• Title/Summary/Keyword: Screen Concentration

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Establishment of High Throughput Screening System Using Human Umbilical Cord-derived Mesenchymal Stem Cells

  • Park, Eu-Gene;Cho, Tae-Jun;Oh, Keun-Hee;Kwon, Soon-Keun;Lee, Dong-Sup;Park, Seung-Bum;Cho, Jae-Jin
    • International Journal of Oral Biology
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    • v.37 no.2
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    • pp.43-50
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    • 2012
  • The use of high throughput screening (HTS) in drug development is principally for the selection new drug candidates or screening of chemical toxicants. This system minimizes the experimental environment and allows for the screening of candidates at the same time. Umbilical cord-derived stem cells have some of the characteristics of fetal stem cell and have several advantages such as the ease with which they can be obtained and lack of ethical issues. To establish a HTS system, optimized conditions that mimic typical cell culture conditions in a minimal space such as 96 well plates are needed for stem cell growth. We have thus established a novel HTS system using human umbilical cord derived-mesenchymal stem cells (hUC-MSCs). To determine the optimal cell number, hUC-MSCs were serially diluted and seeded at 750, 500, 200 and 100 cells per well on 96 well plates. The maintenance efficiencies of these dilutions were compared for 3, 7, 9, and 14 days. The fetal bovine serum (FBS) concentration (20, 10, 5 and 1%) and the cell numbers (750, 500 and 200 cells/well) were compared for 3, 5 and 7 days. In addition, we evaluated the optimal conditions for cell cycle block. These four independent optimization experiments were conducted using an MTT assay. In the results, the optimal conditions for a HTS system using hUC-MSCs were determined to be 300 cell/well cultured for 8 days with 1 or 5% FBS. In addition, we demonstrated that the optimal conditions for a cell cycle block in this culture system are 48 hours in the absence of FBS. In addition, we selected four types of novel small molecule candidates using our HTS system which demonstrates the feasibility if using hUC-MSCs for this type of screen. Moreover, the four candidate compounds can be tested for stem cell research application.

The Effects of Child Cardiopulmonary Resuscitation Education for Childcare Teachers with a Video Self-Instruction Program (Video Self-Instruction Program을 이용한 보육교사의 소아심폐소생술 교육의 효과)

  • Kim, Geon-Hee
    • The Korean Journal of Emergency Medical Services
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    • v.13 no.2
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    • pp.87-98
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    • 2009
  • Purpose : This study set out to compare the educational effects of a video self-instruction program for child CPR education on childcare teachers by applying the 2006 KACPR Guideline. By adopting the nonequivalent control group posttest quasi-experimental design, the study examined the educational effects on a group that did not receive instructions from the instructor, another group that received his instructions, and the other group that received an extra three-minute practice training session in addition to instructions. Methods : Data were gathered from August 6 to 18, 2008. As for research tools, the Knowledge Instrument of CPR by Connolly (2006) was used along with the National Practice Test Protocol for C1ass 1 Emergency Medical Technicians (2007) and Common Protocol for CPR (2006) to examine the performance of child CPR. By shooting the guide screen of $Resusci^{(R)}$ Junior CPR Manikin of Leardal with a video camera and using the Skill Guide Checklist of the Common Protocol for CPR (2006), the subjects' technical accuracy of chi1d CPR was evaluated. There were three subject groups: 29 childcare teachers randomly assigned to received the video self-instruction program training for chi1d CPR and no instructions from the instructor made up the control group; 22 childcare teachers randomly assigned to received the program training and instructions from the instructor made up experiment group I; 23 childcare teachers randomly assigned to received an extra three-minute practice training session in addition to the program training and the instructions made up experiment group II. The gathered data were analyzed with SPSS/PC+ (Version 14.0) in frequency, percentage, $X^2$-test, ANOVA, Scheffe test. Results : 1) There were no statistically significant differences (F=1.030, p=.362) among the groups in terms of knowledge scores after the child CPR education. 2) There were statistically significant differences (F=13.625, p=.000) among the groups in terms of performance abilities after the child CPR education. 3) There were no statistically significant differences (F=1.610, p=.207) among the groups in terms of technical accuracy of mouth-to-mouth resuscitation after the child CPR education 4) There were no statistically significant differences (F=1.484, p=.234) among the groups in terms of technical accuracy of chest compression after the child CPR education. Conclusion : The results indicate that childcare teachers can improve their performance abilities in child CPR when the instructors are active with their instructions and extra practice hours are secured through a VSI program. It's also needed to provide education with increasing concentration ratio about the items of lower knowledge points in order to help the teachers learn the accurate theory of child CPR. And there should be VSI programs of diverse conditions to increase the effects of child CPR training among childcare teachers.

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In vitro Cytotoxin Activity of Urushiol in the Sap of Rhus verniciflua STOKES (옻나무 칠액성분(漆液成分) 중 Urushiol의 암세포(癌細胞) 증식억제(增殖抑制) 효과(效果) - in vitro 세포독성효과(細胞毒性效果) -)

  • Na, Chun-Soo;Jung, Nam-Chul;Oh, Kwang-In
    • Journal of Korean Society of Forest Science
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    • v.87 no.2
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    • pp.260-269
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    • 1998
  • This study was conducted to screen the biological activity of urushiol in the sap of lac tree(Rhus verniciflua STOKES) which has been used in traditional folk remedies. Cytotoxic activity of urushiol was screened with L1210(mouse luekemia cell), PC-9(human lung adenocarcinoma cell), A427(human lung adenocarcinoma cell) and KATO III (human stomach adenocarcinoma cell) The stepwise hexane : acetone eluent fractions of the urushiol were obtained by the silica gel adsorption column chromatography and added to the culture media containing L1210, PC-9. A427, and KATO III, respectively. A hexane : acetone(90 : 10, v/v) eluent fraction of them showed the lowest 50% inhibition concentration($IC_{50}$) of $0.018{\mu}g/m{\ell}$ for the cell line of A427. Much lower level of $IC_{50}$ of the hexane : acetone(90 : 10, v/v) eluent fraction of the urushiol showed the equal inhibition effect with tetraplatin(i.e., anti-cancer drug of platinum complexes) on the cancer cell lines as follows ; 3.4 times lower for L1210, 3.9 times lower for PC-9, and 105.5 times lower for A427. However, $IC_{50}$ of the hexane : acetone(90 : 10 v/v) eluent fraction for KATO III was exceptionally 3.9 times higher than that of tetraplatin.

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Analysis of metabolomic patterns in thoroughbreds before and after exercise

  • Jang, Hyun-Jun;Kim, Duk-Moon;Kim, Kyu-Bong;Park, Jeong-Woong;Choi, Jae-Young;Oh, Jin Hyeog;Song, Ki-Duk;Kim, Suhkmann;Cho, Byung-Wook
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.11
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    • pp.1633-1642
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    • 2017
  • Objective: Evaluation of exercise effects in racehorses is important in horseracing industry and animal health care. In this study, we compared metabolic patterns between before and after exercise to screen metabolic biomarkers for exercise effects in thoroughbreds. Methods: The concentration of metabolites in muscle, plasma, and urine was measured by $^1H$ nuclear magnetic resonance (NMR) spectroscopy analysis and the relative metabolite levels in the three samples were compared between before and after exercise. Subsequently, multivariate data analysis based on the metabolic profiles was performed using orthogonal partial least square discriminant analysis (OPLS-DA) and variable important plots and t-test was used for basic statistical analysis. Results: From $^1H$ NMR spectroscopy analysis, 35, 25, and 34 metabolites were detected in the muscle, plasma, and urine. Aspartate, betaine, choline, cysteine, ethanol, and threonine were increased over 2-fold in the muscle; propionate and trimethylamine were increased over 2-fold in the plasma; and alanine, glycerol, inosine, lactate, and pyruvate were increased over 2-fold whereas acetoacetate, arginine, citrulline, creatine, glutamine, glutarate, hippurate, lysine, methionine, phenylacetylglycine, taurine, trigonelline, trimethylamine, and trimethylamine N-oxide were decreased below 0.5-fold in the urine. The OPLS-DA showed clear separation of the metabolic patterns before and after exercise in the muscle, plasma, and urine. Statistical analysis showed that after exercise, acetoacetate, arginine, glutamine, hippurate, phenylacetylglycine trimethylamine, trimethylamine N-oxide, and trigonelline were significantly decreased and alanine, glycerol, inosine, lactate, and pyruvate were significantly increased in the urine (p<0.05). Conclusion: In conclusion, we analyzed integrated metabolic patterns in the muscle, plasma, and urine before and after exercise in racehorses. We found changed patterns of metabolites in the muscle, plasma, and urine of racehorses before and after exercise.

Optimum Formulation of Starch and Non-muscle Protein for Alkali Surimi Gel from Frozen White Croaker (냉동 백조기의 알칼리 수리미 겔 제조를 위한 전분 및 비근육 단백질의 최적화)

  • 박주동;김진수;조영제;최종덕;최영준
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.32 no.7
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    • pp.1026-1031
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    • 2003
  • The two-level full factorial and mixture design were used to screen ingredient type and to investigate effects of ingredients on properties of alkali surimi gel from frozen white croakers using measurements of a breaking force, deformation and color. The addition of starch decreased a breaking force and deformation of gel regardless of starch type. The breaking force was decreased, but a deformation was not significantly changed (p<0.05) with increasing starch level. The potato starch was more resonable than com and wheat starch for a breaking force and deformation. The bovine plasma protein (BPP) greatly improved a breaking force and deformation. The breaking force and deformation of gel were increased with concentration of BPP. The whiteness of gel was slightly improved with adding starch and non-muscle for all treatments. At 78% moisture, the optimum ratios of ingredients were 89.4∼90.0% for surimi, 5.9∼6.3% for potato starch and 5.0∼5.4% for BPP to obtain above 100g for a breaking force, 4.6 mm for a deformation, and 25.5 for a whiteness.

Antioxidative and Antimicrobial Activities of Sargassum muticum Extracts (경단구슬모자반(Sargassum muticum) 추출물의 항산화 및 항균활성)

  • Kim, Ji-Young;Lee, Jung-A;Kim, Kil-Nam;Yoon, Weon-Jong;Lee, Wook-Jae;Park, Soo-Yeong
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.6
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    • pp.663-669
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    • 2007
  • The solvent extracts of Sargassum muticum, which were extracted by using several solvents with different polarities, were prepared for use as natural preservatives. The S. muticum extract with 80% ethanol was sequentially fractionated with n.hexane, dichloromethane, ethylacetate, and butanol. In order to effectively screen for natural preservatives agents, we first investigated the antioxidant activities such as DPPH radical scavenging capacity, superoxide radical scavenging capacity, and xanthine oxidase inhibitory activity of the S. muticum extracts. Through the screening system, we found that dichloromethane and ethylacetate fraction had high antioxidant activity with increments of the extract concentration. The antimicrobial activities and cell growth inhibition were investigated for each strain with the different concentrations of S. muticum extracts. Antimicrobial activities were shown in ethanol, dichloromethane, and n.hexane fractions of S. muticum. However, butanol, ethylacetate and water fractions showed weak antimicrobial activity against the tested microorganisms. Among the five fractions, dichloromethane fraction showed the highest antimicrobial activities against microorganisms tested, such as Bacillus sublitis, Listeria monocytogenes, Staphylococcus aureus, Escherichia coli, Salmonella Enteritidis and Pseudomonas aeruginosa. The polyphenolic compounds from ethanol, n.hexane, dichloromethane, ethylacetate, butanol, and water fractions were 63.96 mg/g, 8.49 mg/g, 28.11 mg/g, 172.64 mg/g, 114.56 mg/g, and 34.91 mg/g, respectively. The dichloromethane fraction could be suitable for development as a food preservative.

Antioxidant Activity and Physiological Function of the Anomala albopilosa Extracts (청동풍탱이(Anomala albopilosa)추출물의 항산화성 및 생리기능)

  • Yoon, Weon-Jong;Lee, Jung-A;Kim, Ji-Young;Kim, Sang-Bum;Park, Soo-Yeong
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.6
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    • pp.670-677
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    • 2007
  • We analyzed antioxidant and physiological activities to investigate the functional effects of ethanol extracts of Anomala albopilosa imago and A. albopilosa larva. In order to effectively screen for anti-inflammatory agents, we first investigated the antioxidant activities such as DPPH radical scavenging capacity, superoxide radical scavenging capacity, xanthine oxidase inhibitory activity, and nitric oxide scavenging capacity of the A. albopilosa extracts. By the screening system, we found that A. albopilosa extracts had antioxidant activity which increased with increments of the extract concentration. Moreover, we examined the inhibitory effect of the A. albopilosa extracts on the production of anti-inflammatory factors that the nitric oxide (NO), inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2) and prostaglandin $E_2\;(PGE_2)$ production activated with LPS ($1{\mu}g/mL$) in murine macrophage cell line RAW 264.7. A. albopilosa extracts potentially inhibited the iNOS and COX-2 in a dose-dependent manner. The inhibition of iNOS activity was correlated with the decrease in nitrite levels. Additionally, the $PEG_2$ production is markedly inhibited after a treatment with the A. albopilosa extracts.

Assessment of In Vitro Assay System for Thyroid Hormone Disruptors Using Rat Pituitary GH3 Cells

  • Kim, Hee-Jin;Park, Hae-Young;Kim, Jeong-A;Kang, Il-Hyun;Kim, Tae-Sung;Han, Soon-Young;Kang, Tae-Seok;Park, Kui-Lea;Kim, Hyung-Sik
    • Toxicological Research
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    • v.22 no.4
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    • pp.307-313
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    • 2006
  • The development of in vitro assays has been recommended to screening and testing the potential endocrine disruptors (EDs). These assay systems focus only on identifying the estrogenic or antiestrogenic activity of EDs, whereas a few studies have been carried out to screen the thyroid hormone (TH) disruptors. The aim of this study was to evaluate a test system to detect TH disruptors using rat pituitary tumor $GH_3$ cells. The test system is based on the TH-dependent increase in growth rate. As expected, L-3,5,3-triiodothyronine ($(T_3)$ markedly induced a morphological change in $GH_3$ cells from flattened fibroblastic types to rounded or spindle-shaped types. $T_3$ stimulated $GH_3$ cell growth in a dose-dependent manner with the maximum growth-stimulating effect being observed at a concentration $1{\times}10^9M$. In addition, $T_3$ increased the release of growth hormone and prolactin into the medium of the $GH_3$ cells culture. Using this assay system, the TH-disrupting activities of bisphenol A (BPA) and its related compounds were examined. BPA, dimethy/bisphenol A (DMBPA), and TCI-EP significantly enhanced the growth of $GH_3$ cells in the range of $1{\times}10^{-5}M\;to\;1{\times}10^{-6}M$ concentrations. In conclusion, this in vitro assay system might be useful for identifying potential TH disruptors. However, this method will require further evaluation and standardization before it can be used as a broad-based screening tool.

Establishment of ethanol-pretreating animal model to study Helicobacter pylori infection (Helicobacter pylori의 in vivo 연구를 위한 ethanol-pretreating animal model의 개발)

  • Lee, Jin-Uk;Kim, Seung-Hee;Park, Tan-Woo;Kim, Okjin
    • Korean Journal of Veterinary Research
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    • v.46 no.4
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    • pp.327-335
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    • 2006
  • A stable and reliable Helicobacter pylori (H. pylori) infection animal model would be necessary for evaluating vaccine efficacy and helpful for understanding the pathological mechanism of the organism. The aim of the present study is to investigate the effect of ethanol treatment prior to H. pylori inoculation on associated gastric mucosal injury and to establish ethanol-pretreating animal model to study H. pylori infection. Male Mongolian gerbils were used for the study. H. pylori was orally inoculated after 12 h fasting. 3 h prior to H. pylori inoculation, a group of gerbils was orally treated with absolute ethanol, 60% and 40% ethanol respectively. Another group of animals was treated either with H. pylori culture media alone or with different concentrations of ethanol plus culture media. Gerbils were killed 4 or 8 weeks after H. pylori inoculation. The colonization of H. pylori was confirmed by both histological examination and rapid urease test. Mucosal damage was evaluated grossly and histologically according to the criteria. The colonization of H. pylori and pathological changes in gastric mucosa of the animals were also observed. Although no significant change to the gastric mucose was observed in the animals treated either with H. pylori culture media alone or with different concentrations of ethanol plus culture media, persistent H. pylori infection was seen in the mucosa and mucosal leucocyte infiltration and severe epithelial damage was observed in the Helicobacter and ethanol + Helicobacter groups after 4 weeks. The gross and histological scores were higher in the ethanol + Helicobacter than in the Helicobacter alone group. As the results, ethanol-pretreatment with 60% concentration induced severe pathogenic changes by H. pylori infection in 5 weeks-old Mongolian gerbils. These results suggested that ethanol-pretreatment before H. pylori inoculation could increase the severity of gastric mucosal inflammation and enhance the colonization of H. pylori. The established ethanol-pretreating animal model would contribute to screen new drugs against H. pylori and be used as an useful tool for various animal experiments with H. pylori strains.

Construction of Environmental Friendly Special-Purpose Ship for the Removal of Blue-green Algae (친환경적 녹조 제거용 특수선박 건조)

  • Shin, Jae-Ki;Yi, Hye-Suk;Jeong, Sun-A;Hwang, Soon-Jin
    • Korean Journal of Ecology and Environment
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    • v.42 no.3
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    • pp.404-406
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    • 2009
  • This study note wished to introduce special-purpose ship for algae removal that is developed by core technology of our country. The ship is consisted of main frame and adjuvant that can attach and detach as cross (+) shape of a character. The characteristics of ship are super light weight and low draft. That is consisted of four devices as suction, collection, filtration and recovering units. Among these, filtration used screen filter (mesh size 30 ${\mu}m$). Also, can separate and remove water and algae by compression air participle notion. Percentage of moisture content of concentrated algal particle was 85%. Water parted with algae finally is exhausted to water area. Removal efficiency that compare by chlorophyll-$\alpha$ concentration was about 57% (inflow: 83.2 ${\mu}g\;L^{-1}$, outflow: 35.8 $[\mu}g\;L^{-1}$) without physical and chemical pretreatment. Forward, need to achieve effect test in various conditions (algal biomass, flow etc.) for efficiency and technological elevation of exclusion device. We wished to contribute in presuppression system construction of massive algal development that manage blue-green algae occurrence area effectively, and prevents spread as lower part of reservoir.