• Journal of Oriental Neuropsychiatry
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• v.21 no.4
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• pp.41-51
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• 2010

Objectives : Samul-tang(SM). Sagunja-tang(SG). Pamul-tang(PM) and Sipjeondaebo-tang(SJ) was used many diseases such as sterility. menstrual disorder. general prostration. recruitment in Korea. We investigated the immune stimulatory activities of SG. SM. PM and SJ in in vitro. Methods: For comparision for effective of SM. SG. PM and SJ. this study examined anti-inflammation(NO. PGE2 assay). anti-oxidation(DPPH assay) and immune response in in vitro assay. For immune response activities. this study used NO synthesis on RAW 264.7 cells, splenocyte proliferation and cytokine assay(IL-2, IL-4) in splenocyte. Results: The results showed that SG. SM. PM and SJ were no significant effect anti-oxidation and anti-inflammatory effects. For immune response. they showed the splenocyte proliferation and macrophage proliferation. We confirmed that they synthesised NO a dose-dependent manner significantly and secreted the IL-4. Conclusions : These results suggested the SG, SM, PM and SJ had immune stimulatory activity. A efficacy of immune response in them had produced similar results.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.34 no.3
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• pp.183-188
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• 2008

Initial screening assay for depigmenting agents includes in vitro mushroom tyrosinase assay and antioxidant assay. Based on this screening result, melanin synthesis in melanocyte, in screened samples, is further measured. Measuring cellular melanin needs time, human resource, and skills. Therefore initial screening method should be reliable. We examined, 34 Chinese herbs, correlated the screening assay methods with cellular melanin. No reliable relationship was observed between factors, indicating the limitation in the use of these assays, probably due to the complexicity of melanogenesis.

• Environmental Mutagens and Carcinogens
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• v.19 no.1
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• pp.7-13
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• 1999

The mouse lymphoma thymidine kinase (tk+/-) gene assay (MOLY) using L5178Y tk+/- mouse lymphoma cell line is one of the mammalian forward mutation assays. It is well known that MOLY has many advantages and more sensitive than the other mammalian forward mutation assays such as x-linked hyposanthine phosphoribosyltransferase (hprt) gene assay. The target gene of MOLY is a heterozygous tk+/- gene located in 11 chromosome of L5178Y tk+/- cell, so it is able to detect the wide range of genetic changes like point mutation, deletion, rearrangement, and mitotic recombination within tk gene or deletion of entire chromosome 11. MOLY has relatively short expression time (2-3 days) compared to 1 week of hprt gene assay. MOLY can also induce relatively high mutant frequency so a large number of events can be recorded. The bimodal distribution of colony size which may indicate gene mutation and chromosome breakage potential of chemicals according to mutation scale such as large normal-growing mutants and small slow-growing mutants can be observed in this assay. The statistical analysis of data can be performed using the MUTANT program developed by York Electronic Research in association with Hazelton as recommended by the UKEMS (United Kingdom Environmental Mutagen Society) guidelines. This report reviewed MOLY using the microtiter cloning technique (microwell assay).

• Journal of Food Hygiene and Safety
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• v.10 no.3
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• pp.133-138
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• 1995

In vitro antimutagenic activity of methanol extract from brrwn rice and its physico-chemical characteristics were investigated using Salmonella typhimurium reversion assay and SOS chromotest. Methanol extracts of brown rice were not mutagenic compared with direct and indirect, mutagenicities of 4NQO (4-nitroquinoline oxide), 2NF(2-nitrofluorene), Trp-p-1(3-Amino-1,4-dimethyl-5H-pyrido-[4,3-b]indole), and Trp-p-2(3-Amino-1-methy-5H-pyrido-[4,3-b]indole). Antimutagenic activity against the indirect mutagenicties induced by Trp-p-1, Trp-p-2 and AFB1 (aflatoxin B1) was found in methanol extract. Even though antimutagenic activity showed dose-dependent, it remained constant at inhibition rate ranging 60~90% when the concentration was abov 3mg/plate in the S. typhimurium reversion assay and 0.2~0.6 mg/assay in the SOS chromotest. The antimutagenic activity of the methanol extracts was stable at various pH (2, 7 and 10), temperatures (60, 80 and 10$0^{\circ}C$)and heation times (2, 4, 6, 8, 10 min at 10$0^{\circ}C$).

• Journal of Food Hygiene and Safety
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• v.20 no.1
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• pp.18-21
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• 2005

These observations were performed to investigate the safety of the natural herbs (Rhus-II) in respect of genotoxicity. This substance was examined in two in-vitro tests: (1) Salmonella typhimurium reversion assay (Ames test) in strain TA 98, TA 100, TA 1535 and TA 1537, (2) in vitro chromosome aberration test in cultured Chinese hamster ovary (CHO) cells. In the reverse mutation test, Rhus-II did not induced mutagenicity in Salmonella typhimurium reversion assay(Ames test) with or without metabolic activation. In the chromosome aberration assay using CHO cells, there was no increased incidence of structural and numerical aberrations with or without metabolic activation. These results indicated that, the Rhus-II had no genotoxicity.

• Journal of Microbiology and Biotechnology
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• v.16 no.10
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• pp.1646-1649
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• 2006

Protein synthesis is the ultimate outcome of gene expression which, in turn, is regulated by several translation factors. We attempted to identify substances that can inhibit the translation process in vitro when the outcome protein is luciferase. To this end, we developed a sensitive cell-free protein synthesis assay using luciferase as the reporter. The synthesis of luciferase increased proportionately as mRNA was added to a $15-{\mu}l$reaction medium in concentrations raging from 5 ng to 500 ng. The maximum amount of luciferase was synthesized when the media were incubated at $25^{\circ}C$ for 40 min. The concentration of each compound that inhibited luciferase production by 50% ($IC_{50}$) was calculated. Hygromycin, puromycin, and cycloheximide yielded an $IC_{50}$ of 0.008, 0.8, and $0.7{\mu}g/ml$, respectively. A filtrate of Streptomyces spp. isolates inhibited protein synthesis up to S-fold when added to the in vitro translation assay mixture.

• Environmental Analysis Health and Toxicology
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• v.14 no.1_2
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• pp.1-12
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• 1999

Recently, several new methods for the detection of genetic damages in vitro and in vivo based on molecular biological techniques were introduced according to the rapid progress in toxicology combined with cellular and molecular biology. Among these methods, mouse lymphoma thymidine kanase (tk) gene forward mutation assay, single cell gel electrophoresis (comet assay) and transgenic animal and cell line model as a target gene of lac I (Big Blue) and lac Z (Muta Mouse) gene mutation are newly introduced based on molecular toxicological approaches. The mouse lymphoma tk$\^$+/-/ gene assay (MOLY) using L5178Y tk$\^$+/-/ mouse lymphoma cell line is one of the mammalian forward mutation assays, and has many advantages and more sensitive than hprt assay. The target gene of MOLY is a heterozygous tk$\^$+/-/ gene located in 11 chromosome, so it is able to detect the wide range of genetic changes like point mutation, deletion, rearrangement, and mitotic recombination within tk gene or deletion of entire chromosome 11. The comet assay is a rapid, simple, visual and sensitive technique for measuring and analysing DNA breakages in mammalian cells, Also, transgenic animal and cell line models, which have exogenous DNA incorporated into their genome, carry recoverable shuttle vector containing reporter genes to assess endogenous effects or alteration in specific genes related to disease process, are powerful tools to study the mechanism of mutation in vivo and in vitro, respectively. Also in vivo acridine orange supravital staining micronucleus assay by using mouse peripheral reticulocytes was introduced as an alternative of bone marrow micronucleus assay. In this respect, there was an International workshop on genotoxicity procedure (IWGTP) supported by OECD and EMS (Environmental Mutagen Society) at Washington D. C. in March 25-26, 1999. The objective of IWGTP is to harmonize the testing procedures internationally, and to extend to finalization of OECD guideline, and to the agreement of new guidelines under the International Conference of Harmonization (ICH) for these methods mentioned above. Therefore, we introduce and review the principle, detailed procedure, and application of MOLY, comet assay, transgenic mutagenesis assay and supravital staining micronucleus assay.

• BMB Reports
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• v.28 no.1
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• pp.12-16
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• 1995

The branched-chain ${\alpha}$-keto acid dehydrogenase (BCKAD) complex is a rate limiting enzyme which catalyzes the oxidative decarboxylation of branched-chain ${\alpha}$-keto acids. Numerous studies have suggested that BCKAD is subject to covalent modification in vitro via phosphorylation and dephosphorylation, which are catalyzed by a specific kinase and phosphatase, respectively. The biggest difficulty in the assay of BCKAD activity is to arrest the interconversion between the active and inactive forms. BCKAD activity was determined from fresh rat heart and liver tissues using homogenizing and assay buffers containing inhibitors of phosphatase and kinase. The results suggest that a radiochemical assay using ${\alpha}$-keto[1-$^{14}C$]-isovalerate as a substrate for the enzyme can be applied as a reliable method to determine in vitro enzyme activity with arrested interconversion between the active and inactive forms of the BCKAD complex.

• Journal of Pharmaceutical Investigation
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• v.34 no.5
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• pp.393-399
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• 2004

For the efficient determination of activity against solid tumors, an in vitro tumor model that resembles the condition of in vivo solid tumors, is required. The purpose of this study was to establish a rapid culture method and viability assay for an in vitro 3-dimensional tumor model, multicellular spheroid (MCS). Among 12 human cancer cell lines, a few cell lines including DLD-1 (human colorectal carcinoma cells) formed fully compact MCS which was adequate for in vitro viability assay. DLD-1 MCS showed steady growth reaching $700\;{\mu}m$ diameter after 11 day culture. DLD-1 cells grown as MCS showed significant increase in $G_0/G_1$ phase compared to the monolayer cells (73.9% vs 45.7%), but necrotic regions or apoptotic cells were not observed. The cells cultured as MCS showed resistance to 5-FU (10.3 fold higher $IC_{50}$) compared to monolayers, however, tirapazamine (a hypotoxin) showed similar activity in both culture systems. In summary, MCS may be a valid in vitro model for activity screening of anticancer agents against human solid tumors and also exploitable for studying molecular markers of drug resistance in human solid tumors.

• YAKHAK HOEJI
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• v.44 no.5
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• pp.416-423
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• 2000

There is a growing concern that a wide variety of chemicals released into the environment can disrupt the endocrine system of fish, wildlife and humans. Endocrine disrupting chemicals (EDCs) include pesticides such as DDT lindane and atrazine, the food packaging chemicals, phthalates and bisphenol A, alkylphenol ethoxylate detergents and the chemical industry by-products, dioxins. Xenoestrogens in the environment have been argued about health risk, because of estrogen mimetic chemicals are exposed only small amounts to human. A number of in vivo and in vitro assays are now in use to assess the activity of xenoestrogens in the environment. A human breast cancer cell line (MCF-7) was used to develop in vitro screening assay for the detection of xenoestrogenic environmental pollutants. The E-SCREEN (MCF7-BUS) assay is proposed as a reliable, easy and rapid-to-perform method. To optimize and validate this method before it can be used routinely, several phenol compounds and pesticides suspected to be estrogenic were tested using I-SCREEN assay. The results showed that this method is a valuable tool for screening potential estrogen-mimicking environmental pollutants and quantitative determination of estrogeniciy.