• Title/Summary/Keyword: Nitric oxide synthase 3

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Anti-inflammatory Activity of Sargassum micracanthum Water Extract (잔가시 물 추출물의 항염증 효과)

  • Jeong, Da Hyun;Kang, Bo Kyeong;Kim, Koth Bong Woo Ri;Kim, Min Ji;Ahn, Dong Hyun
    • Journal of Applied Biological Chemistry
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    • v.57 no.3
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    • pp.227-234
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    • 2014
  • The anti-inflammatory effect of Sargassum micracanthum water extract (SMWE) was investigated using lipopolysaccharide (LPS)-induced inflammatory response in this study. The murine macrophage cell line RAW 264.7 cells were used and MTT assay was performed to measure the cell proliferation ability. The secretion of nitric oxide (NO), tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), interleukin-6 (IL-6), and IL-$1{\beta}$ was measured in LPS-induced RAW 264.7 cells by ELISA. The expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and nuclear transcription factor-kappa B p65 protein was studied by immunoblotting. The Balb/c mice were used for an acute toxicity test, and imprinting control region mice were purchased to evaluate a croton oil-induced ear edema. As a result, there was no cytotoxicity in the macrophage proliferation treated with SMWE compared to the control. NO levels decreased with increasing concentration of SMWE and were inhibited over 50%. Moreover, the secretion of IL-6, TNF-${\alpha}$, and IL-$1{\beta}$ was suppressed in a dose-dependent manner, especially, IL-$1{\beta}$ inhibition activity was over 50% at 50 ${mu}g$/mL. The formation of ear edema of mice was reduced at the highest dose tested compared to that in the control. Moreover, in acute toxicity test, no moralities occurred in mice administered 5,000 mg/kg body weight of SMWE over 2 weeks observation period. These results suggested that SMWE may have significant effects on inflammatory factors and be potential anti-inflammatory therapeutic materials.

Anti-Inflammatory Effect of Wheat Germ Oil on Lipopolysaccharide-stimulated RAW 264.7 Cells and Mouse Ear Edema (LPS로 유도한 RAW 264.7 세포 및 귀부종 동물 모델에 대한 밀배아유의 항염증 효과)

  • Kang, Bo-Kyeong;Kim, Min-Ji;Jeong, Da-Hyun;Kim, Koth-Bong-Woo-Ri;Bae, Nan-Young;Park, Ji-Hye;Park, Sun-Hee;Ahn, Dong-Hyun
    • Microbiology and Biotechnology Letters
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    • v.44 no.3
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    • pp.236-245
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    • 2016
  • This study investigated the anti-inflammatory effects of wheat germ oil (WGO) on RAW 264.7 cells. It was shown that WGO had no cytotoxicity against the treated cells or negative effect on their proliferation. WGO suppressed nitric oxide (NO) secretion considerably and had inhibitory effects on the production of LPS-induced NO and pro-inflammatory cytokines (IL-6, TNF-α, and IL-1β). In particular, the IL-6 and TNF-α inhibition activities were over 90% at 100 μg/ml concentration of the oil. WGO also inhibited the LPS-induced expression of cyclooxygenase-2, inducible nitric oxide synthase, and nuclear factor-kappa B (NF-κB), and reduced the expression of phosphorylated ERK and JNK. Moreover, the croton-oil-induced edema in mouse ears was reduced by WGO, and no mortalities occurred in mice administered 5,000 mg/kg body weight of WGO over a 2-week observation period. In conclusion, these results provide evidence for the anti-inflammatory effect of WGO that likely occurs via modulation of NF-κB and the JNK/ERK MAPK signaling pathway.

Anti-Inflammatory Effect of Chondrus ocellatus Holmes Ethanol Extract on Lipopolysaccharide-induced Inflammatory Responses in RAW 264.7 Cells (Lipopolysaccharide로 유도된 RAW 264.7 세포와 마우스모델에 대한 진두발 에탄올 추출물의 항염증 효과)

  • Bae, Nan-Young;Kim, Min-Ji;Kim, Koth-Bong-Woo-Ri;Park, Ji-Hye;Park, Sun-Hee;Sung, Nak-Yun;Byun, Eui-Hong;Ahn, Dong-Hyun
    • Microbiology and Biotechnology Letters
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    • v.44 no.3
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    • pp.268-277
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    • 2016
  • This study aimed to investigate the anti-inflammatory effect of the ethanol extract from Chondrus ocellatus Holmes (COHEE) in RAW 264.7 cells and in a mouse ear edema model, by measuring the production of lipopolysaccharide-induced inflammatory response mediators. There were no cytotoxic effects on the proliferation of macrophages treated with COHEE compared with the control. COHEE inhibited the production of nitric oxide and pro-inflammatory cytokines [interleukin (IL)-6, tumor necrosis factor-α, and IL-1β]. The extract also reduced the expression of inducible nitric oxide synthase, cyclooxygenase-2, nuclear factor-κB p65, and phosphorylated mitogen-activated protein kinase in a dose-dependent manner. In the croton-oil-induced ear edema model, COHEE decreased the formation of mouse ear edema at the highest dose compared with the control, and histological analysis revealed that the epidermal/dermal tissue thickness and mast cell numbers were reduced. Therefore, these results suggest that COHEE may be a promising topical anti-inflammatory therapeutic material through its action of modulating NF-κB and the MAPK signaling pathway.

Anti-oxidative and Anti-inflammatory Activities of Fermented Turmeric (Curcuma longa L.) by Rhizopus oryzae (Rhizopus oryzae으로 발효한 울금의 항산화 및 항염효과)

  • Kim, Eun-Ju;Song, Bit-Na;Jeong, Da-Som;Kim, So-Young;Cho, Yong-Sik;Park, Shin-Young
    • Journal of Life Science
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    • v.27 no.11
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    • pp.1315-1323
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    • 2017
  • Turmeric is a rhizomatous herbaceous perennial plant (Curcuma longa (CL)) of the ginger family, Zingiberaceae. A yellow-pigmented fraction isolated from the rhizomes of CL contains curcuminoids belonging to the dicinnamoyl methane group. Curcumin is an important active ingredient responsible for the biological activity of CL. However, CL is not usually used as a food source due to its bitter taste. The present study was designed to determine the effect of the CL fermented by Rhizopus oryzae (FCL) on pro-inflammatory factors such as nuclear factor ${\kappa}B$ ($NF-{\kappa}B$), tumor necrosis factor alpha ($TNF-{\alpha}$), interleukin-6 (IL-6), nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$), inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in lipopolysaccharide (LPS)-induced RAW 264.7 cell line. The cell viability was determined by MTT assay. To evaluate the anti-inflammatory effect of FCL 80% EtOH extracts, IL-6 and $TNF-{\alpha}$ were measured by ELISA kit. Also, the amount of $NO/PGE_2/NF-{\kappa}B$ was measured using the $NO/PGE_2/NF-{\kappa}B$ detection kit and the iNOS/COX-2 expression was measured by Western blotting. The results showed that the FCL reduced NO, $PGE_2$, iNOS, COX-2, $NF-{\kappa}B$, IL-6 and $TNF-{\alpha}$ production without cytotoxicity. These results suggest that FCL extracts may be a developed the functional food related to anti-inflammation due to the significant effects on inflammatory factors.

Immunomodulating activity of Sargassum horneri extracts in RAW264.7 macrophages (RAW264.7 대식세포에서 괭생이 모자반 추출물의 면역활성 증진 효과)

  • Kim, Dong-Sub;Sung, Nak-Yun;Park, Sang-Yun;Kim, Geon;Eom, Ji;Yoo, Jin-Gon;Seo, In-Ra;Han, In-Jun;Cho, Young-Baik;Kim, Kyung-Ah
    • Journal of Nutrition and Health
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    • v.51 no.6
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    • pp.507-514
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    • 2018
  • Purpose: Sargassum horneri (S. horneri) is a species of brown macroalgae that is common along the coast of Japan and Korea. The present study investigated the immuno-modulatory effects of different types of S. horneri extracts in RAW264.7 macrophages. Methods: S. horneri was extracted by three different methods, hot water extraction, 50% ethanol extraction, and supercritical fluid extraction. Cell viability was then measured by MTT assay, while the production levels of tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), interleukin-6 (IL-6), and nitric oxide (NO) were measured by enzyme-linked immunosorbent assay and Griess assay, respectively. The expression and activation levels of inducible NO synthase (iNOS), mitogen-activated protein kinase (MAPK) and nuclear factor ${\kappa}B$ ($NF-{\kappa}B$) were examined by western blot analysis. Results: The three different S. horneri extracts were nontoxic against RAW 264.7 cells up to $50{\mu}g/mL$, among which treatment with hot water extract (HWE) of S. horneri significantly enhanced the production of TNF-${\alpha}$, IL-6, and NO in a dose-dependent manner. Hot water extract of S. horneri also increased the expression level of iNOS, suggesting that up-regulation of iNOS expression by HWE of S. horneri was responsible for the induction of NO production. In addition, treatment of RAW 264.7 macrophages with HWE of S. horneri increased the phosphorylation levels of ERK, p38 and JNK. Furthermore, the activation and subsequent nuclear translocation of $NF-{\kappa}B$ was enhanced upon treatment with HWE of S. horneri, indicating that HWE of S. horneri activates macrophages to secrete TNF-${\alpha}$, IL-6 and NO and induces iNOS expression via activation of the $NF-{\kappa}B$ and MAPKs signaling pathways. Conclusion: Taken together, these findings suggest that HWE of S. horneri possesses potential as a functional food with immunomodulatory activity.

Two new triterpenoid saponins derived from the leaves of Panax ginseng and their antiinflammatory activity

  • Li, Fu;Cao, Yufeng;Luo, Yanyan;Liu, Tingwu;Yan, Guilong;Chen, Liang;Ji, Lilian;Wang, Lun;Chen, Bin;Yaseen, Aftab;Khan, Ashfaq A.;Zhang, Guolin;Jiang, Yunyao;Liu, Jianxun;Wang, Gongcheng;Wang, Ming-Kui;Hu, Weicheng
    • Journal of Ginseng Research
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    • v.43 no.4
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    • pp.600-605
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    • 2019
  • Background: The leaves and roots of Panax ginseng are rich in ginsenosides. However, the chemical compositions of the leaves and roots of P. ginseng differ, resulting in different medicinal functions. In recent years, the aerial parts of members of the Panax genus have received great attention from natural product chemists as producers of bioactive ginsenosides. The aim of this study was the isolation and structural elucidation of novel, minor ginsenosides in the leaves of P. ginseng and evaluation of their antiinflammatory activity in vitro. Methods: Various chromatographic techniques were applied to obtain pure individual compounds, and their structures were determined by nuclear magnetic resonance and high-resolution mass spectrometry, as well as chemical methods. The antiinflammatory effect of the new compounds was evaluated on lipopolysaccharide-stimulated RAW 264.7 cells. Results and conclusions: Two novel, minor triterpenoid saponins, ginsenoside $LS_1$ (1) and 5,6-didehydroginsenoside $Rg_3$ (2), were isolated from the leaves of P. ginseng. The isolated compounds 1 and 2 were assayed for their inhibitory effect on nitric oxide production in LPS-stimulated RAW 264.7 cells, and Compound 2 showed a significant inhibitory effect with $IC_{50}$ of $37.38{\mu}M$ compared with that of NG-monomethyl-L-arginine ($IC_{50}=90.76{\mu}M$). Moreover, Compound 2 significantly decreased secretion of cytokines such as prostaglandin $E_2$ and tumor necrosis factor-${\alpha}$. In addition, Compound 2 significantly suppressed protein expression of inducible nitric oxide synthase and cyclooxygenase-2. These results suggested that Compound 2 could be used as a valuable candidate for medicinal use or functional food, and the mechanism is warranted for further exploration.

Anti-inflammatory Effect of Ethanol Extract from Eupatorium japonicum (등골나물 추출물의 항염증 효과)

  • Lee, Han-Na;Lim, Do-Young;Lim, Soon-Sung;Kim, Jong-Dai;Yoon, Jung-Han
    • Korean Journal of Food Science and Technology
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    • v.43 no.1
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    • pp.65-71
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    • 2011
  • Eupatorium japonicum belongs to a family of Asteraceae plants and flowers of E. japonicum have been consumed as a tea. In this study, we investigated whether E. japonicum extract inhibits lipopolysaccharide (LPS)-induced inflammatory responses in Raw264.7 macrophages. The cells were treated with various concentrations (0, 1, 2.5, 5, or 10 mg/L) of 70% ethanol extract from E. japonicum flowers (EJE) in Raw264.7 cells. LPS-induced nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) production were inhibited by EJE up to 67% and 49% of these productions, respectively without any reduction of viable cell numbers. EJE reduced LPS-induced expression of inducible NO synthase (iNOS) and cyclooxygenase (COX)-2 proteins and their corresponding mRNA levels. Additionally, EJE decreased the levels of interleukin (IL)-6, IL-1${\beta}$, and tumor necrosis factor (TNF)-${\alpha}$ mRNA. EJE was further fractionated with water, butanol, ethylacetate (EA), hexane, or methylene chloride (MC). Among the resulting five fractions, EA and MC, respectively from EJE significantly inhibited LPS-induced NO production (each inhibition rate was 85.3% of 10 mg/L EA fraction and 97.2% of 10 mg/L MC fraction) without significant cytotoxicity in Raw264.7 cells. These results indicate that EJE exhibits powerful effects of anti-inflammation and can be developed as a potential anti-inflammatory agent.

Effect of acupuncture treatment on carrageenan-induced inflammation and NADPH-diaphorase reactivity in the hypothalamus of Stat4 knockout mice (Carrageenan 염증 유도된 Stat 4 유전자 제거생쥐의 시상하부에서 NADPH-diaphorase 반응에 대한 침의 영향)

  • Hong, Mee-Suk;Kim, Mi-Ja;Kim, Jin-Ju;Park, Hi-Joon;Chung, Joo-Ho
    • Korean Journal of Acupuncture
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    • v.24 no.4
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    • pp.221-231
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    • 2007
  • 목 적 : 본 연구의 목적은 시상하부에서 침처치에 대한 nitric oxide synthase (NOS)발현을 nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d)를 이용한 조직화학 염색법으로 관찰하였다. 실험방법 : 동물은 Balb/c (wild type) 와 Stat 4 knockout (KO) 생쥐를 사용하였다. 염증유도는 1% carrageenan 용액 (20ul/마리)을 발 뒤꿈치 표피에 주사하였고, 침 처치는 족삼리 (ST36)에 시침하였다. 침 처치 후 5시간까지 부종율을 부종측정기로 측정하였으며, 마지막으로 부종을 측정한 후 동물을 희생하여 뇌를 적출하여 고정하였다. 침에 대한 효과를 확인하기 위하여 NADPH-d 반응의 조직염색을 실시하였다. 염증유도와 그룹간의 유의성 검증은 one-way ANOVA를 사용하였다. 결 과 : 대조군인 Balb/c와 실험군인 stat4 KO 생쥐를 carrageenan으로 염증을 유도시에 대조군은 90%이상 유도된 반면, Stat4 KO 그룹은 50% 정도의 염증만이 유도되었다. 염증을 유도한 생쥐의 족삼리에 침 처치시 대조군은 1시간에서 약 40%정도 감소하였고 (P<0.05), Stat4 KO 실험군은 유의한 염증 감소율을 보이지 않았다. 시상하부의 lateral hypothalamic area (LHA)와 paraventricular nucleus (PVN)부위의 침에 대한 효과를 NADPH-d 에 양성으로 반응하는 세포수로 비교하여 다음과 같은 결과를 얻었다. (1) 대조군에서 염증 유도시 시상하부의 PVN는 NADPH-d 양성세포수가 감소하였고, LHA에서는 증가하였다. (2) 염증을 유도한 대조군에 침을 처치시 PVN은 세포수가 증가하였고, LHA에서는 감소하는 경향을 보였다. (3) 염증을 유도한 Stat4 KO 군에서는 시상하부의 PVN과 LHA부위 모두에서 NADPH-d 양성세포수가 감소하였고, 염증유도그룹에 침을 처치시 PVN과 LHA부위 모두에서 세포수가 증가함을 관찰 할 수 있었다. (4) 대조군과 실험군 모두에 salicylic acid로 비교하였더니 염증유도 효과 및 NADPH-d 세포 수에서 침 처치와 비슷한 결과를 나타내었다. 결 론 : 침은 염증을 유도한 생쥐에서 염증 감소에 유의한 효과가 있다. 염증을 유도한 Balb/c 와 Stat4 KO 생쥐에 침을 처치 시 시상하부의 NADPH-d 발현이 LHA부위와 PVN에서 서로 다르게 나타나는 것으로 나타난다. 이러한 현상은 침 효과가 시상하부의 위치에 대한 작용이 다르기 때문이라고 생각된다.

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Effect of Sunghyangchungisan on Contractile Reactivity and $Ca^{2+}$ metabolism in Isolated Rabbit Carotid Artery (성향정기산(星香正氣散)이 가토의 경동맥(頸動脈) 평활근(平滑筋) 긴장(緊張) 및 $Ca^{2+}$ 대사(代謝)에 미치는 영향(影響))

  • Kim, Young-Gyun;Kweon, Jung-Nam;Kim, Jong-Hoon
    • The Journal of Internal Korean Medicine
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    • v.21 no.3
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    • pp.377-388
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    • 2000
  • Objective : This study was undertaken to evaluate the effect of Sunghyangchungisan (SHCS) on the regulation of vascular tone and $Ca^{2+}$ metabolism in arterial tissues. Vascular rings isolated from rabbit carotid artery were myographed isometrically in isolated organ baths and the effect of SHCS on contractile activities, endothelial function and $Ca^{2+}$ metabolism were determined. Methods : In phentobarbital sodium-anesthetized rabbits, SHCS administered through ear vein (100 mg/Kg body wt.) or intragastric dwelling tube (300 mg/Kg body wt.) attenuated phenylephrine (PE, 10 ${\mu}g$/Kg, i.v.)-induced increases in both systolic and diastolic cartoid arterial blood pressure. Results : In experiments with isolated arterial strips, SHCS relaxed arterial rings which were pre-contracted by phenylephrine (PE, 1 ${\mu}M$). The responses to SHCS were partially dose-dependent at concentrations lower than 0.5 mg/ml. When SHCS was applied prior to the exposure to PE, it inhibited the PE-induced contraction by a similar magnitude which was comparable to the relaxation of pre-contracted arterial rings. Washout of SHCS after observing its relaxant effect resulted in a full recovery of PE-induced contractions, indicating that the action mechanism is reversible. The observation that SHCS did not change the $ED_{50)$ of PE oh its dose-response curve ruled out the possible interaction of SHCS with ${\alpha}$-receptors. The relaxant effect of SHCS was not affected by removal of endothelium or a nitric oxide synthase inhibitor, L-NAME. Methylene blue, an inhibitor of the soluble guanylate cyclase, did not affect the relaxant effect of SHCS. These results suggest that the action of SHCS is not mediated by the endothelium nor soluble guanylate cyclase. Constant cGMP production determined in arterial strips in the presence or absence of SHCS is consistent with this conclusion. When contraction was induced by additive application of $Ca^{2+}$ in arterial rings which were pre-depolarized by high $K^+$ in a $Ca^{2+}$-free solution, the relaxant effect of SHCS was attenuated by increasing the $Ca^{2+}$ concentration. SHCS, when applied to the arterial rings pre-contracted by PE and then relaxed by nifedipine, a $Ca^{2+}$ channel blocker, did not show additive relaxation. SHCS partially blocked $Ca^{2+}$ influx stimulated by PE and high $K^+$ which was determined by 5-min ^{45}Ca$ uptake, while it did not affect $Ca^{2+}$ efflux. Conclusions : From above results, it is suggested that SHCS relax PE-induced contraction of rabbit carotid artery in an endothelium independent manner, andinhibition of $Ca^{2+}$ influx may contribute to the underling mechanism.

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KISS Korea Computer Congress 2007 (이동 객체의 패턴 탐사를 위한 시공간 데이터 일반화 기법)

  • Ko, Hyun;Kim, Kwang-Jong;Lee, Yon-Sik
    • Proceedings of the Korean Information Science Society Conference
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    • 2007.06c
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    • pp.153-158
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    • 2007
  • 사용자들의 특성에 맞게 개인화되고 세분화된 위치 기반 서비스를 제공하기 위해서는 방대한 이동 객체의 위치 이력 데이터 집합으로부터 유용한 패턴을 추출하여 의미 있는 지식을 탐사하기 위한 시공간 패턴 탐사가 필요하다. 현재까지 다양한 패턴 탐사 기법들이 제안되었으나 이동 패턴들 중 단순히 시공간 제약이 없는 빈발 패턴만을 추출하기 때문에 한정된 시간 범위와 제한적인 영역 범위 내에서의 빈발 패턴을 탐사하는 문제에는 적용하기 어렵다. 또한 패턴 탐사 수행 시 데이터베이스를 반복 스캔하여 탐사 수행시간이 많이 소요되는 문제를 포함하거나 메모리상에 탐사 대상인 후보 패턴 트리를 생성하는 방법을 통해 탐사 시간을 줄일 수는 있으나 이동 객체 수나 최소지지도 등에 따라 트리를 구성하고 유지하는데 드는 비용이 커질 수 있다. 따라서 이러한 문제를 해결하기 위한 효율적인 패턴 탐사 기법의 개발이 요구됨으로써 선행 작업으로 본 논문에서는 상세 수준의 객체 이력 데이터들의 시간 및 공간 속성을 의미 있는 시간영역과 공간영역 정보로 변환하는 시공간 데이터 일반화 방법을 제안한다. 제안된 방법은 공간 개념 계층에 대한 영역 정보들을 영역 Grid 해쉬 테이블(AGHT:Area Grid Hash Table)로 생성하여 공간 인덱스트리인 R*-Tree의 검색 방법을 이용해 이동 객체의 위치 속성을 2차원 공간영역으로 일반화하고, 시간 개념 계층을 생성하여 이동 객체의 시간적인 속성을 시간 영역으로 일반화함으로써 일반화된 데이터 집합을 형성하여 효율적인 이동 객체의 시간 패턴 마이닝을 유도할 수 있다.의 성능을 기대할 수 있을 것이다.onium sulfate첨가배지(添加培地)에서 가장 저조(低調)하였다. vitamin중(中)에서는 niacin과 thiamine첨가배지(添加培地)에서 근소(僅少)한 증가(增加)를 나타내었다.소시켜 항이뇨 및 Na 배설 감소를 초래하는 작용과, 둘째는 신경 경로를 통하지 않고, 아마도 humoral factor를 통하여 신세뇨관에서 Na 재흡수를 억제하는 작용이 복합적으로 나타내는 것을 알 수 있었다.으로 초래되는 복합적인 기전으로 추정되었다., 소형과와 기형과는 S-3에서 많이 나왔다. 이상 연구결과에서 입도분포가 1.2-5mm인 것이 바람직한 것으로 나타났다.omopolysaccharides로 확인되었다. EPS 생성량이 가장 좋은 Leu. kimchii GJ2의 평균 분자량은 360,606 Da이었으며, 나머지 두 균주에 대해서는 생성 EPS 형태와 점도의 차이로 미루어 보아 생성 EPS의 분자구조와 분자량이 서로 다른 것으로 판단하였다.TEX>개로 통계학적으로 유의한 차이가 없었다. Heat shock protein-70 (HSP70)과 neuronal nitric oxide synthase (nNOS)에 대한 면역조직화학검사에서 실험군 Cs2군의 신경세포가 대조군 12군에 비해 HSP70과 nNOS의 과발현을 보였으며, 이는 통계학적으로 유의한 차이를 보였다(p<0.05). nNOS와 HSP70의 발현은 강한 연관성을 보였고(상관계수 0.91, p=0.000), nNOS를 발현하는 세포가 동시에 HSP70도 발현함을 확인할 수 있었다. 결론: 우리는

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