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PLZF+ Innate T Cells Support the TGF-β-Dependent Generation of Activated/Memory-Like Regulatory T Cells

  • Kang, Byung Hyun;Park, Hyo Jin;Park, Hi Jung;Lee, Jae-Il;Park, Seong Hoe;Jung, Kyeong Cheon
    • Molecules and Cells
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    • v.39 no.6
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    • pp.468-476
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    • 2016
  • PLZF-expressing invariant natural killer T cells and CD4 T cells are unique subsets of innate T cells. Both are selected via thymocyte-thymocyte interaction, and they contribute to the generation of activated/memory-like CD4 and CD8 T cells in the thymus via the production of IL-4. Here, we investigated whether $PLZF^+$ innate T cells also affect the development and function of $Foxp3^+$ regulatory CD4 T cells. Flow cytometry analysis of the thymus and spleen from both CIITA transgenic C57BL/6 and wild-type BALB/c mice, which have abundant $PLZF^+$ CD4 T cells and invariant natural killer T cells, respectively, revealed that $Foxp3^+$ T cells in these mice exhibited a $CD103^+$ activated/memorylike phenotype. The frequency of $CD103^+$ regulatory T cells was considerably decreased in $PLZF^+$ cell-deficient $CIITA^{Tg}Plzf^{lu/lu}$ and $BALB/c.CD1d^{-/-}$ mice as well as in an IL-4-deficient background, such as in $CIITA^{Tg}IL-4^{-/-}$ and $BALB/c.IL-4^{-/-}$ mice, indicating that the acquisition of an activated/ memory-like phenotype was dependent on $PLZF^+$ innate T cells and IL-4. Using fetal thymic organ culture, we further demonstrated that IL-4 in concert with TGF-${\beta}$ enhanced the acquisition of the activated/memory-like phenotype of regulatory T cells. In functional aspects, the activated/ memory-like phenotype of Treg cells was directly related to their suppressive function; regulatory T cells of $CIITA^{Tg}PIV^{-/-}$ mice more efficiently suppressed ovalbumin-induced allergic airway inflammation compared with their counterparts from wild-type mice. All of these findings suggest that $PLZF^+$ innate T cells also augmented the generation of activated/memory-like regulation via IL-4 production.

Reactive oxygen species-dependent apoptosis induction by water extract of Citrus unshiu peel in MDA-MB-231 human breast carcinoma cells

  • Kim, Min Yeong;Choi, Eun Ok;HwangBo, Hyun;Kwon, Da He;Ahn, Kyu Im;Kim, Hong Jae;Ji, Seon Yeong;Hong, Su-Hyun;Jeong, Jin-Woo;Kim, Gi Young;Park, Cheol;Choi, Yung Hyun
    • Nutrition Research and Practice
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    • v.12 no.2
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    • pp.129-134
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    • 2018
  • BACKGROUND/OBJECTIVES: Although several recent studies have reported the anti-cancer effects of extracts or components of Citrus unshiu peel, which has been used for various purposes in traditional medicine, the molecular mechanisms for their effects remain unclear. In the present study, the anti-cancer activity of a water-soluble extract of C. unshiu peel (WECU) in MDA-MB-231 human breast carcinoma cells at the level of apoptosis induction was investigated. MATERIALS/METHODS: Cytotoxicity was evaluated using the MTT assay. Apoptosis was detected using DAPI staining and flow cytometry analyses. Mitochondrial membrane potential, reactive oxygen species (ROS) assay, caspase activity and Western blotting were used to confirm the basis of apoptosis. RESULTS: The results indicated that WECU-induced apoptosis was related to the activation of caspase-8, and -9, representative initiator caspases of extrinsic and intrinsic apoptosis pathways, respectively, and caspase-3 accompanied by proteolytic degradation of poly(ADP-ribose) polymerase and down-regulation of the inhibitors of apoptosis protein family members. WECU also increased the pro-apoptotic BAX to anti-apoptotic BCL-2 ratio, loss of mitochondrial membrane potential and cytochrome c release from mitochondria to cytoplasm. Furthermore, WECU provoked the generation of ROS, but the reduction of cell viability and induction of apoptosis by WECU were prevented when ROS production was blocked by antioxidant N-acetyl cysteine. CONCLUSIONS: These results suggest that WECU suppressed proliferation of MDA-MB-231 cells by activating extrinsic and intrinsic apoptosis pathways in a ROS-dependent manner.

Numerical Model for Spreading of Cochlodinium Bloom in the Southern Coastal Waters in Korea (한국 남해안에서 Cochlodinium적조 확산모델)

  • Kwon Chul Hui;Cho Ku Dae
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.35 no.6
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    • pp.568-577
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    • 2002
  • The spreading Cocuoainim polykikoides bloom in the southern coastal waters of Korea was simulated using numerical model including the physical processes of water flow and the chemical processes of increasing cell of C. polykikoides by uptake of dissolved nutrients. The circulation of sea water was simulated by two dimensional tide model reflecting the main four tidal components of $M_2,\;S_2,\;K_1,\;O_1$, and permanent current was driven by inflow/outflow across open boundaries. According to the result of model which tidal and permanent current were reflected simultaneously, eastward flows entering the southern waters from the western waters of Korea are dominant but westward flows are weak relatively. These result suggest that it is difficult for initial C. polykikoides bloom generated in the coastal waters of Goheung to move to the western coast of Korea through Jeju Strait. For spreading model of C. poiyhikoides, the range of generating distribution and the generating time of C. polykikoides bloom in coastal area are similar to those of observation data in the field. Wind is the most important factor in moving and distribution of red tide. Permanent current flowing eastward is also considered to be important factor and tidal current was a little influenced.

Application of nightlight satellite imagery for assessing flooding potential area in the Mekong river basin (메콩강 홍수위험분석을 위한 나이트라이트 위성영상 적용성 검토)

  • Try, Sophal;Lee, Daeup;Lee, Giha
    • Journal of Korea Water Resources Association
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    • v.51 no.7
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    • pp.565-574
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    • 2018
  • High population density in deltaic settings, especially in Asia, tends to increase and causes coastal flood risk because of lower elevations and significant subsidence. Large flood annually causes numerous deaths and huge economic losses. In this paper, an innovative technology of spatial satellite imagery has been used as tool to analyze the socio-economic flood-related damage in Mekong river basin. The relationship between nightlight intensity and flood damages has been determined for the period of 1992-2013 with a spatial resolution of 30 arc sec ($0.0083^{\circ}$), which is nearly one kilometer at the equator. Flow path distance was calculated to identify the distance of each cell to river network and to examine how nightlight intensity correlate to the area close to and far from river network. Statistical analysis results highlight the significant correlation between nocturnal luminosity intensity and flood-related damages in countries along the Mekong river (i.e., Cambodia, China, Lao PDR, Thailand, and Vietnam). This result reveals that the areas close to the river network correspond to high human distribution and causes huge damage during flooding. The result may provide key information to the region with respect to decisions, attentions, and mitigation strategies.

Synthesis and Characteristics of Partially Fluorinated Poly(vinylidene fluroide)(PVDF) Cation Exchange Membrane via Direct Sulfonation (직접술폰화반응에 의한 부분불소화 Poly(vinylidene fluroide)(PVDF) 양이온교환막의 합성 및 특성)

  • Kang, Ki Won;Hwang, Taek Sung
    • Membrane Journal
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    • v.25 no.5
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    • pp.406-414
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    • 2015
  • In this study, partially fluorinated cation exchange membranes were prepared by direct sulfonation of Poly(VDF-co-hexafluoropropylene) copolymers (PVDF-co-HFP) followed by a casting method for application in the Membrane capacitive deionization (MCDI). The structure of sulfonated PVDF-co-HFP (SPVDF) was confirmed by Fourier-transform infrared (FT-IR) and $^1H$ Nuclear magnetic resonance ($^1H$ NMR) analysis. For quantitative analysis of the chemical composition, the X-ray Photoelectron Spectroscopy (XPS) was used. The membrane properties such as water uptake, ion exchange capacity and electrical resistance were measured. It was suggested that the optimum direct sulfonation condition of PVDF-co-HFP ion exchange membranes was $60^{\circ}C$ and 7 hours for temperature and duration of sulfonation, respectively. The water uptake of the SPVDF ion exchange membrane was 21.5%. The ion exchange capacity and electrical resistance were 0.89 meq/g and $3.70{\Omega}{\cdot}cm^2$, respectively. It was investigated that if it is feasible to apply these membranes in MCDI at various cell potentials (0.9~1.5 V) and initial flow rates (10~40 mL/min). In the MCDI process, the maximum salt removal rate was 62.5% in repeated absorption-desorption cycles.

Micro-Spot Atmospheric Pressure Plasma Production for the Biomedical Applications

  • Hirata, T.;Tsutsui, C.;Yokoi, Y.;Sakatani, Y.;Mori, A.;Horii, A.;Yamamoto, T.;Taguchi, A.
    • Proceedings of the Korean Vacuum Society Conference
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    • 2010.02a
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    • pp.44-45
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    • 2010
  • We are currently conducting studies on culturing and biocompatibility assessment of various cells such as neural stem cells and induced pluripotent stem cells(IPS cells) on carbon nanotube (CNT), on nerve regeneration electrodes, and on silicon wafers with a focus on developing nerve integrated CNT based bio devices for interfacing with living organisms, in order to develop brain-machine interfaces (BMI). In addition, we are carried out the chemical modification of carbon nanotube (mainly SWCNTs)-based bio-nanosensors by the plasma ion irradiation (plasma activation) method, and provide a characteristic evaluation of a bio-nanosensor using bovine serum albumin (BSA)/anti-BSA binding and oligonucleotide hybridization. On the other hand, the researches in the case of "novel plasma" have been widely conducted in the fields of chemistry, solid physics, and nanomaterial science. From the above-mentioned background, we are conducting basic experiments on direct irradiation of body tissues and cells using a micro-spot atmospheric pressure plasma source. The device is a coaxial structure having a tungsten wire installed inside a glass capillary, and a grounded ring electrode wrapped on the outside. The conditions of plasma generation are as follows: applied voltage: 5-9 kV, frequency: 1-3 kHz, helium (He) gas flow: 1-1.5 L/min, and plasma irradiation time: 1-300 sec. The experiment was conducted by preparing a culture medium containing mouse fibroblasts (NIH3T3) on a culture dish. A culture dish irradiated with plasma was introduced into a $CO_2$-incubator. The small animals used in the experiment involving plasma irradiation into living tissue were rat, rabbit, and pick and are deeply anesthetized with the gas anesthesia. According to the dependency of cell numbers against the plasma irradiation time, when only He gas was flowed, the growth of cells was inhibited as the floatation of cells caused by gas agitation inside the culture was promoted. On the other hand, there was no floatation of cells and healthy growth was observed when plasma was irradiated. Furthermore, in an experiment testing the effects of plasma irradiation on rats that were artificially given burn wounds, no evidence of electric shock injuries was found in the irradiated areas. In fact, the observed evidence of healing and improvements of the burn wounds suggested the presence of healing effects due to the growth factors in the tissues. Therefore, it appears that the interaction due to ion/radicalcollisions causes a substantial effect on the proliferation of growth factors such as epidermal growth factor (EGF), nerve growth factor (NGF), and transforming growth factor (TGF) that are present in the cells.

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Effect of sun ginseng potentiation on epirubicin and paclitaxel-induced apoptosis in human cervical cancer cells

  • Lin, Yingjia;Jiang, Dan;Li, Yang;Han, Xinye;Yu, Di;Park, Jeong Hill;Jin, Ying-Hua
    • Journal of Ginseng Research
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    • v.39 no.1
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    • pp.22-28
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    • 2015
  • Background: Sun ginseng (SG), a specific formulation of quality-controlled red ginseng, contains approximately equal amounts of three major ginsenosides (RK1, Rg3, and Rg5), which reportedly has antitumor-promoting activities in animal models. Methods: MTT assay was used to assess whether SG can potentiate the anticancer activity of epirubicin or paclitaxel in human cervical adenocarcinoma HeLa cells, human colon cancer SW111C cells, and SW480 cells; apoptosis status was analyzed by annexin V-FITC and PI and analyzed by flow cytometry; and apoptosis pathway was studied by analysis of caspase-3, -8, and -9 activation, mitochondrial accumulation of Bax and Bak, and cytochrome c release. Results: SG remarkably enhances cancer cell death induced by epirubicin or paclitaxel in human cervical adenocarcinoma HeLa cells, human colon cancer SW111C cells, and SW480 cells. Results of the mechanism study highlighted the cooperation between SG and epirubicin or paclitaxel in activating caspase-3 and -9 but not caspase-8. Moreover, SG significantly increased the mitochondrial accumulation of both Bax and Bak triggered by epirubicin or paclitaxel as well as the subsequent release of cytochrome c in the targeted cells. Conclusion: SG significantly potentiated the anticancer activities of epirubicin and paclitaxel in a synergistic manner. These effects were associated with the increased mitochondrial accumulation of both Bax and Bak that led to an enhanced cytochrome c release, caspase-9/-3 activation, and apoptosis. Treating cancer cells by combining epirubicin and paclitaxel with SG may prove to be a novel strategy for enhancing the efficacy of the two drug types.

Immunomodulatory Properties of Lactobacillus plantarum NC8 Expressing an Anti-CD11c Single-Chain Fv Fragment

  • Liu, Jing;Yang, Guilian;Gao, Xing;Zhang, Zan;Liu, Yang;Yang, Xin;Shi, Chunwei;Liu, Qiong;Jiang, Yanlong;Wang, Chunfeng
    • Journal of Microbiology and Biotechnology
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    • v.29 no.1
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    • pp.160-170
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    • 2019
  • The lactic acid bacteria species Lactobacillus plantarum (L. plantarum) has been used extensively for vaccine delivery. Considering to the critical role of dendritic cells in stimulating host immune response, in this study, we constructed a novel CD11c-targeting L. plantarum strain with surface-displayed variable fragments of anti-CD11c, single-chain antibody (scFv-CD11c). The newly designed L. plantarum strain, named 409-aCD11c, could adhere and invade more efficiently to bone marrow-derived DCs (BMDCs) in vitro due to the specific interaction between scFv-CD11c and CD11c located on the surface of BMDCs. After incubation with BMDCs, the 409-aCD11c strain harboring a eukaryotic vector pValac-GFP could lead to more efficient expression of GFP compared with wild-type strains shown by flow cytometry analysis, indicating the enhanced translocation of pValac-GFP from L. plantarum to BMDCs. Similar results were also observed in an in vivo study, which showed that oral administration resulted in efficient expression of GFP in both Peyer's patches (PP) and mesenteric lymph nodes (MLNs) within 7 days after the last administration. In addition, the CD11c-targeting strain significantly promoted the differentiation and maturation of DCs, the differentiation of $IL-4^+$ and $IL-17A^+$ T helper (Th) cells in MLNs, as well as production of $B220^+$ $IgA^+$ B cells in the PP. In conclusion, this study developed a novel DC-targeting L. plantarum strain which could increase the ability to deliver eukaryotic expression plasmid to host cells, indicating a promising approach for vaccine study.

Supplementation of Microalgae (Tetradesmus sp.) to Pre-Starter Diet for Broiler Chickens (초기 육계 사료 내 미세조류(Tetradesmus sp.) 첨가에 따른 장관환경 변화)

  • Rim, Chae Yun;Jung, Hui-su;An, Su Hyun;Joo, Sang Seok;Kim, Z-Hun;Kong, Changsu;Kim, Myunghoo
    • Korean Journal of Poultry Science
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    • v.49 no.2
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    • pp.125-137
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    • 2022
  • This study aimed to determine the effects of dietary microalgae (Tetradesmus sp. (TO)) on intestinal immunity and microbiota of pre-starter broilers. One hundred and twenty 1-day-old birds (Ross 308) were allocated to two dietary treatment groups with six blocks in a randomized complete block design. The two experimental diets consisted of a corn-soybean meal-based basal diet and a diet with 0.5% TO powder instead of cornstarch in the basal diet. After feeding the experimental diets for ten days, all birds' body weight and feed intake were measured, and representative eight birds were selected from each treatment group. Small intestinal lamina propria cells were isolated using flow cytometry to examine the frequency of immune cells. Cecal feces were harvested for 16s rRNA gut microbiota analysis and fecal IgA levels. Here, we found that 0.5% TO supplementation increased CD3+CD4+ T cells in the small intestine, but decreased CD3+CD8+ T cells in the small intestine. Gut microbial analysis showed that TO supplementation significantly increased the alpha diversity of the gut microbiome. Taxonomic analysis showed that TO treatment increased the abundance of Firmicutes and decreased that of Bacteroidetes at the phylum level. The distribution of Enterobacteriaceae containing many harmful bacteria at the family level, was lower in the TO group. In the LEfSe analysis, the TO group had a significantly enriched abundance of Agathobaculum at the genus level. Overall, results show that Tetradesmus sp. supplementation influences intestinal T-cell immunity and induces the expansion of beneficial gut microbes in pre-starter broiler chickens.

Restorative effects of Rg3-enriched Korean Red Ginseng and Persicaria tinctoria extract on oxazolone-induced ulcerative colitis in mice

  • Ullah, H.M. Arif;Saba, Evelyn;Lee, Yuan Yee;Hong, Seung-Bok;Hyun, Sun-Hee;Kwak, Yi-Seong;Park, Chae-Kyu;Kim, Sung Dae;Rhee, Man Hee
    • Journal of Ginseng Research
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    • v.46 no.5
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    • pp.628-635
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    • 2022
  • Background: Ulcerative colitis (UC) is the large intestine disease that results in chronic inflammation and ulcers in the colon. Rg3-enriched Korean Red Ginseng extract (Rg3-RGE) is known for its pharmacological activities. Persicaria tinctoria (PT) is also used in the treatment of various inflammatory diseases. The aim of this study is to investigate the attenuating effects of Rg3-RGE with PT on oxazolone (OXA)-induced UC in mice. Methods: A total of six groups of mice including control group, OXA (as model group, 1.5%) group, sulfasalazine (75 mg/kg) group, Rg3-RGE (20 mg/kg) group, PT (300 mg/kg) group, and Rg3-RGE (10 mg/kg) with PT (150 mg/kg) group. Data on the colon length, body weight, disease activity index (DAI), histological changes, nitric oxide (NO) assay, Real-time PCR of inflammatory factors, ELISA of inflammatory factors, Western blot, and flow cytometry analysis were obtained. Results: Overall, the combination treatment of Rg3-RGE and PT significantly improved the colon length and body weight and decreased the DAI in mice compared with the treatment with OXA. Additionally, the histological injury was also reduced by the combination treatment. Moreover, the NO production level and inflammatory mediators and cytokines were significantly downregulated in the Rg3-RGE with the PT group compared with the model group. Also, NLR family pyrin domain containing 3 (NLRP3) inflammasome and nuclear factor kappa B (NF-𝛋B) were suppressed in the combination treatment group compared with the OXA group. Furthermore, the number of immune cell subtypes of CD4+ T-helper cells, CD19+ B-cells, and CD4+ and CD25+ regulatory T-cells (Tregs) was improved in the Rg3-RGE with the PT group compared with the OXA group. Conclusion: Overall, the mixture of Rg3-RGE and PT is an effective therapeutic treatment for UC.