• Journal of Korean Society of Water and Wastewater
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• v.19 no.5
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• pp.639-646
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• 2005

In this research, we tried to develop the application method to water management and treatment using toxicity test method. When we measure the toxicity of environmental samples, we have to decide whether we take some countermeasures to reduce the toxicity or not. The first issue is how to set these action levels in each bioassays. A new idea was attempted to authorize indirect approach of each bioassays through the response characteristics against mixture of chemicals in water quality standard. The significant response in the cell-growth-inhibition bioassay was detected for standards-mixture(STDs). For acute toxicity assay, STDs-based implicit correlation between risks to humans and bioassay data showed a rational approach to set action levels in practical management. A simple model was proposed to describe and predict the changes in the total toxicity based on the concentrations of toxic-controlling chemicals during the ozonation of landfill leachates. On the basis of this simple model, toxicity reduction was predicted for pre-aggregation treatment before ozonation and ozone concentration during the ozonation. The method proposed in this study would be useful in optimizing water treatment processes and their running conditions in terms of the toxicity reduction efficacy.

• Archives of Pharmacal Research
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• v.25 no.6
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• pp.903-909
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• 2002

Research in the cytokine field has grown exponentially in recent years, and the validity of such studies relies heavily on the appropriate measurement of levels of cytokines in various biological samples. Transforming growth factor (TGF)-$\beta$, a hormonally active polypeptide found in normal and transformed tissue, is a potent regulator of cell growth and differentiation. The most widely used bioassay for TGF-$\beta$ is the inhibition of the proliferation of mink lung epithelial cells. Though detection of [$^3$H]thymidine incorporation is more sensitive than the MTT assay, it presents some disadvantages due to the safety and disposal problems associated with radioisotopes. In this study, we attempted to ascertain the experimental conditions which could be used for measuring the in vitro biological activity of TGF-$\beta$ in a safer and more sensitive way compared with the currently available methods. We compared the commonly used method, the MTT assay, to the XTT assay using different parameters including cell number, incubation time and the wave length used for detecting the product. We examined the anti-proliferative activities of TGF-$\beta$ in three different cell lines: Mv-1-Lu mink lung epithelial cells, MCF10A human breast epithelial cells and H-ras-transformed MCF10A cells. Herein, we present an experimental protocol which provides the most sensitive method of quantifying the biological activity of TGF-$\beta$, with a detection limit of as low as 10 pg/ml: Mv-1-Lu or H-ras MCF10A cells ($1{\times}10^5/well$) were incubated with TGF-$\beta$ at $37^{\circ}C$ in a humidified $CO_2$ incubator for 24 hr followed by XTT treatment and determination of absorbance at 450 or 490 nm. Our results may contribute to the establishment of an in vitro bioassay system, which could be used for the satisfactory quantitation of TGF-$\beta$.

• Korean Journal of Pharmacognosy
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• v.26 no.1
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• pp.47-50
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• 1995

EtOAc soluble part of MeOH extract of Anemarrhena asphodeloides rhizome was evaluated for the cytotoxicity against the five kinds of human tumor cell lines (A-549, SK-OV-3, SK-MEL-2, XF498 and HCT15) in vitro. Bioassay-guided fractionation of EtOAc soluble part led to the isolation of active compound which was identified as timosaponin A-III showed potent cytotoxic activity, but its genin, sarsasapogenin, did not show cell growth inhibition.

• Journal of Marine Bioscience and Biotechnology
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• v.1 no.4
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• pp.260-267
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• 2006

The Liquid Culture Assay using a recombinant Agrobacterium tumefaciens strain has been developed as a means for quorum sensing autoinducer screening. However, the low aqueous solubility of marine natural product extracts used as potential autoinducers has been a hindrance in the screening process. Although the addition of organic solvents and/or surfactants could increase aqueous solubility, errors in data interpretation including false positive results could be a serious problem. Therefore, determining the best possible solvent and surfactant at the optimum concentration is crucial. Evaluating methanol, ethanol, 1-propanol, DMSO and DMF at concentration ranges of 0~10% revealed < 2% methanol to be most favorable when tested for ${\beta}$-gal activity and growth inhibition of the recombinant A. tumefaciens strain. On the other hand, among surfactants tested, Triton X-100 was similarly effective in increasing the delivery of autoinducers for activity at less than 0.05% concentration.

• Journal of Microbiology and Biotechnology
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• v.30 no.12
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• pp.1885-1895
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• 2020

Rumex japonicus Houtt (RJH) is a valuable plant used in traditional medicine to treat several diseases, such as scabies and jaundice. In this study, Jurkat cell growth inhibitory extracts of R. japonicus roots were subjected to bioassay-guided fractionation, resulting in the isolation of three naphthalene derivatives (3-5) along with one anthraquinone (6) and two phenolic compounds (1 and 2). Among these compounds, 2-methoxystypandrone (5) exhibited potent anti-proliferative effects on Jurkat cells. Analysis by flow cytometry confirmed that 2-methoxystypandrone (5) could significantly reduce mitochondrial membrane potential and promote increased levels of mitochondrial reactive oxygen species (ROS), suggesting a strong mitochondrial depolarization effect. Real-time quantitative polymerase chain reaction (qPCR) analysis was also performed, and the results revealed that the accumulation of ROS was caused by reduced mRNA expression levels of heme oxygenase (HO-1), catalase (CAT), glutathione peroxidase (GPx), and superoxide dismutase (SOD). In addition, 2-methoxystypandrone (5) triggered strong apoptosis that was mediated by the arrest of the G0/G1 phase of the cell cycle. Furthermore, 2-methoxystypandrone (5) downregulated p-IκB-α, p-NF-κB p65, Bcl2, and Bcl-xl and upregulated BAX proteins. Taken together, these findings revealed that 2-methoxystypandrone (5) isolated from RJH could potentially serve as an early lead compound for leukemia treatment involving intracellular signaling by increasing mitochondrial ROS and exerting anti-proliferative effects.

• Biomolecules & Therapeutics
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• v.12 no.2
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• pp.122-128
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• 2004

Since reactive oxygen species, prostaglandins, and nitric oxide are closely involved in various pathological conditions anti play important roles in the initiation, promotion, and progression of carcinogenesis, agents that modulate the production or activity of them might be considered as cancer chemopreventive agents. In the present study, we evaluated chemopreventive potential of some grape shoot extracts and their refined materials using various in vitro assay systems. As a result, both grape shoot extracts and refined materials possessed effective radical scavenging activities about 70~80% at the concentration of 500 ${\mu}$g/ml, and especially the Sheridan shoot extract showed the most potent 1, 1-diphenyl-2-picrylhydrazyl radical scavenging activity that was similar to resveratrol. In addition, refined materials from grape shoot extracts suppressed lipopolysaccharide-induced nitric oxide production in macrophage cells, anti refined materials from Kyoho and Campbell shoot extracts exhibited similar inhibitory activities with $IC_{50}$ value of 224 ${\mu}$g/ml and 285 ${\mu}$g/ml, respectively. In addition, at the concentration of 50 ${\mu}$g/ml, all of refined materials inhibited cell proliferation against various human cancer cells about 30~40% compared to control. These findings suggest that grape shoot extract and their refined materials might be useful sources for the development of chemopreventive agents and/or functional foods.

• Biomolecules & Therapeutics
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• v.30 no.2
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• pp.184-190
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• 2022

Targeting the cystine/glutamate exchange transporter, system x_c^-, is a promising anticancer strategy that induces ferroptosis, which is a distinct form of cell death mediated by iron-dependent lipid peroxidation. The concentration of ⳑ-cystine in culture medium is higher than the physiological level. This study was aimed to evaluate the effects of ⳑ-cystine concentration on the efficacy of ferroptosis inducers in hepatocellular carcinoma cells. This study showed that treatment with sulfasalazine or erastin, a system x_c^- inhibitor, decreased the viability of Huh6 and Huh7 cells in a dose-dependent manner, and the degree of growth inhibition was greater in medium containing a physiological ⳑ-cystine concentration of 83 µM than in commercial medium with a concentration of 200 µM ⳑ-cystine. However, RSL3, a glutathione peroxidase 4 inhibitor, decreased cell viability to a similar extent in media containing both ⳑ-cystine concentrations. Sulfasalazine and erastin significantly increased the percentages of propidium iodide-positive cells in media with 83 µM ⳑ-cystine, but not in media with 200 µM ⳑ-cystine. Sulfasalazine- or erastin-induced accumulation of lipid peroxidation as monitored by C11-BODIPY probe was higher in media with 83 µM ⳑ-cystine than in media with 200 µM ⳑ-cystine. In contrast, the changes in the percentages of propidium iodide-positive cells and lipid peroxidation by RSL3 were similar in both media. These results showed that sulfasalazine and erastin, but not RSL3, were efficacious under conditions of physiological ⳑ-cystine concentration, suggesting that medium conditions would be crucial for the design of a bioassay for system x_c^- inhibitors.