• The Korean Journal of Physiology and Pharmacology
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• v.24 no.4
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• pp.363-372
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• 2020

Gardenia jasminoides (GJ) is a widely used herbal medicine with anti-inflammatory properties, but its effects on the ORAI1 channel, which is important in generating intracellular calcium signaling for T cell activation, remain unknown. In this study, we investigated whether 70% ethanolic GJ extract (GJ_EtOH) and its subsequent fractions inhibit ORAI1 and determined which constituents contributed to this effect. Whole-cell patch clamp analysis revealed that GJ_EtOH (64.7% ± 3.83% inhibition at 0.1 mg/ml) and all its fractions showed inhibitory effects on the ORAI1 channel. Among the GJ fractions, the hexane fraction (GJ_HEX, 66.8% ± 9.95% at 0.1 mg/ml) had the most potent inhibitory effects in hORAI1-hSTIM1 co-transfected HEK293T cells. Chemical constituent analysis revealed that the strong ORAI1 inhibitory effect of GJ_HEX was due to linoleic acid, and in other fractions, we found that genipin inhibited ORAI1. Genipin significantly inhibited I_ORAI1 and interleukin-2 production in CD3/CD28-stimulated Jurkat T lymphocytes by 35.9% ± 3.02% and 54.7% ± 1.32% at 30 μM, respectively. Furthermore, the same genipin concentration inhibited the proliferation of human primary CD4⁺ T lymphocytes stimulated with CD3/CD28 antibodies by 54.9% ± 8.22%, as evaluated by carboxyfluorescein succinimidyl ester assay. Our findings suggest that genipin may be one of the active components of GJ responsible for T cell suppression, which is partially mediated by activation of the ORAI1 channel. This study helps us understand the mechanisms of GJ in the treatment of inflammatory diseases.

• The Journal of Korean Society of Virology
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• v.30 no.2
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• pp.125-130
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• 2000

Infection with human cytomegalovirus (HCMV) is of considerable clinical relevance after placental transmission and in immunosuppressed patients such as transplant recipients or patients with AIDS. The rapid detection method of HCMV has been required to overcome the time-consuming methods such as classical plaque assay or other immunological methods. This study was performed to establish the in situ ELISA, in which human lung fibroblasts infected with HCMV were fixed and used directly as antigen in 96 well culture plate. Expressed HCMV antigens were detected with HCMV-specific monoclonal antibodies. This method could detect HCMV dose-dependently upto $3{\times}10^2\;pfu/ml$. Antiviral activity of ganciclovir could be assayed within the known range of effective dose. This result showed that HCMV could be quantitated by in situ ELISA. The chemical, which was selected on the basis of component analysis in natural product, was tested to have the anti-HCMV activity by in situ ELISA, and three among five samples were found to have anti-HCMV activity with the dose-dependent manner. Conclusively in situ ELISA could be useful method for quantitation of HCMV and screening antiviral activity of samples to HCMV.

• Korean Journal of Pharmacognosy
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• v.43 no.2
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• pp.107-121
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• 2012

Twenty-five compounds were isolated from the methanolic extract of Geum japonicum (Rosaceae), and their structures were identified as eleven triterpenoids [ursolic acid 3-acetate (2), cecropiacic acid 3-methyl ester (3), pomolic acid 3-acetate (5), ursonic acid (6), ursolic acid (7), pomolic acid (8), corosolic acid (9), euscaphic acid (11), arjunic acid (16), tormentic acid (18), 23-hydroxytormentic acid (21)], two saponins [rosamultin (22) and kaji-ichigoside $F_1$ (23)], two megastigmanes [blumenol A (14) and (+)-dehydrovomifoliol (15)], three flavonoids [apigenin (13), isoquercitrin (17) and tiliroside (24)], two ellagic acid derivatives [3,3'-di-O-methylellagic acid (12) and ducheside B (25)] and five others [eugenol (1), emodin (4), vanillic acid (10), gallic aldehyde (19), salidroside (20)]. The chemical structures of these compounds were identified on the basis of spectroscopic methods and comparison with literature values. This is the first report of the eleven compounds, 2~6, 10, 15, 16, 20, 23, and 25 from the genus Geum, as well as the first report of apigenin (13) and 3,3'-di-O-methylellagic acid (12) from G. japonicum. The antioxidant properties of 22 isolates (1~11, 14, 16~25) were evaluated by the intracellular reactive oxygen species (ROS) radical scavenging using 2',7'-dichlorodihydrofluorescein diacetate (DCF-DA) assay. Among them, isoquercitrin (17) showed significant scavenging activity, and gallic aldehyde (19) and ducheside B (25) showed weak scavenging activity.

• Applied Biological Chemistry
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• v.42 no.2
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• pp.170-175
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• 1999

On the purpose of development of novel antioxidative compounds from natural sources, 38 plants expected to show antioxidant activity have been examined concerning DPPH radical scavenging activity. Among them, thirteen plants, including Paeoniae radix, the root of Paeonia lactiflora, exhibited the activity. In order to isolate active component, the root was extracted in 80% aqueous MeOH and solvent fractionated with EtOAc, n-BuOH and water, successively. Silica gel column chromatographies of the EtOAc and n-BuOH fraction exhibiting antioxidant activity. were repeatedly carried out with monitoring by DPPH assay to afford three active compounds. On the basis of spectral data and the chemical characteristics, the structures of the compounds were determined as (+)-catechin, $1,2,3,4-tetragalloyl-6-digalloyl-{\beta}-D-glucose$ and $1,2,3,4,6-penta-galloyl-{\beta}-D-glucose$.

• Korean Journal of Pharmacognosy
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• v.42 no.2
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• pp.110-116
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• 2011

As part of our ongoing study focused on the discovery of antioxidants from natural products by measuring the DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging activity, methanol extract of flowers of Carthamus tinctorius L. was found to show potent antioxidant activity. Activity-guided fractionation of the methanol extract lead to the isolation of twenty compounds including two flavonol glycosides, quercertin-3-O-${\beta}$-D-glucopyranoside (12) and kaempferol-3-O-${\alpha}$-L-rhamnopyranosyl-${\beta}$-D-glucopyranoside (18), two flavanone glycosides, (2S)-4',5,6,7-tetrahydroxyflavanone 6-O-${\beta}$-D-glucopyranoside (15) and (2R)-5,7,8',4-tetrahydroxyflavanone 8-O-${\beta}$-D-glucopyranoside (16), and two acetylenic glycosides, 8Z-decaene-4,6-diyne-1-O-${\beta}$-D-glucopyranoside (13) and 4,6-decadiyne-1-O-${\beta}$-D-glucopyranoside (14). Their chemical structures were identified by using spectroscopic analysis. Among them, compounds 12-18 were tested in DPPH assay. Compounds 13-16 were first reported to their antioxidant activity. Quercertin-3-O-${\beta}$-D-glucopyranoside (12) showed the most potent inhibitory effect on DPPH with $IC_{50}$ value of 56.7 ${\mu}M$.

• The Plant Pathology Journal
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• v.29 no.2
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• pp.193-200
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• 2013

Trichoderma asperellum SKT-1 is a microbial pesticide that is very effective against various diseases. Our study was undertaken to evaluate T. asperellum SKT-1 for induction of resistance against yellow strain of Cucumber mosaic virus (CMV-Y) in Arabidopsis plants. Disease severity was rated at 2 weeks post inoculation (WPI). CMV titre in Arabidopsis leaves was determined by indirect enzyme-linked immunosorbent assay (ELISA) at 2 WPI. Our results demonstrated that among all Arabidopsis plants treated with barley grain inoculum (BGI) of SKT-1 NahG and npr1 plants showed no significant reduction in disease severity and CMV titre as compared with control plants. In contrast, disease severity and CMV titre were significantly reduced in all Arabidopsis plants treated with culture filtrate (CF) of SKT-1 as compared with control plants. RT-PCR results showed increased expression levels of SA-inducible genes, but not JA/ET-inducible genes, in leaves of BGI treated plants. Moreover, expression levels of SA- and JA/ET-inducible genes were increased in leaves of CF treated plants. In conclusion, BGI treatment induced systemic resistance against CMV through SA signaling cascade in Arabidopsis plants. While, treatment with CF of SKT-1 mediated the expression of a majority of the various pathogen related genes, which led to the increased defense mechanism against CMV infection.

• Toxicological Research
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• v.31 no.2
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• pp.97-104
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• 2015

The skin exposure to solar irradiation and photoreactive xenobiotics may produce abnormal skin reaction, phototoxicity. Phototoxicity is an acute light-induced response, which occurs when photoreacive chemicals are activated by solar lights and transformed into products cytotoxic against the skin cells. Multifarious symptoms of phototoxicity are identified, skin irritation, erythema, pruritis, and edema that are similar to those of the exaggerated sunburn. Diverse organic chemicals, especially drugs, are known to induce phototoxicity, which is probably from the common possession of UV-absorbing benzene or heterocyclic rings in their molecular structures. Both UVB (290~320 nm) and UVA (320~400 nm) are responsible for the manifestation of phototoxicity. Absorption of photons and absorbed energy (hv) by photoactive chemicals results in molecular changes or generates reactive oxygen species and depending on the way how endogenous molecules are affected by phototoxicants, mechanisms of phototoxcity is categorized into two modes of action: Direct when unstable species from excited state directly react with the endogenous molecules, and indirect when endogeneous molecules react with secondary photoproducts. In order to identify phototoxic potential of a chemical, various test methods have been introduced. Focus is given to animal alternative test methods, i.e., in vitro, and in chemico assays as well as in vivo. 3T3 neutral red uptake assay, erythrocyte photohemolysis test, and phototoxicity test using human 3-dimensional (3D) epidermis model are examples of in vitro assays. In chemico methods evaluate the generation of reactive oxygen species or DNA strand break activity employing plasmid for chemicals, or drugs with phototoxic potential.

• Journal of Applied Biological Chemistry
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• v.62 no.4
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• pp.355-359
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• 2019

In vivo micronucleus tests were performed to investigate the mutagenic potential of 4-butylaniline and N-butylaniline, which are used in dye intermediates and organic intermediates respectively. Groups of 5 male ICR mice were treated with vehicle or 4-butylaniline for 2 consecutive days by oral gavage at concentrations of 0 (control), 64, 160, 400, and 1000 mg/kg. Statistically significant and dose-dependent increases were found for micronuclei frequencies in male mice (p <0.05). These results suggest that 4-butylaniline can induce genetic effects in the micronuclei of male mouse bone marrow cells. Based on the positive results obtained in cytogenetic analyses of somatic cells in vivo, Globally Harmonized System of Classification and Labelling of Chemicals Category 2 was assigned. N-butylaniline was administered for 2 consecutive days by oral gavage to male ICR mice at dose of 0 (control), 64, 160, 400, and 800 mg/kg. N-butylaniline tested negative for micronuclei induction in mice, although N-butylaniline was associated with micronucleus induction at the highest dose. Based on the negative results obtained for cytogenetic analyses of somatic cells in vivo, "Not Classified" was assigned.

• Journal of Microbiology and Biotechnology
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• v.11 no.3
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• pp.399-405
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• 2001

Urushiol, a natural pro-electrophilic quinone compound, has potential structural characteristics as antitumor chemotherapeutic agents. However, urushiol's use as an antitumor drug has some problems, because it is hardly miscible with an aqueous solution. Purified urushiol is highly viscous and soluble only in strong solvents. for this study, we prepared an urushiol-ethanol micro-emulsion with a unimodal size distribution by high-speed homogenization. This generated effective delivery of urushiol to its action wites, so that we could investigate its cytotoxic activity against cancer cells. Using a colony-forming assay, we were able to show that urushiol selectively inhibited the growth of the ovarian cancer cells PA-1 and 2774 at a concentration of $10^{-6}$, whereas it had only a negligible effect on normal CHO cells at the same concentration. The data suggest that urushiol may have potential as an effective antitumor agent in the treatment of ovarian cancer. In addition, we addressed the question of whether the specific cytotoxic effect of urushiol is linked to apoptosis, by DNA fragmentation and DAPI staining assays. The inhibitory effects of urushiol on the growth of ovarian cancer cells was found to be associated with DNA fragmentation and the fragmented nuclei formation, both of which represent markers for the induction of apoptosis. Therefore, the results suggested that urushiol affected its profound cytotoxicity by triggering apoptosis in ovarian cancer cells.

• KSBB Journal
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• v.20 no.5 s.94
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• pp.371-375
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• 2005

In the course of our search for compounds effective to multidrug-resistant cancer cells from myxobacteria with the adriamycin-resistant cancer cell line CL02, we found cytotoxic activity against the CL02 cells in culture extract of Sorangium cellulosum JW1045. Activity-guided fractionation of the culture extract led to the isolation of an active principle, chivosazole F, This compound showed high cytotoxic activity against cultured human cancer cells. The $IC_{50}$ values, measured by a SRB assay with different cell lines, ranged from 0.1 to 10 ng/ml. Furthermore chivosazole F was as active against drug-resistant cancer cells CL02 and CP70 as against the corresponding sensitive cells.