• Title/Summary/Keyword: ex vitro

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Improvement of ex vitro acclimatization of mulberry plantlets by supplement of abscisic acid to the last subculture medium

  • Huh, Yoon Sun;Lee, Joung Kwan;Nam, Sang Young
    • Journal of Plant Biotechnology
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    • v.44 no.4
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    • pp.431-437
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    • 2017
  • Mulberry (Morus sp.) of the family Moraceae is very economically important in Asian countries including Korea, because its leaf and fruit have been commercially used in sericulture and horticultural industries. Therefore it is necessary to develop the optimal production system for rapid and cost-effective propagation of mulberry. Our studies focused on establishing an acclimatization method for the successful plantlet production of new cultivar 'Cheongsu' which was transferred ex vitro after in vitro culture. In particular, effect of abscisic acid (ABA) addition into the last subculture medium on plantlet response to subsequent ex vitro transfer and its growth was investigated. During acclimatization, stomatal conductance and transpiration rate of ABA-pretreated plantlets were significantly lower than those of non-treated plantlets. Net photosynthetic rate of ABA-pretreated plantlets decreased after ex vitro transfer but increased after 14 days, and it was mostly higher than that of non-treated plantlets. Moreover, relative water content as well as chlorophyll contents and its ratio were also higher in ABA-pretreated plantlets. On the other hand, proline was considerably higher than in control plantlets. After 1 month of ex vitro transfer, survival rate of ABA-pretreated plantlets was 85.6%, which increased by 29.1% in comparison with control (56.5%). More vigorous growth was also observed in ABA-pretreated plantlets. From these results, it was found that application of ABA to the last subculture medium could improve acclimatization and promote survival of mulberry plantlets after ex vitro transfer, inducing water stress tolerance and alleviating abiotic stresses.

Comparison of In Vitro, Ex Vivo, and In Vivo Antibacterial Activity Test Methods for Hand Hygiene Products (손 위생 제품에 대한 in vitro, ex vivo, in vivo 항균 시험법 비교)

  • Daeun Lee;Hyeonju Yeo;Haeyoon Jeong
    • Journal of Food Hygiene and Safety
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    • v.39 no.1
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    • pp.35-43
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    • 2024
  • Numerous methods have been applied to assess the antibacterial effectiveness of hand hygiene products. However, the different results obtained through various evaluation methods have complicated our understanding of the real efficacy of the products. Few studies have compared test methods for assessing the efficacy of hand hygiene products. In particular, reports on ex vivo pig skin testing are limited. This study aimed to compare and characterize the methodologies applied for evaluating hand hygiene products, involving in vitro, ex vivo, and in vivo approaches, applicable to both leave-on sanitizers and wash-off products. Our further aim was to enhance the reliability of ex vivo test protocols by identifying influential factors. We performed an in vitro method (EN1276) and an in vivo test (EN1499 and ASTM2755) with at least 20 participants, against Serratia marcescens or Escherichia coli and Staphylococcus aureus. For the ex vivo experiment, we used pig skin squares prepared in the same way as those used in the in vivo test method and determined the optimal treated sample volumes for sanitizers and the amount of water required to wash off the product. The hand sanitizers showed at least a 5-log reduction in bacterial load in the in vitro test, while they showed little antibacterial activity in the in vivo and ex vivo tests, particularly those with a low alcohol content. For the hand wash products, the in vitro test was limited because of bubble formation or the high viscosity of the products and it showed low antibacterial activity of less than a 1-log reduction against E. coli. In contrast, significantly higher log reductions were observed in ex vivo and in vivo tests, consistently demonstrating these results across the two methods. Our findings revealed that the ex vivo and in vivo tests reflect the two different antibacterial mechanisms of leave-on and wash-off products. Our proposed optimized ex vivo test was more rapid and more precise than the in vitro test to evaluate antibacterial results.

Effect of Harvesting Time and Storage Duration of Viscum album Seeds on in vitro and ex vitro Germination on the Branch of Prunus mume

  • Lee, Hyun Woo;Ghimeray, Amal Kumar;Lee, Bo-Duk;Sharma, Pankaja;Shim, Ie Sung;Park, Cheol Ho
    • Korean Journal of Plant Resources
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    • v.27 no.3
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    • pp.256-262
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    • 2014
  • Viscum album var. coloratum (mistletoe) is considered as one of the endangered plant species in Korea. Our objective is to restore its population and multiplication of plant by ex situ method. In this research we explored the maximum germination (in vitro) of freshly harvested and stored seeds of mistletoe collected in different time intervals. Ex vitro germination after artificial inoculation on the branches of Prunus mume in different physiological conditions was also monitored. The research revealed that the lately harvested seeds (Feb. and March 2014) were superior over early harvested seeds (Nov. 2013 and Jan. 2014) of mistletoe due to the higher percentage of germination (above 93%). According to the data, it is also revealed that the survival and germination rate of mistletoe seeds decreased with the increase in storage duration. In ex vitro germination, the fluctuated temperature of a glass house in natural condition enhanced (four fold) the rate of germination on the branches of Prunus mume than the constant temperature condition in the glass house.

In vitro Propagation and Ex vitro Rooting of Tectona grandis (L.f ), APNBV-1 Clone

  • Ramesh, Kommalapati;Chandra, Mouli Kalla;Vijaya, Tartte
    • Journal of Forest and Environmental Science
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    • v.25 no.2
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    • pp.119-126
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    • 2009
  • An efficient in vitro plant regeneration system was developed through shoot proliferation from axillary buds of Tectona grandis (L.f), APNBV-1 (Andhra Pradesh North Badrachalam Venkatapuram-1) clone. Multiple shoots of high quality were produced in vitro from axillary bud explants. An average of 4.39 shoots/explant were obtained on Murashige and Skoog's (MS) medium supplemented with plant growth regulators (PGRs) benzyl amino purine (BA), kinetin (KN), indole acetic acid (IAA), gibberillic acid ($GA_3$), growth adjuvants casein hydrolysate (CH), adenine sulphate (Ads) and antioxidants ascorbic acid, polyvinyl pyrrollidine (PVP). Eighty five percent of rooting was observed in ex vitro rooting media containing IBA and vermiculite. In ex vitro rooting, single shoots with 2 to 3 nodes were subjected to IBA of different concentrations at different periods of time intervals. Direct rooting in vermiculite at 500 ppm concentration of IBA resulted in 4.3 number of roots with 2 cm length. Minimum response of rooting and length of roots were recorded at 100 ppm concentration of IBA. Planlets were transferred to plastic bags for short acclimatization stage in green house where they survived at 95%.

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Anti-platelet Aggregation Effect of Ginkgolide B and Ginkgoflavonoids, Extracted from Ginkgo biloba, in Vitro, ex Vivo and in Clinic. (Ginkgolide B 및 ginkgoflavonoids의 in vitro와 ex vivo 및 임상에서의 항혈전 작용)

  • 권광일;이영신
    • YAKHAK HOEJI
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    • v.39 no.3
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    • pp.337-345
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    • 1995
  • The effects of ginkgolides(natural mixture of ginkgolides, ginkgolide B, ginkgolide C) and flavonoids(quercetin, kaempferol, myricetin), extracted from Ginkgo biloba, on ADP and PAF-induced platelet aggregation in vitro and ex vivo were investigated. In these experiments, both of ginkgolides and ginkgoflavonoids did not affect the ADP(5 $\mu{M}$) induced platelet aggregation in vitro. The IC$_{50}$ value on PAF (0.3 $\mu{M}$) induced platelet aggregation were 2.52 $\mu{M}$ (ginkgolide B) and 6.35 $\mu{M}$ (natural mixture of ginkgolides) and 2.80 $\mu{M}$ (mixture of ginkgolide B and quercetin). Oral administration of ginkgolide B (1 and 3 mg/kg) and quercetin (3 and 9 mg/kg) to rabbits inhibited ex vivo PAF induced platelet aggregation in a dose-dependent manner. Ginkomin-F tablets administered to the diabetic patients showed inhibitory activities on the ADP and PAF induced platelet aggregation in a dose and time dependent manner.

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Protective effect of Korean diet food groups on lymphocyte DNA damage and contribution of each food group to total dietary antioxidant capacity (TDAC) (한식 식품군의 in vitro 총 항산화능 (TDAC)과 ex vivo DNA 손상 보호효과와의 관련성)

  • Lee, Min Young;Han, Jeong-Hwa;Kang, Myung-Hee
    • Journal of Nutrition and Health
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    • v.49 no.5
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    • pp.277-287
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    • 2016
  • Purpose: This study was performed to compare total phenolic contents, in vitro antioxidant capacity, and reduction effect of Korean food groups on ex vivo DNA damage in human cells and analyze correlations between each indicator. Methods: Vegetable foods in the Korean diet based the results of the KNHANES V-2 (2011) were classified into 10 food groups: cereals, fruits, vegetables, nuts, kimchi, seaweeds, potatoes, mushrooms, legumes, and oils. Eighty-four foods constituted more than 1% of the total intake in each food group and finally designated as vegetable foods in the Korean diet. Total phenolic content of each food group was measured. Further, in vitro antioxidant capacity was measured based on DPPH radical scavenging assay, TEAC assay, and $ORAC_{ROO{\cdot}}$ assay. Ex vivo DNA damage in human lymphocytes was assessed using comet assay. Results: Total phenolic contents of food groups of the Korean diet increased in the order of mushrooms, fruits, vegetables, seaweeds, and kimchi. Meanwhile, antioxidant rankings of food groups as mean values from the three in vitro test methods increased in the order of mushrooms, seaweeds, vegetables, kimchi, and fruits. Protection against ex vivo DNA damage in human lymphocytes was highest in mushrooms, followed by vegetables, fruits, seaweeds, and kimchi. The rankings of the food groups for total phenolic content, in vitro DAC, and ex vivo DNA protection activity were similar, and correlations between each indicator were significantly high. Conclusion: Mushrooms, fruits, vegetables, and seaweeds among the tested food groups in the Korean diet showed high total phenolic contents, in vitro antioxidant capacities, and protection against DNA damage. Correlations between each indicator in terms of total phenolic content, in vitro antioxidant capacity, and ex vivo DNA protection between each food group were found to be particularly high.

Effects of Codonopsis lanceolata Extracts on bFGF-induced Angiogenesis in vitro (더덕추출물에 의한 bFGF-유도 시험관내 혈관신생의 억제)

  • 소준노;김종화
    • KSBB Journal
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    • v.18 no.1
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    • pp.25-29
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    • 2003
  • In this study, we examined the effects of the methanolic extract(CL-ex) of Codonopsis lanceolata on the angiogenesis stimulated with basic fibroblast growth factor(bFGF) in vitro, using porcine pulmonary arterial endothelial cells(PPAECs). In addition, we investigated the endothelial functions involved in angiogenesis, such as proliferation, migration and secretion of matrix metalloproteinases(MMPs), using human umbilical vein endothelial cells(HUVECS). CL-ex inhibited FGF-induced sprout formation in vitro at concentrations of 0.1-100 ug/ml. Although CL-ex did not affect the proliferation of endothelial cells, CL-ex strongly inhibited the FGF-induced migration of HUVECS at concentrations of 0.1-1 ug/ml; the degree of inhibition of endothelial cells by C-ex was 49.4% at 0.1 ug/ml and 71.9 % at 1.0 ug/ml. Moreover, CL-ex inhibited the secretion of MMPs from HUVECS stimulated with FGF. Therefore, the inhibitory effect of CL-ex on angiogenesis in vitro could be explained by the inhibition of endothelial cell migration. From these results, we suggest that Codonopsis lanceolata is a useful herb for the development of therapeutics or preventive food factors for angiogenesis related diseases, such as tumors.

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.

PREVENTION OF MURAL THROMBUS IN POROUS INNER TUBE OF DOUBLE-LAYERED TUBE BY SALINE PERFUSION (생리식염수의 재료표면에의 분출에 의한 이중튜브의 응혈 방지)

  • Kim, Sung-S.;Park, Joon-B.
    • Proceedings of the KOSOMBE Conference
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    • v.1993 no.11
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    • pp.110-113
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    • 1993
  • An in vitro experiment under laminar non-pulsatile blood flow and an acute canine ex vivo femoral A-V series shunt experiment were undertaken to investigate the effectiveness of saline perfusion through pores of porous tubes to prevent formation of mural thrombus. PS/SBR porous tubes were used for the in vitro experiment. Commercially obtained ePTFE porous tubes were etched by sodium naphthalenide, and the etched tubes were used for the ex vivo experiment. According to the results of the in vitro experiment, mural thrombus on the surface of the porous tribe could be prevented by the saline perfusion. Adhered blood cells decreased semi-logarithmically with increased perfusion rate (up to $0.022\;ml/min-cm^2$) of isotonic saline solution. According to results of the ex vivo experiment, mural thrombus decreased with increased perfusion rate (upto $0.060\;ml/min-cm^2$).

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Plant regeneration through multiple-shoot induction and ex vitro rooting in Vaccinium oldhamii Miq. (정금나무(Vaccinium oldhamii Miq.)의 다신초 유도 및 기외발근을 통한 식물체 재분화)

  • Yun, Ayoung;Kim, Tae Dong;Kim, Ji Ah;Lee, Na Nyum;Cheong, Eun Ju;Kim, Yong Wook
    • Journal of Plant Biotechnology
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    • v.49 no.1
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    • pp.82-89
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    • 2022
  • In vitro techniques were developed for propagating Vaccinium oldhamii using shoots with apical buds. Explants having an apical bud were cultured on Murashige and Skoog (MS) medium supplemented with 1.0, 2.0, and 5.0 mg/L of each zeatin, thidiazuron, 6-benzylaminopurine (BA), and 6-(γ,γ-dimethylallylamino)purine (2-iP) in order to induce multiple shoots. Among the tested treatments, the 2.0 mg/L of 2-iP proved to be most suited for the multiplication and growth of shoots; the multiple shoot induction rate was 100.0%, the average number of shoots was 7.4 per explant, and the average shoot length was 51.7 mm. The in vitro elongated shoots were rooted on half-strength MS medium containing various concentrations of indole-3-butyric acid (IBA) and 1-naphthaleneacetic acid (NAA). However, overall callus overgrowth was observed in all treatments and resulted in necrosis and abnormal shoot growth in root formation. A low concentration (0.5 mg/L) of IBA was appropriate for normal root development and the in vitro rooting rate was 30%. Ex vitro treatments on root formation using various concentrations of IBA with Talc powder and two types of rooting substrates (Flexi-Plugs or Horticultural soil) were examined. The ex vitro rooting rate (80%) and length of roots (32.9 mm) were obtained when the cut ends of the shoots were treated with 1.0 mg/L IBA and cultivated in Horticultural soil for 2 months. These findings suggest that ex vitro rooting is the more effective method for improving root formation in Vaccinium oldhamii than in vitro rooting.