Youn Seon Min;Oh Young Kee;Kim Joo Heon;Park Mi Ja;Seong In Ock;Kang Kimun;Chai Gyuyong
Radiation Oncology Journal
/
v.23
no.1
/
pp.51-60
/
2005
Purpose : Selective inhibition of multiple molecular targets may improve the antitumor activity of radiation. Two specific inhibitors of selective cyclooxygenase-2 (COX-2) and epidermal growth factor receptor (EGFR) were combined with radiation on the HeLa cell line. To investigate cooperative mechanism with selective COX-2 inhibitor and EGFR blocker, in vitro experiments were done. Materials and Methods : Antitumor effect was obtained by growth inhibition and apoptosis analysis by annexin V-Flous method. Radiation modulation effects were determined by the clonogenic cell survival assay. Surviving fractions at 2 Gy ($SF_2$) and dose enhancement ratio at a surviving fraction of 0.25 were evaluated. To investigate the mechanism of the modulation of radiosensitivity, the cell cycle analyses were done by flow cytometry. The bcl-2 and bax expressions were analyzed by western blot. Results : A cooperative effect were observed on the apoptosis of the HeLa ceil line when combination of the two drugs, AG 1478 and NS 398 with radiation at the lowest doses, apoptosis of $22.70\%$ compare with combination of the one drug with radiation, apoptosis of $8.49\%$. In cell cycle analysis, accumulation of cell on $G_0/G_l$ phase and decrement of S phase fraction was observed from 24 hours to 72 hours after treatment with radiation, AG 1478 and NS 398. The combination of NS 398 and AG 1478 enhanced radiosensitivity on a concentration-dependent manner in HeLa cells with dose enhancement ratios of 3.00 and $SF_2$ of 0.12 but the combination of one drug with radiation was not enhanced radlosensitivity with dose enhancement ratios of 1.12 and SF2 of 0.68 (p=0.005). The expression levels of bcl-2 and bax were reduced when combined with AG 1478 and NS 398. Conclusion : Our results indicate that the selective COX-2 inhibitor and EGFR blocker combined with radiation have potential additive or cooperative effects on radiation treatment and may act through various mechanisms including direct inhibition of tumor cell proliferation, suppression of tumor cell cycle progression and inhibition of anti-apoptotic proteins.
Objective: This study was performed to compare the clinical outcome of elective single embryo transfer (eSET) performed at the cleavage stage to that of elective double embryo transfer (eDET). Methods: Of the women less than 36 years old who visited Daegu Maria from January 2008 to April 2009, the only women (n=330) with more than 8 mm of endometrial thickness and at least one good quality embryo, who were treated with GnRH agonist long protocol, were included in this study. After information about complications that can arise by multiple embryo transfer, either eSET or eDET was conducted by their request (167 and 163, respectively).Results: The implantation rate of eSET group was significantly higher than that of eDET group (53.9% vs. 40.2%, p<0.01). The twin pregnancy rate of eSET group was significantly lower than that of eDET group (1.1% vs. 32.3%, p<0.001). However, there were no significant differences between two groups in the clinical pregnancy (53.3% vs. 60.7%, p=0.172), ongoing pregnancy (47.3% vs. 54.6%, p=0.185) and live birth rates (44.9% vs. 50.9%, p=0.275). The number of the surplus embryos which developed to the blastocyst stage and cryopreserved at that stage was significantly higher in eSET group than that of eDET group ($3.2{\pm}2.6$ vs. $2.1{\pm}2.4$, p<0.001). Conclusion: These results suggest that eSET should reduce significantly the multiple baby pregnancy without decreasing the whole pregnancy rate in women with less than 36 years old.
Journal of the Korean Society of Food Science and Nutrition
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v.36
no.8
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pp.1055-1061
/
2007
Caesalpinia sappan L. is an indeciduous tree distributed in China, India, Miyanmar and Vietnam. Its heartwood has long been used in oriental folk medicines to treat diseases. In this study, antioxidative activities of Caesalpinia sappan L. and the effect of gamma irradiation on its chemical and biological properties were investigated. The ethyl acetate fraction (EtOAc fr.) of Caesalpinia sappan L. was irradiated with 100 kGy of gamma ray. The dark red color of EtOAc fr. was significantly (p<0.05) removed by irradiation (Hunter L and b values increased and a value decreased). The total phenolic content of EtOAc fr. was 865 mg/g and it was increased to 1195 mg/g by gamma irradiation. DPPH radical and superoxide anion radical scavenging activities and lipid peroxidation inhibitory activity of EtOAc fr. were very high and its activities were also increased by gamma irradiation. EtOAc fr. also inhibited the irradiation-induced DNA damage of lymphocyte as determined by comet assay. In conclusion, EtOAc fr. of Caesalpinia sappan L. extract showed high antioxidative activities in vitro. Furthermore, gamma irradiation on EtOAc fr. ameliorated the color and antioxidative properties. Therefore, it can be suggested that Caesalpinia sappan L. may be a good material for antioxidant function and gamma irradiation may be applied for the improvement of chemical and biological properties of Caesalpinia sappan L.
Journal of the Korean Society of Food Science and Nutrition
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v.40
no.4
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pp.500-508
/
2011
This study was carried out to investigate the effects of jujube extracts on intestinal microflora, along with their antioxidant activities, according to extraction method. The antimicrobial activities of the extracts were measured using the agar diffusion method with a jujube extract concentration of 50 mg/mL. Neither the first nor second jujube extracts were inhibitory against the tested intestinal bacteria. However, water extracts of jujube significantly enhanced the growth of lactic acid bacteria, especially Bifidobacterium bifidum and Bifidobacterium adolescentis. Total phenol compounds and flavonoid compounds were higher in the 1st than in the 2nd water extracts. The EDA values of both water and ethanol extracts increased in proportion to the extract concentration. The 1st water extract showed the highest value among all the others, which was 85.60% at the concentration of 0.05 mg/mL. Furthermore, the 1st water extract showed stronger antioxidant activity than the other samples with an activity of 679.91 mg AA eq/g. These results support the potential use of jujube water extracts as a functional food component and a valuable resource for the development of nutraceutical foods, to increase the growth of Bifidobacterium spp. in the human intestine.
Background : Tumor necrosis factor(TNF) has been considered as an important candidate for cancer gene therapy based on its potent anti-tumor activity. However, since the efficiency of current techniques of gene transfer is not satisfactory, the majorities of current protocols is aiming the in vitro gene transfer to cancer cells and re-introducing genetically modified cancer cells to hoot. In previous study, it was shown that TNF-sensitive cancer cells transfected with TNF-$\alpha$ cDNA would become highly resistant to TNF. Understanding the mechanisms of TNF-resistance in TNF-$\alpha$ gene transfected cancer cells would be an important step for improving the efficacy of cancer gene therapy as well as for better understandings of tumor biology. This study was designed to evaluate the role of new protective protein synthesis in the acquired resistance to TNF of TNF-$\alpha$ gene transfected cancer cells. Method : We transfected TNF-$\alpha$ c-DNA to WEHI164, a murine fibrosarcoma cell line, using retroviral vector(pLT12SN(TNF)) and confirm the expression of TNF with PCR, ELISA, MIT assay. Then we determined the TNF resistance of TNF gene transfected cells(WEHI164-TNF) and the changes of TNF sensitivities after treatments with actinomycin D(transcription inhibitor) and cycloheximide ( translation inhibitor). Results : WEHI164 which was sensitive to TNF became resistant to TNF after being transfected with TNF-$\alpha$ gene and the resistance to TNF was partially reversed after treatment with actinomycin D, but not with cycloheximide. Conclusion : The acquired resistance to TNF after TNF-$\alpha$ gene transfection may be associated with synthesis of some protective proteins.
Journal of the korean academy of Pediatric Dentistry
/
v.37
no.1
/
pp.1-12
/
2010
The aim of this study was to evaluate the preventive effect of commercially available anticariogenic products, specifically, the tooth cream containing Casein phosphopeptide-amorphous calcium phosphate(CPP-ACP), fluoride varnish and low-level fluoride mouthrinse on enamel erosion induced by carbonated beverage in a short period of time. Enamel specimens were treated as follows and were then kept in artificial saliva for 24 hours followed by further processing by alternately soaking them in Cola beverage and in distilled water for 1 minute each five times. Group 1: control group (no treatment) Group 2: tooth cream with CPP-ACP Group 3: fluoride varnish (1,000 ppm F) Group 4: low-level fluoride mouthrinse (227 ppm F) Group 5: fluoride varnish + tooth cream with CPP-ACP Group 6: low-level fluoride mouthrinse + tooth cream with CPP-ACP Microhardness and erosion depth were measured and the mineral loss of each specimen was evaluated by measuring the volumetric fluorescence change(${\Delta}Q$) against the stable fluorescent grid using quantitative light-induced fluorescence(QLF). The experiment lasted for 6 days repeated each day. The results were as follows: 1. The microhardness was increased as follows: Group $1{\leq}2{\leq}4$<6<$3{\fallingdotseq}5$. 2. The mean erosion depth was increased as follows: Group $5{\fallingdotseq}3$<6<$4{\fallingdotseq}2{\fallingdotseq}1$. 3. The ${\Delta}Q$ was increased as follows: Group $1{\fallingdotseq}2{\leq}4{\leq}6{\leq}3{\fallingdotseq}5$. The decrement of ${\Delta}Q$ was similar between group 1 and 2, group 4 and 6 and group 3 and 5. 4. The ${\Delta}Q$ showed positive correlation with microhardness (r=0.96, p<0.05), while it was negatively correlated to erosion depth (r=-0.96, p<0.05).
Kim, Kyung-Hee;Kim, Sang-Soo;Kim, Goo-Young;Rhim, Hyang-Shuk
Journal of Life Science
/
v.16
no.7
s.80
/
pp.1133-1140
/
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.
Kim, Min Soo;Oh, In Taek;Jun, Do Youn;Lee, Ji Young;Sohn, Ho-Yong;Kwak, Do Yeon;Seo, Myung Chul;Woo, Koan Sik;Ko, Jee Yeon;Jung, Tae Wook;Nam, Min Hee;Woo, Mi Hee;Kim, Young Ho
Journal of Life Science
/
v.23
no.12
/
pp.1460-1470
/
2013
To examine whether miscellaneous cereal grains have an antithrombotic effect, we investigated the anticoagulant activity of 80% ethanol extracts from eleven selected miscellaneous cereal grains. The 80% ethanol extract of hwanggeumchal sorghum (Sorghum bicolor) showed the highest anticoagulant activity, followed by that of green foxtail millet grains, in terms of thrombin time (TT). When the ethanol extract of hwanggeumchal sorghum was sequentially fractionated with n-hexane, methylene chloride, ethyl acetate, and n-butanol, the majority of the TT-inhibitory activity was detected in the hexane and methylene chloride fractions. Whereas aspirin (final conc. 480 ${\mu}g/ml$) prolonged TT by 2-fold, the ethanol extract, hexane fraction, and methylene chloride fraction in the same dose prolonged TT by 2.2-fold, 2.9-fold, and 2.5-fold, respectively. The ethanol extract of hwanggeumchal sorghum could delay activated partial thromboplastin time (APTT) as well as prothrombin time (PT). Although the APTT-inhibitory activity of the ethanol extract was mainly partitioned into the hexane and methylene chloride fractions, the PT-inhibitory activity of the ethanol extract was solely partitioned into the hexane fraction. The APTT- and PT-inhibitory activities of these organic solvent fractions were more potent than those of the control warfarin (final conc. 3.13 mg/ml). The TT-inhibitory activity of the ethanol extract was heat-stable and acid-stable. The ethanol extract, hexane fraction, and methylene chloride fraction of hwanggeumchal sorghum appeared to possess a direct fibrinolytic activity toward fibrin clotting. These results show that hwanggeumchal sorghum can exert anticoagulant and fibrinolytic effects and, thus, have the potential to be applicable as antithrombotic dietary sources.
Two hundred bacterial strains were isolated from the soil around healthy tomato plants in a polyvinyl house, where most of the other plants showed bacterial wilt symptoms. The strains were screened in vitro for their antibacterial activity. Among them, a strain, KPB3 showed strong bactericidal activity against bacterial wilt pathogen, Ralstonia solanacearum. The strain KPB3 was identified using physiological and biochemical tests, and 16S rRNA analyses. Based on these tests, the strain was found to be closer to genus Paenibacillus. To control the bacterial wilt caused by R. solanacearum, greenhouse experiments were conducted to determine the effectiveness of the Paenibacillus strain KPB3. Drench application of this strain ($4{\times}10^8$ CFU $mL^{-1}$) into the pots containing tomato plants, post-inoculated with the pathogen, R. solanacearum could drastically reduce the disease severity, compared to the non-treated plants. To evaluate effectiveness of this strain under field conditions, experiments were carried out in polyvinyl houses infested with R. solanacearum, during spring and autumn of the year 2006. It was observed that, during spring, bacterial wilt was more prevalent compared to the autumn. During spring, 50.9% disease incidences occurred in non-treated controls, while, Paenibacillus strain KPB3 treated plants showed 24.6% disease incidences. Similarly, during autumn, around 17.2% plants were infected with bacterial wilt in non- treated polyvinyl houses, compared to the Paenibacillus strain KPB3 treated plants, which showed 7.0% disease incidences. These results demonstrated that, Paenibacillus strain KPB3 is a potential biological control agent against bacterial wilt caused by R. solanacearum, effective under greenhouse as well as field conditions. This is the first report showing biocontrol of R. solanacearum using a Paenibacillus spp. under field conditions.
The biological effects of the iminoctadine tris (albesilate) and kresoxim-methyl for the protection of citrus postharvest diseases caused by penicillium spp. were assayed. In vitro tests, $EC_{50}$ values of iminoctadine tris(albesilate) were $0.01{\sim}0.02\;and\;0.01{\mu}g$ a.i./mL against mycelial growth of P. italicum and P. digitatum, respectively, but iminoctadine tris(albesilate) at $0.64{\mu}g$ a.i. /mL inhibited a little mycelial growth of unknown Penicillium sp. which produced another symptom different to blue and green mold caused by P. italicum and P. digitatum, respectively. And against germination and growth of germ tube of P. italicum and P. digitatum, $EC_{50}$ value of iminoctadine tris(albesilate) was $0.0013{\sim}0.0025{\mu}g$ a.i./mL. But spore germination of unknown Penicillium spp. was not nearly inhibited at $0.2{\mu}g$ a.i./mL. $EC_{50}$ values of kresoxim-methyl were $0.08{\sim}0.16$, 0.04 and $0.16{\mu}g$ a.i./mL against mycelial growth of P. italicum, P. digitatum and unknown Penicillium sp., respectively, and $0.04{\sim}0.08{\mu}g$ a.i./mL and $0.01{\sim}0.02{\mu}g$ a.i./mL against germination and growth of germ tube of P. italicum and unknown Penicillium sp., and P. digitatum, respectively. Iminoctadine tris(albesilate) and kresoxim-methyl were markedly effective to control the postharvest disease by 7 days spray prior to harvest. When the fruits were sprayed with iminoctadine-tris(albesilate) ($200{\mu}g$ a.i./mL) and kresoxim-methyl ($155{\mu}g$ a.i./mL) 7 days prior to harvest and subsequently stored for 90 days, the percentage of diseased fruit by Penicillium spp. was $3.6{\pm}1.8%$ in treatment of kresoxim-methyl and $5.9{\pm}1.8%$ in iminoctadine-tris(albesilate), respectively. On the other hand, tile percentage of diseased fruit was relatively high, $20.3{\pm}10.0%\;and\;19.5{\pm}9.6%$ in thiophanate-methyl ($700{\mu}g$ a.i./mL) and non-treatment, respectively. Maximum residue amount (ppm) among fruits (flesh and peel) assayed 0, 30, 60 and 90 days after storage was 0.45 and 0.10 ppm in treatment of kresoxim-methyl and iminoctadine, respectively.
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