• Title/Summary/Keyword: cell-cell interaction

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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.

Impacts on Water Quality to an Artificial Lake Due to Sudden Disturbance of Sediments (급격한 저니토 교란이 인공 하구호 수질에 미치는 영향)

  • 서승원;김정훈
    • Journal of Korean Society of Coastal and Ocean Engineers
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    • v.15 no.1
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    • pp.39-50
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    • 2003
  • This study aims to figure out water quality impacts due to sudden disturbances of sediments during dike construction in land reclamation for the northern part of the Siwhaho Lake where heavily deteriorated settlements from upstream loadings are outstanding. We exploit a 3-D water quality model CE-QUAL-ICM combined with a hydrodynamic model TIDE3D. Simulations are done accounting water-sediment interaction in a 4-$\sigma$ layers. Long-term simulation for 1-year shows that bottom layers around the disturbance location are only affected and marks very high concentration. Complete vertical mixing appears at least 5km apart to downward due to complex effects of geometry, bathymetry and river inflows. It should be addressed that existing condition of the Siwhaho Lake stands for high concentration of COD and TP in winter and spring due to relatively high incoming loadings, however the effect of sediment disturbances yields reverse phenomena, i.e., impacts of dike construction arise greatly in summer and fall. Refined grid system consisting of 150m${\times}$150m rectangular grid, which is doubled system compared to previous study (Suh et al.,2002), gives affordable results by reducing flux differences through a cell especially in front of gate.

Treatment of Aplasia Cutis Congenita on Scalp using Hyalomatix$^{(R)}$: A Case Report (하이알로매트릭스를 이용한, 두개골결손을 동반한 선천성피부무형성증의 치료: 1례 보고)

  • Rhee, Suk-Hyun;Hong, Jong-Won;Roh, Tai-Suk;Kim, Young-Seok;Rah, Dong-Kyun
    • Archives of Plastic Surgery
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    • v.37 no.4
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    • pp.469-472
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    • 2010
  • Purpose: Aplasia Cutis Congenita (ACC) is a rare disease characterized by the focal defect of the skin at birth, frequently involving scalp, but it may affect any region of the body. There are no etiology known but some conditions such as intrauterine vascular ischemia, amniotic adherences and viral infections are associated. The ideal treatment for the ACC is not known. Superficial and relatively small sized defects (< $3{\times}5\;cm$) may heal spontaneously and large defects related with risks of infection and bleeding may require aggressive surgical treatment. Hyalomatrix$^{(R)}$ is a bilayer of an esterified hyaluronan scaffold beneath a silicone membrane. It has been used as a temporary dermal substitute to cover deep thickness skin defect and has physiological functions derive from the structural role in extracellular matrix and interaction with cell surface receptor. This material has been used for the wound bed pre-treatment for skin graft to follow and especially in uncooperative patient, like a newborn, this could be a efficient and aseptic way of promoting granulation without daily irritative wound care. For this reason, using Hyalomatrix$^{(R)}$ for the treatment of ACC was preferred in this paper. Methods: We report a case of a newborn with ACC of the vertex scalp and non-ossified partial skull defect. The large sized skin and skull defect ($6{\times}6\;cm$) was found with intact dura mater. No other complications such as bleeding or abnormal neurologic sign were accompanied. Escharectomy was performed and Hyalomatrix$^{(R)}$ was applied for the protection and the induction of acute wound healing for 3 months before the split-thickness skin graft. During the 3 months period, the dressing was renewed in aseptic technique for every 3 weeks. The skin graft was achieved on the healthy granulation bed. Results: The operative procedure was uneventful without necessity of blood transfusion. Postoperative physical examination revealed no additional abnormalities. Regular wound management was performed in out-patient clinic and the grafted skin was taken completely. No other problems developed during follow-up. Conclusion: Hyalomatrix$^{(R)}$ provides protective and favorable environment for wound healing. The combination of the use of Hyalomatrix$^{(R)}$ and the skin graft will be a good alternative for the ACC patients with relatively large defect on vertex.

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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Characterization of the Immune Regulation Function of Fibroblastic Reticular Cells Originating from Lymph Node Stroma (림프절 스트로마 유래 fibroblastic reticular cell의 면역조절 기능에 대한 특성 규명)

  • Lee, Jong-Hwan
    • Journal of Life Science
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    • v.26 no.7
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    • pp.789-795
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    • 2016
  • A lymph node (LN) is one of the secondary lymphoid organs. An LN consists of a complicated 3 dimensional frame structure and several stromal cells. Fibroblastic reticular cells (FRC) are distributed in the T zone for interaction with T cells. FRC secrete homing chemokines such as CCL19 and CCL21. Moreover, FRC play a pivotal role in the production of extracellular matrix (ECM) into LN for ECM reorganization against pathogen infections. However, not much is known about the involvement of the immune reaction of FRC. The present report is for the characterization of FRC on immune response. For this, FRC were positioned in several infected situations such as co-culture with macrophage, lipopolysaccharide (LPS), and TNFα stimulation. When a co-culture between FRC and macrophage was performed, a morphological change in FRC was observed, and empty space between FRCs was created by this change. The soluble ICAM-1 protein level was up-regulated by co-culturing with Raw264.7 and the treatment of the ROCK inhibitor Y27632. The activity of matrix metalloproteinase (MMP) was up-regulated by LPS onto FRC. Furthermore, the inflammatory cytokine TNFα regulated the expression of ECM in FRC by a gene chip assay. Collectively, it suggests that FRC are involved in immune reactions.

Simple Mass-screening Methods for Resistance of Tomato to Fusarium oxysporum f. sp. lycopersici (토마토 시들음병에 대한 간편 대량 저항성 검정법)

  • Park, Myung Soo;Jang, Kyoung Soo;Choi, Yong Ho;Kim, Jin-Cheol;Choi, Gyung Ja
    • Horticultural Science & Technology
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    • v.31 no.1
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    • pp.110-116
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    • 2013
  • This study was carried out to establish the simple mass-screening methods for resistant tomato to Fusarium wilt caused by Fusarium oxysporum f. sp. lycopersici (FOL). Root dip inoculation method has been used in many studies on the resistance of tomato to disease. On the other hand, in mass-screening for resistant tomato to Fusarium wilt, the inoculation method is time-consuming and laborious procedure. Disease development of two FOL isolates on two cultivars of tomato according to inoculation method including root dip, tip and scalpel methods were investigated. In compatible interaction, tomato seedlings of each cultivar inoculated by tip method showed the lower and more variable disease severities than by root dip method. Whereas the seedlings by scalpel method represented clear resistant and susceptible responses to Fusarium wilt as root dip method. The resistance degree of each cultivar inoculated with FOL isolates by scalpel method was hardly affected by the tested incubation temperature and inoculum concentration. On the basis of the results, we suggest scalpel inoculation method as an efficient mass-screening method for resistant of tomato cultivars to Fusarium wilt. Roots of tomato seedlings at two-leaf stage grown in plastic cell tray were injured with scalpel and then spore suspension (more than $1{\times}10^7\;conidia{\cdot}mL^{-1}$) of FOL was poured directly on the roots. The infected plants were cultivated in a growth room at $25-30^{\circ}C$ for 4 weeks with 12-hours light a day.

Evaluation of the Cytotoxic Effects of Resveratrol Treatment with Over-the-counter Drugs on the Hepatic and Intestinal Cells (식이 폴리페놀 성분 resveratrol과 일반의약품의 복합처리에 의한 간 및 장관계 세포독성 평가)

  • Kim, Da-Ram;Kim, Mi-Ri;Hong, Jung-Il
    • Korean Journal of Food Science and Technology
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    • v.42 no.2
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    • pp.217-222
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    • 2010
  • Resveratrol is a natural polyphenolic compound frequently found in grapes. The biological actions of resveratrol have been extensively investigated both in vitro and in vivo. The interactions of resveratrol with commonly-consumed drugs, however, have rarely been studied. In this study, the cytotoxic properties of resveratrol on the hepatic and intestinal cells in the presence of over-the-counter (OTC) drugs, including acetaminophen (AAP), aspirin (Asp), and ibuprofen (Ibu), were evaluated. The cytotoxic effects of resveratrol on hepatic HepG2 and colonic HCT 116 cells were not markdely changed in the presence of AAP, Asp, or Ibu. Conversely, the cytotoxicity of OTC drugs was not affected by resveratrol either. Concentrations of resveratrol below 10 mM significantly increased HepG2 cell growth after 48 or 72 hr incubation; however, the growth-stimulating effect was not observed in the presence of AAP. When HCT 116 cells were treated with OTC drugs before or after resveratrol, the cytotoxic effects were not significantly altered. The present study provides basic information for the potential health effects of the interactions between resveratrol and commonly-consumed OTC drugs.

Whole genome MBD-seq and RRBS analyses reveal that hypermethylation of gastrointestinal hormone receptors is associated with gastric carcinogenesis

  • Kim, Hee-Jin;Kang, Tae-Wook;Haam, Keeok;Kim, Mirang;Kim, Seon-Kyu;Kim, Seon-Young;Lee, Sang-Il;Song, Kyu-Sang;Jeong, Hyun-Yong;Kim, Yong Sung
    • Experimental and Molecular Medicine
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    • v.50 no.12
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    • pp.1.1-1.14
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    • 2018
  • DNA methylation is a regulatory mechanism in epigenetics that is frequently altered during human carcinogenesis. To detect critical methylation events associated with gastric cancer (GC), we compared three DNA methylomes from gastric mucosa (GM), intestinal metaplasia (IM), and gastric tumor (GT) cells that were microscopically dissected from an intestinal-type early gastric cancer (EGC) using methylated DNA binding domain sequencing (MBD-seq) and reduced representation bisulfite sequencing (RRBS) analysis. In this study, we focused on differentially methylated promoters (DMPs) that could be directly associated with gene expression. We detected 2,761 and 677 DMPs between the GT and GM by MBD-seq and RRBS, respectively, and for a total of 3,035 DMPs. Then, 514 (17%) of all DMPs were detected in the IM genome, which is a precancer of GC, supporting that some DMPs might represent an early event in gastric carcinogenesis. A pathway analysis of all DMPs demonstrated that 59 G protein-coupled receptor (GPCR) genes linked to the hypermethylated DMPs were significantly enriched in a neuroactive ligand-receptor interaction pathway. Furthermore, among the 59 GPCRs, six GI hormone receptor genes (NPY1R, PPYR1, PTGDR, PTGER2, PTGER3, and SSTR2) that play an inhibitory role in the secretion of gastrin or gastric acid were selected and validated as potential biomarkers for the diagnosis or prognosis of GC patients in two cohorts. These data suggest that the loss of function of gastrointestinal (GI) hormone receptors by promoter methylation may lead to gastric carcinogenesis because gastrin and gastric acid have been known to play a role in cell differentiation and carcinogenesis in the GI tract.

The Effects of Co-cultivation Medium and Culture Conditions on Rice Transformation Efficiency (공동배양과정의 배지조성과 배양조건이 벼 형질전환효율에 미치는 영향)

  • Kim, Yul-Ho;Park, Hyang-Mi;Choi, Man-Soo;Yun, Hong-Tai;Choi, Im-Soo;Shin, Dong-Bum;Kim, Chung-Kon;Lee, Jang-Yong
    • Korean Journal of Breeding Science
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    • v.41 no.3
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    • pp.252-260
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    • 2009
  • Rice is the most important cereal crop not only in supplying the basic staple food for more than half of the world's population but also as a model plant for functional genomic studies of monocotyledons. Although rice transformation method using A. tumefaciens has already been widely used to generate transgenic plants, the transformation rate is still low in most Korean elite cultivars. We made several modifications of the standard protocol especially in the co-cultivation step to improve the efficiency of the rice transformation. The co-culture medium was modified by the addition of three antioxidant compounds (10.5 mg/L L-cysteine, 1 mM sodium thiosulfate, 1 mM dithiothreitol) and of Agrobacterium growth-inhibiting agent (5 mg/L silver nitrate). Co-cultivation temperature ($23.5^{\circ}C$ for 1 day, $26.5^{\circ}C$ for 6 days) and duration (7 days) were also changed. The plasmid of pMJC-GB-GUS carrying the GUS reporter gene and the bar gene as the selectable marker was used to evaluate the efficiency of the transformation. After co-cultivation, a high level of GUS gene expression was observed in calli treated with the modified method. It is likely that those newly added compounds helped to minimize the damage due to oxidative bursts during plant cell-Agrobacterium interaction and to prevent necrosis of rice cells. And the transformation rate under the modified method was also remarkably increased approximately 8-fold in Heungnambyeo and 2-fold in Ilmibyeo as compared to the corresponding standard method. Furthermore, we could produce the transgenic plants stably from Ilpumbyeo which is a high-quality rice but its transformation rate is extremely low. Transformation and the copy number of transgenes were confirmed by PCR, bar strip and Southern blot analysis. The improved method would attribute reducing the effort and the time required to produce a large number of transgenic rice plants.

Identification of genes involved in inbreeding depression of reproduction in Langshan chickens

  • Xue, Qian;Li, Guohui;Cao, Yuxia;Yin, Jianmei;Zhu, Yunfen;Zhang, Huiyong;Zhou, Chenghao;Shen, Haiyu;Dou, Xinhong;Su, Yijun;Wang, Kehua;Zou, Jianmin;Han, Wei
    • Animal Bioscience
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    • v.34 no.6
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    • pp.975-984
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    • 2021
  • Objective: Inbreeding depression of reproduction is a major concern in the conservation of native chicken genetic resources. Here, based on the successful development of strongly inbred (Sinb) and weakly inbred (Winb) Langshan chickens, we aimed to evaluate inbreeding effects on reproductive traits and identify candidate genes involved in inbreeding depression of reproduction in Langshan chickens. Methods: A two-sample t-test was performed to estimate the differences in phenotypic values of reproductive traits between Sinb and Winb chicken groups. Three healthy chickens with reproductive trait values around the group mean values were selected from each of the groups. Differences in ovarian and hypothalamus transcriptomes between the two groups of chickens were analyzed by RNA sequencing (RNA-Seq). Results: The Sinb chicken group showed an obvious inbreeding depression in reproduction, especially for traits of age at the first egg and egg number at 300 days (p<0.01). Furthermore, 68 and 618 differentially expressed genes (DEGs) were obtained in the hypothalamus and ovary between the two chicken groups, respectively. In the hypothalamus, DEGs were mainly enriched in the pathways related to vitamin metabolism, signal transduction and development of the reproductive system, such as the riboflavin metabolism, Wnt signaling pathway, extracellular matrix-receptor interaction and focal adhesion pathways, including stimulated by retinoic acid 6, serpin family F member 1, secreted frizzled related protein 2, Wnt family member 6, and frizzled class receptor 4 genes. In the ovary, DEGs were significantly enriched in pathways associated with basic metabolism, including amino acid metabolism, oxidative phosphorylation, and glycosaminoglycan degradation. A series of key DEGs involved in folate biosynthesis (gamma-glutamyl hydrolase, guanosine triphosphate cyclohydrolase 1), oocyte meiosis and ovarian function (cytoplasmic polyadenylation element binding protein 1, structural maintenance of chromosomes 1B, and speedy/RINGO cell cycle regulator family member A), spermatogenesis and male fertility (prostaglandin D2 synthase 21 kDa), Mov10 RISC complex RNA helicase like 1, and deuterosome assembly protein 1) were identified, and these may play important roles in inbreeding depression in reproduction. Conclusion: The results improve our understanding of the regulatory mechanisms underlying inbreeding depression in chicken reproduction and provide a theoretical basis for the conservation of species resources.