• Title/Summary/Keyword: LDH release assay

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Sulforhodamine B Assay to Determine Cytotoxicity of Vibrio vulnificus Against Human Intestinal Cells

  • Lee, Byung-Cheol;Choi, Sang-Ho;Kim, Tae-Sung
    • Journal of Microbiology and Biotechnology
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    • v.14 no.2
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    • pp.350-355
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    • 2004
  • Sulforhodamine B (SRB) assay is a rapid, sensitive, and inexpensive method for measuring cell proliferation and chemosensitivity. However, the lactate dehydrogenase (LDH) release assay is generally used to measure cytototoxicity of infectious microorganisms against host cells. In this study, we investigated the possibility of applying the SRB assay to determine cytotoxicity for infectious microorganisms, and compared the results with those obtained by the LDH release assay. We used Vibrio vulnificus as a model of infectious microorganisms. The SRB assay showed that V vulnificus strongly induced cytotoxic activity against human intestinal cells, Caco-2 and INT-407 cells. The degree of cytotoxicity closely correlated with infection time and number ratios of V. vulnificus to intestinal cells (MOI, multiplicity of infection). Furthermore, cytotoxicity values obtained by SRB assay correlated well with results obtained by the LDH release assay, and both assays gave a linear response with respect to MOI Heat-inactivation of V. vulnificus for 35 min at $60^{\circ}C$ did not induce cytotoxic activity, indicating that viability of V. vulnificus is crucial for cytotoxic activity against intestinal cells. Although both assays are suitable as cytotoxicity endpoints, the SRB assay is recommended for measuring cytotoxicity of infectious microorganisms against host cells because of its significantly lower cost and more stable endpoint than the LDH release assay.

In vitro cytotoxicity of Acanthamoeba spp. isolated from contact lens containers in Korea by crystal violet staining and LDH release assay

  • Shin, Ho-Joon;Cho, Myung-Soo;Jung, Suk-Yul;Kim, Hyung-Il;Im, Kyung-Il
    • Parasites, Hosts and Diseases
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    • v.38 no.2
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    • pp.99-102
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    • 2000
  • In order to observe the cytotoxicity of Acanthamoeba spp., which were isolated from contact lens containers as ethiological agents for the probable amoebic keratitis in Korea, the crystal violet staining method and LDH release assay were carried out. In the crystal violet staining method, among eight contact lens container isolates, isolate 3 (Acanthauloeba KA/LS5) showed 83.6% and 81.8% of cytotoxicity, and isolate 7 (Acanthamoeba KA/LS37) showed 28.2% and 25.1% of cytotoxicity, in 1 mg/ml and 0.5 mg/ml Iysate treatments, respectively. Acanthamoeba cutbertsoni and A. healyi showed 84.0% and 82.8% of cytotoxicity. Similar results were observed in A. costellunii and A. hafchefti which showed 83.6% and 75.5% or cytotoxicity. Acanthamoeba roureba and A. polyphaga showed 9.0% and 1.7% of cytotoxicity. In the LDH release assay, isolate 3 (20.4%) showed higher cytotoxicity than other isolates in 1 mg/ml Iysate treatment. The results provide that at least isolate 3 has the cytotoxic effect against CHO cells and seems to be the pathogenic strain.

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In Vitro Magnetometry, LDH Activity and Apoptosisas Indices of Cytotoxicity in Alveolar Macrophages Exposed to Cadmium Chloride (카드뮴에 폭로된 폐포된 폐포대식세포의 세포독성 평가를 위한 세포자계측정, LDH활성도 및 Apoptosis)

  • 조영채
    • Journal of Environmental Health Sciences
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    • v.26 no.4
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    • pp.115-121
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    • 2000
  • To evaluate the cytotoxicity of cadmium compounds, this study was conducted to measure the in vitro magnetometry, LDH release and cellular apoptosis using alveolar macrophages of hamsters. A series of magnetometric measurements in cadmium-added groups showed a significant dose-dependent decay of the relaxation curves. The LDH release rates showed a dose-dependently increasing tendency as the dose gradually increased. The positive rates of apoptosis were significantly higher in cadmium-added groups than the control groups. Conclusively, the cytotoxicity increased in a dose dependent way as the concentration of cadmium added increased, which reflected in the decay of relaxation curve in magnetometry, and increased LDH release rate and positive rate of apoptosis.

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Effects of Taeumin Chungsimyeunjatang on the Cerebral neurons injured by Hydrogen Peroxide (태음인(太陰人) 청심연자탕(淸心蓮子湯)이 Hydrogen Peroxide에 손상(損傷)된 백서(白鼠)의 대뇌신경세포(大腦神經細胞)에 미치는 영향(影響))

  • Ok, Yun-young;Ryu, Do-gon;Kim, Kyung-yo
    • Journal of Sasang Constitutional Medicine
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    • v.11 no.2
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    • pp.251-266
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    • 1999
  • 1. Purpose : The purpose of this study was to determine the effects of Chungsimyeunjatang on the cerebral neurons injured by hydrogen peroxide($H_2O_2$). 2. Methods : I observed cell viability in mouse cerebral neurons exposed to hydrogen peroxide by NR assay and MTT assay and determined lipid peroxidation and amounts of LDH release in mouse cerebral neurons exposed to hydrogen peroxide. After administration of Chungsimyeunjatang water extracts, I observed significant changes of cell viability, lipid peroxidation and amounts of LDH release in mouse cerebral neurons exposed to hydrogen peroxide. 3. Results : Hydrogen peroxide showed neurotoxicity. Cell viability in mouse cerebral neurons exposed to hydrogen peroxide decreased in NR assay and MTT assay. Lipid peroxidation and amounts of LDH release in mouse cerebral neurons exposed to hydrogen peroxide increased. Chungsimyeunjatang was very effective in blocking hydrogen peroxide-induced neurotoxicity.

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Effects of Traditional Drugs on $CCl_4-induced$ Cytotoxicity in Primary Cultured Rat Hepatocytes (수종의 전통약제가 일차 배양 간세포에서 $CCl_4$ 유발 세포독성에 미치는 영향)

  • Kim, Young-Sook;Park, Ki-Hyun
    • Korean Journal of Pharmacognosy
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    • v.25 no.4 s.99
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    • pp.388-394
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    • 1994
  • 80% Methanol extracts of 44 traditional drugs used for the treatment of liver diseases or tonic effects were screened for anti-hepatotoxic activity by in vitro assay using $CCl_4-induced$ cytotoxicity in primary cultured rat hepatocytes. $CCl_4-induced$ cytotoxicity was evaluated by determination of LDH, GOT or GPT activity in the medium. Rehmaniae Radix Preparata and Gelantina nigra inhibited the release of LDH, GOT or GPT from $CCl_4-treated$ hepatocytes. Gibotii Rhizoma and Eucommiae Cortex showed inhibitory effect on release of LDH from normal hepatocytes as well as $CCl_4-treated$ hepatocytes. Eucommiae Cortex and Lili Bulbus decreased release of GOT and LDH from normal hepatocytes, respectively. Astragali Radix inhibited release of GPT in $CCl_4-treated$ hepatocytes. Phlomidis Radix, Imperatae Rhizoma, Cistanchis Herba, Broussonetiae Fructus, Asparagi Tuber, Trigonellae Semen and Polgonati Rhizoma inhibited release of LDH from $CCl_4-treated$ hepatocytes. Among 44 traditional drugs, most of them released LDH, GOT or GPT at the dose of 1 mg/ml in normal hepatocytes, and Drynariae Rhizoma, Acanthopanacis Cortex, Longanae Arillus, Atratylodis Rhizoma and Ecliptae Herba increased $CCl_4-induced$ cytotoxicity.

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Protective Effect of Coptidis Rhizoma on SNAP-Induced Cytotoxicity in Pancreatic RINm5F Cells (SNAP 유도성 RINm5F 세포 독성에 대한 황연 추출물의 방어효과)

  • 류도곤;권강범;양정예;김은경;김강산
    • The Journal of Korean Medicine
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    • v.24 no.2
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    • pp.159-165
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    • 2003
  • Objectives : Coptidis rhizoma (CR) is an oriental medicine that has been used in many traditional prescriptions against diabetes mellitus in Korea for centuries. Our purpose was to determine the protective effect and its action mechanism of CR on the cytotoxicity of pancreatic -cell line (RINm5F cell). Methods : In this experiment, we used methods such as MTT assay for detection of cytotoxicity, DNA fragmentation assay for detection of apoptotic cell death, LDH activity assay for detection of necrotic cell death, and measurement of $DiOC_{6}$ (3) retention for detection of mitochondrial membrane potential (MMP). Background : Nitric oxide (NO) is believed to playa key role in the process of pancreatic -cell destruction leading to insulin-dependent diabetes mellitus (IDDM). Results : Exposure of RINm5F cells to chemical NO donor such as S-nitroso-N-acetylpenicillamine (SNAP) induced cytotoxic events such as DNA fragmentation and lactate dehydrogenase (LDH) release into medium. However, pretreatment of RINm5F cells with CR extract ($10~50{\mu\textrm{g}}/ml$) for 3 hours prevented SNAP-induced DNA fragmentation and LDH release into medium through the inhibition of MMP disruption. Conclusions : These results suggest that CR may be a candidate for a therapeutic or preventing agent against IDDM.

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CYTOTOXIC EFFECT OF RETROGRADE FILLING MATERIALS INCLUDING GLASS IONMER CEMENT ACCORDING TO CELL LINES AND ASSAY METHODS (광중합형 glass ionomer cement를 포함한 수종 역충전재의 세포주와 검사법에 따른 독성 효과)

  • Im, Mi-Kyung;Koo, Dae-Hoi
    • Restorative Dentistry and Endodontics
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    • v.21 no.1
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    • pp.403-424
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    • 1996
  • Cell culture methods have been used to assess the cytotoxicity of dental materials. Different paramaters are used to monitor cytotoxic effects. But it is difficult to compare each investigator's results with different methods. The objective of this study was to investigate cytotoxic effect of several retrograde filling materials according to cell lines and assay methods. Cytotoxicity of Bestalloy (Dogmyung, Korea), Prisma APH(Densply International Inc., U.S.A.), Clearfil FII (Kuraray Co., Japan), Fuji II (GC Co., Japan), Fuji II LC (GC Co., Japan) and IRM (Densply Co., U.S.A.) on L929, 3T3 and KB permanent cell lines was measured. Radiochromium, Lactate dehydrogenase (LDH) release method and colorimetric assays, namely neutral red (NR) and MTT were used. Each material was mixed according to the manufacturer's instruction. They were tested as solid and extracted state. Cell culture media were added to each mixed or solid materials then the solution was collected and used as extract solutions. Solid Fuji II showed mild cytotoxicity on three cell lines using radiochromium release method. There was no difference in cytotoxicity of extract solution group using radiochromium release method. In colorimetric assay immediate Fuji II group and all the IRM groups showed severe cytotoxic effect. Difference in cyctotoxicity was due to rather kinds of cell lines than assay methods. Solid Fuji II and IRM showed mild cytotoxicity on three cell lines. But extract solutions had different cytotoxic effect according to cell lines using LDH release assay. Light-cured glass ionomer had mild to moderate degree of cytotoxicity on three cell lines. Cytotoxicity was affected by specimen prepaton. Susceptibility of each cell ines were also affected by assay emthods. It was suggested that cytotoxicity study using only one cell line and/or assay method might not accurately reflect the real toxic nature of dental biomaterials.

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Neuroprotective Effect of Extracts from Root Bark of Morus alba on Glutamate-induced Cytotoxicity in Neuronal Cells. (Glutamate가 유도하는 세포독성으로부터 신경세포를 보호하는 상백피 추출물의 효과)

  • Kim, Hyun-Jung;Kim, Ji-Hyun;Son, Eun-Soon;Lee, Jeung-Min;Park, Hae-Ryong
    • Journal of Life Science
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    • v.19 no.7
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    • pp.963-967
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    • 2009
  • This study evaluated the neuroprotective effect of extracts from the root bark of Morus alba (MA) against glutamate-induced cytotoxicity in neuronal cells. Glutamate-induced cytotoxicity was shown by MTT reduction assay. The neuroprotective effects of methanol, ethanol, and acetone extracts from MA against glutamate-induced cytotoxicity were measured. Among the three extracts, the methanolic extracts showed the highest protective effect, as determined by the results of an morphological assay, a lactate dehydrogenase release assay. Furthermore, the methanol extracts were fractionated sequentially with hexane, diethyl ether, ethyl acetate, and water layer according to degree of polarity. The hexane fractions exhibited a neuroprotective effect against glutamate-stressed N18-RE-105 cells. Therefore, these results suggest that extracts of MA could be a new potential candidate as a protective substance against glutamate-induced cytotoxicity.

Anti-stress Effects of Ethanol Extract of Ziziphus jujuba Against Corticosterone-Induced Apoptosis in PC12 Cells

  • Da Hye Song;Yu Jin Choi
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2022.09a
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    • pp.119-119
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    • 2022
  • The coronavirus disease 2019 (COVID-19) pandemic may be stressful for people. Public health actions, such as social distancing, can make people feel isolated and lonely and can increase stress and anxiety. As a result, there is a growing interest towards various materials to relieve stress. Thus, the present study aimed to investigate the anti-stress effects of ethanol extract of Ziziphus jujuba in PC12 cells treated with corticosterone and its underling mechanisms. Furthermore, the viability of the cells, the apoptosis of the cells, the level of phosphorylation of extracellular signal-regulated kinases (p-ERKs) expression were measured by MTT assay, LDH assay, Hoechst staining assay and western blotting. Our results showed that the extract of Ziziphus jujuba reversed corticosterone-induced damage in PC12 cells, which increased cell viability, decreased LDH release, and attenuated corticosterone-induced apoptosis as compared with the corticosterone-treated group. Therefore, these data suggest that the extract of Ziziphus jujuba could be a good candidate for development as a functional food supplement in the improve the anti-stress effect.

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Protective Effect of Methanolic Extracts from Dendrobium nobile Lindl. on $H_{2}O_{2}$-induced Neurotoxicity in PC12 cells (석곡 MeOH 추출물이 $H_{2}O_{2}$에 의한 신경세포 보호효과에 미치는 영향)

  • Yoon, Mi-Young;Kim, Ju-Young;Hwang, Ji-Hwan;Cha, Mi-Ran;Lee, Mi-Ra;Jo, Kyung-Jin;Park, Hae-Ryong
    • Applied Biological Chemistry
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    • v.50 no.1
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    • pp.63-67
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    • 2007
  • The neuroprotective effect of methanolic extracts from Dendrobium nobile Lindl. (DME) against $H_{2}O_{2}$-induced neurotoxicity in PC12 cells was investigated. The treatment of PC12 cells with various DME concentrations under $H_{2}O_{2}$ resulted in the induction of protective effect in a dose-dependent manner, as determined by the results of an MTT reduction assay, an LDH release assays, and a morphological assay. Interestingly, we also detected reduction of apoptotic bodies and inhibition of caspase-3 activity by DME in $H_{2}O_{2}$-indeced PC12 cells. These data show that the neuroprotective effect of DME against PC12 cells might be related to the suppression of caspase-3 activation. Therefore, these results suggest that DME could be a new potential candidate as chemotherapeutic agents against neuronal diseases.