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Characterization of Cigarette Smoke Extract (CSE)-induced Cell Death in Lung Epithelial Cells (폐상피세포에서 흡연추출물-유도성 세포사에 관한 연구)

  • Choi, Eun Kyung;Kim, Yun Seup;Park, Jae Seuk;Jee, Young Koo;Lee, Kye Young
    • Tuberculosis and Respiratory Diseases
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    • v.58 no.1
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    • pp.43-53
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    • 2005
  • Emphysema is characterized by air space enlargement and alveolar destruction. The mechanism responsible for the development of emphysema was thought to be protease/antiprotease imbalance and oxidative stress. A very recent study shows that alveolar cell apoptosis causes lung destruction and emphysematous changes. Thus, this study was performed to support the evidence for the role of apoptosis in the development of emphysema by characterizing cigarette smoke extract (CSE)-induced apoptosis in A549 (type II pneumocyte) lung epithelial cells. CSE induced apoptosis at low concentration (10% or less) and both apoptosis and necrosis at high concentration (20%). Apoptosis was demonstrated by DNA fragmentation using FACScan for subG1 fraction. Discrimination between apoptosis and necrosis was done by morphologic analysis using fluorescent microscopy with Hoecst 33342/propium iodide double staing and electron microscopy. Cytochrome c release was confirmed by using immunofluorescence with monoclonal anti-cytochrome c antibody. However, CSE-induced cell death did not show the activation of caspase 3 and was not blocked by caspase inhibitors. This suggests that CSE-induced apoptosis might be caspase-independent apoptosis. CSE-induced cell death was near completely blocked by N-acetylcystein and bcl-2 overexpression protected CSE-induced cell death. This results suggests that CSE might induce apoptosis through intracellular oxidative stress. CSE also activated p53 and functional knock-out of p53 using stable overexpression of HPV-E6 protein inhibited CSE-induced cell death. The characterization of CSE-induced cell death in lung epithelial cells could support the role of lung cell apoptosis in the pathogenesis of emphysema.

Anti-inflammatory Activity of Peel Extracts in Color-fleshed Potatoes (컬러감자외피 추출물의 항염활성)

  • Nam, Jung-Hwan;Jeong, Jin-Cheol;Kwon, Oh-Keun;Hong, Su-Young;Kim, Su-Jeong;Soh, Hwang-Bae;Lee, Jong-Nam;Lee, Kyung-Tea;Park, Hee-Jhun
    • Korean Journal of Plant Resources
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    • v.26 no.5
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    • pp.533-538
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    • 2013
  • Potatoes were first introduced outside the Andes region four centuries ago, and have become an integral part of much of the world's food. Potatoes were first introduced into Europe in the 16th century and Korea in the early 19th century. In the nutritional aspects, potatoes contain abundant vitamins and minerals, as well as an assortment of phytochemicals such as carotenoids and natural phenols. Chlorogenic acid constitutes up to 90% of potato natural phenols. Due to the high content of potato functional compounds, it has known that potatoes are effective in the prevention of various human diseases. Recently, color-fleshed potatoes 'Hongyoung' and 'Jayoung' were developed by RDA, and it has reported that they have high content of anthocyanin. Additionally they show higher radical scavenging activity compared to white or yellow fleshed potatoes. So it will be expected that the consumption of color-fleshed potatoes grandually increase by pre-peeled potatoes and color potato chips. This study was conducted to enhance the peel of color-fleshed potatoes utilization and to determine the biological activity of peel of color-fleshed potatoes extract. The anti-inflammatory effects on ethanol extract and its solvent fraction were also evaluated. The anti-inflammatory activities of $CHCl_3$ fraction was evaluated for inhibitory activities against lipopolysacchride(LPS) induced nitric oxide(NO) and prostaglandin $E_2(PGE_2)$ production as well as inducible nitric oxide synthase(iNOS) and cyclo oxygenase-2(COX-2) protein expressions in RAW264.7 cell lines. The fraction inhibitory activity for both tests with $IC_{50}$ values showed in the ranges of $25{\sim}50{\mu}g/ml$. This result revealed that $CHCl_3$ fraction of Jayoung's peel is expected to be good candidate for development into source of anti-inflammatory agent.

Effect of Feeding Prunus mume By-products on Productivity and Blood Composition in Semi-Broiler Chicks (매실 부산물 급여가 준육용계의 생산 능력과 혈액 성상에 미치는 영향)

  • Jeong, Y.D.;Cho, I.K.
    • Korean Journal of Poultry Science
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    • v.34 no.4
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    • pp.237-243
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    • 2007
  • The objective of this experiment was to investigate the feeding influence of Prunus memu by-products (PMB) on productivity, blood composition, meat quality and intestinal microflora in semi-broiler chicks. Four hundred one day old chicks(Hanhyup 3) were raised in the floor pen with five treatments(0, 0.5, 1.0 2.0 3.0%) of four replicate for ten weeks. Basal diets contained 3,000, 3,100, 3,100, 3,200 kcal/kg ME and 22, 21, 19, 17% CP, respectively. There were no different performance for the first two weeks, but showed significantly different weight gain, feed intake in PMB 2.0, 3.0% addition treatments compared with control from three to five weeks of age. Weight gain of birds fed 2.0% PMB was significantly higher from nine to 10 weeks of age than control(P<0.05). Total weight gain tended to increase in PMB 1.0, 2.0, 3.0% addition treatments. Feed intake also showed increase in PMB 2.0, 3.0% addition treatments, but there were no different feed conversion. Total protein, albumin, cholesterol, neutral fat, glucose was significantly decreased in PMB 0.5% addition treatments compared with control. There were no different physico-chemical characteristics in breast meat. The number of yeast was significantly improved in PMB 3.0% compared with control(P<0.05), but found no difference in number of E. coli between control and PMB treatments. ND titer of birds fed PMB was not statistically different, but tended to decrease as dietary PMB increased. As the result of this experiment, PMB would be available as a feed additives in semi-broiler chicks.

Anti-inflammatory effect of Polygonum multiflorum extraction in activated RAW 264.7 cells with lipopolysaccharide (Lipopolysaccharide로 활성화된 RAW 264.7 세포에서 적하수오(Polygonum multiflorum) 추출물의 항염증 효과 검증)

  • Lee, Eunsu;Kim, Hyeongjeong;Yu, Jae-Myo;Cho, Yong-Hun;Kim, Dong-In;Shin, Yuhyeon;Cho, Yeongje;Kwon, O-Jun;An, Bongjeon
    • Food Science and Preservation
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    • v.21 no.5
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    • pp.740-746
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    • 2014
  • The anti-inflammatory effects of Polygonum multiflorum water extracts (PMWs) and Polygonum multiflorum 70 % ethanol extracts(PMEs) were investigated using lipopolysaccharide-induce by inflammatory response. The inhibitory effects of PMWs and PMEs on the production of nitric oxide (NO) and pro - inflammatory cytokines in LPS - activated Raw 264.7 cells were investigated. The effects were examined after reducing production of Nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$), interleukin-6 (IL-6), tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), interleukin-$1{\beta}$ (IL-$1{\beta}$), nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) protein levels. RAW 264.7 cells were cultured with LPS ($1{\mu}g/mL$) in the presence or absence of PMWs and PMEs for 24 h to determine their NO, iNOS, COX-2 levels. During the entire experimental period 10, 25, 50 and $100{\mu}g/mL$ of PMWs and PMEs showed no cytotoxicity. At these concentrations, PMWs and PMEs concentration dependently reduced the production of nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$), interleukin-6 (IL-6), tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) and interleukin-$1{\beta}$ (IL-$1{\beta}$). PMWs and PMEs were inhibited the activittion of iNOS, COX-2 by 89%, 54%, 91% and 57% respectively, at $100{\mu}g/mL$. These results indicate that PMWs and PMEs significantly reduces the effect of oxidative and inflammatory cytokines.

Hepatic Cirrhosis Secondary to chronic Hepatitis in an English Cocker Spaniel (ECS) Dog (잉글리쉬 코커스파니엘 견에서 발생한 만성 간염 및 간경화 증례)

  • Park Chul;Yoo Jong-Hyun;Jung Dong-In;Kim Ha-Jung;Kang Byeong-Teck;Lim Chae-Young;Yoon Hun-Young;Jeong Soon-Wuk;Sur Jung-Hyang;Park Hee-Myung
    • Journal of Veterinary Clinics
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    • v.23 no.1
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    • pp.72-76
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    • 2006
  • A 1-year-old, female English cocker spaniel (ECS) dog was presented with 3-month history of vomiting and retaking of the vomitus, and chronic weight loss. The client had noticed mild abdominal distension 10 days before. The dog was diagnosed as chronic hepatitis with hepatic cirrhosis based on complete blood count (CBC), serum chemistry profiles, radiography, ascites assessment, bile acid evaluation, and liver biopsy through exploratory laparotomy and necropsy. CBC and serum chemistry profiles revealed mild anemia, slightly elevated hepatic enzymes (ALT and AST), increased creatinine kinase (CK), hyperammonemia, and hypoproteinemia with hypoalbuminemia. Ascites was transudate according to analysis of components. Bile acid assessment (fasting; $174.4{\mu}mol/L$ and postprandial; $198.4{\mu}mol/L$) showed strongly suspected hepatic insufficiency. On radiological findings, ascites was evident. Atrophied liver (especially left side lobes) and distended mesenteric vasculatures were observed by exploratory laparotomy. Histopathological examination of marginal lesion of left lateral lobe of liver by biopsy revealed the necrosis of hepatic cells, dilation of sinusoids, infiltration of neutrophils in sinusoids, and vacuolation of hepatic cytoplasm. The patient had been managed with careful low protein diet and specific supportive therapy (ursodeoxycholic acid, prednisolone, vitamine E, and interferon). Vomiting and ascites disappeared with medical management. The dog was monitored periodically by CBC, serum chemistry and radiographic examination. The dog survived more 18 months with medical therapy. After spontaneous death, necropsy and histopathologic examination were performed.

Construction of Web-Based Database for Anisakis Research (고래회충 연구를 위한 웹기반 데이터베이스 구축)

  • Lee, Yong-Seok;Baek, Moon-Ki;Jo, Yong-Hun;Kang, Se-Won;Lee, Jae-Bong;Han, Yeon-Soo;Cha, Hee-Jae;Yu, Hak-Sun;Ock, Mee-Sun
    • Journal of Life Science
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    • v.20 no.3
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    • pp.411-415
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    • 2010
  • Anisakis simplex is one of the parasitic nematodes, and has a complex life cycle in crustaceans, fish, squid or whale. When people eat under-processed or raw fish, it causes anisakidosis and also plays a critical role in inducing serious allergic reactions in humans. However, no web-based database on A. simplex at the level of DNA or protein has been so far reported. In this context, we constructed a web-based database for Anisakis research. To build up the web-based database for Anisakis research, we proceeded with the following measures: First, sequences of order Ascaridida were downloaded and translated into the multifasta format which was stored as database for stand-alone BLAST. Second, all of the nucleotide and EST sequences were clustered and assembled. And EST sequences were translated into amino acid sequences for Nuclear Localization Signal prediction. In addition, we added the vector, E. coli, and repeat sequences into the database to confirm a potential contamination. The web-based database gave us several advantages. Only data that agrees with the nucleotide sequences directly related with the order Ascaridida can be found and retrieved when searching BLAST. It is also very convenient to confirm contamination when making the cDNA or genomic library from Anisakis. Furthermore, BLAST results on the Anisakis sequence information can be quickly accessed. Taken together, the Web-based database on A. simplex will be valuable in developing species specific PCR markers and in studying SNP in A. simplex-related researches in the future.

Comparison of Anti-inflammatory Activities among Ethanol Extracts of Sophora flavescens, Glycyrrhiza uralensis and Dictamnus dasycarpus, and their Mixtures in RAW 246.7 Murine Macrophages (RAW 246.7 대식세포 모델에서 고삼, 감초, 백선피 에탄올 추출물 및 추출복합물의 항염증 효능 비교)

  • Han, Min Ho;Lee, Moon Hee;Hong, Su Hyun;Choi, Yung Hyun;Moon, Ju Sung;Song, Myung Kyu;Kim, Min Ju;Shin, Su Jin;Hwang, Hye Jin
    • Journal of Life Science
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    • v.24 no.3
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    • pp.329-335
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    • 2014
  • Sophora flavescens, Glycyrrhiza uralensis and Dictamnus dasycarpus have been widely used in folk medicine for several inflammatory disorders in Korea and China. In this study, we compared the anti-inflammatory effects of the ethanol extracts of S. flavescens (EESF), G. uralensis (EEGU) and D. dasycarpus (EEDS), and their mixtures (medicinal herber mixtures, MHMIXs) on production of inflammatory mediators and cytokines in lipopolysaccharide (LPS)-stimulated RAW 264.7 murine macrophages. Our data indicated that treatment with EESF, EEGU and EEDD significantly inhibited the excessive production of pro-inflammatory mediators such as nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) in LPS-stimulated RAW 264.7 cells. The ethanol extracts and MHMIXs also attenuated the production of pro-inflammatory cytokines, including interleukin-$1{\beta}$ ($IL-1{\beta}$) and tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$) by suppressing their protein expression, respectively. Interestingly, MHMIX-1, which basic ingredients are EESF, EEGU and EEDS in the proportion 3:1:1, more safely and effectively inhibits the LPS-induced inflammatory status in LPS-stimulated RAW 264.7 macrophages compared to ethanol extracts of each medicinal herb and other MHMIXs without causing any cytotoxic effects. Our study provides scientific evidence to support that a berbal mixture, MHMIX-1 may be useful in the treatment of inflammatory diseases by inhibiting inflammatory regulator responses in activated macrophages.

Anti-Inflammatory Effects of Picrasma Quassioides (D.DON) BENN Leaves Extracts (소태나무 잎 추출물의 항염증 효과)

  • Jung, Yeon Seop;Eun, Cheong Su;Jung, Young Tae;Kim, Hyun Jeong;Yu, Mi Hee
    • Journal of Life Science
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    • v.23 no.5
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    • pp.629-636
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    • 2013
  • This study was performed to evaluate the anti-inflammatory and antioxidant activities of methanol extract from the leaves of Picrasma quassioides BENNET (PLME). The antioxidant effects of PLME were measured based on polyphenol and flavonoid contents. PLME was found to have $367.52{\mu}g/mg$ and $46.61{\mu}g/mg$ high polyphenol and flavonoid contents. Cell viability was determined by MTT assay. The production of nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) was measured by Griess assay and enzyme-linked immunosorbent assay (ELISA). In order to effectively anti-inflammatory agents, we examined the inhibitory effects on the production of lipopolysaccharide (LPS)-induced NO and $PGE_2$ in RAW 264.7 cells. PLME significantly decreased the production of NO and $PGE_2$ in a dose-dependent manner, and also reduced the expression of iNOS, a COX-2 protein. In addition, PLME reduced the NF-${\kappa}B$, $I{\kappa}B$ phosphorylation in RAW 264.7 cells upon stimulation with LPS (100 ng/ml) for 24 h. These results provide evidence for the anti-inflammatory and antioxidant effects of Picrasma quassioides leaves.

Anti-oxidative and Neuroprotective Activities of Pig Skin Gelatin Hydrolysates (돈피젤라틴 효소분해물의 항산화 활성 및 신경세포보호효과)

  • Kim, Dong Wook;Park, Kimoon;Ha, Goeun;Jung, Ju Ri;Chang, Ounki;Ham, Jun-Sang;Jeong, Seok-Geun;Park, Beom-Young;Song, Jin;Jang, Aera
    • Food Science of Animal Resources
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    • v.33 no.2
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    • pp.258-267
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    • 2013
  • This study was conducted to determine the antioxidative and neuroprotective effect of pig skin extracts (PS) and pig skin gelatin hydrolysates (LPS) using a human neuroblastoma cell line (SH-SY5Y). The extraction yield of PS was 3 fold higher than that of LPS. The protein content of PS was about 10 fold higher than that of LPS (p<0.05). Also LPS increased antioxidative activity dose dependently, and the activity was significantly higher than PS at all concentration (p<0.05). DPPH radical scavenging activity of LPS at 50 mg/mL was 92.97%, which was similar to $1{\mu}M$ vitamin C as a positive control. ABTS radical scavenging activity of LPS (20 mg/mL) was 89.83% and oxygen radical absorbance capacity of LPS at 1 mg/mL was $141.39{\mu}M$ Trolox Equvalent/g. No significant change of human neuroblastoma cells was determined by MTT test. Cell death by oxidative stress induced by $H_2O_2$ and amyloid beta 1-42 ($A{\beta}_{1-42}$) was protected by LPS rather than PS. Acetylcholine esterase was significantly inhibited, by up to 33.62% by LPS at 10 mg/mL. Therefore, these results suggest that pig skin gelatin hydrolysates below 3 kDa have potential to be used as anti-oxidative and neuroprotective functional additives in the food industry, while further animal test should be determined in the future.

The Effects of Prostaglandin and Dibutyryl cAMP on Osteoblastic Cell Activity and Osteoclast Generation (Prostaglandin과 Dibutyryl cAMP가 조골세포의 활성과 파골세포 형성에 미치는 영향)

  • Mok, Sung-Kyu;You, Hyung-Keun;Shin, Hyung-Shik
    • Journal of Periodontal and Implant Science
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    • v.26 no.2
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    • pp.448-468
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    • 1996
  • To maintain its functional integrity, bone is continuously remodelled by a process involving resorption by osteoeclasts and formation by osteoblasts, In order to respond to changes in the physical environment or to trauma with the relevant action, this process is strictly regulated by locally synthesized or systemic fators, Prostaglandin $E_2(PGE_2$) is perhaps one of the best studied factors, having been known to affect bone cell function for several decades.$PGE_2$ has both anabolic and catabolic activities. Excess of $PGE_2$ has been implicated in a number of pathological states associated with bone loss in a number of chronic inflammatory conditions such as periodontal disease and rheumatoid arthritis. $PGE_2$ and other arachidonic acid metabolites have been shown to be potent stimulators of osteoclastic bone resorption in organ culture. The anabolic effects of $PGE_2$ were first noticed when an increase in periosteal woven bone formation was seen after the infusion of $PGE_2$ into infants in order to prevent closure of the ductus arteriosus. The cellular basis for the catabolic actions of $PGE_2$ has been well characterized. $PGE_2$increases osteoclast recruitment in bone marrow cell cultures. Also $PGE_2$ has a direct action on osteoclast serving to inhibit activity and can also indirectly activate osteoclast via other cells in the vicinity, presumably osteoblast. The cellular mechanisms for the anabolic actions of $PGE_2$ are not nearly so well understood. The purpose of this paper was to study the effects of $PGE_2$ and dibutyl(DB)cAMP on osteoblastic clone MC3T3El cells and on the generation of osteoclasts from their precursor cells. The effect of $PGE_2$ and DBcAMP on the induction of alkaline phoaphatase(AlP) was investigated in osteoblastic clone MC3T3El cells cultured in medium containing 0.4% fetal bovine serum. $PGE_2$ and DBcAMP stimulated ALP activity and MTT assay in the cells in a dose-dependent manner at concentrations of lO-SOOng/ml. Cycloheximide, protein synthesis inhibitor, inhibited the stimulative effect of $PGE_2$ and DBcAMP on ALP activity in the cells. $PGE_2$also increased the intracellular cAMP content in a dose-dependent fashion with a maximal effect at 500ng/ml. The effect of $PGE_2$ on the generation of osteoclasts was investigated in a coculture system of mouse bone marrow cells with primary osteoblastic cells cultured in media containing 10% fetal bovine serum.After cultures, staining for tartrate-resistant acid phosphatase(TRAP)-marker enzyme of osteoclast was performed. The TRAP(+) multinucleated cells(MNCs), which have 3 or more nuclei, were counted. More TRAP(+) MNCs were formed in coculture system than in control group. $PGE_2(10^{-5}10^{-6}M)$ stimulated the formation of osteoclast cells from mouse bone marrow cells in culture. $PGE_2(10^{-6}M)$ stimulated the formation of osteoclast cells from mouse bone marrow cells in coculture of osteoblastic clone MC3T3E1 cells This results suggest that $PGE_2$ stimulates the differentiation of osteoblasts and generation of osteoclast, and are involved in bone formation, as well as in bone resorption.

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