• Proceedings of the Korean Society of Applied Pharmacology
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• 1994.04a
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• pp.129-130
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• 1994

The science of toxicology is the understanding of the mechanisms by which exogenous agents produce deleterious effects in biological systems. The actions of chemicals such as drugs are ultimately exerted at the cellular and gene levels. Over the past decade. several in vitro alternative methods such as cultured cells for assessing the toxicity of various xenobiotics have been proposed to reduce the use of animals. In this workshop three advanced methods will be presented. These methods are novel important models for toxicologic studies. Dr. Tabuchis group has establishcd two immortalized gastric surface mucosa cell lines from the pminary cultore of gastric fundic mucosal cells of adult transgenic mice harboring a temperature sensitive simian virus 40 large T-anugen gene. As the immortalized cell lines of various tissues possess unique characteristics to maintain their normal functions for several months, these cell lines are extremely useful for not only toxicity testing but also pharmacological screening in new drug development. Professor Funatsu have studied the formation of spherical multicelluar aggregates of adult rat hepatocytes(spheroid) having tissue like structure. The sphcroid shown thre is a prototype module of an artificial liver support system. Thus, the urea synthesis activity of the artificial liver was maintained at least to days in 100％ rat blood plasma. Dr. Takezawa and his coworkers have developed a novel culture system of multicellular spheroids considered 〃organoids〃 by utilizing a thermo-responsive polymer as a substratum of anchorage dependent cells. His final goal is to reconstitute the organoids of various normal organs, e.g., liver, skin etc. and also abnormal deseased organs such as tumor.

• Journal of Environmental Health Sciences
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• v.31 no.4 s.85
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• pp.254-259
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• 2005

Permethrin, a synthetic pyrethroid insecticide, has been widely used to protect domestic animals and the public health, as well as in agriculture against a variety of pests, which provides potential for environmental exposure. Permethrin is classified as possible human carcinogen and endocrine disrupting chemical by many international authorities. However, its developmental effects have been rarely studied. This study investigated the effects of permethrin during embryo-genesis. Developmental toxicity of permethrin was evaluated using short-term in vitro battery system. Gestation day 9.5 rat embryos (organogenesis) were cultured with permethrin (0.1,0.4 and 0.8 mg/ml) for 48 hours using whole embryo culture system. All the treatments exhibited significant decreases in the total morphological score. Permethrin induced significant growth retardation and the developmental abnormality at doses of 0.4 and 0.8 mg/ml. Moreover, the DNA and protein contents of embryos decreased in dose-dependent manner. These observations suggest that permethrin contributes to toxicity on embryonic developments in rats.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.15 no.1
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• pp.37-50
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• 1989

The in Vitro chemosensitivity of fibroblast cell strains was determined using a semiautomated tetrazolium-based colorimetric assay(MTT assay) to 16 cosmetic materials. This assay is useful method to evaluate toxic effects of the chemicals. From assay results, we determined that the preservatives are more toxic than moisteurizers. The chemicals in the same group have a different toxicity. That is, in preservatives, Germall -115 is more toxic than Danisol -M, -p, and in surfactant sodium laurel sulfate than Myrj 52, and in moisteurizers, 1, 3-butylene glycol is more safe than the others. When the results from this assay for preservatives were compared with patch test results, good correlation was observed. Forthemore, this assay method can be used together with Patch test for the evaluation of the chemical toxicity, particularly in cosmetic field.

• Journal of Korean Society of Occupational and Environmental Hygiene
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• v.23 no.2
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• pp.50-56
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• 2013

Objectives: The present study investigated cytotoxicity and DNA damage in human lung cells in vitro. Methods: Neodymium oxide and lanthanum oxide were dispersed by ultrasonic treatments. The assay was performed with MRC-5 (Human male fetus lung cell). Cytotoxicity and comet assay of lanthanum oxide and neodymium oxide were measured after 24 and 48 hours incubation. Results: After 24 hours of exposure to rare earth metals, the cytotoxicities of lanthanum oxide in more than $1{\mu}M$ concentration groups were significantly increased when compared to the control group, but the cytotoxicities of neodymiun oxide in more than $100{\mu}M$ concentration groups were statistically increased. After 48 hours exposure, cytotoxicities of both materials were statistically increased in $100,000{\mu}M$ concentration groups. Olive tail moments of the lanthanum oxide treated group were significantly increased when compared to the control group. Conclusions: The cytotoxicity of lanthanum oxide was higher than that of neodymium oxide. The DNA of MRC-5 cells treated with lanthanum oxide for 48 hours were significantly damaged.

• Korean Journal of Pharmacognosy
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• v.27 no.4
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• pp.309-315
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• 1996

As part of our search for less toxic antimicrobial agents from natural resources, antimicrobial activity of rutin isolated from Sophora japonica was tested in vitro against four kinds of gram positive bacteria and four kinds of gram negative bacteria by serial broth dilution method. Among eight kinds of bacteria tested, the antimicrobial activity of rutin was the most potent against Mycobacterium smegmatis showing MIC of $375\;{\mu}g/ml$. The acute toxicity of rutin was examined in mice and rats. and $LD_{50}$ value administered intraperitoneally in mice was 650 mg/kg(Confidence limit: 894-1,473 mg/kg). There were no significant changes in serum biochemical values and histopathological changes as compared with those of control after intraperitoneal administration with rutin in rats.

• Environmental Analysis Health and Toxicology
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• v.13 no.3_4
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• pp.87-94
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• 1998

The use of cisplatin is limited by severe side effects such as renal toxicity. Our platinum-base drug discovery is aimed at developing drugs capable of diminishing toxicity and improving antitumor activity. We synthesized new Pt (II) complex analogue [Pt (cis-DACH)(DPPP)]. 2NO$_3$ (PC) containing cis-1,2-diaminocyclohexane as a carrier ligand and 1,3-bis(diphenylphosphino) propane as a leaving group. Furthermore, nitrate was added to improved the solubility. In this study, its structure was determined and its antitumor activity against SKOV-3 and NIH-OVCAR-3 human ovarian adenocarcinoma, and in vitro cytotoxicity was determined against primary cultured rabbit kidney proximal tubular and renal cortical cells of human kidney using colorimetric MTT assay. PC demonstrated acceptable antitumor activity against SKOV-3 and NIH-OVCAR-3 human ovarian adenocarcinoma and significant activity as compared with that of cisplatin. The toxicity of PC was found quite less than that of cisplatin using MTT and $^3$H-thymidine uptake tests in rabbit proximal tubular cells and human kidney cortical cells. PC was used for human cortical tissue in 7 weeks hitoculture by the glucose-consumption tests. We determined that the new platinum drug has lower nephrotoxicity than cisplatin. Based on these results, this novel platinum (II) complex compound (PC) represent a valuable lead in the development of a new anticancer chemotherapeutic agent capable of improving antitumor activity and low nephrotoxicity.

• Biomolecules & Therapeutics
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• v.28 no.4
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• pp.302-310
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• 2020

Botulinum toxins are neurotoxic modular proteins composed of a heavy chain and a light chain connected by a disulfide bond and are produced by Clostridium botulinum. Although lethally toxic, botulinum toxin in low doses is clinically effective in numerous medical conditions, including muscle spasticity, strabismus, hyperactive urinary bladder, excessive sweating, and migraine. Globally, several companies are now producing products containing botulinum toxin for medical and cosmetic purposes, including the reduction of facial wrinkles. To test the efficacy and toxicity of botulinum toxin, animal tests have been solely and widely used, resulting in the inevitable sacrifice of hundreds of animals. Hence, alternative methods are urgently required to replace animals in botulinum toxin testing. Here, the various alternative methods developed to test the toxicity and efficacy of botulinum toxins have been briefly reviewed and future perspectives have been detailed.

• Clinical and Experimental Reproductive Medicine
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• v.25 no.2
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• pp.109-113
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• 1998

Objective: To ananlyze the direct effect of nitric oxide (NO), generated from sodium prusside (SNP) on the embryo developments in reproductive process. Design: Ova from mouse were treated to allow fertilization in in vitro culture. And the samples of fertilized ova were alloted into five alliqutos. Each alliquot was cultured in media treated with either concentration at 0 (n=92), $25{\mu}M$ (n=84), $50{\mu}M$ (n=80), $100{\mu}M$ (n=77), $500{\mu}M$ (n=54) of SNP. Main Outcome Measure: Rates of embryonal cell cleavages, viability and cell morphology were assessed during in vitro fertilization and culture. Results: As analyse the cell cleavage at 24 hours after in vitro culture of fertilised egg in variuos NO concentration, all of egg cells of each alliquot were developed to $2\sim4$ cell stage. But the alliquot of egg cells treated with $50{\mu}M$, which were totally degenerated. And also all embryonal cells of each alliquot were developed to 8 cell stage and morula stage on culture continuosly. And the embryonal cells of each alliquot were analysed at 24 and 48 hours following the in vitro culture. The rates of cell fragmentation and fusion were $4.2{\pm}3.4%$ in control group which is not treated with NO, while experimental groups was high, as rated $23.4{\pm}6.2%$ in $25{\mu}M$, $28.2{\pm}5.7%$ in $50{\mu}M$ and $32.1{\pm}6.4%$ in $100{\mu}M$ concentration of NO. Accordingly the rate of abnormal morphology of embryonal cell in control was lower significantly than that in each alliquot of experimental groups (p<0.05). And the degenerated rates of embryonal cells were 0% in control, $17.8{\pm}6.7%$ in $25{\mu}M$, $23.6{\pm}4.7%$ in $50{\mu}M$ and $26.8{\pm}11.2%$ in $100{\mu}M$ at 8 cells and morula on culture of 48 and 72 hours. On the examination of embryonal cells developed to blastocyst through in vitro culture, the rates of degenerated cells were $16.8{\pm}7.2%$ in control, $37.5{\pm}6.2%$ in $25{\mu}M$, $73.4{\pm}4.6%$ in $50{\mu}M$, 100% in $100{\mu}M$. Conclusion: This results suggeted that the NO in any concentrations is harmful on embryos in view of morphology as well as viability of cell, and the toxicity of NO on embryo is stronger at condition in higher concentration of NO.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1995.04a
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• pp.123-123
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• 1995

To evaluate the genetic toxicity of NP-77A which is selected as the candidate of anti-HBV agent, we performed ames test, micronucleus test, and chromosome aberration test on the CHL cell in vitro. The Ames test was carried out with 5 fold diluted 5 concentrations from 25mg/plate using S. typhimurium and E.coli. After 48hrs incubation, revertant colony numbers was calculated with and without metabolic activation system. In vivo micronucleus test, we investigated the rate of the occurrence of micronucleus after I.P. administration to mice. Andalso, we observed the incidence rate of cells with chromosomal aberration by NP-77A treatment using CHL cell line.

• Molecular & Cellular Toxicology
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• v.4 no.2
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• pp.157-163
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• 2008

Formation of ${\beta}$-amyloid $(A{\beta})$ fibrils has been identified as one of the major characteristics of Alzheimer's disease (AD). Inhibition of $A{\beta}$ fibril formation in the CNS would be attractive therapeutic targets for the treatment of AD. Several small compounds that inhibit amyloid formation or amyloid neurotoxicity in vitro have been known. Citrate has surfactant function effect because of its molecular structure having high anionic charge density, in addition to the well-known antibacterial and antioxidant properties. Therefore, we hypothesized that citrate might have the inhibitory effect against $A{\beta}$ fibril formation in vitro and have the protective effect against $A{\beta}$-induced neurotoxicity in PC12 cells. We examined the effect of citrate against the formation of $A{\beta}$ fibrils by measuring the intensity of fluorescence in thioflavin-T (Th-T) assay of between $A{\beta}_{25-35}$ groups treated with citrate and the control with $A{\beta}_{25-35}$ alone. The neuroprotective effect of citrate against $A{\beta}$-induced toxicity in PC12 cells was investigated using the WST-1 assay. Fluorescence spectroscopy showed that citrate inhibited dose-dependently the formation of $A{\beta}$ fibrils from ${\beta}$-amyloid peptides. The inhibition percentages of $A{\beta}$ fibril formation by citrate (1, 2.5, and 5 mM) were 31%, 60%, and 68% at 7 days, respectively in thioflavin-T (Th-T) assay. WST-1 assay revealed that the toxic effect of $A{\beta}_{25-35}$ was reduced, in a dose-dependent manner to citrate. The percentages of neuroprotection by citrate (1, 2.5, and 5 mM) against $A{\beta}-induced$ toxicity were 19%, 31 %, and 34%, respectively. We report that citrate inhibits the formation of $A{\beta}$ fibrils in vitro and has neuroprotective effect against $A{\beta}$-induced toxicity in PC12 cells. Neuroprotective effects of citrate against $A{\beta}$ might be, to some extent, attributable to its inhibition of $A{\beta}$ fibril formation. Although the mechanism of anti-amyloidogenic activity is not clear, the possible mechanism is that citrate might have two effects, salting-in and surfactant effects. These results suggest that citrate could be of potential therapeutic value in Alzheimer's disease.