• Title/Summary/Keyword: ex vitro

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Rapid Micropropagation by Stem Node Culture of Japanese Yew (주목의 줄기절간 조직배양에 의한 급속 대량증식)

  • 선정훈
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.6
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    • pp.335-337
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    • 1995
  • The effect of plant growth regulators on proliferation of shoot from stem node culture of Japanese yew (Taxus cuspidata Sieb. et Zucc.) was studied using Quoirin and Lepoivre (1977) medium. Among the cytokinin tested, BAP, kinetin, and thidiazuron at various concentrations had no effect on shoot multiplication However when zeatin at 5$\times$10$^{-5}$ M was added to the medium, an average of 6 shoots were regenerated per explant after 8 weeks of culture. The ratio of rooting ex vitro was remarkably increased up to 34% by dipping the basal end in 0.5 to 1.0% IBA on talc compared with 3% in vitro rooting. Rooted plantlets were acclimated in greenhouse conditions for one month and successfully transplanted to the field.

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Antiplatelet Action of Ilexoside D, a Triterpenoid Saponin from Ilex pubescens

  • Lee, Dug-Keun;Lee, Hye-Sun;Huh, Min-Do;Lee, Chul-Hoon;Lee, Young-Su;Kim, Hyun-Su;Han, Yong-Nam
    • Archives of Pharmacal Research
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    • v.14 no.4
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    • pp.352-356
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    • 1991
  • The anti-platelet activity of ilexoside D isolated from the roots of Ilex pubescens Hook. et Arn. was investigated in in vitro and ex vivo models of platelet aggregation induced by ADP, thrombin or collagen in rats. In vitro ilexoside D inhibited more effectively platelet aggregation induced by ADP and thrombin than by collagen as compared with aspirin. Ex vivo ilexoside D also inhibited platelet aggregation induced by ADP and collagen, but not by thrombin, and the inhibitory action of ilexoside D was more effective than that of aspirin. However, in vitro ilexoside D inhibited very poorly the generation of malonyldialdehyde, which is known to be concomitantly released with thromboxane $A_2$ during platelet aggregation. These results suggest that the anti-platelet activity of ilexoside D may not be responsible for prostaglandin synthesis in platelets.

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Depression of L-type $Ca^{2+}$ and Transient Outward $K^+$ Currents in Endotoxin-treated Rat Cardiac

  • Park, Kyu-Sang;Lee, Boo-Soo;Kong, In-Deok;Lee, Joong-Woo
    • The Korean Journal of Physiology and Pharmacology
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    • v.3 no.6
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    • pp.623-630
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    • 1999
  • Decreased cardiac contractility occurs in endotoxicosis, but little is known about the ionic mechanism responsible for myocardial dysfunction. In this study, we examined the changes in $Ca{2+}$ and $K^+$ currents in cardiac myocytes from endotoxin-treated rat. Ventricular myocytes were isolated from normal and endotoxemic rats (ex vivo), that were treated for 10 hours with Salmonella enteritidis lipopolysaccharides (LPS; 1.5 mg/kg) intravenously. Normal cardiac myocytes were also incubated for 6 hours with 200 ng/ml LPS (in vitro). L-type $Ca{2+}$ current $(I_{Ca,L})$ and transient outward $K^+$ current $(I_{to})$ were measured using whole cell patch clamp techniques. Peak $I_{Ca,L}$ was reduced in endotoxemic myocytes (ex vivo; 6.00.4 pA/pF, P<0.01) compared to normal myocytes (control; 10.90.6 pA/pF). Exposure to endotoxin in vitro also attenuated $I_{Ca,L}$ (8.40.4 pA/pF, P<0.01). The amplitude of $(I_{to})$ on depolarization to 60 mV was reduced in endotoxin treated myocytes (16.51.5 pA/pF, P<0.01, ex vivo; 20.00.9 pA/pF, P<0.01 , in vitro) compared to normal myocytes (control; 24.71.0 pA/pF). There was no voltage shift in steady-state inactivation of $I_{Ca,L}$ and $(I_{to})$ between groups. These results suggest that endotoxin reduces $Ca{2+}$ and $K^+$ currents of rat cardiac myocytes, which may lead to cardiac dysfunction.

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Ginsenoside Rh2 inhibiting HCT116 colon cancer cell proliferation through blocking PDZ-binding kinase/T-LAK cell-originated protein kinase

  • Yang, Jianjun;Yuan, Donghong;Xing, Tongchao;Su, Hongli;Zhang, Shengjun;Wen, Jiansheng;Bai, Qiqiang;Dang, Dongmei
    • Journal of Ginseng Research
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    • v.40 no.4
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    • pp.400-408
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    • 2016
  • Background: Ginsenoside Rh2 (GRh2) is the main bioactive component in American ginseng, a commonly used herb, and its antitumor activity had been studied in previous studies. PDZ-binding kinase/T-LAK cell-originated protein kinase (PBK/TOPK), a serine/threonine protein kinase, is highly expressed in HCT116 colorectal cancer cells. Methods: We examined the effect of GRh2 on HCT116 cells ex vivo. Next, we performed in vitro binding assay and in vitro kinase assay to search for the target of GRh2. Furthermore, we elucidated the underlying molecular mechanisms for the antitumor effect of GRh2 ex vivo and in vivo. Results: The results of our in vitro studies indicated that GRh2 can directly bind with PBK/TOPK and GRh2 also can directly inhibit PBK/TOPK activity. Ex vivo studies showed that GRh2 significantly induced cell death in HCT116 colorectal cancer cells. Further mechanistic study demonstrated that these compounds inhibited the phosphorylation levels of the extracellular regulated protein kinases 1/2 (ERK1/2) and (H3) in HCT116 colorectal cancer cells. In vivo studies showed GRh2 inhibited the growth of xenograft tumors of HCT116 cells and inhibited the phosphorylation levels of the extracellular regulated protein kinases 1/2 and histone H3. Conclusion: The results indicate that GRh2 exerts promising antitumor effect that is specific to human HCT116 colorectal cancer cells through inhibiting the activity of PBK/TOPK.

Effects of Host Tree Species, Temperature and Humidity on ex vitro Seed Germination in Endangered Species of Loranthus tanakae (기주목, 온도 및 습도가 멸종위기 종 꼬리겨우살이 종자의 기외발아에 미치는 영향)

  • Lee, Su-Gwang;Lee, Song-Hee;Park, Kwang-Woo;Kwon, Yeong-Han;Kang, Ho-Duck
    • Journal of Korean Society of Forest Science
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    • v.99 no.6
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    • pp.871-877
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    • 2010
  • This study was conducted to investigate the effects of host tree species, temperature and humidity on ex vitro for seed germination in endangered species of Loranthus tanakae. In addition, we compared seed shapes between Loranthus tanakae and Viscum album that we could have easily shown in ex vitro condition. Seeds were germinated after one week inoculation and followed to develop radicles. Seed germination rates of Loranthus tanakae were 80~95% in most of the experimental conditions. The highest rate of holdfast penetrated to host plants was 72% in Populus alba var. pyramidalis among 13 different species tested at $20^{\circ}C$. Also the rates of their penetration were 57% in Morus bombycis, 55% in Acer palmatum and 42% in Castanea crenata at $20^{\circ}C$. Seeds were germinated under condition without irrigation and followed to withered in 12 weeks later. Stages of seed germination of the Loranthus tanakae were followed by radicle induction, holdfast development, haustorium formation and penetration in order in total period of 14 weeks.

Lipid A of Salmonella typhimurium Suppressed T-cell Mitogen-Induced Proliferation of Murine spleen Cells in the Presence of Macrophage (Salmonella typhimurium lipid A를 처리한 식세포 존재 조건에서 mitogen에 유도되는 이자 세포의 증식억제)

  • Kang, Gyong-Suk;Chung, Kyung-Tae
    • Journal of Life Science
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    • v.17 no.1 s.81
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    • pp.31-38
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    • 2007
  • Infection with virulent or attenuated Salmonella typhimuriumhas known to induce reduction in proliferative responses of spleen cells. We investigated a role of lipid A from S. typhimurium, a B cell mitogen, on proliferation of spleen cells by T cell mitogens such as concanavaline A and phytohemagglutinin under in vitro and ex vivo conditions. Lipid A alone induced proliferation of spleen cells in vitroin a dose-dependent manner. However, subsequent treatment of concanavaline A or phytohemagglutin in after lipid A treatment induced proliferation suppression of murine spleen cells in vitro and ex vivo. Removal of macrophages from spleen cells, which were obtained from a lipid A-injected mouse, restored proliferation by concanavaline A and phytohemagglutinin, indicating that macrophages appeared to play a role in lipid A-induced suppression. Secreted molecules from macrophages did not accounted for the suppression because suppressive effect was not achieved when the supernatant from macrophage-containing spleen cell culture was conditoned to macrophage-depleted spleen cell culture. Co-culture of spleen cells from lipid A-treated and - untreated mice showed proliferation suppression as increasing cell numbers of lipid A-treated mouse. These data suggested that the cell-to-cell contact of macrophage with splenic lymphocyte cells is responsible for immune responses against lipid A, which is applicable to the case of human S. typhi infection.

Gametophyte Propagation and Sporophyte Formation of Asian Chain Fern [Woodwardia japonica (L. f.) Sm.] Under Various Medium Conditions In Vitro and Ex Vitro

  • Cho, Ju Sung;Jang, Bo Kook;Park, Kyungtae;Lee, Ha Min;Lee, Cheol Hee
    • Korean Journal of Plant Resources
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    • v.32 no.6
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    • pp.735-742
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    • 2019
  • This study investigated a suitable method that could be applied for Asian chain fern [Woodwardia japonica (L. f.) Sm.] to propagate gametophytes and promote sporophyte formation. The gametophytes used in all experiments were obtained from germinated spores in vitro and were subcultured at 8-week intervals. The most appropriate media for gametophyte propagation was identified by culturing 300 mg of gametophyte in Murashige and Skoog (MS) basal medium (1/8, 1/4, 1/2, 1, 2), and Knop medium for 8 weeks. As a result, fresh weight of the gametophyte was increased by 56.7-fold on MS medium. Moreover, antheridium formation as well as gametophyte growth was improved on MS medium, especially. To improve the sporophyte formation ex vitro, 1.0 g of gametophyte was ground with distilled water and spread on eight combinations onto four different culture mediums, such as bed soil, peat moss, perlite and decomposed granite. Then generation and growth of sporophytes were investigated after cultivation for 10 weeks. As a result of this experiment, peat moss had a promotive effect of sporophyte formation at single-use and mixed culture soils. In particular, a mixture of bed soil, peat moss and perlite in a 1:1:1 ratio (v/v/v) led to the accelerated formation (782.5 ea/pot) and the frond growth of sporophytes. This included increases in length and width of fronds. However, promotive effect of gametophyte growth and sporophyte formation was not found at single-use and treatment with high ratio of bed soil.

Several Factors Affecting on In vitro Culture of Prothallus and Ex Vitro Sporophyte Formation from Prothallus of Dyropteris varia (L.) O. Kuntze (족제비고사리 전엽체의 기내배양 및 기외 포자체 형성에 미치는 제요인)

  • Jeong Jin-A;Lee Cheol-Hee
    • Korean Journal of Plant Resources
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    • v.19 no.2
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    • pp.252-258
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    • 2006
  • The most effective conditions of In vitro culture and ex vitro sporophyte formation from prothallus were studied for mass propagation of Dyropteris varia. The most effective medium of prothallus proliferation was Murashige and Skoog's basal medium supplemented with 10:50mM of $NH_4^+:NO_3^-$ and 2% sucrose. The optimum pH level was 5.8 and prothallus growth was promoted on medium containing $0.6{\sim}0.8%$ agar. Almost of the tested growth regulators (NAA, IAA, 2,4-D, BAP, kinetin and 2ip) were inhibitory in prothallus proliferation as the concentration of growth regulators became higher. The highest number of sporophytes was obtained by transplanting prothallus on compost only than on any other soil compositions. Sporophyte formation was promoted remarkably by soaking prothallus with $100{\mu}M\;GA_3$ for 3 hours.

Toxicity Assessment of Gas Phase in Cigarette Smoke Using Cell-free Assay

  • Park, Chul-Hoon;Sahn, Hyung-Ok;Shin, Han-Jae;Lee, Hyeong-Seok;Min, Yaung-Keun;Hyun, Hak-Chul
    • Journal of the Korean Society of Tobacco Science
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    • v.29 no.2
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    • pp.110-117
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    • 2007
  • In vitro toxicity tests such as cytotoxicity, mutagenicity and genotoxicity assay are useful for evaluating the relative toxicity of smoke or smoke condensates obtained from different cigarette configurations. A major disadvantage of these tests is relatively time-consuming, complicated and expensive. Recently, a cell-free glutathione consumption assay (GCA) as a rapid and simple screening method for the toxicity assessment of smoke has been reported by Cahours et al. (CORESTA, 2006). This study was carried out to assess the GCA application capable of predicting the toxicity of gas/vapor phase (GVP) of cigarette smoke and to identify individual compounds responsible for the glutathione (GSH) consumption in smoke. Each GVPs from 2R4F, standard cigarette, carbon filter cigarette (ExC) and new carbon filter cigarette (ExN), test cigarettes were collected by automatic smoking machine and evaluated the relative toxicity by GCA and neutral red uptake (NRU) assay. Toxic compounds existed in smoke were also chosen, relative toxicities of these compounds were screened by using two methods and compared individually. The overall order of toxicity by GCA was 2R4F > ExC > ExN, which was consistent with the result of Neutral Red Uptake assay. The levels of carbonyl compounds of ExN were lower than those of 2R4F and ExC, indicating that GSH consumption was associated with carbonyl compound yields. A major toxicant under current study is acrolein, which contributed to more than half of the GSH consumption. Collectively, the toxicity of GVP determined by GCA method may be mainly attributed to acrolein.

The Effects of Anti-Thrombotic Activities and Cardiovascular Improvement of Fermented Garlic Extracts (발효마늘 추출물의 항혈전 및 심혈관개선 효과)

  • Kim, Hyun Kyoung
    • The Journal of the Convergence on Culture Technology
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    • v.6 no.2
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    • pp.567-572
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    • 2020
  • The purpose of this study was to investigate the effects of Anti-Thrombotic Activities and Cardiovascular Improvement of Fermented Garlic Extracts. The incidence of cardiovascular diseases (CVDs) is increasing rapidly in developed countries, with CVDs now representing the leading cause of morbidity and mortality. Natural products and ethnomedicines have been shown to reduce the risk of CVDs. Garlic is a medicinal plant used throughout the world for its anti-inflammatory, antioxidant, and antiplatelet activities. We hypothesized that fermented preparations of these products may possess stronger antiplatelet effects than the non-fermented forms owing to the increased bioavailability of the bioactive compounds produced during fermentation. Therefore, we compared these compounds via in vitro and ex vivo platelet aggregation assays by using standard light transmission aggregometry and ex vivo granule secretions from rat platelets. We found that fermented preparations exerted more potent and significant inhibition of platelet aggregation both in vitro and ex vivo. Likewise, ATP release from dense granules of platelets was also significantly inhibited in fermented preparation-treated rat platelets compared to that in non-fermented preparation-treated ones. We concluded that fermented preparations exerted more potent effects on platelet function both in vitro and ex vivo, possibly as a result of the increased bioavailability of active compounds produced during fermentation. We therefore suggest that fermented products may be potent therapeutics against platelet-related CVDs and can be used as antiplatelet and antithrombotic agents.