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Neuroprotective Effects of Cheongnoemyeongsin-hwan against Hydrogen Peroxide-induced DNA Damage and Apoptosis in Human Neuronal-Derived SH-SY5Y Cells (인체 신경세포에서 청뇌명신환(淸腦明神丸)의 산화적 스트레스에 대한 세포보호 효과)

  • Pi, Guk Hyun;Hwang, Won Deuk
    • Herbal Formula Science
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    • v.25 no.1
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    • pp.51-68
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    • 2017
  • Objectives : Oxidative stress due to excessive accumulation of reactive oxygen species (ROS) is one of the risk factors for the development of several chronic diseases, including neurodegenerative diseases. Methods : In the present study, we investigated the protective effects of cheongnoemyeongsin-hwan (CNMSH) against oxidative stress‑induced cellular damage and elucidated the underlying mechanisms in neuronal-derived SH-SY5Y cells. Results : Our results revealed that treatment with CNMSH prior to hydrogen peroxide (H2O2) exposure significantly increased the SH-SY5Y cell viability, indicating that the exposure of the SH-SY5Y cells to CNMSH conferred a protective effect against oxidative stress. CNMSH also effectively attenuated H2O2‑induced comet tail formation, and decreased the phosphorylation levels of the histone ${\gamma}H2AX$, as well as the number of apoptotic bodies and Annexin V‑positive cells. In addition, CNMSH exhibited scavenging activity against intracellular ROS generation and restored the mitochondria membrane potential (MMP) loss that were induced by H2O2, suggesting that CNMSH prevents H2O2‑induced DNA damage and cell apoptosis. Moreover, H2O2 enhanced the cleavage of caspase-3 and degradation of poly (ADP-ribose)-polymerase, a typical substrate protein of activated caspase-3, as well as DNA fragmentation; however, these events were almost totally reversed by pretreatment with CNMSH. Furthermore, CNMSH increased the levels of heme oxygenase-1 (HO-1), which is a potent antioxidant enzyme, associated with the induction of nuclear factor-erythroid 2-related factor 2 (Nrf2). According to our data, CNMSH is able to protect SH-SY5Y cells from H2O2-induced apoptosis throughout blocking cellular damage related to oxidative stress through a mechanism that would affect ROS elimination and activating Nrf2/HO-1 signaling pathway. Conclusions : Therefore, we believed that CNMSH may potentially serve as an agent for the treatment and prevention of neurodegenerative diseases caused by oxidative stress.

Investigation of Lichen Species as a Biomonitor of Atmospheric Ozone in 'Backwoon' Mountain, Korea (백운산(白雲山) 서식(棲息) 지의류(地衣類)를 이용(利用)한 오존 민감성(敏感性) 지표종(地表種) 선발(選拔))

  • Hur, Jae-Seoun;Kim, Pan-Gi
    • Journal of Korean Society of Forest Science
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    • v.89 no.1
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    • pp.65-76
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    • 2000
  • Lichen flora of 'Backwoon' mountain, Korea, was investigated during the summer of 1998 and 1999 to screen out lichen species which can be used as a biomonitor for atmospheric ozone. The identified foliose or fructicose lichens in the area were classified into 9 families, 20 genera and 34 species. The dominant lichen species in the area were found to be Leptogium sp., Parmelia sp., Parmotrema sp., Phaeophyscia sp. and Cladonia sp. It was also found that lichen species showing a wide range of sensitiveness to air pollution were distributed in 'Backwoon' mountain. The Parmotrema austrosinense, P. tinctorum, Certrelia braunsiana and Ramalina yasudae collected in the area were exposed to ozone at the level of 200ppb, 8hr/day in controlled growth chamber for 2 weeks. Malondialdehyde(MDA), hydroperoxy conjugated dienes(HPCD), soluble protein content and OD435nm/OD415nm ratio for phaeophytinization of chlorophyll were measured and a pollution index(PI) was calculated for each lichen. Ozone exposure severely damaged to the lichens in the order of P. austrosinense>P. tinctorum>C. braunsiana>R. yasudae. P. austrosinens easily identified and widely distributed in Korea is likely to be a very useful biomonitor of air pollution, especially for ozone. These results indicate that Korean lichen species can be used as a biomonitor for air pollution to evaluate air quality contaminated with ozone.

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Ishige sinicola Extracts Induce Apoptosis via Activation of a Caspase Cascade in Human HeLa Cells (넓패 추출물이 HeLa 자궁암세포의 세포사멸에 미치는 영향)

  • Cho, Byoung-Ok;Ryu, Hyung-Won;So, Yang-Kang;Jin, Chang-Hyun;Byun, Myung-Woo;Kim, Wang-Geun;Jeong, Il-Yun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.7
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    • pp.901-906
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    • 2012
  • The purpose of this study was to elucidate the anti-proliferative effect and the mechanisms underlying apoptosis induced by a methanol extracts from Ishige sinicola (ISE) in HeLa cells. ISE treatment for 24 hr significantly inhibited cell viability in a dose-dependent manner. Apoptosis was detected by Hoechst 33258 staining and an annexin V/PI assay after 24 hr treatment. Moreover, ISE treatment triggered the cleavage of caspase-8, -9, -3, and poly(ADP-ribose) polymerase (PARP) in dose-dependent and time-dependent manners. In addition, z-VAD-fmk, a general caspase inhibitor, blocked ISE-induced cell death. Taken together, these results suggest that ISE-induced apoptosis is mediated by the activation of a caspase cascade in HeLa cells.

Effects of dietary energy sources on early postmortem muscle metabolism of finishing pigs

  • Li, Yanjiao;Yu, Changning;Li, Jiaolong;Zhang, Lin;Gao, Feng;Zhou, Guanghong
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.12
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    • pp.1764-1772
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    • 2017
  • Objective: This study investigated the effects of different dietary energy sources on early postmortem muscle metabolism of finishing pigs. Methods: Seventy-two barrow ($Duroc{\times}Landrace{\times}Yorkshire$, DLY) pigs ($65.0{\pm}2.0kg$) were allotted to three iso-energetic and iso-nitrogenous diets: A (44.1% starch, 5.9% crude fat, and 12.6% neutral detergent fibre [NDF]), B (37.6% starch, 9.5% crude fat, and 15.4% NDF) or C (30.9% starch, 14.3% crude fat, and 17.8% NDF). After the duration of 28-day feeding experiment, 24 pigs (eight per treatment) were slaughtered and the M. longissimus lumborum (LL) samples at 45 min postmortem were collected. Results: Compared with diet A, diet C resulted in greater adenosine triphosphate and decreased phosphocreatine (PCr) concentrations, greater activity of creatine kinase and reduced percentage bound activities of hexokinase (HK), and pyruvate kinase (PK) in LL muscles (p<0.05). Moreover, diet C decreased the phosphor-AKT level and increased the hydroxy-hypoxia-inducible $factor-1{\alpha}$ ($HIF-1{\alpha}$) level, as well as decreased the bound protein expressions of HK II, PKM2, and lactate dehydrogenase A (p<0.05). Conclusion: Diet C with the lowest level of starch and the highest levels of fat and NDF could enhance the PCr utilization and attenuate glycolysis early postmortem in LL muscle of finishing pigs.

Prognostic Evaluation of Categorical Platelet-based Indices Using Clustering Methods Based on the Monte Carlo Comparison for Hepatocellular Carcinoma

  • Guo, Pi;Shen, Shun-Li;Zhang, Qin;Zeng, Fang-Fang;Zhang, Wang-Jian;Hu, Xiao-Min;Zhang, Ding-Mei;Peng, Bao-Gang;Hao, Yuan-Tao
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.14
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    • pp.5721-5727
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    • 2014
  • Objectives: To evaluate the performance of clustering methods used in the prognostic assessment of categorical clinical data for hepatocellular carcinoma (HCC) patients in China, and establish a predictable prognostic nomogram for clinical decisions. Materials and Methods: A total of 332 newly diagnosed HCC patients treated with hepatic resection during 2006-2009 were enrolled. Patients were regularly followed up at outpatient clinics. Clustering methods including the Average linkage, k-modes, fuzzy k-modes, PAM, CLARA, protocluster, and ROCK were compared by Monte Carlo simulation, and the optimal method was applied to investigate the clustering pattern of the indices including platelet count, platelet/lymphocyte ratio (PLR) and serum aspartate aminotransferase activity/platelet count ratio index (APRI). Then the clustering variable, age group, tumor size, number of tumor and vascular invasion were studied in a multivariable Cox regression model. A prognostic nomogram was constructed for clinical decisions. Results: The ROCK was best in both the overlapping and non-overlapping cases performed to assess the prognostic value of platelet-based indices. Patients with categorical platelet-based indices significantly split across two clusters, and those with high values, had a high risk of HCC recurrence (hazard ratio [HR] 1.42, 95% CI 1.09-1.86; p<0.01). Tumor size, number of tumor and blood vessel invasion were also associated with high risk of HCC recurrence (all p< 0.01). The nomogram well predicted HCC patient survival at 3 and 5 years. Conclusions: A cluster of platelet-based indices combined with other clinical covariates could be used for prognosis evaluation in HCC.

Mucosal Mast Cell Responses in the Small Intestine of C3H/HeN and BALB/c Mice Infected with Echinostoma hortense

  • Ryang, Yong-Suk;Im, Jee-Aee;Kim, In-Sik;Kim, Keun-Ha
    • Biomedical Science Letters
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    • v.9 no.3
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    • pp.145-150
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    • 2003
  • In the intestinal mucosa, mast cells are thought to be responsible for the expulsion of parasites. We investigated the relationship of worm expulsion and mast cells in C3H/HeN and BALB/c mice infected with Echinostoma hortense. In addition, we examined whether the worm recovery rate was associated with the strain of mice, and whether a toluidine stain and immunohistochemistry using the c-kit antibody was effective in the detection of mast cells. In order to investigate the mucosal immune response of C3H/HeN and BALB/c mice, each mouse was infected orally with 30 E. hortense metacercariae. Then, the number of mucosal mast cells and worm recovery rates was observed in experimentally infected mouse strains between 1 week and 8 weeks post infection (PI). Mucosal mast cells were increased in 3 weeks P.I. in C3H/HeN and BALB/c mice. On the other hand, only mucosal goblet cells and worm recovery rates correlated in C3H/HeN mice (P=0.0482). Worm recoveries in C3H/HeN mice were 65.7$\pm$5.6, 53.3$\pm$5.4 and 6.7$\pm$0.6 in week 1, 2, and 3 P.I. and strongly decreased in week 3 P.I. Worm recoveries in BALB/c mice were 23.0$\pm$2.5, 10.0$\pm$1.0, and 6.7$\pm$0.6% in week 1, 2, and 3 P.I. and gradually decreased from week 1 P.I. to week 3 P.I. Worm recoveries in C3H/HeN mice were significantly higher than in BALB/c mice (P<0.00l). The number of mast cells in C3H/HeN and BALB/c mice using the anti-c-kit antibody reached to a peak in week 3 P.I. and recovered as normal level in week 5 P.I. and 6 P.I. The number in E. hortense-infected C3H/HeN mice (P=0.0015) was higher than in E. hortense-infected BALB/c mice (P=0.01) compared with the control group. There were significant differences in the number of mast cells among regions of the intestine in in C3H/HeN mice (P<0.05) but not in BALB/c mice (P>0.05). Immunohistochemistry using the anti-c-kit antibody was significant method as an examination of the number of mast cells (P=0.0002). In conclusion, the present study demonstrated that mast cells play an important role in worm recovery, and immunohistochemistry using the anti-c-kit antibody was superior to toluidine stain as an examination of mast cells.

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Robust Design for Showerhead Thermal Deformation

  • Gong, Dae-Wi;Kim, Ho-Jun;Lee, Seung-Mu;Won, Je-Hyeong
    • Proceedings of the Korean Vacuum Society Conference
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    • 2014.02a
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    • pp.150.1-150.1
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    • 2014
  • Showerhead is used as a main part in the semiconductor equipment. The face plate flatness should remain constant and the cleaning performance must be gained to keep the uniformity level of etching or deposition in chemical vapor deposition process. High operating temperature or long period of thermal loading could lead the showerhead to be deformed thermally. In some case, the thermal deformation appears very sensitive to showerhead performance. This paper describes the methods for robust design using computational fluid dynamics. To reveal the influence of the post distribution on flow pattern in the showerhead cavity, numerical simulation was performed for several post distributions. The flow structure appears similar to an impinging flow near a centered baffle in showerhead cavity. We took the structure as an index to estimate diffusion path. A robust design to reduce the thermal deformation of showerhead can be achieved using post number increase without ill effect on flow. To prevent the showerhead deformation by heat loading, its face plate thickness was determined additionally using numerical simulation. The face plate has thousands of impinging holes. The design key is to keep pressure drop distribution on the showerhead face plate with the holes. This study reads the methodology to apply to a showerhead hole design. A Hagen-Poiseuille equation gives the pressure drop in a fluid flowing through such hole. The assumptions of the equation are the fluid is viscous-incompressible and the flow is laminar fully developed in a through hole. An equation can be expressed with radius R and length L related to the volume flow rate Q from the Hagen-Poiseuille equation, $Q={\pi}R4{\Delta}p/8{\mu}L$, where ${\mu}$ is the viscosity and ${\Delta}p$ is the pressure drop. In present case, each hole has steps at both the inlet and the outlet, and the fluid appears compressible. So we simplify the equation as $Q=C(R,L){\Delta}p$. A series of performance curves for a through hole with geometric parameters were obtained using two-dimensional numerical simulation. We obtained a relation between the hole diameter and hole length from the test cases to determine hole diameter at fixed hole length. A numerical simulation has been performed as a tool for enhancing showerhead robust design from flow structure. Geometric parameters for the design were post distribution and face plate thickness. The reinforced showerhead has been installed and its effective deposition profile is being shown in factory.

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Divergence of the cbp Genes in 4-Chlorobiphenyl Catabolizing Bacteria (4-Chlorobiphenyl 분해 세균에서 cbp 유전자군의 상이성)

  • 윤덕중;한재진;김치경;김영수
    • Korean Journal of Microbiology
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    • v.30 no.1
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    • pp.53-59
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    • 1992
  • Four bacterial strains capable of catabolizing 4-chlorobiphen!;l (4CB) were isolated from the industrial waste waters. The bacterial isolates designated as PO$. P20, P27, and P1242. respectively, were examined for their catabolic activities. And in order to examine molecular homology of the 4CB catabolizing genes of these bacterial isolates. Southern hybridization was conducted with bphABC genes of P. p.srudoalculigrnrs KF707 as a DNA probe. The metabolites of 2-hydroxy-6-0x0-6-(4'-chlorophenyl)hexa-2 .4-dienoic acid and Cchlorobenzoate were detected to be produced by the isolatc:~ in the MM2 liquid cultures. But Cchlorobenzoate was further catabolized to produce 4.-hydroxybenzoate by DJ-12, P08. and P27. but not by P20 and P1242. As results of hybridization, homologous regions were commonly observed in Xhol fragments of 2.2 and 1.8 kb and in EcoRl fragment of 11 kb in the DJ- 12. P08, and P27 isolates. But in any restriction enzyme digests ot the P20 and PI242 isolates. homologous region was not detected. The cbp genes of the bactcria capable of catabolizing 4CB in nature could be divided into two groups by divergence<

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Distribution of Virulence Factors of Vibrio cholerae non-O1 and non-O139 Isolated from Korea (한국에서 분리된 Vibrio cholerae serovar non-O1 및 non-O139 병독 인자의 분포)

  • 성희경
    • Korean Journal of Microbiology
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    • v.35 no.3
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    • pp.248-252
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    • 1999
  • The PI-oduction of virulence factors such as cholera toxin, heinolysin and hemagglutinin in V cliolerae non-01 and non-0139 were examined. Among 65 strains isolated from environmental and clinical blood sources, 29 (14.6%) strains produced hemolysin only, 35(53.9%) sh.ains produced both hemolysin and hemagglutinin. From one 037 slrain isolated from environmenl, cholera toxin, ctx gene, hemolysin, and hemagglutinin were detected. All of the strains isolated from clinical and environmental sources showed hemolytic activity against human 0 group e~ythrocytes. In inhibition patterns of heinagglotination, 5 of 18 clinical strains (27.8%) were inhibited by less than 1% mannose and galactose, while, among the 47 environmental isolates. hose paltems by less than 1% mannose and galactose 55.4% wel-e inhibited. Thel-ehre, exohamagglutinin positive rate was high in clinical blood isolates but in environnlental sources, the rate was almost similar lo ihe rate or endohemagglutinin positive. These results indicaled that V cholerae non-01 and non-0139 produced various virnlence factors such as cholera toxin, hemolysin, and hemagglutinin but not a single factor. Further studies are need for epidemiological or bacteriological shtdies of V cholerae 037 isolated from environment.

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Shear Performance of Board-type Two-way Voided Slab (일체형 중공재의 중공부 내부형상에 따른 이방향 중공슬래브의 전단성능 평가)

  • Choi, Hyeon-Min;Park, Tae-Won;Paik, In-Kwan;Kim, Je-Sub;Han, Ju-Yeon
    • Journal of the Korea Concrete Institute
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    • v.27 no.6
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    • pp.651-659
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    • 2015
  • Currently, social demands for long span building structures are increasing due to architectural planning purposes and economic efficiency. As a result, lighter board-type voiding materials were suggested. With the use of board-type voiding materials, a slab is able to become light weight and convenient. This process efficiently eliminates concrete where it is not required; considerably diminishing dead weight while maintaining the flexural strength of the slab. The reduction in concrete also allows for overall cost reductions and design flexibility. Also it can be ease with fixing the voided material that is composed of one body form. Although board-type voiding materials are ideal, the top and bottom concrete plates lack integrity. Because of this, test results show horizontal cracking towards the tops and bottoms of the concrete columns, or webs, connecting the slabs. The key to correcting this problem is to increase the shear strength. In order to increase the shear strength of the structure, horizontal shear area must increase. R70(100)-D-F has the largest horizontal shear area as it also shows stronger strength. As a result, shear strength ($V_{nh}$) is dependent on the horizontal shear area (N). $V_{nh}={\alpha}{\times}0.16{\sqrt{f_{ck}}}{\frac{{\pi}D^2}{4}}{\times}N({\alpha}=1.8125)$. The web columns have a shear span to depth ratio (a/d) that is less than 2; which classifies it as a deep beam. In this case, however, the shear strength of the deep beams may be as much as 2 to 3 times greater than that predicated conventional equations developed for members of normal proportions. As a result, ${\alpha}$ is suggested as an extra coefficient in the equation for shear strength ($V_{nh}$).