• Title/Summary/Keyword: in vitro cultivation

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Cytopathic Change and Inflammatory Response of Human Corneal Epithelial Cells Induced by Acanthamoeba castellanii Trophozoites and Cysts

  • Sohn, Hae-Jin;Seo, Ga-Eun;Lee, Jae-Ho;Ham, A-Jeong;Oh, Young-Hwan;Kang, Heekyoung;Shin, Ho-Joon
    • Parasites, Hosts and Diseases
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    • v.57 no.3
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    • pp.217-223
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    • 2019
  • Acanthamoeba castellanii has ubiquitous distribution and causes primary acanthamoebic keratitis (AK). AK is a common disease in contact lens wearers and results in permanent visual impairment or blindness. In this study, we observed the cytopathic effect, in vitro cytotoxicity, and secretion pattern of cytokines in human corneal epithelial cells (HCECs) induced by A. castellanii trophozoites and/or cysts. Morphological observation revealed that panked dendritic HCECs co-cultured with amoeba cysts had changed into round shape and gradually died. Such changes were more severe in co-culture with cyst than those of co-cultivation with trophozoites. In vitro cytotoxicity assay revealed the highest cytotoxicity to HCECs in the co-culture system with amoeba cysts. A. castellanii induced the expression of $IL-1{\alpha}$, IL-6, IL-8, and CXCL1 in HCECs. Secreted levels of $IL-1{\alpha}$, IL-6, and IL-8 in HCECs co-cultured with both trophozoites and cysts were increased at an early incubation time (3 and 6 hr). These results suggested that cytopathic changes and pro-inflammatory cytokines release of HCECs in response to A. castellanii, especially amoebic cysts, are an important mechanism for AK development.

Effect of harvesting time and night temperature on tuber production of calla (Zantedeschia) (수확시기와 야간온도가 유색칼라(Zantedeschia)의 구근생산에 미치는 영향)

  • Nam, Chun-Woo;Yoo, Dong-Lim;Kim, Su-Jeong;Suh, Jong-Teak;Paek, Kee-Yoeup;Lee, Sang Gyu;Yoon, Moo Kyung
    • Korean Journal of Agricultural Science
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    • v.40 no.4
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    • pp.271-276
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    • 2013
  • This experiment was carried out to determine optimal culture conditions for the production of tubers of Calla (Zantedeschia elliottiana 'Golden Affair' and 'Black Magic') in Korea highland. In vitro produced plantlets and tuberlets of Calla 'Golden Affair' and 'Black Magic' were planted plastic film greenhouse and grown for 100, 120, 140 days, with different night temperature treatments ($0{\sim}10^{\circ}C$ : no heating, 10, $15^{\circ}C$). In both cultivars, tuber size(tuber diameter, tuber height) and tuber weight increased with increasing cultivation period when the night temperature was maintained at $10^{\circ}C$. The largest tuber diameter in vitro produced plantlets was 5.8cm in 'Black Magic' and 3.2cm in 'Golden Affair', and daily tuber growth rate was 1.110g in 'Black Magic' and 0.092g in 'Golden Affair' under the culture conditions. Consequently we think that tuber harvest date was Oct. 30 and night temperature was $10^{\circ}C$ and no heating that was proper method of tuber production. However we had selection of $10^{\circ}C$ treatment for tuber production because it appeared freezing damage occasionally in highland late in October.

Evaluation of a Fungal Strain, Myrothecium roridum F0252, as a Bioherbicide Agent

  • Lee, Hyang-Burm;Kim, Jin-Cheol;Hong, Kyung-Sik;Kim, Chang-Jin
    • The Plant Pathology Journal
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    • v.24 no.4
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    • pp.453-460
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    • 2008
  • In the course of in vitro and in vivo screening for bioherbicidal agents, a hyphomycete fungus, Myrothecium sp. F0252 was selected as a candidate for the biocontrol of weeds. The isolate was identified as Myrothecium roridum Tode ex. Fries based on the morphological characteristics and 18S ribosomal DNA sequence analysis and registered as Myrothecium roridum F0252. In order to evaluate the in vitro effect of M. roridum F0252 on germination of ladino clover and white clover (Trifolium repens L.) seeds, spore solution of the fungus was employed in two concentrations, $6.5{\times}10^6$ and $2.5{\times}10^7$ spores per mL and then inoculated to the seeds. The fungal spores inhibited the seed germination, infected the seedlings, and caused an abnormal withering and inhibition of seedling growth. In addition, when the herbicidal activity of crude ethyl acetate extract from the liquid culture was assessed on a mini-plant, duck-weed (Lemna paucicostata (L.) Hegelm.), the extract showed high inhibitory effect at the level of $12.5{\mu}g$ per mL. On the other hand, in vivo herbicidal activity of M. roridum F0252 was evaluated by a whole plant spray method. M. roridum F0252 exhibited strong and broad-spectrum herbicidal activity. The herbicidal values ranged from 95-100% against 7 weeds, including Abutilon avicennae and Xanthium strumarium, and 70-80% against Digitaria sanguinalis and Sagittaria pygmaea. When the nutritional utilization (95 carbon sources) pattern of M. roridum F0252 was investigated, it varied with water activity ($a_w$) and temperature conditions, supplying good, basic information in regard to nutritional utilization for proper cultivation and formulation. Our results showed that M. roridum F0252 might be used as a potential biocontrol agent against weedy plants.

Effects of inorganic salts on biomass production, cell wall components, and bioethanol production in Nicotiana tabacum

  • Sim, Seon Jeong;Yong, Seong Hyeon;Kim, Hak Gon;Choi, Myung Suk;Choi, Pil Son
    • Journal of Plant Biotechnology
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    • v.48 no.4
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    • pp.278-288
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    • 2021
  • The development of bioenergy through biomass has gained importance due to the increasing rates of fossil fuel depletion. Biomass is important to increase the productivity of bioethanol, and production of biomass with high biomass productivity, low lignin content, and high cellulose content is also important in this regard. Inorganic salts are important in the cultivation of biomass crops for the production of biomass with desirable characteristics. In this study, the roles of various inorganic salts in biomass and bioethanol production were investigated using an in vitro tobacco culture system. The inorganic salts evaluated in this study showed dramatic effects on tobacco plant growth. For example, H2PO4 substantially improved plant growth and the root/shoot (R/S) ratio. The chemical compositions of tobacco plants grown in media after removal of various inorganic salts also showed significant differences; for example, lignin content was high after Mg2+ removal treatment and low after K+ treatment and H2PO4 removal treatment. On the other hand, NO3- and H2PO4 treatments yielded the highest cellulose content, while enzymatic hydrolysis yielded the highest glucose concentration ratio 24 h after NH4+ removal treatment. The ethanol productivity after H2PO4 removal treatment was 3.95% (w/v) 24 h after fermentation and 3.75% (w/v) after 36 h. These results can be used as the basis for producing high-quality biomass for future bioethanol production.

Establishing porcine jejunum-derived intestinal organoids to study the function of intestinal epithelium as an alternative for animal testing

  • Bo Ram Lee;Sun A Ock;Mi Ryung Park;Min Gook Lee;Sung June Byun
    • Journal of Animal Reproduction and Biotechnology
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    • v.39 no.1
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    • pp.2-11
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    • 2024
  • Background: The small intestine plays a crucial role in animals in maintaining homeostasis as well as a series of physiological events such as nutrient uptake and immune function to improve productivity. Research on intestinal organoids has recently garnered interest, aiming to study various functions of the intestinal epithelium as a potential alternative to an in vivo system. These technologies have created new possibilities and opportunities for substituting animals for testing with an in vitro model. Methods: Here, we report the establishment and characterisation of intestinal organoids derived from jejunum tissues of adult pigs. Intestinal crypts, including intestinal stem cells from the jejunum tissue of adult pigs (10 months old), were sequentially isolated and cultivated over several passages without losing their proliferation and differentiation using the scaffold-based and three-dimensional method, which indicated the recapitulating capacity. Results: Porcine jejunum-derived intestinal organoids showed the specific expression of several genes related to intestinal stem cells and the epithelium. Furthermore, they showed high permeability when exposed to FITC-dextran 4 kDa, representing a barrier function similar to that of in vivo tissues. Collectively, these results demonstrate the efficient cultivation and characteristics of porcine jejunum-derived intestinal organoids. Conclusions: In this study, using a 3D culture system, we successfully established porcine jejunum-derived intestinal organoids. They show potential for various applications, such as for nutrient absorption as an in vitro model of the intestinal epithelium fused with organ-on-a-chip technology to improve productivity in animal biotechnology in future studies.

Proteomic Changes in Chick Brain Proteome Post Treatment with Lathyrus Sativus Neurotoxin, β-N-Oxalyl-L-α,β-Diaminopropionic Acid (L-ODAP): A Better Insight to Transient Neurolathyrism

  • Anil Kumar, D;Natarajan, Sumathi;Omar, Nabil A M Bin;Singh, Preeti;Bhimani, Rohan;Singh, Surya Satyanarayana
    • Toxicological Research
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    • v.34 no.3
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    • pp.267-279
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    • 2018
  • Neurolathyrism is a neurodegenerative disorder characterized by spastic paraplegia resulting from the excessive consumption of Lathyrus sativus (Grass pea). ${\beta}$-N-Oxalyl-L-${\alpha},{\beta}$-diaminopropionic acid (L-ODAP) is the primary neurotoxic component in this pea. The present study attempted to evaluate the proteome-wide alterations in chick brain 2 hr and 4 hr post L-ODAP treatment. Proteomic analysis of chick brain homogenates revealed several proteins involved in cytoskeletal structure, signaling, cellular metabolism, free radical scavenging, oxidative stress and neurodegenerative disorders were initially up-regulated at 2 hr and later recovered to normal levels by 4 hr. Since L-ODAP mediated neurotoxicity is mainly by excitotoxicity and oxidative stress related dysfunctions, this study further evaluated the role of L-ODAP in apoptosis in vitro using human neuroblastoma cell line, IMR-32. The in vitro studies carried out at $200{\mu}M$ L-ODAP for 4 hr indicate minimal intracellular ROS generation and alteration of mitochondrial membrane potential though not leading to apoptotic cell death. L-ODAP at low concentrations can be explored as a stimulator of various reactive oxygen species (ROS) mediated cell signaling pathways not detrimental to cells. Insights from our study may provide a platform to explore the beneficial side of L-ODAP at lower concentrations. This study is of significance especially in view of the Government of India lifting the ban on cultivation of low toxin Lathyrus varieties and consumption of this lentil.

An efficient in vitro shoot regeneration system for Atractylodes ovata (Thunb.) DC. (삽주(Atractylodes ovata (Thunb.) DC.)의 효율적인 기내 줄기 재분화)

  • Hui Yeong Jeong;Ji Ah Kim
    • Journal of Plant Biotechnology
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    • v.49 no.4
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    • pp.325-330
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    • 2022
  • In this study, a plant tissue culture system was established for Atractylodes spp., an economically valuable medicinal crop in Korea that has low domestic production and is increasingly imported. In particular, Atractylodes ovata was treated with four types of cytokinins, 6-benzylaminopurine (BA), zeatin, kinetin, and thidiazuron (TDZ), in two different concentrations (0.5 and 1.0 mg/L). Among the four types of cytokinins, the BA treatment was effective for the shoot and root growth of A. ovata. Both the 0.5 mg/L and 1.0 mg/L concentrations of BA showed similar results; however, the 1.0 mg/L concentration of BA was more effective in promoting shoot and root growth. The treatments showed that the TDZ treatment was not effective for the shoot and root growth, except for the number of shoots and the fresh weight (FW) of the root; therefore, it was unsuitable for this species. In this study, we established a mass production system of A. ovata. Our results showed that direct in vitro regeneration may make a significant contribution to improving the cultivation of the medicinal plant A. ovata.

Optimal Culture Conditions for the Shoot Growth of Freesia 'Shiny Gold' during Bioreactor Culture

  • Kang, Ji Su;Jeong, Kyeong Jin;Choi, Youn Jung;Yun, Jae Gill
    • Korean Journal of Plant Resources
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    • v.30 no.6
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    • pp.699-706
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    • 2017
  • For rapid production of freesia 'Shiny Gold' shoots by using a bioreactor, several culture conditions were investigated. Young shoots (< 1 cm) obtained from freesia corm section in vitro were used as plant materials for this experiment. As a basic experimental environment, 20 young shoots were inoculated into a 5 L balloon type bubble reactor which contained 1 L 1/2 strength MS medium supplemented with 30 g sucrose (3%), and the aeration was 0.1 vvm (vessel volumes per minute). The bioreactors were placed in a growth room with $23^{\circ}C$ temperature, 60% relative humidity and $60{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$ light condition (16 h/8 h, day/night). The concentrations of MS media were set with 1/4, 1/2, 1 strength, medium volume 10, 20, 40%, sucrose concentration 3, 6, 9%, and aeration 0.1, 0.2, 0.4 vvm. After 4 weeks of cultivation, the growth indexes including the fresh and dry weight, and plant height were evaluated. At the same time, the consumption, pH, and EC of medium were estimated 4 weeks after incubating. The best results were achieved when 40 young shoots were incubated in a bioreactor in which 1 L of 1/2 strength MS medium supplemented with 6% sucrose was used for the rapid production of freesia shoots. The shoots were 17 cm in plant height and 1.0 g in fresh weight only 4 weeks after incubation which could be a good plant material suitable for corm enlargement in vitro. No correlation was observed between the growth of freesia shoots and the consumption, pH or EC of medium.

Effects of Thiabendazole on Green Mold, Trichoderma spp. during Cultivation of Oyster Mushroom, Pleurotus spp. (느타리버섯 푸른곰팡이병에 대한 Thiabendazole의 방제효과)

  • Jhune, Chang-Sung;You, Chang-Hyun;Cha, Dong-Yeol;Kim, Gwang-Po
    • The Korean Journal of Mycology
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    • v.18 no.2
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    • pp.89-96
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    • 1990
  • This study was conducted to find out the effects of Thiabendazole on controlling green mold causing serious damage to oyster mushroom, Pleurotus spp. during the cultivation. In vitro, the strains of oyster mushroom such as ASI 2018, 2072 and 2016 were inhibited by 500 ppm of the fungicide, but the strain of ASI 2001 and ASI 2070 was inhibited by 100 and 500 ppm on oatmeal agar, respectively. The mycelial growth of the oyster mushroom started to be inhibition by soak treatment at a 0.2g/1000 ml aqueous solution of the fungicide. When the oyster mushroom and green mold inoculated both or separately on the substrates of soak treatment, the green mold did not grow at all, but the oyster mushroom grown well. The maximum control effect of the green mold showed when $2g/m^2\;and\;5g/m^2$ of the fungicide was sprayed on the surface of substrates before pasteurization. The highest yield of the sporophores of oyster mushroom was obtained from $5g/m^2$ treatment.

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In vitro Characterization of Bacteriocin Produced by Lactic Acid Bacteria Isolated from Nem Chua, a Traditional Vietnamese Fermented Pork

  • Pilasombut, Komkhae;Rumjuankiat, Kittaporn;Ngamyeesoon, Nualphan;Duy, Le Nguyen Doan
    • Food Science of Animal Resources
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    • v.35 no.4
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    • pp.473-478
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    • 2015
  • The aim of this study was to screen and In vitro characterize the properties of bacteriocin produced by lactic acid bacteria isolated from Vietnamese fermented pork (Nem chua). One hundred and fifty LAB were isolated from ten samples of Nem chua and screened for bacteriocin-producing lactic acid bacteria. Antimicrobial activity of bacteriocin was carried out by spot on lawn method against both gram positive and gram negative bacteria. One isolate, assigned as KL-1, produced bacteriocin and showed inhibitory activity against Lactobacillus sakei, Leuconostoc mesenteroides and Enterococcus faecalis. To characterize the bacteriocin-producing strain, optimum temperature, incubation period for maximum bacteriocin production and identification of bacteriocin-producing strain were determined. It was found that the optimum cultivation temperature of the strain to produce the maximum bacteriocin activity (12,800 AU/mL) was obtained at 30℃. Meanwhile, bacteriocin production at 6,400 AU/mL was found when culturing the strain at 37℃ and 42℃. The isolate KL-1 was identified as L. plantarum. Antimicrobial activity of cell-free supernatant was completely inhibited by proteolytic enzyme of trypsin, alpha-chymotrypsin and proteinase K. Bacteriocin activity was stable at high temperature up to 100℃ for 10 min and at 4℃ storage for 2 d. However, the longer heating at 100℃ and 4℃ storage, its activity was reduced.