• Title/Summary/Keyword: 독성평가

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β-Galactosidase-catalyzed Synthesis of 1, 2-Hexanediol Galactoside and its Purification using Ethyl Acetate Extraction followed by Silica Gel Chromatography (대장균 β-Galactosidse를 이용한 1, 2-Hexanediol galactoside의 합성과 Ethyl Acetate 추출 및 Silica Gel Chromatography를이용한 정제)

  • Kim, Yi-Ok;Jung, Kyung-Hwan
    • Journal of the Korean Applied Science and Technology
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    • v.33 no.3
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    • pp.498-506
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    • 2016
  • 1, 2-Hexanediol galactoside (HD-gal) has been previously synthesized from 1, 2-hexanediol (HD), in which recombinant ${\beta}$-galactosidase (${\beta}$-gal) of Escherichia coli (E. coli) was used for transgalactosylation reaction. In this study, a method for HD-gal purification from the reaction mixture was particularly investigated. Using ${\beta}$-gal-containing E. coli, HD-gal was synthesized from 75 mM HD for 48 hr under 300 g/l lactose concentration. Then, HD-gal synthesis from HD was confirmed by TLC analysis, and the existence of E. coli ${\beta}$-gal during 48 hr-reaction was also confirmed by Western blotting, in which the conversion yield of HD to HD-gal reached about 94% during 48 hr. To establish an efficient method for HD-gal purification, we carried out the solvent extraction of the reaction mixture, followed by silica gel chromatography, particularly in order to remove the residual HD. Two water-immiscible solvents, such as methylene chloride and ethyl acetate, were investigated comparatively to find out appropriate solvent. Then, it was found that residual HD was almost removed when ethyl acetate extraction of water phase of reaction mixture was carried out four times. Subsequently, silica gel chromatography was carried out, and purified HD-gal could be finally obtained. The production yield for HD-gal from 75 mM HD was $8.9{\pm}0.6%$ (n=3) (mole basis) or $21.1{\pm}1.4%$ (n=3) (weight basis). For further study, using purified HD-gal, we will investigate the minimum inhibitory concentrations (MICs) of HD-gal against bacteria. In addition, cytotoxicity to human skin cells of HD-gal will be examined.

The Effect of Rubus coreanum Miquel Against Lipopolysaccharide-induced Oxidative Stress and Lipid Metabolism (복분자 추출물이 LPS로 유도된 산화적 스트레스와 지질대사에 미치는 영향)

  • Kim, In-Deok;Kang, Kum-Suk;Kwon, Ryun-Hee;Yang, Jeong-Ok;Lee, Joong-Sook;Ha, Bae-Jin
    • Journal of Food Hygiene and Safety
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    • v.22 no.3
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    • pp.213-217
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    • 2007
  • LPS induces the synthesis of several inflammatory cytokine, chemokine, NO and inflammation in the liver of rats. The purpose of this study was to investigate the preventive effects of Rubus coreanum Miquel (RCM) In lipid metabolism. RCM of 100 mg/kg concentration was intraperitoneally administered into rats at dose of 1.5ml/kg for 20 days. On the day 21, 1.5ml/kg of LPS was injected 4 hours before anesthetization. We examined the lipid-related functions by measuring the levels of triglyceride (TG), total cholesterol (TC), total lipid (TL), high-density lipoprotein cholesterol (HDL-C) in serum and malondialdehyde(MDA) in liver tissue. The results showed that LPS treatment increased the values of TG, TC, TL and MDA, decreasing that of HDL-C. But RCM pretreatment decreased the high values of TG, TC, TL and MDA to the low values and increased the low value of HDL-C to the high value. These results suggested that RCM could be used as the potential candidate for the lipid metabolism natural supplement.

Antioxidant Effects of Eriodictyol on Hydrogen Peroxide-Induced Oxidative Stress in HepG2 Cells (산화스트레스가 유도된 HepG2 세포에서 Eriodictyol의 항산화 효과)

  • Joo, Tae-Woo;Hong, Sung-Hyun;Park, Sun-Young;Kim, Gur-Yoo;Jhoo, Jin-Woo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.4
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    • pp.510-517
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    • 2016
  • This study was conducted to investigate the antioxidant and hepatoprotective effects of eriodictyol compound against hydrogen peroxide-induced oxidative stress in HepG2 cells by measuring expression levels of antioxidant enzymes, liver function index enzyme activities, and inhibitory effects against reactive oxygen species (ROS) production. HepG2 cell viability was assessed using 3-(4,5-dimethyl thiazole-2-yl)-2,5-diphenyl tetrazolium bromide assay. In the concentration range of $10{\sim}50{\mu}g/mL$, eriodictyol displayed over 98% cell viability in HepG2 cells. The effects of increased gene expression on hydrogen peroxide-induced oxidative stress were analyzed by monitoring antioxidant enzyme (superoxide dismutase, SOD; catalase, CAT; glutathione peroxidase, GPx) gene expression levels using real-time PCR. Eriodictyol compound significantly increased gene expression levels of SOD, CAT, and GPx in a dose-dependent manner ($10{\sim}50{\mu}g/mL$). Hepatoprotective effects against hydrogen peroxide-induced oxidative stress were analyzed by monitoring glutamic oxaloacetic transaminase (GOT), lactate dehydrogenase (LDH), and gamma-glutamyl transferase (GGT) activities in HepG2 cell culture medium using a biochemistry analyzer. Eriodictyol compound significantly reduced GOT, LDH, and GGT activities in a dose-dependent manner in HepG2 cells. ROS level in HepG2 cells was analyzed by 2',7'-dichlorofluorescein fluorescence diacetate assay, and eriodictyol compound effectively reduced the intracellular ROS level in HepG2 cells. The results reveal that eriodictyol compound can be useful for development of effective antioxidant and hepatoprotective agents.

Heavy Metal Contamination around the Abandoned Au-Ag and Base Metal Mine Sites in Korea (국내 전형적 금은 및 비(base)금속 폐광산지역의 중금속 오염특성)

  • Chon Hyo-Taek;Ahn Joo Sung;Jung Myung Chae
    • Economic and Environmental Geology
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    • v.38 no.2 s.171
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    • pp.101-111
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    • 2005
  • The objectives of this study we to assess the extent and degree of environmental contamination and to draw general conclusions on the fate of toxic elements derived from mining activities in Korea. 인t abandoned mines with four base-metal mines and four Au-Ag mines were selected and the results of environmental surveys in those areas were discussed. In the base-metal mining areas, the Sambo Pb-Zn-barite, the Shinyemi Pb-Zn-Fe, the Geodo Cu-Fe and the Shiheung Cu-Pb-Zn mine, significant levels of Cd, Cu, Pb and Zn were found in mine dump soils developed over mine waste materials, tailings and slag. Furthermore, agricultural soils, stream sediments and stream water near the mines were severely contaminated by the metals mainly due to the continuing dispersion downstream and downslope from the sites, which was controlled by the feature of geography, prevailing wind directions and the distance from the mine. In e Au-Ag mining areas, the Kubong, the Samkwang, the Keumwang and the Kilkok mines, elevated levels of As, Cd, Cu, Pb and Zn were found in tailings and mine dump soils. These levels may have caused increased concentrations of those elements in stream sediments and waters due to direct dis-charge downstream from tailings and mine dumps. In the Au-Ag mines, As would be the most characteristic contaminant in the nearby environment. Arsenic and heavy metals were found to be mainly associated with sulfide gangue minerals, and mobility of these metals would be enhanced by the effect of oxidation. According to sequential extraction of metals in soils, most heavy metals were identified as non-residual chemical forms, and those are very susceptible to the change of ambient conditions of a nearby environment. As application of pollution index (PI), giving data on multi-element contamination in soils, over 1.0 value of the PI was found in soils sampled at and around the mining areas.

Characteristics of Geochemical Behaviors of Trace Metals in Drainage from Abandoned Sechang Mine (세창 폐금속광산 수계에서 미량원소의 지구화학적 거동특성 규명)

  • Kang Min-Ju;Lee Pyeong-Koo;Youm Seung-Jun
    • Economic and Environmental Geology
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    • v.39 no.3 s.178
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    • pp.213-227
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    • 2006
  • The geochemical evolution of mine drainage and leachate from waste rock dumps and stream water in Pb-As-rich abandoned Sechang mine area was investigated to elucidate mechanisms of trace metals. Total and sequential extractions were applied to estimate the distribution of trace metals in constituent phases of the waste rocks and to assess the mobility of trace metals according to physicochemical conditions. These discharged waters varied largely in chemical composition both spatially and temporally, and included cases with significant]y low pH (in the range 2.1-3.3), and extremely sulphate (up to 661 mg/l and metal contents (e.g. up to 169 mg/l for Zn, 27 mg/l for As, 3.97 mg/l for Pb, 2.99 mg/l for Cu, and 1.88 mg/l for Cd). Arsenic and heavy metal concentrations at the down-stream of Sechang mine have been decreased nearly to the background level in downstream sites (sites 8 and 16) without any artificial treatments. The oxidation of Fe-sulfides and the subsequent hydrolysis, of Fe(II), with precipitation of poorly crystallized minerals, constituted an efficient mechanism of natural attenuation which reduces considerably the transference of trace metals (i.e. Fe and As) to rivers. The dilution of drainage by mixing with pristine waters provoked an additional decrease of trace metal concentrations and a progressive pH increase. On the other hand, the most soluble cations (i.e. Zn) remained significantly as dissolved solutes until the pH was raised to approximately neutral values. With respect to ecotoxicity, it is likely that the Zn pollution is of particular concern in Sechang mine area. This was confirmed by the sequential extraction experiment, where Zn in wet waste-rock samples occurred predominantly in the exchangeable fraction (65-89% of total), while Pb was the highest in the reducible and carbonate fractions, and Cd, Cu and As in the residual fraction. Pb concentration in the readily available exchangeable fraction (34-48% of total) was dominated for dried waste rock samples. Considering the proportion of metals bound to the exchangeable and carbonate fractions, the comparative mobility of metals probably decreased in the order of Zn>Pb>Cd>As=Cu.

Neuroprotective effects of cultured and fermented wild ginseng extracts on oxidative stress induced by hydrogen peroxide in PC12 cells (발효산삼배양근농축액의 산화방지 효과 및 과산화수소로 유발된 PC12 세포독성 보호효과)

  • Choi, Yeo Ok;Kim, Yu-Ri;Shin, Seung-Yong;Lee, Jae Geun;Kim, Chul Joong;Lee, Ye ji;Kang, Byeongju;Kim, Gwansu;Choi, Jee Eun;Han, Beom-Seok
    • Korean Journal of Food Science and Technology
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    • v.50 no.4
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    • pp.383-390
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    • 2018
  • Most neurodegenerative diseases are known to be influenced by oxidative stress. We investigated the anti-oxidative activity of the concentrate of fermented wild ginseng root culture (HLJG0701) containing ginsenosides Rg5 and Rk1. HLJG0701 showed effective DPPH and ABTS radical scavenging ability ($IC_{50}$: 16- and 4-fold dilution, respectively) and was inhibited dose-dependently by the $FeSO_4$-induced lipid peroxidation group (8- and 4-fold dilution: 2.3 and 1.5 nM, respectively). In MTT and LDH assays, 8-, 16-, 32- and 64-fold diluted HLJG0701 significantly increased cell viability by 70, 53, 35, and 26%, respectively. LDH released by HLJG0701 was reduced 1.3-fold with 8-fold diluted HLJG0701 compared to the $H_2O_2$-treated control. In addition, the inhibitory effect of HLJG0701 on oxidative stress in PC12 cells was confirmed by DCF-DA analysis (16-, 4-fold diluted HLJG0701: 50 and 68% ROS inhibition, respectively), TBARS (16- and 4-fold diluted HLJG0701: 50.7 and 46.5% inhibition, respectively), GPx (16- and 4-fold diluted HLJG0701: 133.3 and 227.3% release, respectively), and SOD analysis (16- and 4-fold diluted HLJG0701: 118.2 and 218.2% release, respectively). These results suggested that HLJG0701 protects neuronal cells by its anti-oxidative effects and hence can be a potential preventive material against neurodegenerative diseases.

Estimation of Anti-proliferative Activity of Saccharin against Various Cancer Cell Lines and MSCs (다양한 암세포 주와 MSCs에 대한 Saccharin의 항증식성 평가)

  • Choi, Jeong Su;Park, Sang Yong;Yang, Man Gil;Lee, Dong Beom;Lee, Tae Bok;Heo, Ji Hye;Lee, Min Woo;Kim, Suhng Wook
    • Korean Journal of Clinical Laboratory Science
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    • v.48 no.3
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    • pp.169-175
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    • 2016
  • Saccharin (o-benzoic sulfimide) is the first artificial and non-caloric sweetener that was first synthesized in 1879. In this study, we examined the biological activity of saccharin against various human cancer cell lines and human bone marrow-derived mesenchymal stem cells. A viability assay based on the conversion of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was performed to test for the cytotoxicity of saccharin about the four human cancer cell lines (H460, H157, A549 and SKOV3), one murine cancer cellline (Raw264.7), and MSCs. In order to find the differentially expressed gene in saccharin-treated MSCs against untreated MSCs, we performed annealing control primer (ACP)-based differential display reverse transcriptionp-olymerase chain reaction (DDRT-PCR). All tested cells were treated with saccharin at various concentrations (0.0, 4.8, 7.2, 9.6, 12.0, 14.4 mg/mL) for 48 hr. The number of metabolically active cancer cells decreased when treated with the saccharin at various concentrations for 48 hr as compared with the untreated cells. The decrease in cell survival was more evident with increasing concentrations of saccharin. Moreover, novel candidate genes, which were differentially expressed in MSCs in response to saccharin, were identified in 16 bands on 2% agarose gel. This revealed 16-7 up-regulated and 9 down-regulated-differentially expressed genes indicated by arrows. One of these candidate genes was a FK506-binding protein gene. The functional roles of FK506 binding proteins, with respect to the activities of stem cell proliferation, were not characterized. Further studies are required to get a better understanding of FK506-binding proteins in its roles in increasing stem cell proliferative activities from using saccharin.

Experimental Models of Schizophrenia (정신분열병의 실험적 모델)

  • Cheon, Jin-Sook
    • Korean Journal of Biological Psychiatry
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    • v.6 no.2
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    • pp.153-160
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    • 1999
  • Animal models can provide a useful tool for the study of some aspects of psychiatric disorders and their treatment. The four criteria for the evaluation of animal models of psychiatric disorders are as following : 1) similarity of inducing conditions 2) similarity of behavioral state 3) common underlying neurobiological mechanisms 4) reversal by clinically effective treatment techniques. Several animal models have been proposed for schizophrenia : phenylethylamine model, L-dopa model, hallucinogen model, cocaine model, amphetamine model, phencyclidine model, noradrenergic reward system lesion model, reticular stimulation model, social isolation model, conditioned avoidance reaction, catalepsy test, paw test, self-stimulation paradigms, latent inhibition paradigms, blocking paradigms, prepulse inhibition of the startle reflex, rodent interaction, social behavior in monkeys, hippocampal damage, high ambient pressure, and models using selective breeding. Among them, animals with bilateral lesion of the hippocampus may provide an adequate animal model for several symptoms of schizophrenia, and ketamine model can reproduce negative symptoms and cognitive deficits as well as positive symptoms of schizophrenia. In conclusion, no model of schizophrenia is entirely representative of the disease, and findings gleaned from model systems must be cautiously interpreted. Furthermore, the process of developing and validating animal models must work in concert with the process to identify reliable measures of human phenomenology.

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Design of Aerosol Generator for Inhalation Toxicology Study of Lead and Evaluation with Real Time Monitoring (납의 흡입독성 연구를 위한 에어로졸 발생장치의 고안 및 실시간 모니터링을 이용한 성능평가)

  • Jeung Jae Yeal;Kim Jung Man;Kim Tae Hyeung;Chong Myoung Soo;Ko Kwang Jae;Kim Sang Duck;Kang Sung Ho;Song Young Sun;Lee Ki Nam
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.16 no.2
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    • pp.373-379
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    • 2002
  • This paper was the design of aerosol generator for inhalation toxicology study of lead and evaluation with real time monitoring, and applied several engineering methodology to classical aerosol generator to cope with it's disadvantages. According to the testing conditions, source temperature 50℃ and inlet-duct band heater temperature 150℃, aerosol generation results for sodium chloride and lead acetate were as followings: CPM(Count Per Minute) for Sodium chloride that used for the testing material in aerosol generation and inhalation system was decreased in the 2nd and the 3rd hour's serial trials, but CVs(coefficient of variation) were maintained within 10%. CPMs for 5 and 2.5 gram of lead acetate that used for aerosol generation and inhalation exposure of lead showed similar results because of the sedimentation of lead acetate on piezoelectric crystal with time. For that reason, heating and mixing of nebulizing solution will be needed to generate lead aerosol with stable profile and maximum generation efficiency. Fluctuations of 10 and 5 gram lead acetate were low but 2.5gram was high. However, CVs for 10, 5, and 2.5gram lead acetate were within 10%. Considering the theoretical efficiencies for sodium chloride and lead acetate, 5gram sodium chloride and 2.5gram lead acetate were appropriate choice. Aerosol generation characteristics for two materials with 1 hour interval were different with respect to the fluctuation of CPM and the decrease to 10gram in it's material. For that reason, sodium chloride can not be used to estimate the aerosol generation and it's related parts for lead acetate. According to the testing conditions, source temperature 20, 50, 70℃, and inlet-duct band heater temperature 20, 50, 100, 150, 200℃, aerosol generation results for sodium chloride and lead acetate were as followings: Excluding inlet-duct band temperature 200℃, maximum CPM for sodium chloride was manifested in source temperature 70℃ with each inlet-duct band temperature conditions. We suggest that this condition was the optimum in the design of aerosol generator, inhalation system, and the testing. Maximum CPMs for 10, 5, and 2.5gram sodium chloride were from source temperature 70℃ and inlet-duct band temperature 20℃. Excluding inlet-duct band temperature 50, 200℃, maximum CPMs for lead acetate were indicated in source temperature 50℃ with each inlet-duct band temperature conditions. We suggest that this condition was the optimum in the design of aerosol generator, inhalation system, and the testing for lead inhalation study. Source and inlet-duct band temperatures for 10, 5, 2.5gram lead acetate were 50 and 100℃, 50 and 100℃, 50 and 150℃, respectively. In conclusion, considering above 2 paragraphs of results for aerosol generation, 5gram efficiencies for sodium chloride, lead acetate were higher than 2.5gram's. If inlet-duct band temperature was same, aerosol generation was increased with increase of source temperature. To get maximum aerosol generation will be the conditions that set the appropriate inlet-duel band temperature for each materials and increase the source temperature.

Factors Associated with Renal Scar in Children with Vesicoureteral Reflux (방광 요관 역류가 있는 소아에서 신반흔 형성과 관련된 인자들)

  • Kim Kyoung Hee;Jang Sung Hee;Lee Dae-Yeol
    • Childhood Kidney Diseases
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    • v.5 no.1
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    • pp.43-50
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    • 2001
  • Purpose : The urinary tract infection associated with vesicoureteral reflux(VUR) in children may result in serious complications such as renal scarring, hypertension, proteinuria and end stage renal disease. The purpose of this study was to evaluate the factors affecting renal scar such as age, gender, grade of VUR, and ACE gene polymorphism, and body growth in the patients with and those without renal scar associated with VUR Methods : During the period from January 1994 to July 2000, We had 93 children with urinary tract infection associated with VUR who were admitted to the Department of pediatrics of Chonbuk National University Hospital. The patients were divided into two groups according to follow up 99mTc-DMSA renal scan; patients with renal scar group and those with non-scar group. We analyzed and compared the factors associated with renal scarring between the two groups. Results : There were no significant difference in gender, causative organism, ACE gene polymorphism, height and weight at diagnosis between renal scar group and non-scar group. Fifty four patients were in renal scar group and forty seven of them had VUR. The age at diagnosis was significantly higher in renal scar group (2.48${\pm}$2.64yr) than in non renal scar group (1.26${\pm}$1.83yr). Especially, the infants who were less than 1 year of age with VUR developed relatively more renal scar compared with infants older than 1 tear of age. The incidence of renal scarring showed a direct correlation with the severity of VUR. Conclusion : The factors affecting renal scar formation were age at diagnosis, presence and grade of VUR, but the other factors such as gender, causative organism, ACE gene polymorphism were not associated with renal scarring. Therefore, further evaluation about uropathogenic E coli and foflow up study about body growth associated with severity of renal scar would be necessary. (J. Korean Soc Pediatr Nephrol 5 : 43- 50, 2001)

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