• Title/Summary/Keyword: culturing

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Biological Control of Stem Rot of Pepper caused by Sclerotium rolfsii using by Bacillus amyloliquefaciens KBC1009 (길항세균 Bacillus amyloliquefaciens KBC1009를 이용한 고추 흰비단병의 생물학적 방제)

  • Kang, Jae-Gon;Lee, Young-Ui;Park, Jeong-chan;Jeong, Yoon-Woo;Park, Chang-Seuk;Kang, Hoon-Serg
    • Journal of agriculture & life science
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    • v.50 no.4
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    • pp.27-34
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    • 2016
  • Sclerotium rolfsii is a well known broad host range soil borne plant pathogenic fungus and caused serious damage to various vegetable crops. To develop an effective biological control agent for S. rolfsii, an isolate which showed strong inhibitory effect on the mycelial growth of S. rolfsii was selected among the antagonistic bacterial isolates collected from vinyl-house soil. The bacterial isolate was identified as Bacillus amyloliquefaciens KBC1009 based on the morphological, physiological characteristics and by 16S rRNA sequence analysis. The growth conditions for B. amyloliquefaciens KBC1009 were optimized in LB media(pH7) by culturing at 30℃ for 72 hrs. Glucose and yeast extract were confirmed as the best carbon and nitrogen sources, respectively. In order to test the inhibitory effect of B. amyloliquefaciens KBC1009 to stem rot of pepper, green house experiment was conducted. Drench of 1/500 diluted bacterial suspension of B. amyloliquefaciens KBC1009(5×108 cfu/ml) to each pepper plant 3 times with 10 days interval showed 66.7% control effectiveness. These results suggest that B. amyloliquefaciens KBC1009 is one of promising biocontrol agent to control stem rot caused by Sclerotium rolfsii.

Trends in the rapid detection of infective oral diseases

  • Ran-Yi Jin;Han-gyoul Cho;Seung-Ho Ohk
    • International Journal of Oral Biology
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    • v.48 no.2
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    • pp.9-18
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    • 2023
  • The rapid detection of bacteria in the oral cavity, its species identification, and bacterial count determination are important to diagnose oral diseases caused by pathogenic bacteria. The existing clinical microbial diagnosis methods are time-consuming as they involve observing patients' samples under a microscope or culturing and confirming bacteria using polymerase chain reaction (PCR) kits, making the process complex. Therefore, it is required to analyze the development status of substances and systems that can rapidly detect and analyze pathogenic microorganisms in the oral cavity. With research advancements, a close relationship between oral and systemic diseases has been identified, making it crucial to identify the changes in the oral cavity bacterial composition. Additionally, an early and accurate diagnosis is essential for better prognosis in periodontal disease. However, most periodontal disease-causing pathogens are anaerobic bacteria, which are difficult to identify using conventional bacterial culture methods. Further, the existing PCR method takes a long time to detect and involves complicated stages. Therefore, to address these challenges, the concept of point-of-care (PoC) has emerged, leading to the study and implementation of various chair-side test methods. This study aims to investigate the different PoC diagnostic methods introduced thus far for identifying pathogenic microorganisms in the oral cavity. These are classified into three categories: 1) microbiological tests, 2) microchemical tests, and 3) genetic tests. The microbiological tests are used to determine the presence or absence of representative causative bacteria of periodontal diseases, such as A. actinomycetemcomitans, P. gingivalis, P. intermedia, and T. denticola. However, the quantitative analysis remains impossible, and detecting pathogens other than the specific ones is challenging. The microchemical tests determine the activity of inflammation or disease by measuring the levels of biomarkers present in the oral cavity. Although this diagnostic method is based on increase in the specific biomarkers proportional to inflammation or disease progression in the oral cavity, its commercialization is limited due to low sensitivity and specificity. The genetic tests are based on the concept that differences in disease vulnerability and treatment response are caused by the patient's DNA predisposition. Specifically, the IL-1 gene is used in such tests. PoC diagnostic methods developed to date serve as supplementary diagnostic methods and tools for patient education, in addition to existing diagnostic methods, although they have limitations in diagnosing oral diseases alone. Research on various PoC test methods that can analyze and manage the oral cavity bacterial composition is expected to become more active, aligning with the shift from treatment-oriented to prevention-oriented approaches in healthcare.

Study of Plant Growth Inhibition with Edible Sweetener Saccharin and Acesulfame Potassium (식용 감미료 사카린과 에이스셜팜 칼륨의 식물 성장 저해 연구)

  • Donggiun Kim
    • The Journal of the Convergence on Culture Technology
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    • v.9 no.5
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    • pp.661-667
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    • 2023
  • Five commercially available edible sweeteners are used as diet products because they can replace sucrose. In studies on the effects on animals and the human body, stability has been proven by excreting-oriented studies with characteristics of animal cells, and accumulation in small amounts has been ignored. On the other hand, plants can absorb, degrade, and accumulate foreign substances, so the effect of degradability and accumulation potential can be studied using plants. Metabolic effects in plants of commercially available saccharin and acesulfame potassium (Ace K) were tested using germinated barley and bean sprouts. In germinated barley and bean sprouts, saccharin and ace K showed inhibitory effects on plant growth in all organs from low concentrations in leaves, stems and roots. In addition, it can be observed that the symptoms of death appear clearly over time, so it can be seen that they are accumulated in the body of the plant. As the accumulated amount increases, the toxic effect increases and the plant reaches a state where it is unable to metabolize, turning black from the tip of the leaf and reaching a state of death. In order to remove the accumulated artificial sweetener, recovery was attempted by culturing in distilled water, but it acts as a substance that is not degraded and dies without avoiding toxicity. Saccharin and ace K cannot be excreted from the cell. Its toxic effects are thought to be persistent, inhibiting growth and eventually leading to cell death.

Regeneration of adventitious root from Calystegia soldanella L. in Jeju island and mass proliferation method using bioreactor system (제주지역 갯메꽃(Calystegia soldanella L.) 유래 부정근 재분화 및 생물반응기 시스템 이용 대량증식법)

  • Jong-Du Lee;Eunbi Jang;Weon-Jong Yoon;Yong-Hwan Jung
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2021.04a
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    • pp.37-37
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    • 2021
  • Calystegia soldanella L. is a perennial herbaceous halophyte belonging to the convolvulaceae family, which mainly grows in coastal sand dunes in Korea. Shoots and rhizomes are edible, and roots called 'Hyoseon Chogeun' are known to have medicinal effects such as antipyretic, sterilization, and diuretic. In addition, physiological activities of antioxidant, anti-inflammatory, antiviral, antifungal and PTP-1B (protein tyrosine phosphate-1B) inhibition have been reported. In this study, in vitro induction cell lines of C. soldanella L. collected from the coastal sand dunes in Jeju island was redifferentiated into adventitious roots that can be used as medicinal resources. Also the biomass of mass-proliferated adventitious roots using a bioreactor were evaluated. Plants of C. soldanella L. were collected from the crevice of the seashore in the coastal area of Taeheung 2-ri, Namwon-eup, Seogwipo-si. Then, it was separated into leaves, stems, rhizomes, and roots, and surface sterilized with 70% ethyl alcohol and 2% NaOCl (sodium hypochlorite). After washing with sterilized water, each organ section was cultured in Hormone-free MS medium (Murashige & Skoog Medium). As a result, the induction response rates were evaluated at 85% and 55%, respectively, in terms of callus formation and shoot generation in the rhizome segment. In the case of the adventitious roots morphological characteristics induced by single-use treatment of auxin-based plant growth regulators IBA and NAA from redifferentiated shoots were compared. Most efficient adventitious root culture method as a rooting rate, number, length, and biomass proliferation in the bioreactor system was confirmed when treated by culturing in MS salts, Sucrose 30 g·L-1, and IBA 1mg·L-1 for 4 weeks. In this study, the medium composition and culture period were confirmed using a bioreactor system to mass-proliferate adventitious roots derived from C. soldanella L. in Jeju island. Also this adventitious root line developed a new medicinal material could increase value of the bio-industry ingredient through quantitative and qualitative screening of phyto-bioactive compounds.

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Evaluation of Manufacturing and Biological Properties of Y Zone Care Hydrogel Solution (Y존 케어 하이드로젤솔루션의 제조 및 생물학적 특성 평가)

  • Eun-Ji Kim;In-Kyoung Kim
    • Journal of Advanced Technology Convergence
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    • v.3 no.2
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    • pp.25-31
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    • 2024
  • In recent industrialization and development due to information and communication technology, modern women in modern society are exposed to physical and mental health due to numerous stresses. Popular inflammations are attributable to a decrease in lactic acid bacteria, frequent antibiotic use, and a decrease in immunity. It is necessary to develop products that are helpful and reflected. The inner care gel currently introduced on the market can increase beneficial bacteria and maintain a healthy y-zone. The inner gel contains a hydrogel component. 90% is made up of water, and other components act as support for supporting water and are formed through crosslinking between polymer chains. Hydroxyethyl cellulose (HEC) is a hydroxyethyl ethylenetel of cellulose. The purpose of use is to act as a binder, an emulsion stabilizer, a viscosity enhancer (water-soluble), and a film forming agent. CA (crosslinker) is a crosslinking agent and serves to bind. Hydrogel in the beauty field acts as a film forming agent that gently wraps around the skin by forming a thin film and serves as an emulsion stabilizer that helps to prevent separation of other raw materials. It also acts as a thickener by increasing viscosity in cosmetics. In addition, it is used for glucose monitoring, nursing care, cell transplantation, and wound treatment in the bio field. Currently, it is understood that no products using functional hydrogel have been released, so in this study, a Y zone care hydrogel solution was manufactured to find out the antibacterial properties of the functional hydrogel, and a new solution was developed. As a result, it was confirmed that the appropriate Ph was applied to the Y zone, and after culturing Candida albicans in PDB medium, all three products of the Y zone care hydrogel solution showed an antibacterial effect of 0.5-1.0mm

LymphanaxTM Enhances Lymphangiogenesis in an Artificial Human Skin Model, Skin-lymph-on-a-chip (스킨-림프-칩 상에서 LymphanaxTM 의 림프 형성 촉진능)

  • Phil June Park;Minseop Kim;Sieun Choi;Hyun Soo Kim;Seok Chung
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.50 no.2
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    • pp.119-129
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    • 2024
  • The cutaneous lymphatic system in humans plays a crucial role in draining interstitial fluid and activating the immune system. Environmental factors, such as ultraviolet light and natural aging, often affect structural changes of such lymphatic vessels, causing skin dysfunction. However, some limitations still exist because of no alternatives to animal testing. To better understand the skin lymphatic system, a biomimetic microfluidic platform, skin-lymph-on-a-chip, was fabricated to develop a novel in vitro skin lymphatic model of humans and to investigate the molecular and physiological changes involved in lymphangiogenesis, the formation of lymphatic vessels. Briefly, the platform involved co-culturing differentiated primary normal human epidermal keratinocytes (NHEKs) and dermal lymphatic endothelial cells (HDLECs) in vitro. Based on our system, LymphanaxTM, which is a condensed Panax ginseng root extract obtained through thermal conversion for 21 days, was applied to evaluate the lymphangiogenic effect, and the changes in molecular factors were analyzed using a deep-learning-based algorithm. LymphanaxTM promoted healthy lymphangiogenesis in skin-lymphon-a-chip and indirectly affected HDELCs as its components rarely penetrated differentiated NHEKs in the chip. Overall, this study provides a new perspective on LymphanaxTM and its effects using an innovative in vitro system.

The Role of SDF-1𝛼-CXCR4/CXCR7 in Migration of Human Periodontal Ligament Stem Cells

  • Jialei Xu;Fan Yang;Shuhan Luo;Yuan Gao;Dingming Huang;Lan Zhang
    • International Journal of Stem Cells
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    • v.16 no.2
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    • pp.180-190
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    • 2023
  • Background and Objectives: Regenerative endodontic procedures (REPs) are a research hotspot in the endodontic field. One of the biggest problems of REPs is that it is difficult to realize regeneration of pulp-dentin complex and functional reconstruction. The reason is still not clear. We hypothesize that the migration may be different in different dental stem cells. Periodontal ligament stem cells (PDLSCs) may migrate faster than stem cells of apical papilla (SCAPs), differentiating into cementum-like tissue, bone-like tissue and periodontal ligament-like tissue and, finally affecting the outcomes of REPs. Hence, this study aimed to explore the mechanism that regulates the migration of PDLSCs. Methods and Results: After isolating and culturing PDLSCs and SCAPs from human third molars, we compared the migration of PDLSCs and SCAPs. Then we investigated the role of SDF-1𝛼-CXCR4/CXCR7 axis in PDLSC migration. We further investigated the impact of Porphyromonas gingivalis (P. gingivalis) lipopolysaccharide (LPS) on PDLSC migration and the potential mechanism. PDLSCs showed better migration under both noninflammatory and inflammatory conditions than SCAPs. SDF-1𝛼 can promote the migration of PDLSCs by elevating the expression of CXCR4 and CXCR7, increasing the interaction between them, promoting expression of 𝛽-arrestin1 and activating the ERK signaling pathway. P. gingivalis LPS can promote the migration of PDLSCs toward SDF-1𝛼 through increasing the expression of CXCR4 via the NF-𝜅B signaling pathway, promoting the expression of 𝛽-arrestin1, and activating the ERK signaling pathway. Conclusions: This study helped elucidate the potential reason for the difficulty in forming pulp-dentin complex.

A Study on Multiplication Response of "Tricholoma matsutake" (Pine Mushroom) Conidio to Cultural Media Environment (송이균(松茸菌) (Tricholoma matsutake)의 배양환경(培養環境)에 대한 증식반응(增殖反應)에 관한 연구(硏究))

  • Kim, Chang Ho
    • Journal of Korean Society of Forest Science
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    • v.64 no.1
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    • pp.33-41
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    • 1984
  • This study was conducted to examine the physiology of pine mushroom mycelia cultured with various media for artificial culture of pine mushroom. The results obtained were as follows: 1) Among the various media, the medium composed of honey, boiled pine mushroom and soil extract fluid, fibrous root extract fluid, dry yeast, $KH_2PO_4$ inositol, folic acid, and biotin was the best for the growth of pine mushroom mycelium. 2) The optimum temperature for germinating pine mushroom spore and for culturing pine mushroom mycelium, was $24^{\circ}C$ and the optimum pH was 4.5. 3) There was no significant difference in growth between the mycelium separated from the tissue of pine mushroom sporophore and that separated from the spore. 4) No noticeable effect was found on the growth if such salts as $ZnSO_4$, $MnSO_4$, $MgSO_4$, $CaCl_2$ and ferric citrate were added to the Hamada's medium. 5) The addition of fibrous root extract promoted the growth of pine mushroom mycelium. 6) As a carbon source of artificial media, honey was more effective than glucose. 7) The culture infiltration of Mortierlla growing often in Fairy Ring was good for the growth of mycelium compared with the control. 8) The addition of fibrous root extract, inositol, biotin, and folic acid to artificial culture media was greatly effective in growth. When the temperature was lowered $19^{\circ}C$ after mycelium has appeared, the formation of primordium was observed.

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The Effects of Air-borne Particulate Matters on the Alveolar Macrophages for the TNF-α and IL-1β Secretion (미세분진이 흰쥐의 폐포대식세포에서 TNF-α와 IL-1β의 형성에 미치는 효과)

  • Li, Tian Zhu;Lee, Soo-Jin;Park, Se-Jong;Chang, Byung-Joon;Lee, Jong-Hwan;Kim, Kil-Soo;Lee, Myoung-Heon;Choe, Nong-Hoon
    • Tuberculosis and Respiratory Diseases
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    • v.60 no.5
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    • pp.554-563
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    • 2006
  • Background: PM is known to induce various pulmonary diseases, including asthma, cancer, fibrosis and chronic bronchitis. Despite the epidemiological evidence the pathogenesis of PM-related pulmonary diseases is unclear. Methods: This study examined the effects of PM exposure on the secretion of $TNF-{\alpha}$ and $IL-1{\beta}$ in the cultured alveolar macrophages. The cultured primary alveolar macrophages were treated with the medium, PM ($5{\sim}20{\mu}g/cm^2$), LPS (5ng/ml), and PM with LPS for 24h and 48h respectively. ELISA was used to assay the secreted $TNF-{\alpha}$ and $IL-{\beta}$ in the culture medium. Western blotting was used to identify and determine the level of proteins isolated from the culture cells. The cells cultured in the $Lab-Tek^{(R)}$ chamber slides were stained with immunocytochemical stains. Results: PM induced $TNF-{\alpha}$ and $IL-1{\beta}$ secretion in the culturing alveolar macrophages, collected from the SPF and inflammatory rats. However, the effects were only dose-dependent in the inflammatory macrophages. When the cells were co-treated with PM and LPS, there was a significant synergistic effect compared with the LPS in the both cell types. Conclusion: PM might be play an important role in the induction and/or potentiation of various lung diseases by oversecretion of $TNF-{\alpha}$ and $IL-1{\beta}$.

The Optimal Condition of Performing MTT Assay for the Determination of Radiation Sensitivity (방사선 감수성 측정법으로서 MTT 법 시행 시의 최적 조건에 대한 연구)

  • Hong, Se-Mie;Kim, Il-Han
    • Radiation Oncology Journal
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    • v.19 no.2
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    • pp.163-170
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    • 2001
  • Purpose : The measurement of radiation survival using a clonogenic assay, the established standard, can be difficult and time consuming. In this study, We have used the MTT assay, based on the reduction of a tetrazolium salt to a purple formazan precipitate by living cells, as a substitution for clonogenic assay and have examined the optimal condition for performing this assay in determination of radiation sensitivity. Materials and Methods : Four human cancer cell lines - PCI-1, SNU-1066, NCI-H630 and RKO cells have been used. For each cell line, a clonogenic assay and a MTT assay using Premix WST-1 solution, which is one of the tetrazolium salts and does not require washing or solubilization of the precipitate were carried out after irradiation of 0, 2, 4, 6, 8, 10 Gy. For clonogenic assay, cells in $25\;cm^2$ flasks were irradiated after overnight incubation and the resultant colonies containing more than 50 cells were scored after culturing the cells for $10\~14$ days. For MTT assay, the relationship between absorbance and cell number, optimal seeding cell number, and optimal timing of assay was determined. Then, MTT assay was performed when the irradiated cells had regained exponential growth or when the non-irradiated cells had undergone four or more doubling times. Results : There was minimal variation in the values gained from these two methods with the standard deviation generally less than $5\%$, and there were no statistically significant differences between two methods according to t-test in low radiation dose (below 6 Gy). The regression analyses showed high linear correlation with the $R^2$ value of $0.975\~0.992$ between data from the two different methods. The optimal cell numbers for MTT assay were found to be dependent on plating efficiency of used cell line. Less than 300 cells/well were appropriate for cells with high plating efficiency (more than $30\%$). For cells with low plating efficiency (less than $30\%$), 500 cells/well or more were appropriate for assay. The optimal time for MTT assay was after 6 doubling times for the results compatible with those of clonogenic assay, at least after 4 doubling times was required for valid results. In consideration of practical limits of assay (12 days, in this study) cells with doubling time more than 3 days were inappropriate for application. Conclusion : In conclusion, it is found that MTT assay can successfully replace clonogenic assay of tested cancer cell lines after irradiation only if MTT assay was undertaken with optimal assay conditions that included plating efficiency of each cell line and doubling time at least.

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