• Molecules and Cells
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• v.21 no.2
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• pp.261-268
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• 2006

The Drosophila methuselah (mth) mutant has an approximately 35 percent increase in average lifespan, and enhanced resistance to various forms of stress, including starvation, high temperature, and dietary paraquat. To examine the transcriptional regulation of mth, we used luciferase assays employing Drosophila S2 cells. Two positive control elements were found at -542 ~ -272 (PE1) and +28 ~ +217 (PE2), where putative binding sites for transcription factors including Dorsal (Dl) were identified. Cotransfection of a Dl expression plasmid with a mth-luciferase reporter plasmid resulted in decreased reporter activity. PE1 and PE2, the minimal elements for strong promoter activity, were required for maximal repression by Dl protein. The N-terminal Rel homology domain (RHD) of Dl was not sufficient for repression of mth. We demonstrated by chromatin affinity precipitation (ChAP) assays in S2 cells that Dl bound to the putative PE1 binding site. Unexpectedly, semi-quantitative RT-PCR analysis revealed that the level of mth transcripts was reduced in dl flies. However, the in vivo result support the view that mth expression is regulated by dl, since it is well known that Dl functions as both a transcriptional activator and repressor depending on what other transcription factors are present. These findings suggest that both innate immunity and resistance to stress are controlled by Dl protein.

• Molecules and Cells
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• v.40 no.1
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• pp.17-23
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• 2017

Mitogen-activated protein kinase (MAPK) signaling cascades play critical roles in various cellular events in plants, including stress responses, innate immunity, hormone signaling, and cell specificity. MAPK-mediated stress signaling is also known to negatively regulate nitrogen-fixing symbiotic interactions, but the molecular mechanism of the MAPK signaling cascades underlying the symbiotic nodule development remains largely unknown. We show that the MtMKK5-MtMPK3/6 signaling module negatively regulates the early symbiotic nodule formation, probably upstream of ERN1 (ERF Required for Nodulation 1) and NSP1 (Nod factor Signaling Pathway 1) in Medicago truncatula. The overexpression of MtMKK5 stimulated stress and defense signaling pathways but also reduced nodule formation in M. truncatula roots. Conversely, a MAPK specific inhibitor, U0126, enhanced nodule formation and the expression of an early nodulation marker gene, MtNIN. We found that MtMKK5 directly activates MtMPK3/6 by phosphorylating the TEY motif within the activation loop and that the MtMPK3/6 proteins physically interact with the early nodulation-related transcription factors ERN1 and NSP1. These data suggest that the stress signaling-mediated MtMKK5/MtMPK3/6 module suppresses symbiotic nodule development via the action of early nodulation transcription factors.

• Biomolecules & Therapeutics
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• v.23 no.4
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• pp.345-349
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• 2015

Betulinic acid, a pentacyclic triterpene isolated from Jujube tree (Zizyphus jujuba Mill), has been known for a wide range of biological and medicinal properties such as antibacterial, antimalarial, anti-inflammatory, antihelmintic, antinociceptive, and anticancer activities. In the study, we investigated the antiviral activity on influenza A/PR/8 virus infected A549 human lung adenocarcinoma epithelial cell line and C57BL/6 mice. Betulinic acid showed the anti-influenza viral activity at a concentration of $50{\mu}M$ without a significant cytotoxicity in influenza A/PR/8 virus infected A549 cells. Also, betulinic acid significantly attenuated pulmonary pathology including increased necrosis, numbers of inflammatory cells and pulmonary edema induced by influenza A/PR/8 virus infection compared with vehicle- or oseltamivir-treated mice in vivo model. The down-regulation of IFN-${\gamma}$ level, which is critical for innate and adaptive immunity in viral infection, after treating of betulinic acid in mouse lung. Based on the obtained results, it is suggested that betulinic acid can be the potential therapeutic agent for virus infection via anti-inflammatory activity.

• Development and Reproduction
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• v.22 no.4
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• pp.341-350
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• 2018

Chemokines is a small protein that plays a major role in inflammatory reactions and viral infections as a chemotactic factor of cytokines involved in innate immunity. Most of the chemokines belong to the chemokine groups CC and CXC. To investigate the immune system of the olive flounder (Paralichthys olivaceus), an expression pattern specifically induced in the early developmental stages of analysis is examined using qRT-PCR. We also examined tissue-specific expression of both CC and CXC chemokine in healthy olive flounder samples. CC and CXC chemokine shows increased expression after immune-related organs are formed compared to expression during early development. CC chemokine was more highly expressed in the fin, but CXC chemokine showed higher expression in the gills, spleen, intestines, and stomach. Spatial and temporal expression analysis of CC and CXC chemokine were performed following viral hemorrhagic septicemia virus (VHSV) infection. CC chemokine showed high expression in the gills, which are respiratory organs, whereas CXC chemokine was more highly expressed in the kidneys, an immune-related organ. These results suggest that CC and CXC chemokine play an important role in the immune response of the olive flounder, and may be used as basic data for the immunological activity and gene analysis of it as well as other fish.

• Journal of Applied Biological Chemistry
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• v.64 no.3
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• pp.277-284
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• 2021

The emergence and rapid spread of the potentially fatal coronavirus disease 2019, caused due to infection by severe acute respiratory syndrome coronavirus-2, has led to worldwide interest in developing functional bioactive ingredients that act as immunomodulatory agents. In this study, we aimed to characterize Carica papaya extract and explore its potential as an immunomodulator by performing in vitro cell screening. Papaya leaf water extract (PLW) was found to significantly increase the levels of nitric oxide (NO) and prostaglandin E₂ (PGE₂) by upregulating inducible nitric oxide synthase and cyclo-oxygenase-2 activity, respectively. Additionally, PLW increased the production of tumor necrosis factor-α and interleukin 1β in RAW 264.7 cells. Furthermore, PLW activated the expression of c-Jun N-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK) but not that of p38 mitogen-activated protein kinase. These results indicate that PLW increased the production of NO, PGE₂, and pro-inflammatory cytokines by activating the JNK and ERK pathways in macrophages, thus demonstrating immunomodulatory properties. Finally, high-performance liquid chromatography fingerprint analysis indicated the presence of rutin, narirutin, and ρ-coumaric acid in PLW (6.30, 119.76, and 47.25 ppm, respectively). Treating cells with these compounds at non-toxic concentrations had no effect on NO production. Taken together, these results suggest that PLW may have potential as an immunity-enhancing supplement.

• Asian-Australasian Journal of Animal Sciences
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• v.32 no.9
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• pp.1355-1362
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• 2019

Objective: S100A7A, a member of the S100 protein family, is involved in various biological processes, including innate immunity, antimicrobial function, and epithelial tumorigenesis. However, the expression and function of S100A7A in the endometrium during the estrous cycle and pregnancy are not well understood in pigs. Therefore, this study determined the expression and regulation of S100A7A at the maternal-conceptus interface in pigs. Methods: We obtained endometrial tissues from pigs throughout the estrous cycle and pregnancy, conceptus tissues during early pregnancy, and chorioallantoic tissues during midto late pregnancy and analyzed the expression of S100A7A in these tissues. We also determined the effects of steroid hormones, estradiol-$17{\beta}$ ($E_2$) and progesterone, and interleukin-$1{\beta}$ (IL1B) on S100A7A expression in endometrial tissues. Results: We found that S100A7A was expressed in the endometrium during the estrous cycle and pregnancy in a pregnancy status- and stage-dependent manner and was localized to endometrial luminal epithelial (LE) and superficial glandular epithelial cells with strong intensity in LE cells on day 12 of pregnancy. Early stage conceptuses and chorioallantoic tissues from day 30 to term pregnancy also expressed S100A7A. The expression of S100A7A was increased by $E_2$ and IL1B in endometrial tissues. Conclusion: S100A7A was expressed at the maternal-conceptus interface at the initiation of implantation in response to conceptus-derived estrogen and IL1B and could be a unique endometrial epithelial marker for conceptus implantation in pigs. These findings provide an important insight into the understanding of conceptus-endometrial interactions for the successful establishment of pregnancy in pigs.

• Journal of The Korean Society of Integrative Medicine
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• v.7 no.2
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• pp.141-151
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• 2019

Purpose : Hypochlorous acid (HOCl), a major inorganic bactericidal compound of innate immunity, is effective against a broad range of microorganisms. In particular, HOCl is well-known as a non-antibiotic antimicrobial substance. However, effects of HOCl as an antimicrobial agent are still needed to study these functions against various specific type of microorganisms. In this study, we investigated the antimicrobial effect of hypochlorous acid (HOCl) in S. mutans and A. actinomycetemcomitans to cause dental caries and periodontal disease. Experiments were conducted to observe whether HOCl become effective replacement of disinfectant. Methods : To observe antimicrobial effect of HOCl, stabilized HOCl is prepared in the form of a physiologically balanced solution in pre-conditioned and post-conditioned HOCl solution. As a control, commercially available disinfectant MAXCLEAN was used as positive control. Moreover, S. mutans and A. actinomycetemcomitans distribution in gagrin, filtered tap water, and culture media. Cell viability were measured by viable cell count methods and disk diffusion test. Results : Our results showed that treatment of HOCl have no effect against antimicrobial effect compare to control group especially gagrin in disk diffusion test. HOCl tended to reduced viability against S. mutans in group of post-conditioned than pre-conditioned of HOCl solution however, there was no significant difference as well as no effect in A. actinomycetemcomitans. Conclusion : HOCl showed tendency to reduce viability against S. mutans in group of post-conditioned of HOCl solution and no effect of antimicrobial effect. Although HOCl is well known as effective against a broad range of microorganisms, HOCl seems to have diversity following type of species to be used as antimicrobial drug following our results. Therefore, it is necessary to be rigidly controlled and regulated in using HOCl solution clinically.

• Journal of Ginseng Research
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• v.43 no.4
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• pp.666-675
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• 2019

Background: Korean Red Ginseng (KRG) has been widely used as an herbal medicine to normalize and strengthen body functions. Although many researchers have focused on the biological effects of KRG, more studies on the action mechanism of red ginseng are still needed. Previously, we investigated the proteomic changes of the rat spleen while searching for molecular signatures and the action mechanism of KRG. The proteomic analysis revealed that differentially expressed proteins (DEPs) were involved in the increased immune response and phagocytosis. The aim of this study was to evaluate the biological activities of KRG, especially the immune-enhancing response of KRG. Methods: Rats were divided into 4 groups: 0 (control group), 500, 1000, and 2000 mg/kg administration of KRG powder for 6 weeks, respectively. Isobaric tags for relative and absolute quantitation was performed with Q-Exactive LC-MS/MS to compare associated proteins between the groups. The putative DEPs were identified by a current UniProt rat protein database search and by the Gene Ontology annotations. Results: The DEPs appear to increase the innate and acquired immunity as well as immune cell movement. These results suggest that KRG can stimulate immune responses. This analysis refined our targets of interest to include the potential functions of KRG. Furthermore, we validated the potential molecular targets of the functions, representatively LCN2, CRAMP, and HLA-DQB1, by Western blotting. Conclusion: These results may provide molecular signature candidates to elucidate the mechanisms of the immune response by KRG. Here, we demonstrate a strategy of tissue proteomics for the discovery of the molecular function of KRG.

• Journal of Microbiology and Biotechnology
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• v.29 no.5
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• pp.704-712
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• 2019

Although nanometric dead Lactobacillus plantarum has emerged as a potentially important modulator of immune responses, its underlying mechanism of action has not been fully understood. This study aimed to identify the detailed biochemical mechanism of immune modulation by micronized and heat-treated L. plantarum LM1004 (MHT-LM1004, <$1{\mu}m$ in size). MHT-LM1004 was prepared from L. plantarum LM1004 via culture in a specifically designed membrane bioreactor and heat treatment. MHT-LM1004 was shown to effectively induce the secretion of $TNF-{\alpha}$ and IL-6 and the mRNA expression of inducible nitric oxide synthase (iNOS). MHT-LM1004 enhanced the expression of TLR-2, phosphorylation of MAPKs (ERK), and nuclear translocation of $NF-{\kappa}B$ in a dose-dependent manner. Oral administration of MHT-LM1004 ($4{\times}10^9$ or $4{\times}10^{11}cells/kg$ mouse body weight) increased the splenocyte proliferation and serum cytokine levels. These results suggested that MHT-LM1004 effectively enhances early innate immunity by activating macrophages via the TLR-2/MAPK/$NF-{\kappa}B$ signalling pathway and that this pathway is one of the major routes in immune modulation by the Lactobacillus species.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.53 no.6
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• pp.900-908
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• 2020

This study was conducted to partially replace fish meal with black soldier fly (BSF) larvae or mealworm (MW) larvae in the diet for Pacific white shrimp Litopenaeus vannamei. A tuna by-product meal (27%) was used as the protein source in a control (Con) diet, which was replaced with 20%, 40%, or 60% BSF (designated as BSF20, BSF40, and BSF60, respectively), or MW (MW20, MW40, and MW60, respectively). The shrimp (average body weight, 0.09 g) were randomly stocked in 28 acrylic tanks and quadruplicate groups were fed one of the experimental diets six times daily. After 57 days of the feeding trial, growth and feed efficiency were significantly higher in shrimp that were fed the BSF40 and BSF60 diets than those in shrimp fed the Con diet. Phenoloxidase, superoxide dismutase, glutathione peroxidase and gene expression of crustin were significantly higher in shrimp fed BSF or MW diets than those in shrimp fed the Con diet. The results indicate that BSF or MW could be used as a fish meal replacement or as a functional protein source in diets and can help improve the growth, feed utilization, innate immunity and antioxidant capacity of Pacific white shrimp.