• Title/Summary/Keyword: 생체 외 실험

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Adherence-induced gene expression in human alveolar macrophages (표면부착에 의한 사람 폐포대식세포의 유전자 발현에 관한 연구)

  • Chung, Man Pyo;Yoo, Chul Gyu;Han, Sung Koo;Shim, Young-Soo;Rhee, Chong H.;Han, Yang Chol;Kim, Young Whan
    • Tuberculosis and Respiratory Diseases
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    • v.43 no.6
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    • pp.936-944
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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. Adhesion molecules and gene transcription of the inflammatory mediators are known to be associated in this process. To evaluate whether adhesion molecule and transcriptional activation of the inflammatory substances are also involved in the activation of human alveolar macrophage by the adherence procedure, we designed this experiment. Method : Bronchoalveolar lavage was performed in the person whose lung of either side was confirmed to be nonnal by chest cr and alveolar macrophage was harvested. To measure the expression of Interleukin-8(IL-8) mRNA, manganese superoxide dismutase(SOD) mRNA and CD11/CD18 mRNA in human alveolar macrophage of both adherence state and suspension state, Northern blot analysis was done at 0, 2, 4, 8 and 24hrs after the adherence to plastic surface and during suspension state. Then, phorbol myristate acetate(pMA) and N-formyl-methionyl-leucyl-phenylalanine(fMLP) were added respectively in the same experimental condition. Result : 1) Human alveolar macrophages in the adherent state induced IL-8 mRNA and SOD mRNA expression which was maximal at 8 hours after the adherence to plastic surface. But we could not observe the upregulation of CD18 mRNA by surface adherence. 2) PMA induced these mRNA expression both in the adherent cell and the nonadherem cells, but the induction of mRNA expression by fMLP occurred only in the adherent cells. Conclusion: These results suggest that adherence of huamn alveolar macropahge is an important cell-activating event that may play a critical role in the modulation of lung inflammatory respones.

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Adhesion-induced generation of oxygen free radical from human alveolar macrophages and its mechanisms (폐포대식세포의 부착에 의한 산소유리기 분비능 활성화 및 그 기전)

  • Chung, Man-Pyo;Yoo, Chul-Gyu;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Han, Yong-Chol
    • 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.

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Selectivity of Bleaching Herbicides Caused by Physiological Differences between Rice and Barnyardgrass (벼와 피의 생리적(生理的) 차이(差異)에 의한 백화형(白化型) 제초제(除草劑)의 선택성(選擇性))

  • Na, J.Y.;Kim, J.S.;Kim, T.J.;Cho, K.Y.;Pyon, J.Y.
    • Korean Journal of Weed Science
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    • v.12 no.2
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    • pp.89-101
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    • 1992
  • In this study, various physiological and biochemical experiments were conducted to know whether the selectivity between rice and barnyardgrass treated with bleaching herbicides containing diphenyl ether compounds was also partly based on their basic physiological proterties such as peroxidation ability, membrane stability or antioxidant system. It seemed to be partly based on the differences of their physiological characteristics that barnyardgrass was commonly more susceptible to most of the bleaching herbicides than rice. The scenescence of intact leaf segment was more rapid in barnyardgrass than in rice, indicating that barnyardgrass is weak at early stage. Also pigment metabolic ability, antioxidant enzyme activities(peroxidase, catalase, superoxide dismutase, glutathione reductase) and antioxidant content (tocopherol, ascorbic acid, glutathione, carotenoids) were lower in barnyardgrass on the basic of fresh weight. However, lipoxygenase activity and the content of unsaturated fatty acid which is vulnerable to oxygen radicals were higher in barnyardgrass, suggesting that barnyardgrass seedling bave a properties easy to be peroxidized. The differences of PPIX (protoporphyrin IX) or carotenoid content, which are the primary substances inducing herbicide activity, were not related to the selectivity between rice and barnyardgrass.

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Growth of 'Nokkwang' Hot Pepper Plug Seedlings as Influenced by Various Ratios of Pre-planting NH4+:NO3- in Root Substrate (상토에 기비로 혼합된 NH4+:NO3- 비율에 따른 '녹광' 고추 플러그 묘의 생장)

  • Oh, Sang Se;Park, Myong Sun;Kim, Hyun Cheul;Choi, Jong Myung
    • Journal of Bio-Environment Control
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    • v.28 no.2
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    • pp.110-116
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    • 2019
  • This study was conducted to determine the effects of a pre-planting fertilizers with various $NH_4{^+}:NO_3{^-}$ ratios in a coir dust:peatmoss:perlite (3.5:3.5:3.0, v/v/v) medium on the growth of hot pepper (Capsicum annuum L. cv. Nokkwang) plug seedling. Nitrogen levels were fixed to $300mg{\cdot}L^{-1}$ and the $NH_4{^+}:NO_3{^-}$ ratios were varied to 0:100, 27:73, 50:50, 73:27, and 100:0. The 50-cell trays were filled with treatment media containing pre-plant fertilizers, then seeds were sown. After seeds were germinated, the trays were moved to greenhouse and seedlings were feed with 13-2-13 and 20-9-20 fertilizers, alternatively. The changes in pH and EC were measured every week and soil solution for nutrient concentrations were analyzed in week 0, 3, and 7. The measurements of seedling growths as well as analysis of tissue nutrient contents were also conducted in week 7. The varied $NH_4{^+}:NO_3{^-}$ ratios did not influence on the pHs of root media after incorporation of pre-planting fertilizers, but the ECs were heightened as proportion of $NH_4{^+}$ to $NO_3{^-}$ were elevated. During the raising of seedlings, the pHs rose over time in the treatments of 0:100 and 27:73 ($NH_4{^+}:NO_3{^-}$). The concentrations of all macro-elements in root media decreased gradually as seedlings grew in all treatments. The seedling growths 7 weeks after seed sowing were the highest in the treatments of 27:73 and 50:50 ($NH_4{^+}:NO_3{^-}$) and those became worse in the treatments of higher $NH_4{^+}$ ratios than 73%. In terms of inorganic element contents based on the dry weight of above ground tissue, the treatment of 0:100 showed the lowest content of Ca, Mg, Na, Cu, Mn, and Zn. Based on the results, it is desired that $NH_4{^+}$ ratio in pre-planting fertilization is maintained to be 50% or less for the raising of hot pepper plug seedlings.

The effect of reduced thickness in different regions on the fracture resistance of monolithic zirconia crowns (다양한 부위에서의 감소된 두께가 지르코니아 크라운의 파절 저항에 미치는 영향)

  • Abukabbos, Layla;Park, Je Uk;Lee, Wonsup
    • The Journal of Korean Academy of Prosthodontics
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    • v.60 no.2
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    • pp.135-142
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    • 2022
  • Purpose. This study aims to evaluate the combined effect of reduced thickness in different regions on the fracture resistance of monolithic zirconia crowns. Materials and methods. Seven nickel-chromium dies were generated from a 3D model of mandibular first molar using the digital scanner with the following geometries: 1.5 mm occlusal reduction, 1.0 mm deep chamfer. Based on the abutment model, Zirconia blocks (Luxen Zirconia) were selected to fabricate Sixty-three zirconia crowns with occlusal thicknesses of 0.3 mm, 0.5 mm, and 1.5 mm, and different axial thicknesses of 0.3 mm, 0.5 mm, and 1.0 mm. All crowns were cemented by resin cement. Next, the crowns were subjected to load-to-fracture test until fracture using an electronic universal testing machine. In addition, fracture patterns were observed with a scanning electron microscope (SEM). Two-way ANOVA and the Tuckey HSD test for post hoc analysis were used for statistical analysis (P < .05). Results. The mean values of fracture resistancerecorded was higher than the average biting force in the posterior region. The two-way ANOVA showed that the occlusal and axial thickness affected the fracture resistance significantly (P < .05). However, the effect of axial thickness on fracture resistance did not show a statistical difference when thicker than 0.5 mm. The observed failure modes were partial or complete fracture depending on the severity of crack propagation. Conclusion. Within the limitations of the present study, the CAD-CAM monolithic zirconia crown with extremely reduced thickness showed adequate fracture resistance to withstand occlusal load in molar regions. In addition, both occlusal and axial thickness affected the fracture resistance of the zirconia crown and showed different results as combined.

Primary Culture of Chicken Tracheal Epithelial Cells and Study on Those Characters for Recombinant Virus Infection (닭 기도 상피세포의 1차 배양과 유전자 재조합 바이러스의 감염 특성 연구)

  • Park, Mi Na;Jang, Hyun-Jun;Keum, Dae Ho;Choi, Jin Ae;Yoo, Jae Gyu;Byun, Sung June;Park, Jong Ju;Ji, Ju Young;Lee, Kyung-Tai;Kim, Tae-Hun;Lee, Hyun-Jeong
    • Korean Journal of Poultry Science
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    • v.40 no.4
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    • pp.299-304
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    • 2013
  • Tracheal epithelial cells (TECs) are an important tool for studies of viral respiratory diseases. Primary TECs have been cultured from human, mouse and hamster. It is also necessary to diagnose viral respiratory disease and reveal infection mechanisms in chicken. In this study, we isolated tracheal epithelial layers from tracheal of 20-day-old chicks and cultured primary TECs from the isolated layers. Ciliated cells which were a typical morphology of TECs were observed in cultured primary TECs and maintained until cell passage 5 (15 to 20 days). When we analyzed expression patterns of epithelial marker genes (retinoic acid responder, FGF-binding protein, virus activating protease (VAP) in TECs compared to immortalized chicken embryonic fibroblast cell line (DF-1), all the marker genes are highly expressed in TECs than in DF-1. When TECs were cultured with 0.1 and 1 MOI of ND virus (rNDV-GFP strain) to test the susceptibility of TECs for ND virus, 12.6% and 48.2% of the incubated TECs were infected respectively. In addition, when DF-1 was incubated with 1 MOI of ND virus, the virus infection rate of DF-1 was three times lower than the virus infection rate of TECs. These data could contribute to study infection mechanisms of viral respiratory diseases and control them in chicken.

Effects of Soil Moisture and Planting Depth on the Growth of 2-year Old Ginseng Plant (Panax ginseng C. A. Meyer) (토양수분(土讓水分) 및 재식심도(栽植深度)가 저년근(低年根) 고려인삼(高麗人蔘)의 생육(生育)에 미치는 영향(影響))

  • Lee, Jong Chul;Mok, Seong Kyun;Lee, Jong Wha;Jo, Jae Seong
    • Korean Journal of Agricultural Science
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    • v.10 no.2
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    • pp.235-241
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    • 1983
  • This experiment was conducted to determine the effect of soil moisture content and planting depth on the growth of 2-year old ginseng plant. The results obtained are summarized as follows; 1. When the ginseng seedlings were planted in soil by 4 to 5cm in depth, the length of leaflet and stem and the number of branch roots were significantly decreased but the stem diameter was increased. 2. Highly significant quadratic regressions were shown between soil moisture content and the growth of the stem, leaf and root of the ginseng plant. 3. Estimated amount of soil moisture for the maximum growth of the stem was 75% of field capacity, and that for length and width of the leaflet was about 65 to 66% of field capacity. Estimated soil moisture for the maximum growth of the root was about 56 to 58% of field capacity and that for increase in root weight was about 60 to 61% of field capacity. 4. Estimated soil moisture content for best growth of ginseng roots was 1 to 5% lower when the seedling was planted in 3cm depth compared with 2cm in depth. And when the amount of soil moisture was 31% of field capacity, the deep planting was adequate for good root growth. 5. Significant correlations were resulted between the dry weight of roots and the leaf length, the leaf width and the dry weight of stem and leaves. And also, significant correlations were obtained between the ratio of root dry weight to root fresh weight and the stem length, the leaf length, the leaf width and the dry weight of stem and leaves.

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Gadolinium-Enhanced Magnetic Resonance Imaging of Atherosclerotic Plaques in Comparison with Histopathology: An In Vivo Study in Aorta of Rabbits (조직병리와 비교한 죽상경화반의 가돌리니움 조영증강 자기공명영상: 토끼 대동맥을 이용한 생체 실험)

  • Choi, Byoung-Wook;Hur, Jin;Lee, Hye-Jeong;Kim, Young-Jin;Kim, Tae-Hoon;Choe, Kyu-Ok
    • Investigative Magnetic Resonance Imaging
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    • v.13 no.1
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    • pp.81-87
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    • 2009
  • Purpose : We sought to evaluate enhancement of plaque with gadolinium-based contrast agent by magnetic resonance imaging (MRI) in comparison with histopathology, namely lipid-rich and macrophage-rich components that were two representative characteristics of plaque vulnerability using atherosclerotic rabbit aorta in order to determine which histopathologic component is relevant to the enhancement. Materials and Methods : New Zealand white rabbit (n=4, weight 3.0 to 3.5 kg, all male) was used for animal model of atherosclerosis. Atherosclerotic aortic lesions were induced by high-cholesterol diet and double balloon injury. T1-weight axial images were acquired before and after gadolinium-based contrast agent using a 3-T MRI. MR images and the matched histopathological sections (n=35) were divided into 4 quadrants or 3 (n=130). Enhancement ratio (ER, ER=SIpost/SIpre) on MRI was calculated for each quadrant and compared with histopathology in regard to lipid-rich and macrophage-rich areas. Results : Lipid-rich quadrants were 72 and fibrous quadrants were 58. The number of quadrants which had macrophage-rich areas was 105 and that of quadrants which did not have macrophage-rich areas was 25. ER was significantly higher in lipid-rich quadrants than in fibrous quadrants (mean ER 2.25c$\pm$0.41 vs. 2.72$\pm$0.65, p=0.013). ER poorly correlated with macrophage-rich areas when lipid-component was controlled (correlation coefficient -0.203, p=0.236). Conclusion : Lipid-rich plaques showed stronger enhancement than fibrous plaques using a standard gadolinium-based extracellular contrast agent. Macrophage infiltration did not correlate with degree of enhancement. Further study is warranted that account for optimal time of imaging after contrast injection using various plaque models from early to advanced stages and all possible parameters associated with contrast enhancement.

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Anti-inflammatory Effects of Pentoxifylline and Neutrophil Elastase Inhibitor on Lipopolysaccharide-Induced Acute Lung Injury In Vitro (In Vitro 내독소 유도성 급성 폐손상에서 Pentoxifylline과 Neutrophil Elastase Inhibitor의 항염효과)

  • Kim, Young-Kyoon;Kim, Seung-Joon;Park, Yong-Keun;Kim, Seok-Chan;Kim, Kwan-Hyoung;Moon, Hwa-Sik;Song, Jeong-Sup;Park, Sung-Hak;Kim, Sang-Ho
    • Tuberculosis and Respiratory Diseases
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    • v.49 no.6
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    • pp.691-702
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    • 2000
  • Background : Acute lung injury (ALI) is a commonly encountered respiratory disease and its prognosis is poor when the treatment is not provided promptly and properly. However no specific pharmacologic treatment is currently available for ALI, although recently several supportive drugs have been under scrutiny. We studied anti-inflammatory effects of pentoxifylline (PF), a methylated xanthine, and ONO-5046, a synthetic neutrophil elastase inhibitor on lipopolysaccharide (LPS)-induced ALI in vitro. Methods : To establish an in vitro model of LPS-induced ALI, primary rat alveolar macrophages and peripheral neutrophils in various ratios (1:0, 5:1, 1:1, 1:5, 0:1) were co-cultured with transformed rat alveolar epithelial cells (L2 cell line) or vascular endothelial cells (IP2-E4 cell line) under LPS stimulation. Each experiment was divided into five groups-control, LPS, LPS+PF, LPS+ONO, and LPS+PF+ONO. We compared LPS-induced superoxide anion productions from primary rat alveolar macrophages and peripheral neutrophils in various ratios, and the resultant cytotoxicity on L2 cells or IP2-E4 cells between groups. In addition we also compared the productions of tumor necrosis factor (TNF)-$\alpha$ interleukin (IL)-$1{\beta}$, monocyte chemotactic protein(MCP)-1, IL-6, and IL-10 as well as mRNA expressions of TNF-$\alpha$ inducible nitric oxide synthetase(iNOS), and MCP-1 from LPS-stimulated primary rat alveolar macrophages between groups. Results : (1) PF and ONO-5046 in each or both showed a trend to suppress LPS-induced superoxide anion productions from primary rat alveolar macrophages and peripheral neutrophils regardless of their ratio, except for the LPS+PF+ONO group with the 1:5 ratio, although statistical significance was limited to a few selected experimental conditions. (2) PF and ONO-5046 in each or both showed a trend to prevent IP2-E4 cells from LPS-induced cytotoxicity by primary rat alveolar macrophages and peripheral neutrophils regardless their ratio, although statistical significance was limited to a few selected experimental conditions. the effects of PF and/or ONO-5046 on LPS-induced L2 cell cytotoxicity varied according to experimental conditions. (3) PF showed a trend to inhibit LPS-induced productions of INF-$\alpha$ MCP-1, and IL-10 from primary rat alveolar macrophages. ONO-5046 alone didnot affect the LPS-induced productions of proinflammatory cytokines from primary rat alveolar macrophages but the combination of PF and ONO-5046 showed a trend to suppress LPS-induced productions of INF-$\alpha$ and IL-10 PF and ONO-5046 in each or both showed a trend to increase LPS-induced IL-$\beta$ and IL-6 productions from primary rat alveolar macrophages. (4) PF and ONO-5046 in each or both showed a trend to attenuate LPS-induced mRNA expressions of TNF-$\alpha$ and MCP-1 from primary rat alveolar macrophages but at the same time showed a trend increase iNOS mRNA expression. Conclusion : These results suggest that PF and ONO-5046 may play a role in attenuating inflammation in LPS-induced ALI and that further study is needed to use these drugs as a new supportive therapeutic strategy for ALI.

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