• Maxillofacial Plastic and Reconstructive Surgery
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• v.32 no.3
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• pp.218-228
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• 2010

Components of dental resin-based restorative materials are reported to leach from the filling materials even after polymerization. Hydroquinone (HQ) is one of the major monomers used in the dental resin and is known as a carcinogen. Thus, carcinogenic risk of HQ leaching from the dental resin becomes a public health concern. The present study attempted to examine the carcinogenic potentials of HQ on the human epithelial cell, which is the target cell origin of the most of oral cancers. Cytotoxicity of HQ was observed above 50${\mu}M$ as measured by LDH assay, indicating a relatively low toxicity of this substance in human epithelial cells. The parameters of neoplastic cellular transformation such as cell saturation density, soft agar colony formation and cell aggregation were analyzed to examine the carcinogenic potential of HQ. The study showed that 2-week exposure of HQ showed the tendency of increase in the saturation density and the significant enhancement of soft agar colony formation at the highest dose, 50 ${\mu}M$ only. It is suggested that HQ has a weak potential of carcinogenicity. When cells were treated with HQ and TPA, a well-known tumor promoter, the parameters of neoplastic cellular transformation was significantly increased. This result indicates that the potential risk of carcinogenicity from HQ is largely dependent upon the presence of promoter. Exposure of 50 ${\mu}M$ HQ increased the time-dependent apoptosis as measured by the ELISA kit. This concentration coincides with a dose of neoplastic transformation, indicating a possible link between apoptosis and HQ-induced cellular transformation. Hydroquinone generated Reactive Oxygen Species (ROS) which was evidenced by the treatment of antioxidants such as trolox and N-acetyl cysteine and the GSH depleting agent, BSO. Antioxidants blocked the generation of ROS and the GSH depleting agent, BSO dramatically increased the ROS production. Since HQ is known to increase ROS production thru activation of transcriptional factor such as c-Myb and Pim-1, it is speculated that ROS generation by HQ plays a role in the activation of oncogene, which may lead to neoplastic transformation. In addition, ROS is involved in the alteration of signal transduction, which regulates the apoptosis in many cellular systems. Thus, ROS-mediated apoptosis may be involved in the HQ-induced carcinogenic processes. Protein kinase C (PKC) is known to play pivotal roles in neoplastic transformation of cells and its high expression is often found in a variety of types of tumors including oral cancer. PKC translocation of PKC-${\alpha}$ was observed following HQ exposure. Altered signaling system may also play a role in the transformation process. Taken together, HQ leached from the dental resin does not pose a significant threat as a cancer causing agent, but its carcinogenic potential can be significantly elevated in the presence of promoter. The mechanism of HQ-induced carcinogenesis involved ROS generation, apoptosis and altered signaling pathway. The present study will provide a valuable data to estimate the potential risk of HQ as a carcinogen and understand mechanism of HQ-induced carcinogenesis in human epithelial cells.

• THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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• v.6 no.1
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• pp.81-97
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• 2000

Objectives: This experimental study was carried out to evaluate the effects of aqueous and methanol extracts of Hedyotis diffusa which has long been used for cancer treatment in oriental medicines on the induction of apoptotic cell death in human lymphoid leukemia cell line, HL-60. Methods: Cells were treated with various concentrations (200 to $0.4{\mu}g$) and periods (6 to 30 hr) of $H_2O$ and methanol extracts of Hedyotis diffusa. Then, cells were tested for viability by MTT assay. Cells wrere treated with $200{\mu}g/ml$ of methanol extract fork various periods. Genomic DNA was isolated, separated, on 1.5% agarose gels, stained with ethidium bromide and visualized under UV light. Cells were treated with $200{\mu}g/ml$ of each extract for 16 hr. Then, cells were treated with Hoechst dye 33342 and observed by fluorescence microscopy. Cells were treated with various doses of each for 12 hr and $100{\mu}g/ml$ of methanol extract for various periods. Lysate from the cells used to measure the activity of Caspase-1 and-3 proteases by using fluorogenic peptide substrates including acetyl-YVAD-AMC and acetyl-DEVD-AMC, respectively. Cells were treated with $200{\mu}g/ml$ of each extract for various periods. Cell lysates were immunoprecipated with anti-JNKl antibodies. The immune complex was reacted with $32^p-ATP$ and c-Jun as a substrate. The phosphotransferase activity of JNKI was measured by using PhosphoImage analyzer (Fuji Co., Japan). Nuclear extracts were isolated and incubated with oligonucleotide probe of $NF-{\kappa}B$. Transcriptional activation of ${\kappa}B$ was measured by using EMSA and visualized by PhosphoImage analyzer (Fuji Co, Japan). Cell lysates were prepared and analyzed by Western blotting with anti-Bc12 antibodies and anti-Bax antibodies. Cells were pretreated with various doses of methanol extract for 2 hr. Then, the extract was removed by centrifugation. Cells were resuspended with RPMI-1640 media containing 0.3% agarose, 10% FBS, overlayred onto bottom layer agarose and incubated at $CO_2$ incubator for 6 days. The number of colony was counted under light microscopy ($\time100$). Results: The death of HL-60 cells was markedly induced by the addition of methanol extract of Hedyotis diffusa in a dose and time-dependent manners. The apoptotic characteristic ladder pattern of DNA strand break was observed in death of HL-60 cells. In addition, it was shown nucleus chromatin condensation and fragmentation under Hoechst staining. Therefore, Hedyotis diffusa extract-induced death of HL-60 cells is mediated by apoptotic signaling processes. The activity of Caspase 3-like proteases remained in a basal level in HL-60 cells treated with aqueous extract of Hedyotis diffusa. However, it was markedly increased in HL-60 cells treated with methanol extract of Hedyotis diffusa. In addition, the phosphotransferase activity of JNKl was increased in HL-60 cells treated with methanol extract of Hedyotis diffusa. Furthermore, the activation of transcriptional activator, $NF-{\kappa}B$ was markedly induced by methanol extract of Hedyotis diffusa. Anti-apoptotic Bc12 was cleaved into 23Kda fragment by treatment of methanol extract of Hedyotis diffusa. However, expression of proapoptotic Bax protein was increased by treatment of methanol extract of Hedyotis diffusa in a time-dependent manner. Furthermore, methanol extract markedly inhibited the colony forming efficiency of HL-60 cells in semisolid agar culture. Conclusions: Above results suggest that methanol extract of Hedyotis diffusa induces the apoptotic death of human leukemic HL-60 cells via activations of Caspase-3 proteases, JNKI, transcriptional activator $NF-{\kappa}B$, In addition, our results also suggest that methanol extract of Hedyotis diffusa reduces the malignant potential of HL-60 cells via down regulation of colony forming effciency through cleavage of Bc12 as well as induction of Bax.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.60 no.4
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• pp.484-490
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• 2015

Thirty-eight Pea (Pisum sativum L.) genotypes were screened to identify varieties to be suitable for sprout. Based on seed yield and sprout qualities such as whole length and sprout yield, five genotypes (PI269803, PI343278, PI343283, PI343300 and PI 343307) were primarily selected as candidates for pea sprouts. In order to determine optimal cultivation condition for pea sprouting, growth characteristics were investigated according to the change of germination temperature and days for sprouting. Whole length and hypocotyl length were observed to increase as a time dependent manner at each tested temperature (20, 23, and $25^{\circ}C$). However, whole length, hypocotyl length, and sprout yield were highly increased at $23^{\circ}C$ compared to 20 and $25^{\circ}C$. Especially, PI269803 and PI343300 showed higher sprout yield than the others. In addition, the effect of the change of germination temperature on antioxidant properties was estimated by measuring total phenolic content (TPC) and free radical scavenging activity (DPPH and ABST activity). TPC and DPPH/ABST activities of PI269803 and PI343300 were higher at $23^{\circ}C$ than at 20 and $25^{\circ}C$, while antioxidant properties of PI343278 and PI343283 were decreased in a temperature-dependent manner. The results show a high degree of correlation between TPC and antioxidant activities and suggest that the temperature change for pea sprouting could be responsible for antioxidant properties. Taken together, these results provide optimal cultivation conditions for pea sprouting and suggest that PI269803 and PI343300 with high sprout yield and antioxidant properties could be used for pea sprouts.

• Journal of Veterinary Clinics
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• v.29 no.2
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• pp.141-147
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• 2012

This work describes the characteristics of $Malassezia$ $pachydermatis$ isolated from dog ear canals and the effect of essential oils on the growth of this organism. Sterile cotton swabs were used to collect specimens from the external ear canal and culture tests were performed to detect the population size of $Malassezia$ yeast. Using three different isolation media, included Sabouraud dextrose agar (SDA) to isolate common $M.$ $pachydermatis$, and SDA supplemented with olive oil (SDAO) and Leeming's medium (LM) to detect lipophilic yeast, $Malassezia$ spp were isolated from 14 of 18 dogs (77.8%); isolation rates were 33.3% in SDA, 72.2% in SDAO and 66.7% in LM media. All $Malassezia$ spp isolates were identified as $M.$ $pachydermatis$ according to results of PCR amplification, but gross colony morphology and SDA growth rates suggested four different subtypes. Large (LC) and medium colony (MC) types respectively describe large colony (diameter > 3 mm) and medium colony (around 2 mm) after 72 hour incubation, and small (SC) type refers to smaller colony (< 1 mm) even after 5 days incubation; lipid dependent colonies did not grow onto SDA. Large Colony type strains were isolated from 4, 11, and 11 samples, MC type strains from 2, 3 and 1 and SC type strains from 1, 2 and 1 in SDA, SDAO and LM, respectively. Lipid-dependent $M.$ $pachydermatis$ (Lipo) were isolated from 3 samples each in SDAO and LM. Anti-$M.$ $pachydermatis$ activity testing was done using disc-diffusion assays and well diffusion tests. Most essential oils inhibited the growth of $M.$ $pachydermatis$ in a range from 0.5% to 1.0% of essential oils. MIC90 and MIC50 were variable depending upon the nature of essential oils. Thyme oil was found to be highly effective in inhibiting the growth of $M.$ $pachydermatis$ in a range from 0.125% to 0.0625% while marjoram and then tea tree oil exhibited lower inhibitory capacity.

• Journal of the Korean Society of Food Science and Nutrition
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• v.43 no.1
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• pp.9-15
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• 2014

To characterize novel biologically-active ingredients in traditional Korean soy sauces, polysaccharide fractions were isolated from two different soy sauces made either commercially (CSP-0) or through a traditional Korean process (KTSP-0), after which their intestinal immune-modulating activities were examined. CSP-0 and KTSP-0 showed enhanced production of interleukine-6 (IL-6) in culture supernatant of Peyer's patch cells. However, KTSP-0 activity was more potent than that of CSP-0. Only KTSP-0 increased in vitro immunoglobulin A (IgA) production by Peyer's patch cells in a dose-dependent manner. KTSP-0 also showed the higher bone marrow cell proliferation activity through Peyer's patch cells than that of the CSP-0 group. To investigate the in vivo effects on the intestinal immune system, CSP-0 and KTSP-0 were administered orally to four experimental groups of mice (0.0, 0.5, 1.0, and 5.0 mg/mouse/day, 30 days). Oral administration of CSP-0 and KTSP-0 induced IgA production by Peyer's patch cells and increased IgA excretion into mouse stools in a dose-dependent manner. Peyer's patch cells from the mice administered both CSP-0 and KTSP-0 showed significantly higher IL-6 production than that of the untreated or CSP-0 groups. However, oral administration of KTSP-0 was more effective at the same dosage. KTSP-0 administration augmented IL-6 content in mouse sera, whereas CSP-0 did not show any effect on IL-6 induction. The above data lead us to conclude that the intestinal immune-stimulating activities of polysaccharides from Korean traditional soy sauce are much better than those of commercial ones.

• Tuberculosis and Respiratory Diseases
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• v.43 no.2
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• pp.210-220
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• 1996

Background : Neutrophils or monocytes separated in vitro by the adherence to plastic surface are known to be activated by surface adherence itself and subsequent experimental data might be altered by surface adherence. In the process of surface adherence, adhesion molecules have a clear role in intracellular signal pathway of cellular activation. Human alveolar macrophages(HAM) are frequently purified by the adherence procedure after bronchoalveolar lavage. But the experimental data of many reports about alveolar macrophages have ignored the possibility of adhesion-induced cellular activation. Method : Bronchoalveolar lavage was performed in the person whose lung of either side was confirmed to be normal by chest CT. With the measurement of hydrogen peroxide release from adherent HAM to plastic surface and non-adherent HAM with or without additional stimulation of phorbol myristate acetate(PMA) or N-formyl-methionyl-leucyl-phenylalanine (fMLP), we observed the effect of the adherence to plastic surface. We also evaluated the effect of various biological surfaces on adhesion-induced activation of HAM. Then, to define the intracellular pathway of signal transduction, pretreatment with cycloheximide, pertussis toxin and anti-CD11/CD18 monoclonal antibody was done and we measured hydrogen peroxide in the culture supernatant of HAM. Results : 1) The adherence itself to plastic surface directly stimulated hydrogen peroxide release from human alveolar macrophages and chemical stimuli such as phorbol myristate acetate(PMA) or N-formyl-methionyl-leucyl-phenylalanine(fMLP) colud not increase hydrogen peroxide release in these adherent macrophages which is already activated. 2) PMA activated human alveolar macrophages irrespective of the state of adhesion. However, fMLP stimulated the release of hydrogen peroxide from the adherent macrophages, but not from the non-adherent macrophages. 3) HAM adherent to A549 cell(type II alveolar epithelium-like human cell line) monolayer released more hydrogen peroxide in response to both PMA and fMLP. This adherence-dependent effect of fMLP was blocked by pretreatment of macrophages with cycloheximide, pertussis toxin and anti-CD18 monoclonal antibody, Conclusion : These results suggest that the stimulatory effect of PMA and fMLP can not be found in adherent macrophage because of the activation of human alveolar macrophage by the adherence to plastic surface and the cells adhered to biologic surface such as alveolar epithelial cells are appropriately responsive to these stimuli. It is also likely that the effect of fMLP on the adherent macrophage requires new protein synthesis via G protein pathway and is dependent on the adhesion between alveolar macrophages and alveolar epithelial cells by virtue of CD11/CD18 adhesion molecules.

• Journal of Life Science
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• v.16 no.7 s.80
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• pp.1133-1140
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• 2006

Human HtrA1 (High temperature requirement protein A1) is a homologue of the E. coli periplasmic serine protease HtrA. A recent study has demonstrated that HtrA1 is a serine protease involved in processing of insulin like growth factor binding protein (ICFBP), indicating that it serves as an important regulator of IGF activity. Additionally, several lines of evidence suggest a striking correlation between proteolytic activity of HtrA1 serine protease and the pathogenesis of several diseases; however, physiological roles of HtrA1 remain to be elucidated. We used the pGEX bacterial expression system to develop a simple and rapid method for purifying HtrA1, and the recombinant HtrA1 protein was utilized to investigate the optimal conditions in executing its proteolytic activity. The proteolytically active HtrA1 was purified to approximately 85% purity, although the yield of the recombinant HtrA1 protein was slightly low $460{\mu}g$ for 1 liter E. coli culture). Using in vitro endoproteolytic cleavage assay, we identified that the HtrA1 serine protease activity was dependent on the enzyme concentration and the incubation time and that the best reaction temperature was $42^{\circ}C$ instead of $37^{\circ}C$. We arbitrary defined one unit of proteolytic activity of the HtrA1 serine protease as 200nM of HtrA1 that cleaves half of $5{\mu}M\;of\;{\beta}-casein$ during 3 hr incubation at $37^{\circ}C$. Our study provides a method for generating useful reagents to investigate the molecular mechanisms by which HtrA1 serine protease activity contributes in regulating its physiological function and to identify natural substrates of HtrA1.

• Journal of Intelligence and Information Systems
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• v.18 no.1
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• pp.39-57
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• 2012

Recently, due to the introduction of high-tech equipment in interactive exhibits, many people's attention has been concentrated on Interactive exhibits that can double the exhibition effect through the interaction with the audience. In addition, it is also possible to measure a variety of audience reaction in the interactive exhibition. Among various audience reactions, this research uses the change of the facial features that can be collected in an interactive exhibition space. This research develops an artificial neural network-based prediction model to predict the response of the audience by measuring the change of the facial features when the audience is given stimulation from the non-excited state. To present the emotion state of the audience, this research uses a Valence-Arousal model. So, this research suggests an overall framework composed of the following six steps. The first step is a step of collecting data for modeling. The data was collected from people participated in the 2012 Seoul DMC Culture Open, and the collected data was used for the experiments. The second step extracts 64 facial features from the collected data and compensates the facial feature values. The third step generates independent and dependent variables of an artificial neural network model. The fourth step extracts the independent variable that affects the dependent variable using the statistical technique. The fifth step builds an artificial neural network model and performs a learning process using train set and test set. Finally the last sixth step is to validate the prediction performance of artificial neural network model using the validation data set. The proposed model is compared with statistical predictive model to see whether it had better performance or not. As a result, although the data set in this experiment had much noise, the proposed model showed better results when the model was compared with multiple regression analysis model. If the prediction model of audience reaction was used in the real exhibition, it will be able to provide countermeasures and services appropriate to the audience's reaction viewing the exhibits. Specifically, if the arousal of audience about Exhibits is low, Action to increase arousal of the audience will be taken. For instance, we recommend the audience another preferred contents or using a light or sound to focus on these exhibits. In other words, when planning future exhibitions, planning the exhibition to satisfy various audience preferences would be possible. And it is expected to foster a personalized environment to concentrate on the exhibits. But, the proposed model in this research still shows the low prediction accuracy. The cause is in some parts as follows : First, the data covers diverse visitors of real exhibitions, so it was difficult to control the optimized experimental environment. So, the collected data has much noise, and it would results a lower accuracy. In further research, the data collection will be conducted in a more optimized experimental environment. The further research to increase the accuracy of the predictions of the model will be conducted. Second, using changes of facial expression only is thought to be not enough to extract audience emotions. If facial expression is combined with other responses, such as the sound, audience behavior, it would result a better result.

• Journal of Physiology & Pathology in Korean Medicine
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• v.20 no.4
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• pp.844-852
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• 2006

This study was to evaluate the effect of Yeongyupaedog-san (YGPDS) on mouse Thl and Th2 cells' differentiation and ovalbumin (OVA)-induced allergic inflammation. The proliferation of mouse CD4 T cells and the secretion of Th1/Th2 cytokines under the influence of YGPDS extract were measured as well as the amount of ${\beta}-hexosaminidase$ in RBL-2H3 cells and the levels of $TNF-{\alpha}$ and 1L-6 secretion in Raw264.7 cells. BALB/c mice were orally administered with YGPDS extract and simultaneously inoculated with OVA to induce allergic reaction and measure the level of total IgE, OVA-specific IgE and the production of IFN- g, IL-4, IL-5 by the spleen cells. When mouse CD4 T cell were stimulated with anti-CD3 and anti-CD28 for 48 hours in various concentrations of YGPDS extract, it increased proliferation of CD4 cells by 11% in $100\;{\mu}g/^{ml}$ concentration but it showed an inhibition by 37% at $200\;{\mu}g/^{ml}$ CD4 T cells under Th1/Th2 polarizing conditions for 3 days with YGPDS resulted in mild decrease of IFN- g in Thl cells and significant decrease of IL-4 in Th2 cells at $500\;{\mu}g/^{ml}\;and\;100\;{\mu}g/^{ml}$ by 18% and 21%, respectively. YGPDS extract had a dose-dependent inhibitory effect on antigen-induced release of ${\beta}-hexosaminidase$ in RBL-2H3 cells. Treatment of YGPDS extract on LPS stimulated Raw 264.7 cells showed dose-dependent decrease in TNF-n production. Oral administration of YGPDS extract on OVA-induced allergic mice showed an inhibitory effect on the levels of total serum IgE and OVA-specific IgE by 25% and 34% , respectively. Culture of spleen cells with OVA resulted in significant increase of IFN- g by 44% and significant decrease of IL-4 and IL-5 by 56%, and 24%, respectively. The results show that YGPDS does not strongly induce mouse T cells to transform into Thl or Th2 but it has an anti-allergic effect in vitro, and that it also corrects the unbalance between the reactions of Th cells in allergic diseases.

• Journal of Physiology & Pathology in Korean Medicine
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• v.20 no.6
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• pp.1467-1476
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• 2006

This study was to evaluate the effect of Bulhwangeumjeonggi-san (BS) on mouse Th1 and Th2 cells' differentiation and ovalbumin (OVA)-induced allergic inflammation. The proliferation of mouse CD4 T cells and the secretion of Th1/Th2 cytokines under the influence of BS extract were measured as well as the amount ${\beta}$-hexosaminidase in RBL-wH3 cells and the levels of TNF-${\alpha}$ and IL-6 secretion in Raw264.7 cells. BALB/c mice were orally administered with BS extract and simultaneously inoculated with OVA to induce allergic reaction and measure the level of total lgE, OVA-specific lgE and the production of IFN- g, IL-4, IL-5 by the spleen cells. When mouse CD4- T cell were stimulated with anti-CD3 and anti-CD28 for 48 hours in various concentrations of BS extract, it increased proliferation of CD4 cells by 14% in 50 ${\mu}g/m{\ell}$ concentration but it showed an inhibition in higher concentrations. CD4 T cells under Th1/Th2 polarizing conditions for 3 days with BS resulted in mild decrease of IFN- g in Th1 cells and mild increase of IL-4 in Th2 cell at 50 ${\mu}g/m{\ell}$ but the level of IL-4 decreased by 18% at 100 ${\mu}g/m{\ell}$. BS extract had a dose-dependent inhibitory effect on antigen-induced release of ${\beta}$-hexosaminidase in RBL-2H3 cells. Treatment of BS extract on LPS stimulated Raw 264.7 cells showed dose-dependent decrease in TNF-${\alpha}$ production. Oral administration of BS extract on OVA-induced allergic mice showed an inhibitory effect on the levels of total serum lgE and OVA-specific lgE by 50% and 55%, respectively. Culture of spleen cells with OVA resulted in significant increase of IFN- g by 25% and significant decrease of IL-4 and IL-5 by 53%, and 38%, respectively. The results show that BS does not strongly induce mouse T cells to transform into Th1 or Th2 but it has an anti-allergic effect in vitro, and that it also corrects the unbalance between the reactions of Th cells in allergic diseases.