken together, this study shows that statins or their derivatives have the potential to be used as anti-ZIKV therapeutic agents.

This study considers the orbit insertion targeting problem for guidance of a multi-stage space launch vehicle. To design guidance algorithm for a particular mission target, the orbit designation method should be determined first, as the target orbit of closed-loop guidance during the upper stage flight is usually specified at the end of the final stage. We investigate some advantages of this single target approach over the one with intermediate targets in addition to the final target by comparing them in various flight conditions using an explicit guidance algorithm. We notice that in some cases application of the single targeting can be rather tricky and thus propose here an extended version of the single targeting algorithm that can be applied to problems where the target orbit plane has to be changed during the upper stages flight. We also demonstrate that this pseudo-single target method can be applied to these kinds of problems smoothly in contrast to the conventional single target algorithms by conducting numerical simulations for disturbed off-nominal conditions as well as for the nominal condition.

As the recent rise of immune checkpoint inhibitors has resulted in durable clinical outcomes in cancer patients, the need for a newer and more widely effective target for immunotherapy has been vigorously sought in the field; T cell immunoreceptor with Ig and ITIM domain (TIGIT) is an immune checkpoint molecule expressed on CD8+ T cells, CD4+ T cells, NK cells, and regulatory T cells (Treg). TIGIT induces exhaustion of effector T cells (Teff) and NK cells in the tumor microenvironment via engagement with its ligand PVR (CD155) which is dominantly expressed on malignant tumor cells. After initial screening processes from a synthetic library, MG1131 clone was chosen based on the strongest binding affinity to human TIGIT and the superior PVR-blocking activity. MG1131 was cross-reactive with the cynomolgus monkey TIGIT, but not with the mouse TIGIT. Our in vitro efficacy data demonstrated that MG1131 activated NF-κB signaling in T cells, and enhanced NK-mediated tumor killing activities in a PVR-dependent manner. In vitro Treg suppression assays showed that MG1131 inhibited immunosuppressive functionality of Treg, leading to restoration of proliferation and IFN-γ secretion capacities of Teff even in the presence of Treg. In addition, expression levels of TIGIT on CD8+ T cells from PBMCs of cancer patient samples were higher compared with other immune inhibitory molecules such as PD-1, Tim-3, CTLA-4, or LAG-3, and the blockade of TIGIT by MG1131 resulted in increased IFN-γ secretion. Thus, our data indicate that MG1131 is a promising candidate for cancer immunotherapy by modulating NK and T cell functions. Citation Format: Jeewon Lee, Munkyung Kim, Hye-mi Nam, Joong Hyuk Sheen, Eun Jung Song, Hye In Yum, So Jung Lim, Hye-Young Park. MG1131, a novel TIGIT-targeting monoclonal antibody, enhances anti-tumor immune responses by modulating NK and T cell activity [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2230.

Eukaryotic cells contain a collection of spatially separated internal organelles embedded in the cytoplasm. Communication between these internal organelles is mediated by trafficking events that are mainly accomplished by vesicular transport. These trafficking events facilitate the targeted delivery of newly synthesized proteins and lipids, uptake of extracellular cargo, and also play a role in regulatory processes [1]. The cargo recognized by coat proteins is stabilized, carried in the vesicle, and delivered to the target membrane. Among the various coat proteins identified to date, coatomer, the coat complex of COPI-coated vesicles composed of seven COPs (α-, β-, β'-, γ-, δ-, e-, and ζ-COP) and the small ras-like GTPase, ADP ribosylation factor, mediates recycling of proteins in Golgi-to-endoplasmic reticulum (ER) interactions and within the Golgi complex [2, 3] In yeast, all COPs, except e-COP, are essential for cell viability. e-COP stabilizes α-COP, and thus coatomer in the elevated temperatures [4]. In addition, α-, γ-, δ-, and ζ-COP mutants, referred to as ret mutants, show inability to retrieve proteins with a di-lysine motif (KKXX) [5, 6, 7]. α- and β'-COPs are involved in the recruitment of dilysine-tagged proteins [6]. Ret1/α-COP of Saccharomyces cerevisiae is known to contribute to the maintenance of cell wall integrity and biogenesis by influencing glycosylation and localization of the secretome including cell wall proteins [8, 9, 10, 11, 12]. Quantitative proteomic analysis has revealed an association between calcineurin, a calcium-calmodulin-associated protein phosphatase, and e-COP during high temperature stress in the human fungal pathogen Cryptococcus neoformans [13]. In Aspergillus nidulans, α-COP is essential for viability. However, like in yeast, a mutation in A. nidulans α-COP gene results in a temperature-sensitive defect in growth, cell-wall construction, and protein secretion [14, 15]. Interestingly, the temperature dependent-osmo-sensitive phenotype of the S. cerevisiae α-COP mutant is suppressed by introduction of the A. nidulans sodVIC (stabilization of disomy) gene for α-COP [15]. In addition, the C-terminus of the A. nidulans e-COP, AneA, including tetratrico peptide repeat (TPR: W4G8Y11G15Y17A20Y24A27P32) interacts with the C-terminus of α-COP [16]. These findings suggest that the action of the coatomer in filamentous fungi may be similar to that in yeast, but the function of e-COP in conjunction with α-COP needs to be elucidated in A. nidulans. In this study, to understand the function of e-COP in A. nidulans during development, we performed phenotypic analyses of the aneA deletion mutant and α-COP mutant with over-expression of the gene encoding e-COP, aneA+.

This paper considers the effects of propellant mixture ratio and loading errors on the performance of a liquid rocket. Propellant residuals generated by error sources are analyzed for a launch vehicle model whose first stage consists of a cluster rocket of four 75-tonf class engines using a statistical Monte-Carlo approach and then the optimal fuel biases minimizing residuals are computed. The results are validated through comparison with analytic method using approximate formula, which have been applied for other space launch vehicles.

This paper analyzes and compares the performance of two prominent explicit guidance algorithms: the iterative guidance mode and the powered explicit guidance. We performed a series of numerical simulations of a space launch vehicle model for both nominal and off-nominal conditions. One of our findings is that if we take into account the originally ignored higher-order terms from the guidance parameters of the iterative guidance mode for a long-range flight, the guidance performance can be enhanced to a level comparable to that of the powered explicit guidance. These higher-order terms can be included by employing an iterative predictor–corrector method like the powered explicit guidance. Also we proposed a remedy of preventing relatively earlier divergence of the guidance commands with the predictor–corrector iteration than that of the linear differential corrector approach by making a better initial guess.

Influenza A viruses must undergo adaptation to acquire virulence in new host species. In mouse models, host adaptation for virulence is generally performed through 5 to 20 lung-to-lung passages. However, highly pathogenic avian influenza viruses (e.g., H5N1 and H7N7 subtypes) have been observed to acquire virulence in mice after only a few in vivo passages. In this study, a low-pathogenic avian influenza H5N2 virus, A/Aquatic Bird/Korea/CN2/2009, which was a prevalent subtype in South Korea in 2009, was serially passaged in mice to evaluate its potential to become highly pathogenic. Unexpectedly, the virus became highly pathogenic in mice after a single lung-to-lung passage, resulting in 100% lethality with a mean death time (MDT) of 6.1 days postinfection (DPI). Moreover, the pathogenicity gradually increased after subsequent in vivo passages with an MDT of 5.2 and 4.2 DPI after the second and third passage, respectively. Our molecular analysis revealed that two amino acid changes in the polymerase complex (a glutamate-to-lysine substitution at position 627 of PB2 and a threonine-to-isoleucine substitution at position 97 of PA) were associated with the increased pathogenicity; the PB2 E627K mutation was responsible for the initial virulence conversion (0 to 100% lethality), while the PA T97I mutation acted as an accessory for the increased virulence.

Background Although Helicobacter pylori have been known to induce vascular endothelial growth factor (VEGF) production in gastric epithelial cells, the precise mechanism for cellular signaling is incompletely understood. In this study, we investigated the role of bacterial virulence factor and host cellular signaling in VEGF production of H. pylori-infected gastric epithelial cells. Materials and methods We evaluated production of VEGF, activation of nuclear factor nuclear factor-kappaB (NF-κB) and mitogen-activated protein kinases (MAPKs) and hypoxia-inducible factor-1α (HIF-1α) stabilization in gastric epithelial cells infected with H. pylori WT or isogenic mutants deficient in type IV secretion system (T4SS). Results H. pylori induced VEGF production in gastric epithelial cells via both T4SS-dependent and T4SS-independent pathways, although T4SS-independent pathway seems to be the dominant signaling. The inhibitor assay implicated that activation of NF-κB and MAPKs is dispensable for H. pylori-induced VEGF production in gastric epithelial cells. H. pylori led to HIF-1α stabilization in gastric epithelial cells independently of T4SS, NF-κB, and MAPKs, which was essential for VEGF production in these cells. N-acetyl-cysteine (NAC), a reactive oxygen species (ROS) inhibitor, treatment impaired H. pylori-induced HIF-1α stabilization and VEGF production in gastric epithelial cells. Conclusion We defined the important role of ROS-HIF-1α axis in VEGF production of H. pylori-infected gastric epithelial cells, and bacterial T4SS has a minor role in H. pylori-induced VEGF production of gastric epithelial cells.

In this study, we developed a further-modified outer membrane vesicle (fmOMV) from the ΔmsbB/ΔpagP mutant of Escherichia coli transformed with the plasmid, pLpxF, in order to use it as an adjuvant for pandemic H1N1 (pH1N1) influenza vaccine. We evaluated the efficacy of the pH1N1 influenza vaccine containing the fmOMV in animal models as compared to the commercial adjuvants, alum or AddaVaxTM. The fmOMV-adjuvanted pH1N1 influenza vaccine induced a significant increase in the humoral immunity; however, this effect was less than that of the AddaVaxTM. The fmOMV-adjuvanted vaccine displayed pronounced an enhanced protective efficacy with increased T cell immune response and reduced the viral load in the lungs of the infected mice after challenging them with a lethal dose of the homologous virus. Moreover, it resulted in a significantly higher cross-protection against heterologous virus challenge than that of the pH1N1 vaccine with alum or with no adjuvants. In ferrets, the fmOMV-adjuvanted vaccine elicited a superior antibody response based on the HI titer and efficiently protected the animals from the lethal viral challenges. Taken together, the nontoxic fmOMV could be a promising adjuvant for inducing robust T cell priming into the pH1N1 vaccine and might be broadly applicable to the development of preventive measures against influenza virus infection.

Corynebacterium glutamicum is one of the well-studied industrial strain that is used for the production of nucleotides and amino acids. Recently, it has also been studied as a possible producer of organic acids such as succinic acid, based on its ability to produce organic acids under an oxygen deprivation condition. In this study, we conducted the optimization of medium components for improved succinate production from C. glutamicum under an oxygen deprivation condition by Plackett-Burman design and applied a response surface methodology. A Plackett-Burman design for ten factors such as glucose, ammonium sulfate, magnesium sulfate, potassium phosphate (K2HPO4 and KH2PO4), iron sulfate, manganese sulfate, biotin, thiamine, and sodium bicarbonate was applied to evaluate the effects on succinate production. Glucose, ammonium sulfate, magnesium sulfate, and dipotassium phosphate were found to have significant influence on succinate production, and the optimal concentrations of these four factors were sequentially investigated by the response surface methodology using a Box-Behnken design. The optimal medium components obtained for achieving maximum concentration of succinic acid were as follows: glucose 10 g/l, magnesium sulfate 0.5 g/l, dipotassium phosphate (K2HPO4) 0.75 g/l, potassium dihydrogen phosphate (KH2PO4) 0.5 g/l, iron sulfate 6 mg/l, manganese sulfate 4.2 mg/l, biotin 0.2 mg/l, thiamine 0.2 mg/l, and sodium bicarbonate 100 mM. The parameters that differed from a normal BT medium were glucose changed from 40 g/l to 10 g/l, dipotassium phosphate (K2HPO4) 0.5 g/l changed to 0.75 g/l, and ammonium sulfate ((NH4)2SO4) 7 g/l changed to 0 g/l. Under these conditions, the final succinic acid concentration was 16.3 mM, which is about 1.46 fold higher than the original medium (11.1 mM) at 24 h. This work showed the improvement of succinate production by a simple change of media components deduced from sequential optimization.

In order to screen interactor(s) of the Aspergillus nidulans -COP of COPI vesicle, we performed the yeast two hybrid screening by using the gene for A. nidulans -COP as a bait and identified -COP of the COPI vesicle as an interacting protein. The A. nidualns gene for the -COP was designated ( idulans psi-lone-COP), which encoded 296 amino acid residues with high level of identity with orthologs from other fungi. Domain analyses with yeast two-hybrid system suggested that the interaction between -COP and -COP relied on the C-terminus of both proteins, and that the N-terminal WD domian of -COP and the TPR region of -COP were not essential but required for the enhancement of the interaction. These results indicate that the interaction mode between -COP and -COP of COPI vesicle is evolutionarily well conserved in eukaryotes.

H3N2 canine influenza virus emerged in South Korea in 2007 and subsequently spread to China and Thailand, causing epidemic or endemic respiratory diseases in dogs. Through intermammalian species transmission, the virus has also infected cats. However, no direct evidence of significant genetic evolution has been reported since its first emergence. Here, we describe in depth the genetic and molecular characteristics of the ancestral strain (i.e., the first virus isolate from South Korea) of the H3N2 canine influenza virus currently circulating in East Asia.

This paper considers one of the explicit guidance algorithms, which has been proposed by Jaggers, to determine the closed-loop guidance algorithm for upper stages of a 3-staged space launch vehicle. Its commanded thrust vector is closer to the optimal solution when compared with that obtained by using the well-known Powered Explicit Guidance (PEG), which has been developed through the Space Shuttle program. Its performance is evaluated here by applying for guidance of the launcher during the second and third stages. Furthermore, to generate more precise guidance commands, it is attempted not to use the approximate formulas for the derivation of the original guidance law, and it is shown that performance is improved in comparison with the original.

In this study, a stability analysis of the explicit guidance laws for space launch vehicles is performed with consideration of varying thrust integrals. Among various forms of explicit guidance, linear tangent guidance in its general form is selected and six different acceleration profiles are considered for this numerical experiment. A linear system modeling which includes all of the significant dynamic elements of a space launcher is performed to analyze the effect of the characteristics of thrust integrals on stability margins. Numerical results show that in an aspect of guidance stability, it is advantageous to have thrust integrals derived from increasing acceleration profiles, such as constant thrust case, which may be considered in the development of propulsion systems. Finally, time-domain simulation with the original nonlinear models is performed to verify the approach and the result shows that the nonlinear dynamics of the system is conserved well in the linear model.

As a loudspeaker's physical size decreases, its ability to reproduce low frequencies is hindered. In this paper we propose a method of creating virtual bass using an efficient frequency tracking algorithm. The proposed algorithm has low computational complexity but excellent performance. The results of computer simulations and subjective tests are presented to validate the algorithm .

As part of a NASA dryden autonomous formation flight program for improved drag reduction of multiple F/A-18 aircraft, a new instrument, the formation flight instrumentation system (FFIS), for the precise estimation of the relative position, velocity, and attitude between two moving aircraft without the aid of ground-based instruments, was developed. The FFIS uses a global position system (GPS) receiver and an inertial navigation sensor (INS) instrumentation package on each aircraft combined with a wireless communication system for sharing measurements between vehicles. An extended Kalman filter structure blends the outputs of each GPS/INS in a distributed manner so as to maximize the accuracy of the relative state estimates. Differential carrier phase GPS measurements are used to provide high accuracy relative range measurements to the filtering algorithm. A multiple hypothesis Wald test for estimating the integer ambiguity between the two moving vehicles was developed as part of this project. The FFIS was tested in a hardware-in-the-loop simulation (HIL Sim) before being tested in actual F-18 flight tests. Test results validated the FFIS performance. Flight test results showed that the Wald test accurately estimates the integer ambiguity and that relative range estimates using least squares provide accurate position estimates with a mean of approximately 7 cm and a standard deviation of 13 cm

Background Since avian-origin H3N2 canine influenza virus (CIV) was first identified in South Korea in 2008, the novel influenza virus has been reported in several countries in Asia. Reverse zoonotic transmission of pandemic H1N1 (2009) influenza virus (pH1N1) has been observed in a broad range of animal species. Viral dominance and characterization of the reassortants of both viruses was undertaken in the present study. Findings Here we describe the viral dominance of 23 CIV reassortants between pH1N1 and canine H3N2 influenza viruses from a naturally co-infected dog. These results indicate that the M gene of pandemic H1N1 and the HA gene of canine H3N2 are predominant in the reassortants. Furthermore, unlike the original canine H3N2 virus, some reassortants showed high pathogenicity in mice. Conclusions This study suggests that continuous monitoring of influenza infection in companion animals may be necessary to investigate the potential of the emergence of novel influenza viruses.

Aims As an alternative strategy to obtain large amounts of ginseng extract with high yield of ginsenosides, we have utilized culture of cambial meristematic cells (CMCs) from wild ginseng. The anti-tumor effects of methanol extract of ginseng CMCs (MEGC) and their action mechanisms were investigated. Main methods Mice were intraperitoneally administered with MEGC, and we explored NK cell activity, suppression of in vivo growth of tumor cells and relevant molecule expression. Key findings MEGC significantly potentiated NK cell activity and suppressed in vivo growth of B16 melanoma cells. However, we observed no increase in NK cell number and unaltered expression of NK cell-activating (NKG2D) and inhibitory (Ly49, CD94/NKG2A) receptors as well as NK cell activation markers (CD25, CD69, CD119, and CD212) in MEGC-treated group compared to the controls. Instead, MEGC significantly enhanced IL-2 responsiveness in the early effector phase and the constitutive expression of granzyme B. Significance Our data indicate that culture of CMCs is an attractive alternative method for sustainable production of ginseng extracts and clinical use. In addition, we have unraveled a novel mechanism underlying the potentiation of NK cell activity and antitumor effect of ginseng extract, in which it upregulates the constitutive expression of cytotoxic mediator(s) and IL-2 responsiveness.

The objective of this study is to enhance neural-network guidance to consider the impact condition. Missile impact angle error, a measure of the degree to which the missile is not steering for a head-on attack, can have a significant influence on the final miss distance. Midcourse guidance using neural networks is employed to reduce the deviation angle from head-on effectively in the three-dimensional space. In addition, a coordinate transformation is introduced to simplify the three-dimensional guidance law and reduce training data for the neural network. Computational results show that the current neural-network guidance law with the coordinate transformation can be used to reduce the impact angle errors.

The objective of this study is to enhance neural-network guidance to consider the impact condition. The optimal impact condition in this study is defined as an head-on attack. Missile impact-angle error, which is a measure of the degree to which the missile is not steering for a head-on attack, can also have an influence on the final miss distance. Therefore midcourse guidance is used to navigate the missile, reducing the deviation angle from head on, given some constraints on the missile g performance. A coordinate transformation is introduced to simplify the three-dimensional guidance law and, consequently, to reduce training data. Computer simulation results show that the neural-network guidance law with the coordinate transformation reduces impact-angle errors effectively.

Models to predict the split growth in notched AS4/3501-6 graphite/epoxy quasi-isotropic laminates under tension-dominated fatigue are presented. First, a power law model and an artificial neural network (ANN) model are developed to describe the split growth under constant-amplitude fatigue. They are then applied in conjunction with a linear damage growth model to predict the split growth under spectrum fatigue. The ANN model is found to work better than the power law model as a predictive tool for split growth.

Decentralized filtering for a formation flight instrumentation system by INS/GPS integration is considered in this paper. An elaborate tuning method of the measurement noise covariance is suggested to compensate modeling errors caused by decentralizing the extended Kalman filter. It does not require large data transfer between formation vehicles. Covariance analysis exhibits the superior performance of the proposed approach when compared with the existent decentralized filter and the global filter, which has the target-filter performance.