• Journal of Korean society of Dental Hygiene
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• v.14 no.4
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• pp.601-608
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• 2014

Objectives : The purpose of the study is to examine the apoptotic effects of Pseudomonas aeruginosa exotoxin A(PEA) in squamous cell carcinoma(SCC) 25 cells. Methods : Cell growth reduction and apoptosis induced by PEA were confirmed by WST-1 assay, Hoechst 33258 staining, flow cytometry analysis, and Western blot assay. Results : The PEA treatment decreased the cell viability in a dose and time dependent manner: control; $100{\pm}0^e$(p<0.01), 0.1875 nM; $87{\pm}4.36^d$(p<0.01), 0.375 nM; $82{\pm}0.58^d$(p<0.01), 0.75 nM; $72{\pm}1.67^c$(p<0.01), 1.5 nM; $51{\pm}1.53^{bc}$(p<0.01), 7.5 nM; $31{\pm}1.20^{ab}$(p<0.01), 15 nM; $26{\pm}0.67^a$(p<0.01), control; $100{\pm}0^a$(p<0.05), 24 h; $51{\pm}1.53^b$(p<0.05), 48 h; $16{\pm}0.5^c$(p<0.05), 72 h; $12{\pm}1.67^d$%(p<0.05). The PEA was observed on SCC 25 cells with the half maximal inhibitory concentration(IC50) value of 1.5 nM at 24 hours. The PEA treated SCC 25 cells demonstrated several types of apoptotic indications, such as nuclear condensation, the increase of sub G1, and the cleavage of PARP-1 and DFF 45. Conclusions : PEA showed anti-cancer activity against SCC 25 cells via apoptosis. PEA may potentially contribute to human oral cancer treatment.

• Reproductive and Developmental Biology
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• v.36 no.1
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• pp.13-19
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• 2012

In order to provide the basis for developing practical mouse embryonic stem cells (mESCs) culture method, how the endogenous level of self-renewal-stimulating factor genes was altered in the mESCs by different extracellular signaling was investigated in this study. For different extracellular signaling, mESCs were cultured in 2 dimension (D), 3D and integrin-stimulating 3D culture system in the presence or absence of leukemia inhibitory factor (LIF) and transcriptional level of $Lif$, $Bmp4$ and $Wnt3a$ was evaluated in the mESCs cultured in each system. The expression of three genes was significantly increased in 3D system relative to 2D system under LIF-containing condition, while only $Wnt3a$ expression was increased by 3D culture under LIF-free condition. Stimulation of integrin signaling in mESCs within 3D system with exogenous LIF significantly up-regulated transcriptional level of $Bmp4$, but did not induce transcriptional regulation of $Lif$ and $Wnt3a$. In the absence of LIF inside 3D system, the expression of $Lif$ and $Bmp4$ was significantly increased by integrin signaling, while it significantly decreased $Wnt3a$ expression. Finally, the signal from exogenous LIF significantly caused increased expression of $Lif$ in 2D system, decreased expression of $Bmp4$ in both 2D and 3D system, and decreased expression of $Wnt3a$ in integrin-stimulating 3D system. From these results, we identified that endogenous expression level of self-renewal-stimulating factor genes in mESCs could be effectively regulated through artificial and proper manipulation of extracellular signaling. Moreover, synthetic 3D niche stimulating endogenous secretion of self-renewal-stimulating factors will be able to help develop growth factor-free maintenance system of mESCs.

• Journal of fish pathology
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• v.22 no.1
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• pp.1-7
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• 2009

To investigate the effect of hydrogen peroxide in israel carp, Cyprinus carpio, toxicity and microbial activities were determined. For hydrogen peroxide toxicity test, the median lethal concentration ($LC_{50}$) to israel carps Cyprinus carpio (average weight 0.44 g) by acute toxicity was determined after 24 hour treatment. All israel carp were alive in 24 hours treatment at 80 $\mu\ell/\ell$ concentration and $LC_{50}$ value was 148.9 $\mu\ell/\ell$. For biocidal activities of hydrogen peroxide, remove of parasite and growth inhibition of pathogenic bacteria were determined. The parasite Trichodina sp. infected on the skin and gills of israel carps (average weight 0.1 g) was completely eliminated at 40 $\mu\ell/\ell$ of hydrogen peroxide treatment for 24 hour. Most of the minimum inhibitory concentrations (MICs) against 30 fish pathogenic bacteria were less than 40 $\mu\ell/\ell$.

• Journal of Microbiology and Biotechnology
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• v.15 no.3
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• pp.652-655
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• 2005

This paper describes the development of an enzymatic assay system for the identification of inhibitors of isocitrate lyase (ICL), one of the key enzymes of the glyoxylate cycle that is considered as a new target for antifungal drugs. A 1.6 kb DNA fragment encoding the isocitrate lyase from Candida albicans ATCC10231 was amplified by PCR, cloned into a vector providing His-Patch-thioredoxin-tag at the N-terminus, expressed in Escherichia coli, and purified by metal chelate affinity chromatography. The molecular mass of the purified ICL was approximately 62 kDa, as determined by SDS-PAGE, and the enzyme activity was directly proportional to incubation time and enzyme concentration. The effects of itaconate-related compounds on ICL activity were also investigated. Among them, itaconic acid, 3-nitropropionate, and oxalate had strong inhibitory activities with $IC_{50}$ values of 5.8, 5.4 and $8.6\;{mu}g/ml$, respectively. These inhibitors also exhibited antifungal activity on YPD agar media containing acetate as a sole carbon source, albeit at high concentration. The results indicate that the C. albicans ICL may be a regulatory enzyme playing a crucial role in fungal growth and is a prime target for antifungal agents.

• Animal cells and systems
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• v.1 no.1
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• pp.115-121
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• 1997

To precisely analyze the role of ovarian steroids in the regulation of laminin chain gene expression in mouse uterine tissues, the ovariectomized mouse model was used. Ovariectomized mice received a single injection of steroid hormones and total RNA was isolated from whole uterine tissues. Messenger RNA levels of each laminin chain (A, 81, and 82) were determined by competitive RT-peR procedures. Estradiol decreased mRNA levels of laminin 81 chain about two-fold, and 82 chain rather moderately. Estradiol-induced inhibition of laminin 81 and 82 chain mRNA levels were completely blocked by pretreatment with estrogen receptor antagonist tamoxifen. Estriol, a short acting estrogen which cannot induce hyperplastic responses of rodent uterine tissues, also showed an inhibitory effect on 81 and 82 chain mRNA levels, while estrone, an inactive estrogen, failed to influence either 8 chain mRNA levels. Effects of steroids on A chain mRNA level were quite different from those on 8 chains. Laminin A chain mRNA level was slightly increased by estradiol treatment, but negatively affected by progesterone. Progesterone treatment greatly increased both 8 chain mRNA levels, but slightly decreased A chain mRNA level compared to the control. The effect of progesterone on laminin chain-specific mRNA levels was further increased by co-injection of estradiol in a time-dependent manner. Progesterone-induced 81 and 82 chain mRNA transcription was inhibited by RU486, a synthetic anti-progesterone /anti-glucocorticoid. The present study demonstrates for the first time that steroids are able to regulate laminin gene expression in mouse uterine tissues, indicating that steroid-regulated laminin gene expression is involved in uterine growth and probably differentiation.

• Food Science and Preservation
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• v.11 no.4
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• pp.461-467
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• 2004

Antioxidant effect of traditional doenjang(TD) was reduced by increasing of maturation period. Methanol fractionate of TD matured for 1 year showed strong antioxidant effect against linoleic acid, following the order of hexane and water layer. Antioxidant effect in lipophilic and hydrophilic extracts of TD were gradually increased according to increasing maturation period, whereas their values or two extracts were lower than those or fractionates from TD. Hydrogen-scavenging effect in hydrophobic extract, methanol and butanol fractionates of TD were much higher than those of the other samples. Chloroform and ethylacetate fractionates were markedly lower in the range of $10.57{\sim}22.84\%\;and\;7.82{\sim}22.58\%$, respectively. Anticarcinogenic effect of extracts and fractionates from TD were higher in water fractionates for A549 cell (human lung carcinoma) and methanol fractionates fur MCF-7 cell (human breast adenocarcinoma). Especially, inhibitory effect for growth of cancer cell was increased by the increasing maturation period of TD.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.17 no.3
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• pp.26-32
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• 2004

Objective: The aim of this work is to investigate the antibacterial activity of the essential oil obtained from Artemisia lavandulaefolia (A. lavandulaefolia), as the development of microbial resistance to antibiotics make it essential to constantly look for new and active compounds effective against pathogenic bacteria. Method: The aerial parts of A. lavandulaefolia (1 kg) were subjected to steam distillation for 3 h, using a modified Clevenger type apparatus in order to obtain essential oil. Diethyl ether was the extracting solvent kept at 25?. The essential oil were analyzed by gas chromatography (GC) and gas chromatography／mass spectrometry (GC／MS). The essential oil and the composition were tested for antimicrobial activities against 15 different genera of oral bacteria. Ninety-nine compounds accounting for 94.74$\%$<／TEX> of the oil were identified. The main compounds in the oil were 1,8-cineole (5.63$\%$), yomogi alcohol (4.49$\%$), camphor (4.92$\%$), a-caryophyllene (16.10$\%$), trans-a-famesene (5.09$\%$), a-terpineol (3.91$\%$), borneol (5.27$\%$), cis-chrysanthenol (6.98$\%$), and a-humulene oxide (3.33$\%$). The essential oil and its compounds were tested for antimicrobial activity against 10 different genera of oral bacteria. Conclusion: The essential oil of A. lavandulaefolia exhibited considerable inhibitory effects against all obligate anaerobic bacteria (MICs, 0.025 - 0.05 ㎎／ml) tested, while their major compounds demonstrated various degrees of growth inhibition

• Korean Journal of Weed Science
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• v.17 no.4
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• pp.421-430
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• 1997

This experiment was conducted to detect the presence of allelopathic substances in the leaves of Ginkgo biloba L. Water extracts from G. biloba leaves which collected at different season markedly inhibited the germination and growth of O. sativa, E. crus-galli, D. sanguinalis, and L. sativa, indicating the presence of biological substances. Linolenic and palmitic acid were the major fatty acids of G. biloba leaves. The biochemical substances such as salicylic arid, p-coumaric acid, catechol, hydroquinone, orchinol, ferulic acid, phloroglucinol, and umbelliferone etc., belonging to the phenolic, compounds were, detected in a large amount, which may be responsible for exhibition inhibitory effects. The common phenolic compounds were detected in the early-harvested and late-harvested G. biloba leaves were salicylic and p-coumaric acid. All these compounds were related to the allelopathic activities in G. biloba leaves.

• Journal of Life Science
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• v.23 no.4
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• pp.495-500
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• 2013

Harmonia axyridis is known to display diverse biological activities, such as growth promotion. However, few studies have investigated the effect of H. axyridis on inflammation of the skin. In this study, we explored the anti-inflammatory effect of the Harmoniasin gene fragment (HaGF) peptide from H. axyridis on macrophage cells. During the entire experimental period, 5, 25, 50, and 100 ${\mu}g/ml$ of HaGF showed no cytotoxicity. However, at these concentrations, HaGF inhibited the activity of iNOS and COX-2 by 51% and 49%, respectively. In addition, the HaGF extract reduced the release of inflammatory cytokines, including TNF-a and IL-6. Therefore, HaGF has been LPS-induced macrophage Raw 264.7 cells could be expected from the inhibitory effects of the inflammatory.

• Environmental Mutagens and Carcinogens
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• v.20 no.1
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• pp.14-20
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• 2000

DHAQ-97, (2-(3-[p-bis(2-chloroethyl)aminophenyl]-2 formylaminopropanoyloxy) methy1-1,4-dihy-droxy-9,10-anthraquinone), is a novel anthraquinone derivative synthesized for use as an anti-neoplastic agent. In the present study, we have evaluated the selective cytotoxicity of DHAQ-97 by comparing its effects on viability and proliferation of human breast cancer cell line (NCF-7) versus normal immortalized breast epithelial cell line (MCF-10A). Thus, DHAQ-97 reduced both viability and proliferation of MCF-7 cells to a much greater extent than did for MCF-10A cells. The growth inhibitory and anti-proliferative properties of DHAQ-97 appear to be attributable to its ability to induce apoptosis as revealed by positive staining after in 냐셔 nick-end labeling (TUNEL), cleavage of poly(ADP-ribose)polymerase, release of mitochondrial cytochrome c into cytoplasm, and increased expression of pro-apoptotic Bax protein. Recent studies have indicated possible involvement of the ubiquitous eukaryotic transcription factor, NF-kappa B (NF-kB) in the regulation of apoptotic cell death. In line induced cytotoxicity in cultured MCF-7 cells. Furthermore, mild activation of NF-kB, as determined by its increased DNA binding capability, was observed 30 min after treatment with 10$\mu\textrm{m}$ DHAQ-97. Taken together, the above findings suggest that DHAQ-97 exerts selective cytotoxicity towards cancer cells through induction of apoptosis, which appears to be regulated by NF-kB.