• Title/Summary/Keyword: Pi-b

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Anti-inflammatory Activities of an Ethanol Extract of Sargassum macrocarpum in Lipopolysaccharide (LPS)-stimulated RAW 264.7 Macrophages (Lipopolysaccaride로 유도된 Raw 264.7 세포에서 큰열매모자반 에탄올 추출물의 항염증 활성)

  • Cheon, Ji Min;Kim, Hyang Suk;Choi, Eun Ok;Kwon, Da Hye;Choi, Yung Hyun;Kim, Byung Woo;Hwang, Hye Jin
    • Journal of Life Science
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    • v.27 no.12
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    • pp.1437-1444
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    • 2017
  • Sargassum macrocarpum is a widely distributed marine brown algae found in the North Pacific. The objective of this study was to evaluate the anti-inflammatory activity of an ethanol extract of S. macrocarpum (EESM). First, we investigated the anti-inflammatory activities of EESM in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. EESM treatment suppressed nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) production and inhibited the expressions of the inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) at the mRNA and protein levels. In addition, the expression of pro-inflammatory cytokines, such as tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$) and interleukin-1 beta ($IL-1{\beta}$), was decreased in a dose dependent manner. Investigation of the signaling pathways of nuclear factor kappa B ($NF-{\kappa}B$), phosphoinositide-3-kinase (PI3K)/Akt, and mitogen-activated protein kinases (MAPKs) revealed suppression of $NF-{\kappa}B$ translocation from the cytosol to nucleus by EESM treatment. The phosphorylation of the Akt and ERK proteins was also inhibited by EESM treatment. EESM treatment also stimulated the expression of the heme oxygenase-1 (HO-1) enzyme and its upstream transcription factor, nuclear factor-E2-related factor 2 (Nrf2). These results suggest that EESM has anti-inflammatory activity and could have potential uses in the field of nutraceuticals.

Viability Assessment of Fresh and Frozen-thawed Dog Spermatozoa by Flow Cytometry (Flow Cytometry에 의한 개 신선정액과 동결정액의 생존성 분석)

  • Hong Y. M.;Kim Y. J.;Yu I.;Ji D. B.;Kim M. S.
    • Reproductive and Developmental Biology
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    • v.28 no.3
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    • pp.167-172
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    • 2004
  • This study was performed to examine the correlations among dog sperm viabilities evaluated by flow cytometry, by microscopic evaluation (ME), by carbo-xifluorescein diacetate and propidium iodide (CFDA/PI) and by hypoosmotic swelling (HOS) test. Semen were collected from 5 dogs ranging in age from 2 to 4 years. Each ejaculate was divided into 3 aliquots and different proportions of freeze-killed cells were added to each aliquot (1:0, 1:1 and 1:3). In the other experiment, semen was extended with Sweden extender containing 5% glycerol and equex STM paste, and frozen using liquid nitrogen vapor. Fresh and frozen-thawed dog sperm viability were assessed by flow cytometry using PI staining method. The accuracy of flow cytometry was evaluated by comparing with other classic assessments, microscopic evaluation, epifluorescence microscopic analysis using CFDA/PI, and HOS test. High correlations of sperm viabilities were found among flow cytometry, epifluorescence evaluation, HOS test (p<0.01) in fresh semen. Especially, sperm viability assessed by HOS test was highly correlated with viability by flow cytometry in all the ratios of live and dead spermatozoa, 1:0, 1:1 and 1:3 (p<0.01). The viability evaluated by ME were significantly correlated with that by flow cytometry in ratios of 1:0 and 1:3 (p<0.05) however, there was no significance in ratio of 1:1. The viability evaluated by C/p were highly correlated with that by flow cytometry in ratio of 1:0 and 1:1 (p<0.01) and significantly correlated in ratio of 1:3 (p<0.05). In frozen-thawed spermatozoa, the viability determined by HOS test was considerably correlated with that by flow cytometry (p<0.01). There was significant correlation between the viabilities by ME and by flow cytometry (p<0.05). But the viability evaluated by CFDA/PI was not correlated with viability by flow cytometry. The result from this study validate the use of flow cytometry as a precise method for assessing the viability of fresh and frozen-thawed dog spermatozoa.

Development of the Line Scan Diffusion Weighted Imaging at Low Tesla Magnetic Resonance Imaging System (저자장 자기공명영상시스템에서 선주사확산강조영상기법 개발)

  • Hong, Cheol-Pyo;Lee, Dong-Hoon;Lee, Do-Wan;Lee, Man-Woo;Paek, Mun-Young;Han, Bong-Soo
    • Journal of the Korean Society of Radiology
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    • v.2 no.2
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    • pp.31-38
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    • 2008
  • Line scan diffusion weighted imaging (LSDI) pulse sequence for 0.32 T magnetic resonance imaging (MRI) system was developed. In the LSDI pulse sequence, the imaging volume is formed by the intersection of the two perpendicular planes selected by the two slice-selective $\pi$/2-pulse and $\pi$-pulse and two diffusion sensitizing gradients placed on the both side of the refocusing $\pi$-pulse and the standard frequency encoding readout was followed. Since the maximum gradient amplitude for the MR system was 15 mT/m the maximum b value was $301.50s/mm^2$. Using the developed LSDI pulse sequence, the diffusion weighted images for the aqueous NaCl solution phantom and triacylglycerol solution phantom calculated from the line scan diffusion weighted images gives the same results within the standard error range (mean diffusivities = $963.90{\pm}79.83({\times}10^{-6}mm^2/s)$ at 0.32 T, $956.77{\pm}4.12({\times}10^{-6}mm^2/s)$ at 1.5 T) and the LSDI images were insensitive to the magnetic susceptibility difference and chemical shift.

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Systematic Identification of Hepatocellular Proteins Interacting with NS5A of the Hepatitis C Virus

  • Ahn, Ji-Won;Chung, Kyung-Sook;Kim, Dong-Uk;Won, Mi-Sun;Kim, Li-La;Kim, Kyung-Shin;Nam, Mi-Young;Choi, Shin-Jung;Kim, Hyoung-Chin;Yoon, Mi-Chung;Chae, Suhn-Kee;Hoe, Kwang-Lae
    • BMB Reports
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    • v.37 no.6
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    • pp.741-748
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    • 2004
  • The hepatitis C virus is associated with the development of liver cirrhosis and hepatocellular carcinomas. Among the 10 polyproteins produced by the virus, no function has been clearly assigned to the non-structural 5A (NS5A) protein. This study was designed to identify the hepatocellular proteins that interact with NS5A of the HCV. Yeast two-hybrid experiments were performed with a human liver cDNA prey-library, using five different NS5A derivatives as baits, the full-length NS5A (NS5A-F, amino acid (aa) 1~447) and its four different derivatives, denoted as NS5A-A (aa 1~150), -B (aa 1~300), -C (aa 300~447) and D (aa 150~447). NS5A-F, NS5A-B and NS5A-C gave two, two and 10 candidate clones, respectively, including an AHNAK-related protein, the secreted frizzled-related protein 4 (SFRP4), the N-myc downstream regulated gene 1 (NDRG1), the cellular retinoic acid binding protein 1 (CRABP-1), ferritin heavy chain (FTH1), translokin, tumor-associated calcium signal transducer 2 (TACSTD2), phosphatidylinositol 4-kinase (PI4K) and $centaurin{\delta}$ 2 ($CENT{\delta}2$). However, NS5A-A produced no candidates and NS5A-D was not suitable as bait due to transcriptional activity. Based on an in vitro binding assay, CRABP-1, PI4K, $CENT{\delta}2$ and two unknown fusion proteins with maltose binding protein (MBP), were confirmed to interact with the glutathione S-transferase (GST)/NS5A fusion protein. Furthermore, the interactions of CRABP-1, PI4K and $CENT{\delta}2$ were not related to the PXXP motif (class II), as judged by a domain analysis. While their biological relevance is under investigation, the results contribute to a better understanding of the possible role of NS5A in hepatocellular signaling pathways.

The Change of c-jun Promoter Activity in TPA-Induced U937 Cells Infected with Human Cytomegalovirus (HCMV) (TPA로 분화된 U937 세포에서 사람 세포거대바이러스에 의한 c-jun Promoter 활성도의 변화)

  • Park, Chung-Gyu;Kim, Dae-Joong;Kim, Jin-Hee;Han, Tae-Hee;Hwan, Eung-Soo;Choi, Myong-Sik;Kook, Yoon-Hoh;Choi, Sung-Bae;Cha, Chang-Yong
    • The Journal of Korean Society of Virology
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    • v.29 no.2
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    • pp.129-136
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    • 1999
  • Transient transfection assay has been done to evaluate whether the c-jun activation would be prerequisite to the induction of permissiveness against human cytomegalovirus using in vitro cell model in which U937 has been induced to express CD11b and CD14 to become potential monocyte/macrophage cells by TPA treatment. U937 cells were treated with $10\;{\mu}M$, $50\;{\mu}M$ or $100\;{\mu}M$ of TPA. The cell morphology change was observed and the expression of the CD11b and CD14 was confirmed by FACS. Differentiated cells were transfected with pJLuc reporter vector which contained the wild type murine c-jun promoter spanning the SP1, CTF, ATF/CREB and MEF-2 binding sites upstream of the firefly luciferase gene. After 48 hrs of transfection, the cells were infected with HCMV Towne strain and the luciferase activity was assessed at 1 hand 4 h pi. The transfection assay showed no activation of the c-jun promoter at 1 h pi, instead, it showed 2 times increase of the its activity at 4 h pi. There was no difference of the c-jun promoter activation between TPA treated and untreated U937 cells, implying that c-jun activation might not be prerequisite for allowing cells to be premissive to HCMV, although HCMV infection itself could activate c-jun promoter.

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The Magnetic Properties of $Co_{84}\;Hf_{16}$ Thin Films by FMR (강자성공명을 이용한 $Co_{84}\;Hf_{16}$ 박막의 자기적 성질 연구)

  • 김기현;장재호;김영호
    • Journal of the Korean Magnetics Society
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    • v.7 no.4
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    • pp.191-195
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    • 1997
  • $Co_{84}Hf_{16}$ (1300$\AA$, 2150$\AA$) thin films were prepared by dc magnetron sputtering method. To investigate the uniaxial anisotrpy of the sample, the saturation and effective magnetization of the thin films were measured by VSM and FMR, respectively. The spectroscopic splitting g factor were estimated from the ferromagnetic resonance curves. For 1300$\AA$, 2150$\AA$, the effective magnetization was measured at the temperatures from T=77K to T=300K. The results were analyzed in terms of Bloch's law $M_s(T)=M_s(0)(1BT^{3/2}CT^{5/2}$. The Bloch coefficient B and C were determined by fitting. $M_{eff}(0)$ was obtained by extrapolating $M_{eff}$ to 0 K. From this result, the spinwave stiffness constants D was also determined and the exchange stiffness constants $A_{eff}$ were calculated by Kittel's resonance conditions.

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Periodontopathogen LPSs Regulate MicroRNA Expression in Human Gingival Epithelial Cells

  • Lee, Hwa-Sun;Na, Hee-Sam;Jeong, So-Yeon;Jeong, Sung-Hee;Park, Hae-Ryoun;Chung, Jin
    • International Journal of Oral Biology
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    • v.36 no.3
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    • pp.109-116
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    • 2011
  • Periodontitis results from the activation of host immune and inflammatory defense responses to subgingival plaque bacteria, most of which are gram-negative rods with lipopoly-saccharides (LPSs) in their cell walls. LPSs have been known to induce proinflammatory responses and recently it was reported also that they induce the expression of microRNAs (miRNAs) in host cells. In our current study therefore, we aimed to examine and compare the miRNA expression patterns induced by the LPSs of major periodontopathogens in the human gingival epithelial cell line, Ca9-22. The cells were treated with 1 ${\mu}g$/ml of E. coli (Ec) LPS or 5 ${\mu}g$/ml of an LPS preparations from four periodontopathogens Porphyromonas gingivalis (Pg), Prevotella intermedia (Pi), Aggregatibacter actinomycetemcomitans (Aa), and Fusobacterium nucleatum (Fn) for 24 h. After small RNA extraction from the treated cells, miRNA microarray analysis was carried out and characteristic expression profiles were observed. Fn LPS most actively induced miRNAs related to inflammation, followed by Aa LPS, Pi LPS, and Ec LPS. In contrast, Pg LPS only weakly activated miRNAs related to inflammation. Among the miRNAs induced by each LPS, miR-875-3p, miR-449b, and miR-520d-3p were found to be commonly up-regulated by all five LPS preparations, although at different levels. When we further compared the miRNA expression patterns induced by each LPS, Ec LPS and Pi LPS were the most similar although Fn LPS and Aa LPS also induced a similar miRNA expression pattern. In contrast, the miRNA profile induced by Pg LPS was quite distinctive compared with the other bacteria. In conclusion, miR-875-3p, miR-449b, and miR-520d-3p miRNAs are potential targets for the diagnosis and treatment of periodontal inflammation induced by subgingival plaque biofilms. Furthermore, the observations in our current study provide new insights into the inflammatory miRNA response to periodontitis.

Molecular Mechanisms Involved in Peptidoglycan-induced Expression of Tumor Necrosis Factor-α in Monocytic Cells (펩티도글리칸에 의한 단핵세포의 Tumor necrosis factor-α 발현 기전 연구)

  • Jeong, Ji-Young;Son, Yonghae;Kim, Bo-Young;Kim, Koanhoi
    • Journal of Life Science
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    • v.29 no.11
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    • pp.1251-1257
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    • 2019
  • Peptidoglycan (PG) is found in atheromatous lesions of arteries, where monocytes/macrophages express inflammatory cytokines, including tumor necrosis factor-alpha ($TNF-{\alpha}$). This study investigated the effects of PG on $TNF-{\alpha}$ expression and examined possible cellular factors involved in $TNF-{\alpha}$ upregulation. The overall aim was to identify the molecular mechanisms underlying inflammatory responses to bacterial pathogen-associated molecular patterns in the artery. Exposure of human THP-1 monocytic cells to PG enhanced the secretion of $TNF-{\alpha}$ and induced its gene transcription. Inhibition of TLR-2/4 with OxPAPC significantly inhibited $TNF-{\alpha}$ gene expression, whereas inhibition of LPS by polymyxin B did not. The PG-induced expression of $TNF-{\alpha}$ was also significantly suppressed by pharmacological inhibitors that modulate activities of cellular signaling molecules; for example, U0126 (an ERK inhibitor), SB202190 (a p38 MAPK inhibitor), and SP6001250 (a JNK inhibitor) significantly attenuated PG-induced transcription of $TNF-{\alpha}$ and secretion of its gene product. $TNF-{\alpha}$ expression was also inhibited by rapamycin (an mTOR inhibitor), LY294002 (a PI3K inhibitor), and Akt inhibitor IV (an Akt inhibitor). ROS-regulating compounds, like NAC and DPI, also significantly attenuated $TNF{\alpha}$ expression induced by PG. These results suggest that PG induces $TNF-{\alpha}$ expression in monocytes/macrophages by multiple molecules, including TLR-2, PI3K, Akt, mTOR, MAPKs, and ROS.

CPW travelling-wave electrodes, Mach-Zehnder type Ti:LiNbO3 optical modulator (CPW 진행파 전극, M-Z형 Ti:LiNbO3 광 변조기)

  • Pi Joong-Ho;Jung Eun-Joo;Choi Jung-Sung;Kim Chang-Min
    • Korean Journal of Optics and Photonics
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    • v.15 no.5
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    • pp.475-482
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    • 2004
  • We have fabricated a LiNbO$_3$ travelling-wave optical modulator. The travelling-wave modulator consisted of M-Z(Mach-Zehnder) interferometer and CPW(Coplanar Waveguide) travelling-wave electrodes. Design of CPW electrodes were performed by SOR(Successive Over Relaxation) to obtain the conditions of Microwave effective index and impedance matching. The fabricated modulator had -3.2 dB insertion loss, and S$_{11}$ below -15 dB up to 12 GHz, S$_{21}$ better than -3 dB upto 7 GHz. It turned out that the optical response showed the 3 dB bandwidth of above 12 GHz.

Searching for Rotationable Vegetables for Paratylenchus projectus in Lettuce Greenhouse (해바라기침선충(Paratylenchus projectus) 피해 경감을 위한 윤작작물 탐색)

  • Kwon, Giyoon;Seo, Jongmin;Park, Sohee;Kang, Heonil;Park, Namsook;Choi, Insoo
    • Research in Plant Disease
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    • v.26 no.4
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    • pp.272-278
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    • 2020
  • The severe lettuce damage caused by Paratylenchus projectus was first reported in 2019 in Korea. To find high-value rotation crops for the control of P. projectus, nine vegetables, Brassica juncea (leaf mustard), B. rapa subsp. nipposinica (kyona), B. oleracea var. italica (broccoli), B. rapa subsp. chinensis (bok choy), B. oleracea var. viridis (kale), B. oleracea var. gongylodes (kohlrabi), Cichorium endivia (endive), C. intybus (chicory), Ipomoea aquatica (morning glory) were planted in d-10-cm clay pots in greenhouse. The growth of vegetables was compared between inoculated with 3,000 P. projectus per 100 ㎤ of soil and non-inoculated. Treatments were replicated 10 times. After 100 days, the reduction of fresh top weight was 30.4% in C. intybus, 35.1% in I. aquatica, 36.9% in B. oleracea var. acephala, 40.5% in C. endivia, 42.1% in B. rapa, 47.5% in B. rapa subsp. nipposinica, 50.4% in B. oleracea var. gonglodes, 56.3% in B. oleracea var. italica, and 66.0% in B. juncea. Nematode multiplication rates (Pf/Pi) were lower in I. aquatica (0.64) and C. endivia (1.1), but higher in B. oleracea var. gongylodes (2.54). Considering these results, I. aquatica is suitable for the rotation crop with lettuce until better rotation crops developed.