• Title/Summary/Keyword: viability test

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The venom of jellyfish, Chrysaora pacifica, induces neurotoxicity via activating Ca2+-mediated ROS signaling in HT-22 cells

  • Yang, Yoon-Sil;Kang, Young-Joon;Kim, Hye-Ji;Kim, Min-Soo;Jung, Sung-Cherl
    • Journal of Applied Biological Chemistry
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    • v.62 no.4
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    • pp.347-353
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    • 2019
  • Stings of jellyfish, which frequently occur in a warm season, cause severe pain, inflammation and sometimes irreversible results such as the death. Harmful venoms from jellyfish, therefore, have been studied for finding the therapeutic agents to relieve pain or to neutralize toxic components. However, it is still unclear if and how jellyfish venom reveal neuronal toxicity even though pain induction seems to result from the activation of nociceptors such as nerve endings. In this study, using HT-22 cell line, we investigated neurotoxic effects of the venom of Chrysaora pacifica (CpV) which appears in South-East ocean of Korea. In 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay, CpV significantly reduced the viability of HT-22 cells in a dose-dependent manner. Additionally, in 2',7'-Dichlorofluorescin diacetate fluorescence test under the culture condition lacking dominant inflammatory factors, CpV remarkably increased the production of intracellular reactive oxygen species (ROS). Reduced responsive fluorescence to Rhodamine123 and increased expression of intracellular cytochrome c were also observed in HT-22 cells treated with CpV. These indicate that CpV-reduced viability of HT-22 cells may be due to the activation of apoptotic signalings mediated with oxidative stress and mitochondrial dysfunction. Furthermore, removing Ca2+ ion or adding N-acetyl-Lcystein remarkably blocked the CpV effect to reduce the viability of HT-22 cells. The findings in this study clearly demonstrate that CpV may activate Ca2+-mediated ROS signalings and mitochondrial dysfunction resulting in neuronal damage or death, and suggest that blocking Ca2+ pathway is a therapeutic approach to possibly block toxic effects of jellyfish venoms.

A novel retentive type of dental implant prosthesis: marginal fitness of the cementless double crown type implant prosthesis evaluated by bacterial penetration and viability

  • Hong, Seoung-Jin;Kwon, Kung-Rock;Jang, Eun-Young;Moon, Ji-Hoi
    • The Journal of Advanced Prosthodontics
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    • v.12 no.4
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    • pp.233-238
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    • 2020
  • PURPOSE. This study aims to compare the marginal fitness of two types of implant-supported fixed dental prosthesis, i.e., cementless fixation (CL.F) system and cement-retained type. MATERIALS AND METHODS. In each group, ten specimens were assessed. Each specimen comprised implant lab analog, titanium abutment fabricated with a 2-degree tapered axial wall, and zirconia crown. The crown of the CL.F system was retained by frictional force between abutment and relined composite resin. In the cement-retained type, zinc oxide eugenol cement was used to set crown and abutment. All specimens were sterilized with ethylene oxide, immersed in Prevotella intermedia culture in a 50 mL tube, and incubated with rotation. After 48 h, the specimens were washed thoroughly before separating the crown and abutment. The bacteria that penetrated into the crown-abutment interface were collected by washing with 500 µL of sterile saline. The bacterial cell number was quantified using the agar plate count technique. The BacTiter-Glo Microbial Cell Viability Assay Kit was used to measure bacterial adenosine triphosphate (ATP)-bioluminescence, which reflects the bacterial viability. The t-test was performed, and the significance level was set at 5%. RESULTS. The number of penetrating bacterial cells assessed by colony-forming units was approximately 33% lower in the CL.F system than in the cement-retained type (P<.05). ATP-bioluminescence was approximately 41% lower in the CL.F system than in the cement-retained type (P<.05). CONCLUSION. The CL.F system is more resistant to bacterial penetration into the abutment-crown interface than the cement-retained type, thereby indicating a precise marginal fit.

Cellular Uptake and Transfection Efficiency of Plasmid DNA Using Low Molecular Weight Polyethylenimine (저분자량 폴리에틸렌이민을 이용한 유전자 송달 및 발현 유효성 연구)

  • Jeong, Gil-Jae;Park, Kui-Lye;Shin, Ji-Young;Choi, Han-Gon;Oh, Yu-Kyoung
    • Journal of Pharmaceutical Investigation
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    • v.34 no.4
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    • pp.263-267
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    • 2004
  • Branched and linear polyethylenimines (PEIs) have been studied as efficient and versatile agents for gene delivery in vitro and in vivo. PEIs exist in a linear or branched topology and are available in a wide range of molecular weight (Mw). Most studies have been done using PEIs with Mw higher than 10Kd. This study was aimed to test the transfection efficiency and the cell viability following gene delivery using PEI of Mw 2Kd, a relatively lower Mw cationic polymer. We used murine interleukin-2(mIL-2) plasmid DNA complexed with branched PEI 2Kd or 25Kd, and transfected them into a myoblast muscle cell line, C2C12. The cellular uptake of mIL-2 plasmid DNA was determined using quantitative polymerase chain reaction. RNA transcript levels were studied in the myoblast cells. Our results show that PEI 2Kd was as effective as PEI 25Kd in celluar gene delivery and transfection efficiency in C2C12 cells. Moreover, MTT assay indicated that PEI 2Kd/DNA complexes did not significantly reduce the cell viability regardless of N/P ratios. These results suggest that PEI of Mw 2Kd might play a role as effective and low toxic nonviral vector systems for muscular cell lines.

Effect of Seed Dehydration and Temperature during Cold-Stratification on the Seed Quality of Panax ginseng C. A. Meyer (인삼 종자의 생리적 휴면타파기간 중 건조처리 및 저장온도가 종자 건전성에 미치는 영향)

  • Suh, Su Jeoung;Jang, In Bae;Yu, Jin;Jang, In Bok;Park, Hong Woo;Seo, Tae Cheol;Kweon, Ki Bum
    • Korean Journal of Medicinal Crop Science
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    • v.25 no.4
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    • pp.209-216
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    • 2017
  • Background: Dehisced ginseng seeds need to be stored at cold temperatures for around 3 months to break their physiological dormancy, and thus, to aid in gemination. In the presence of high moisture in such an environment, seed spoilage and pre-germination may lower seed quality and productivity. To improve seed quality during cold-stratification, the effects of seed dehydration and temperature were tested. Methods and Results: In early December, dehisced ginseng seeds were dehydrated at 4 different levels and stored at $2^{\circ}C$ $-2^{\circ}C$, and $-20^{\circ}C$ for 3 months. Germination was carried out on the filter papers moistened with distilled water; emergence of root, shoot, and seed spoilage were assessed. Seed viability was examined by the tetrazolium test. More than 90% of the seeds stored at $2^{\circ}C$ and $-2^{\circ}C$ without drying or endocarp dehydration germinated, but seeds that were dehydrated to have a moisture content (MC) below 31% showed poor germination and lost their viability. In addition, the seeds stored at $-20^{\circ}C$ failed to show effective germination. Conclusions: Seed storage after endocarp dehydration might help to improve seed quality and increase seedling's ability to stand during the spring-sowing of ginseng.

Nonlinear Rheological Properties of Endothelial Cell Laden-cellulose Nanofibrils Hydrogels (내피세포가 배양된 나노셀룰로오스 하이드로겔의 비선형 유변물성 분석)

  • Song, Yeeun;Kim, Min-Gyun;Yi, Hee-Gyeong;Lee, Doojin
    • Composites Research
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    • v.35 no.3
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    • pp.153-160
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    • 2022
  • Cellulose nanofibrils (CNF) based on wood pulp fibers are gained much attention as part of biocompatible hydrogels for biomedical applications such as tissue engineering scaffolds, biomedicine, and drug carrier. However, CNF hydrogels have relatively poor mechanical properties, impeding their applications requiring high mechanical integrity. In this work, we prepare 2,2,6,6-tetramethylipiperidin-oxyl (TEMPO) oxidated cellulose nanofibrils hydrogels mediated with metal cations, which form the metal-carboxylate coordination bonds for enhanced mechanical strength and toughness. We conduct the large amplitude oscillatory shear (LAOS) test and Live/dead cell assay for obtaining nonlinear viscoelastic parameters and cell viability, respectively. In particular, the cell proliferation and viability change depending on the type of metal salt, which also affected the rheological properties of the hydrogels.

Physical Properties and Cytotoxicity of Dental Pit and Fissure Sealants Containing Cerium Oxide Nano Particles(CNPs) (세륨옥사이드나노입자(Cerium Oxide Nano Particles; CNPs)를 첨가한 치면열구전색재의 물리적 특성 및 세포독성)

  • Jeong, Mi-Ae;Kim, Dong-Ae
    • The Journal of the Korea Contents Association
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    • v.22 no.3
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    • pp.586-592
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    • 2022
  • In this study cerium nano particles(CNPs) with 0-4.0 wt% was incorporated to the conventional dental pit and fissure sealant(ConciseTM) to produce new pit and fissure sealant the physical properties and cytotoxicity. The physical properties were measured for polymerizing depth the degree of water absorption and solubility. The cytotoxicity of cell viability was analyzed by MTT assay using immortalized human oral keratinocyte(IHOK). As a result of this preceding study the polymerizing depth was decreased by the increasing of the amount of CNPs. The solubility degree of the sealant added CNPs with 2.0 wt% showed was the lower and the water absorption showed no significantly difference with the control groups(p>0.05). The cytotoxicity test results showed high survival rates in all experimental groups. Therefore, pit and fissure sealant by the addition of CNPs excellent cell viability be produced without weaken the physical property of the cell viability fissure sealant containing CNPs does not weaken physical properties and has no cytotoxic effects biocompatibility. Considering its properties effect of CNPs, further studies are required for distribution technology application.

The effect of isoflavonoid contents in SH003 and its subfractions on breast cancer (SH003과 분획물의 구성 성분인 이소플라본의 유방암에 대한 효과)

  • Yu-Jeong Choi;Won-Geun Choi;Kangwook Lee;Miso Jeong;Sang Cheol Park;Young Pyo Jang;Seong-Gyu Ko
    • The Journal of Korean Medicine
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    • v.43 no.3
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    • pp.79-93
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    • 2022
  • Objectives: We investigated the isoflavone contained in SH003 and its fractions, and the effect of these components on the inhibition of breast cancer. Methods: The isoflavones in solvent fractions of SH003 extract were identified by UPLC-MS and its contents were quantified using HPLC analysis. The estrogenic activity of SH003 or fractions was assessed by ERE luciferase assay in estrogen receptor (ER)-positive MCF-7 cells. To test the breast cancer inhibitory effect, the cell viability was measured using an MTT assay. Results: In this study, we demonstrated that SH003 and fractions contain 4 isoflavones which are calycosin-7-β-D-glucoside, formononetin-7-β-D-glucoside, calycosin, and formononetin. Despite containing isoflavones, estrogen-dependent transcription activity was not altered by both SH003 and fractions. On the other hand, SH003 and fractions inhibited the cell viability of breast cancer. In addition, its isoflavone components also showed reduced cell viability in various breast cancer cells. Conclusions: Overall, the phytoestrogen included in SH003 and fractions did not influence the estrogenic activity, emphasizing the safety of SH003 and fractions in breast cancer treatment.

Application of Neutral Red Uptake Assay Using EPC Cells as an Alternative to the Fish Acute Toxicity Test for Pesticide (어류급성독성시험 대체법으로서 잉어표피세포를 이용한 Neutral Red Uptake 분석법 적용)

  • Seo, Ji-Hyun;Park, June-Woo;Lee, Sung-Kyu;Kim, Woo-Keun
    • The Korean Journal of Pesticide Science
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    • v.18 no.1
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    • pp.8-13
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    • 2014
  • This study evaluated in vitro cytotoxicity of 5 pesticides, including 2 herbicides, 2 germicides, and an insecticide, as an alternative to the fish acute toxicity test. The in vitro cytotoxicity was tested using a neutral red uptake (NRU) assay with epithelioma papulosum cyprini (EPC) cells that originated from the epidermal tissue of Cyprinus carpio (common carp). An in vivo fish acute toxicity test was conducted according to OECD Test Guideline No. 203 using Aphyocypris chinensis (Chinese bleak), Oryzias latipes (Japanese medaka), and C. carpio. The results showed that the sensitivity of the cell viability assay for the pesticides was similar to the fish acute test in ranking order despite having approximately 10 times less absolute sensitivity. The $r^2$ correlation values were calculated as 0.38 (p = 0.26), 0.76 (p = 0.05) and 0.90 (p = 0.01) for A. chinensis, O. latipes, and C. carpio, respectively. These results suggested that the potential of EPC cell viability assay as an alternative to the fish acute toxicity test due to their good correlation and NRU assay is expected to serve as a useful tool for predicting acute fish lethality for pesticides if further studies with a large set of pesticides are conducted.

Analysis on inclined or rounded tip piles using 3D printing technology and FE analysis

  • Jaehong Kim;Junyoung Ko;Dohyun Kim
    • Geomechanics and Engineering
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    • v.33 no.1
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    • pp.91-99
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    • 2023
  • To test the effect of various pile tip shape series of model scale loading tests were carried out on test piles with special pile tips. Special pile tips were made using the 3D printer and were attached to the bottom end of the test pile for loading tests. The pile tips were made to have 30°, 45°, 60° inclined tips, as well as a rounded tip. The main objective of the test was to observe the effect of various pile tip shapes on settlement and penetrability of the pile. Moreover, a numerical model simulating the pile loading test carried out in this study was established and verified based on the loading test results. From this, the stress concentration around the pile tip was investigated. This will allow us to analyze the decrease of stress concentration around the pile tip which is the main cause of the pile tip damage during pile installation. However, modifying the pile tip shape will eventually increase the settlement of the pile. By estimating the degree of increase in pile settlement, the viability and the efficiency of the pile shape modification was judged. In addition, case studies on the effect of different pile tip shape and ground conditions on pile settlement and stress dispersion was conducted.

Establishment of Days after Anthesis(DAA) and Fruit After-ripening Period(FAP) for High-Quality Seed Production of Watermelon (우량 수박종자 생산을 위한 적정 성숙일수 및 후숙일수 확립)

  • Park, Eun-Ji;Lee, Gyu-Bin;Park, Young-Gil;Suh, Jeong-Min;Kang, Jum-Soon
    • Journal of Environmental Science International
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    • v.24 no.12
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    • pp.1681-1689
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    • 2015
  • This study was conducted to investigate days after anthesis (DAA) and fruit after-ripening period (FAP) for seed-harvesting of high quality watermelon seeds. Fruit weight and number of seed per fruit increased according to DAA, while those did not significant about FAP. Ratio of cotyledon at whole seed was higher about 2 to 4% compared to seed coat irrespective of DAA and FAP. Germinability of watermelon was not a significant effect by DAA, however, it had differences by FAP. Percent of germination (PB) was below 50%, when 30 days maturated fruits after anthesis was omitted ripening, while PB was increased to 92% by ripening. In addition, seeds at DAA 40 and FAP 20 were higher general seedling vigors (hypocotyl length, diameter etc.) in BP test. Results indicated that considering seed productivity, it had maximized seed viability at DAA 40 and FAP 20.