• Title/Summary/Keyword: Node culture

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In vitro Multiplication through Single-Node Culture of Sea-Milkwort (Glaux maritima L.) (갯봄맞이(Glaux maritima L.) 실생의 단마디배양을 통한 기내증식)

  • Bae, Su-Ji;Kang, Beum-Chang;Jeong, Mihye;Kim, Soochong;Kim, Chang Kil;Han, Jeung-Sul
    • Horticultural Science & Technology
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    • v.34 no.3
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    • pp.461-471
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    • 2016
  • This study was conducted to establish an in vitro propagation system for sea-milkwort (Glaux maritima L.), which is an endangered coastal plant species with high horticultural value. Two phenotypes, 'Red type (RT)' and 'Pistachio type (PT)' based on the colors of stem and flower, were obtained from a personal horticulturist in 2009 and used for this study as plant materials. The stock plants showed typical morphologies in flower, capsule, and seed appearances as previously reported. Low temperature treatment at $4^{\circ}C$ for four or more weeks after in vitro sowing maximized seed germination percentage, indicating that imbibition of seed and subsequent low temperature treatment are crucial for its germination. The in vitro seedlings had phenotypic variation, falling into 'RT' and 'PT' classes like the stock plants. Although slight differences depending on genotype and medium were recognized, the fourth or fifth nodes detached from the in vitro seedlings revealed the best multiplication efficacy when estimated on the basis of total number of nodes of newly developed axillary shoots. In addition, the nodes from 'RT' and 'PT' regenerated the most shoots on medium supplemented with $0.5mg{\cdot}L^{-1}$ BA alone and $0.5mg{\cdot}L^{-1}$ BA plus $0.5mg{\cdot}L^{-1}$ IAA, respectively. The node culture-derived plantlets were well acclimatized in a culture room ex vitro and completed the pseudo-annual life cycle coincident with that in the natural salt march habitat with the current cultivation method of applying fresh water-irrigation under an inland environment. This work represents the first report of in vitro propagation of sea-milkwort. Thus, our study will contribute to exo-habitat conservation and natural habitat restoration of this endangered species in addition to development of a horticultural product.

Suppressive Effect of Solanum Iyratum Aqueous Extract Via Down-regulation of $TNF-{\alpha}$ and $IFN-{\gamma}$ Production on Collagen-induced Arthritis in Mice

  • Kim, Seung-Hyung;Seo, Chang-Woo;Kim, Chang-Min;Kim, Yang-Jin;Lee, Boo-Kyun;Choi, Yong-Sun;Oh, Hwang;Yoon, Ho-Sok;Lee, Seon-Goo;Lee, Jang-Cheon;Lee, Young-Cheal
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.21 no.5
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    • pp.1278-1284
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    • 2007
  • Solanum Iyratum Thunb (Solanaceae) has multiple applications in korean traditional medicine because of its cytotoxic, immunological and anti-inflammatory activities. However, no study on the anti-arthritic activity of Solanum Iyratum Thunb has been reported in vivo. Rheumatoid arthritis (RA) is a systemic autoimmune disease with chronic inflammation characterized by hyperplasia of synovial cells in affected joints, which ultimately leads to the destruction of cartilage and bone. Cytokine production were assessed during CIA(collagen-induced arthritis) model mice in lymph node (LN), in knee joint and spleen, using ELISA. DBA/1j mice were immunized with bovine type II collagen. After a second collagen immunization, mice were treated with Solanum Iyratum Thunb (SLT) orally at 400, 200 mg/kg once a day for 4 weeks. The severity of arthritis within the knee joints was evaluated by histological assessment of cartilage destruction and pannus formation. SLT significantly suppressed the progression of CIA and inhibited the production of TNF-alpha and IFN-gamma in serum and spleen cell culture supernatant. The erosion of cartilage was dramatically reduced in mouse knees after treatment with SLT. In conclusion, our results demonstrates that SLT significantly suppressed the progression of CIA. This action was characterized by suppression of arthritis index, cartilage erosion and synovial cell infiltration and the decreased production of $TNF-{\alpha}$, $IFN-{\gamma}$, CD4+, CD19+, CD3+/CD69+ cells (in lymph node), CD11b+/Gr-1 + (in knee joint).

Growth and Tuber Yield of Sweet Potato Slips Grown under Different Light-Emitting Diodes (LED 광질에 따른 고구마의 묘소질 및 괴근 수량성)

  • Lee, Na Ra;Lee, Seung Yeob
    • Journal of Bio-Environment Control
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    • v.23 no.4
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    • pp.356-363
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    • 2014
  • This work was conducted to investigate the field growth and yield of the sweetpotato (Ipomoea batatas) slips grown under different light emitting diodes (LEDs). Sweet potato cuttings of 3 cultivars ('Matnami', 'Shinhwangmi', and 'Yeonhwangmi') were cultivated under fluorescent lamp (FL) and several LEDs (PPF $150{\pm}5{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$ at 20cm distance) in deep flow culture system for 20 days. The plants were acclimatized under sunlight for 10 days, and then cuttings (30cm length) were planted with $75{\times}25cm$ planting density on June 10th, covered with black vinyl film during growth period. Length and diameter of vine, number of root were excellent in the red plus blue (7:3) LED than the other treatments. At 30 days after planting, the survival rate in red plus blue (7:3) LED was significantly higher than that in FL and red LED, and it was not different among cultivars. Vine length, vine diameter, and number of node were not significant among LED light qualities and cultivars. After 120 days in the field cultivation, vine length, vine diameter, number of node, number of branch, and fresh weight of shoot were not significant among LED light qualities, but those except the number of branch showed significant differences among cultivars. Yield characteristics among LED light colors were not significant, but weight of storage root per plant, mean weight of storage root, and yield showed significant differences among cultivars. The yield per 10a in 'Matnami', and 'Yeonhwangmi' was significantly higher than that in 'Shinhwangmi'.

Therapeutic Effects of Yijungtang on Atopic Dermatitis-like Skin Lesions of NC/Nga Mouse Induced by Mite Antigen (이중탕(理中湯)이 Mite Antigen으로 유발된 NC/Nga 생쥐의 아토피 피부염에 미치는 영향)

  • Seo, Hui-Yeon;Han, Jae-Kyung;Kim, Yun-Hee
    • The Journal of Pediatrics of Korean Medicine
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    • v.25 no.1
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    • pp.1-27
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    • 2011
  • Objectives: The purpose of this study is to investigate the effects of Yijungtang(YJT) on atopic dermatitis in an in-vitro and in-vivo experiment using a RBL-2H3 mast cells and a NC/Nga atopic dermatitis mouse. Methods: In-vitro experiment, IL-4, IL-13 mRNA expression were evaluated by a real-time PCR, IL-4, IL-13 production by ELISA and transcription factor as GATA-1, GATA-2, NF-AT1, NF-AT2, AP-1 and NF-kB by western blotting. In-vivo experiment, clinical skin score we evaluated by, hematology and Serum total IgE and IgG1 of NC/Nga atopic dermatitis mouse, cytokine level, total number of cell, Immunohistochemical staining and Histological features of auxiliary lymph node(ALN), draining lymph node(DLN), peripheral blood mononuclear cells(PBMCs) and dorsal skin tissue in NC/Nga mouse. Results: YJT decreased IL-4, IL-13 mRNA expression, IL-4, IL-13 production and prominently decreased the expression of mast cell specific transcription factors including GATA-2, NF-AT2, c-Fos and NF-kB. YJT oral administration reduced the levels of skin severity scores. It also decreased the level of inflammatory cytokines such as IL-5, IL-13, histamine and IgE in the serum. It elevated IFN-gamma level in the spleenocyte culture supernatant but decreased. $CD3e^+$, $CD19^+$, $CD4^+$, $CD8^+$, $CD3e^+CD69^+$, $CD11b^+Gr-1^+$, $CCR3^+$ in the PBMCs, $CD4^+$, $CD8^+$, $CD3e^+CD69^+$, $B220^+CD23^+$ in the ALN, $CD4^+$, $CD3e^+CD69^+$ in the ALN and $CD4^+$, $CD11b^+Gr-1^+$ in the dorsal skin. Histological examination showed that infiltration levels of immune cells in the skin of AD-induced NC/Nga mice were much improved by YJT oral administration. Conclusions: The anti-allergic activities of YJT may be mediated by down-regulation of Th2 cytokines, such as IL-4 and IL-13, through the regulation GATA-2, NF-AT2 and NF-kB transcription factors in mast cells. YJT would be regulate molecular mediators and immune cells which are functionally associated with atopic dermatitis induced in NC/Nga mice, and may play an important role in recovering AD symptoms.

Functional Analysis of Fibroblastic Reticular Cells Derived from Mouse Lymph Node via Bidirectional Crosstalk with T Cells (T세포와 양방향 작용을 통한 마우스 림프절로부터 분리된 fibroblastic reticular cell의 기능적 분석)

  • Park, Sung Hee;Lee, Jong-Hwan
    • Journal of Life Science
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    • v.23 no.10
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    • pp.1199-1208
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    • 2013
  • Fibroblastic reticular cells (FRCs) form the structural backbone of the T zone provide a guidance path for immigrating T cells in the lymph node (LN). FRCs may contribute directly to developing T-cell biology in the LN and allow analyses of fundamental aspects of FRC biology related to T cells. FRCs inhibited T-cell apoptosis, and FRC culture supernatants strongly induced the expression of Bcl-xL in T cells against doxorubicin. Coculture of FRC and T cells resulted in rearrangements of the actin cytoskeleton, as well as global changes in the morphology of the FRCs. In addition, when cocultured, the T cells adhered to the FRC monolayer, and the membrane intercellular adhesion molecule (ICAM)-1 was slightly increased by day-dependent manner. In contrast, the expression of soluble ICAM-1 was dramatically increased in a day-dependent manner. Several chemokines, such as CCL5, CXCL1, CXCL5, CXCL16, CCL8, CXCL13, and ICAM-1, and MMPs were expressed in FRCs sensed by tumor necrosis factor (TNF) families. Nuclear factor kappa B ($NF{\kappa}B$)-RelA of the $NF{\kappa}B$ canonical pathway was translocated into FRC nuclear by $TNF{\alpha}$. In contrast, p52 proteolyzed from p100, a counterpart of RelB of the noncanonical $NF{\kappa}B$ pathway, accumulated in the peripheral FRC nucleus by agonistic anti-$LT{\beta}R$ antibody. In summary, we propose a model in which FRCs engage in bidirectional crosstalk to increase the efficiency of T-cell biology. This cooperative feedback loop may help to maintain tissue integrity and function during immune responses.

Effect of Day/Night Temperatures during Seedling Culture on the Growth and Nodes of Early Flower Cluster Set of 'Seokwang' Tomato (Lycopersicum esculentum Mill.) (육묘시의 주야간 기온이 서광 토마토의 생육 및 초기 착화 절위에 미치는 영향)

  • 김오임;정병룡
    • Journal of Bio-Environment Control
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    • v.8 no.2
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    • pp.75-82
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    • 1999
  • This study was carried out to examine the effect of day/nignt temperatures during seedling culture on the vegetative and reproductive growth of Lycopersicum esculentum ‘Seokwang’. The study was consisted of two culture stages, plug seedling production in the growth chamber and hydroponic culture of the plant in a glasshouse. Experiments were replicated over time. The germinated seedlings were raised for 33 days (experiment 1) and 35 days (experiment 2) in 4 growth chambers, each with day/night temperatures of either $25^{\circ}C$/$25^{\circ}C$, 16$^{\circ}C$/16$^{\circ}C$, 16$^{\circ}C$/$25^{\circ}C$ or $25^{\circ}C$/16$^{\circ}C$. Cool-white fluorescent lamps provided 140$\mu$mol.m$^{-2}$ .s$^{-1}$ light for 12h each day. In the second experiment, all chambers were supplied with 1000$\mu$mol.mol$^{-1}$ CO$_{2}$ during the photoperiod and had an air velocity of 0.3m.s$^{-1}$ and relative humidity of 80%. Plug seedlings raised were transplanted to rockwool slabs in a glasshouse and were grown hydroponically using the same nutrient solutions used for seedling culture for 37 days (experiment 1) and 35 days (experiment 2). Plant height was affected more by mean daily temperature than by interaction of day and night temperatures. Plant height was the highest in 16/16$^{\circ}C$ treatment. Leaf count was not affected by day and night temperatures, and the chlorophyll concentration was the highest in 16/$25^{\circ}C$ treatment. Fresh and dry weights of stem tended to be greater in treatments of constant day and night temperature. The number of node on which first and second flower clusters were set was significantly higher in 25/$25^{\circ}C$ treatment than in the other treatments. Days to flower of the first flower on the first flower cluster were the greatest in 25/$25^{\circ}C$ and the least in 16/$25^{\circ}C$ treatment. Vegetative and reproductive growth, such as height, fresh and dry weights, days to flower, and nodes of the 1st and 2nd flower cluster set were affected by day/night temperatures.

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Performance of Quantitative Real-Time PCR for Detection of Tuberculosis in Granulomatous Lymphadenitis Using Formalin-Fixed Paraffin-Embedded Tissue

  • Munkhdelger, Jijgee;Mia-Jan, Khalilullah;Lee, Dongsup;Park, Sangjung;Kim, Sunghyun;Choi, Yeonim;Wang, Hye-Young;Jeon, Bo-Young;Lee, Hyeyoung;Park, Kwang Hwa
    • Biomedical Science Letters
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    • v.19 no.2
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    • pp.153-157
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    • 2013
  • Although culture is the gold standard method to identify mycobacteria, its use in tuberculous lymphadenitis (TBL) is limited due to formalin fixation of the submitted specimens. We evaluated the performance of quantitative real-time PCR (q-PCR) for Mycobacterium Tuberculosis (MTB) in granulomatous lymphadenitis using formalin-fixed paraffin-embedded (FFPE) tissues. From 2000 to 2010, a total number of 117 cases of lymph node samples with granulomatous inflammation which were surgically removed and fixed in formalin were studied. Hematoxylin & Eosin (H&E) and Ziehl-Neelsen-stained (ZN) slides were reviewed. qPCR using Real TB-Taq$^{(R)}$ was performed for all cases to identify Mycobacterium tuberculosis. Thirteen non-tuberculous lymphadenopathy cases were used as negative control. Cervical lymph nodes were more frequently affected (60%, 70/117) than other sites. ZN stain for acid fast bacilli was positive in 19 (16.24%) cases. qPCR for tuberculosis was positive in 92 (78.63%) cases. Caseous necrosis was found in 103 (88.03%) cases. While the ZN stain and qPCR were both negative in all control cases, the qPCR showed a significantly higher positive rate (78.63% vs. 16.24%) compared to ZN stain in histologically diagnosed TBL. Quantitative real-time PCR proves to be more sensitive than ZN stain for diagnosis of tuberculous lymphadenitis.

Measurement of Mass Flow of Water in the Stem of Musk Melon by Sap Flow Gauge (열목지 경유센서에 의한 멜론 경유양의 측정)

  • 강곡명;양원모
    • Journal of Bio-Environment Control
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    • v.7 no.3
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    • pp.268-274
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    • 1998
  • The mass flow of water in the stem of melon measured by Sap Flow Gauge was compared with the actual flow calculated by the difference between supply and drainage nutrient water to investigate the possibility and accuracy of estimation of melon's transpiration in rockwool culture. The Sap Flow Gauge which was made with copper-constantan theromocouple and nichrome fiber by our research team, was attached to the 3rd node of melon. The outdoor temperature, room temperature, solar radiation and relative humidity were continually measured. The amount of supply and drainage nutrient water were simultaneously measured for calculation of practical consumption of nutrient water to compare with mass flow of sap. The measuring errors of Sap Flow Gauge were 0.3 to 31.8%, which were small at solar radiation of 20MJ.m$^{2}$.d$^{-1}$ . The mass flow of water was lower for the measured value by Sap Flow Gauge than the actual value at higher solar intensity, however it was higher at lower solar intensity The variation of error rate of each Sap Flow Gauge was 0.1 to 13.0%. The measuring error with Sap Flow Gauge was negatively related with solar intensity and temperature. Therefore, to measure more exactly the mass flow of sap for estimation of melon's transpiration, the compensation factor must be calculated.

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Detection of Aspergillus and Penicillium genera by Enzyme-Linked Immunosorbent Assay Using a Monoclonal Antibody

  • Kwak, Bo-Yeon;Shon, Dong-Hwa;Kwon, Byung-Joon;Kweon, Chang-Hee;Lee, Ke-Ho
    • Journal of Microbiology and Biotechnology
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    • v.11 no.1
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    • pp.21-28
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    • 2001
  • Enzyme linked-immunosorbent assay (ELISA) for a rapid detection of fungi, Aspergillus and Penicillium genera in food, were developed and their efficiencies were approved by detecting artificially contaminated agricultural commodities. Mice were immunized with partially purified Aspergillus flavus extracellualr polysaccharide (EPS) and lymph node cells of the mice were fused with the myeloma cells for production of monoclonal antibodies. Mab 1G11, one of the antibodies, was selected and purified. A sandwich ELISA was established and its detection limit toward A. flavus EPS was 1mg/ml. Among the 59 strains tested (including 18 species of Aspergillus, 16 of Penicillium, 11 of Fusarium, 1 of Absidia, 2 of Alternaria, 2 of Candida, 2 of Cladosporium, 2 of Geotrichum, 2 of Mucor, 2 of Rhizopus, 1 of Trichoderma), species of Aspergillus and penicillium had a high reactivity with Mab 1G11 even up to 10,000 times dilution of culture broths. The other genera except Cladosporium resinae showed no reactivity, thus Mab 1G11 was specific to the genera of Aspergillus and Penicillium. The epitope of A. flavus EPS against monoclonal Mab 1G11 was on the carbohydrate moiety when 1 to 100$\mu g/g$ A. flavus EPS were put into rice, potato, and mandarin orange, the average recoveries detected by sandwich ELIA were 123, 59, and 76%, respectively. Correlation was found to be linear between the EPS, and mycelium of A. flavus and Penicillium citrinum grown in a liquid medium (r=0.87 and 0.96), and also between the EPS and colony forming unit in solid media of rice of potato (r=0.91-0.99).

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Effect of carbon source and concentration on in vitro regeneration and propagation in Lycopus lucidus by node culture (쉽싸리의 기내 마디배양 시 탄소급원의 종류와 농도가 식물체 재분화 및 증식에 미치는 영향)

  • Lee, Na-Nyum;Kim, Ji Ah;Kim, Yong Wook;Kim, Tae Dong
    • Journal of Plant Biotechnology
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    • v.45 no.2
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    • pp.131-139
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    • 2018
  • We examined the effect of carbon sources on the regeneration and ex vitro acclimatization of Lycopus lucidus Turcz. ex Benth. Plantlets were regenerated on the 1/2MS medium supplemented with different concentrations (3 ~ 10%) of sucrose and glucose. The sucrose concentrations of 3% and 5% that were supplied enhanced shoot multiplication and rooting but hampered high concentration growth (including the length of the shoot and root). During ex vitro acclimatization, the tuberization of the root, the root length, the shoot length and the survival rate of Lycopus lucidus plantlets grown using 3% and 5% sucrose were found to be better than the other carbon sources and concentrations. Thus a sucrose concentration of 3% and 5% in the 1/2MS medium appeared to be better for both in vitro growth and ex vitro acclimatization of Lycopus lucidus.