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Relationship between porcine miR-20a and its putative target low-density lipoprotein receptor based on dual luciferase reporter gene assays

  • Ding, Yueyun;Zhu, Shujiao;Wu, Chaodong;Qian, Li;Li, DengTao;Wang, Li;Wan, Yuanlang;Zhang, Wei;Yang, Min;Ding, Jian;Wu, Xudong;Zhang, Xiaodong;Gao, Yafei;Yin, Zongjun
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.7
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    • pp.922-929
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    • 2019
  • Objective: Mutations in low-density lipoprotein receptor (LDLR), which encodes a critical protein for cholesterol homeostasis and lipid metabolism in mammals, are involved in cardiometabolic diseases, such as familial hypercholesterolemia in pigs. Whereas microRNAs (miRNAs) can control LDLR regulation, their involvement in circulating cholesterol and lipid levels with respect to cardiometabolic diseases in pigs is unclear. We aimed to identify and analyze LDLR as a potential target gene of SSC-miR-20a. Methods: Bioinformatic analysis predicted that porcine LDLR is a target of SSC-miR-20a. Wild-type and mutant LDLR 3'-untranslated region (UTR) fragments were generated by polymerase chain reaction (PCR) and cloned into the pGL3-Control vector to construct pGL3 Control LDLR wild-3'-UTR and pGL3 Control LDLR mutant-3'-UTR recombinant plasmids, respectively. An miR-20a expression plasmid was constructed by inserting the porcine premiR-20a-coding sequence between the HindIII and BamHI sites in pMR-mCherry, and constructs were confirmed by sequencing. HEK293T cells were co-transfected with the miR-20a expression or pMR-mCherry control plasmids and constructs harboring the corresponding 3'-UTR, and relative luciferase activity was determined. The relative expression levels of miR-20a and LDLR mRNA and their correlation in terms of expression levels in porcine liver tissue were analyzed using reverse-transcription quantitative PCR. Results: Gel electrophoresis and sequencing showed that target gene fragments were successfully cloned, and the three recombinant vectors were successfully constructed. Compared to pMR-mCherry, the miR-20a expression vector significantly inhibited wild-type LDLR3'-UTR-driven (p<0.01), but not mutant LDLR-3'-UTR-driven (p>0.05), luciferase reporter activity. Further, miR-20a and LDLR were expressed at relatively high levels in porcine liver tissues. Pearson correlation analysis revealed that porcine liver miR-20a and LDLR levels were significantly negatively correlated (r = -0.656, p<0.05). Conclusion: LDLR is a potential target of miR-20a, which might directly bind the LDLR 3'-UTR to post-transcriptionally inhibit expression. These results have implications in understanding the pathogenesis and progression of porcine cardiovascular diseases.

A Study on Junghui Kim's Concepts in Seodok(書牘) (서독(書牘)에 나타난 완당(阮堂) 김정희(金正喜)의 사상(思想) 연구(硏究))

  • Kwon, Hyok-Soon
    • (The)Study of the Eastern Classic
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    • no.33
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    • pp.279-304
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    • 2008
  • This paper draws out the contents related to "Yuk(易)" Silhak(實學), and discusses the tendency in order to review the ideas shown in Wandang's Seodok(書牘). Also, it studies Taoism expressed in Seodok in terms of figuring out Wandang's Taoism. The features of his thoughts are following. The first one is the use of "Yuk(易)" for Soogichiin(修己治人). What he considered most important was Eumsiknamnyu(飮食男女) linked directly to the people's life. He maintained that a country must be ruled by doing Soogi(修己) with "Yuk(易)" and by making use of "Yuk(易)". The next one is both a view of Gyungsechiyong(經世致用) of Dongseoboolboon(東西不分) and a natural view of Iyonghooseng(利用厚生), standing on Silsagusi(實事求是). He actively accepted new learning and concepts those days, and he asserted that Western techniques should be even imitated for the sake of the nation. Thirdly, his view of Moowi(無爲) and Boolun(不言). He didn't use to do any Jakwi(作爲) of Jeosool(著述). This kind of view seems to save his life and be connected to Yangshin(養身) even though others tried to keep a jealous eye on and entrap him. Last, his concept of Jayeonsooneung(自然順應) and Jayeonhoigui(自然回歸). It is shown through his wish of farm work and his politics, saying that a king ought not to bind the people with faithfulness and propriety, and that he ought to rule the nation with humanity. In sum, Wandang's ideas shown in Seodok can be divided into two streams. One is Boshin(保身) through Moowijayeon(無爲自然), Boolun(不言), and Jayeonhoigui(自然回歸). The other is a pursuit of his studies for the purpose of Dukchi(德治) followed by Soogichiin, Kyongsechiyong, Iyonghooseng, and Injung. Here, since Boshin can be said to be advanced Soogi, and Soogichiin and others are connected directly to people's comfortable life, they can be integrated to Soogiianbaeksung(修己以安百姓). In other words, his thoughts were based on the fusion of Confucianism and Taoism, and he aimed at Soogiianbaeksung by accepting and using to take a look at the change of period and learning.

Type Variations of 'Stepmother' and 'Sister' in the Novels of Park Kyong-Ni and Their Meanings -Focused on Jaegwiyeol, Eunha, Kimyakgukeue Ddaldeul, Nabiwa Unggungkwi (박경리 장편소설의 '계모'·'자매' 유형 변화와 그 의미 -『재귀열』, 『은하』, 『김약국의 딸들』, 『나비와 엉겅퀴』를 중심으로)

  • Cho, Yun-A
    • Journal of Popular Narrative
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    • v.26 no.4
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    • pp.145-181
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    • 2020
  • This study analyzes type variations of the 'stepmother' and 'sister' in the full-length novels of Park Kyong-Ni and attempted to point out their meanings. The pattern of "negative stepmother" that appeared in classical and new novels also appeared repeatedly in Park Kyong-Ni's full-length novels and this was because a change took place in later full-length novels. Novels analyzed with focus were Jaegwiyeol(1959), Eunha(1960), Kimyakgukeue Ddaldeul(1962), and Nabiwa Unggungkwi(1969). The stepmother that appears in Eunha is a type that appears often in the classic and new novels of Korea. While the stepmother newly gained the role and status of 'mother', she forms a competitive relationship with the daughter of the former wife while still refusing to be a member of the family and she puts the former wife's daughter in critical situations by committing misdeeds. However, the young stepmother in Nabiwa Unggungkwi actually becomes a victim to the malicious and morbid harassment of the former wife's daughter. This stepmother is a good-natured figure who shows a sense of guilt for failing to fulfill her responsibilities of upbringing and education and she eventually dies as a victim to a bomb during the war, leaving her young biological daughter behind. On one hand, the sisters in Jaegwiyeol and Kimyakgukeue Ddaldeul are not strongly bonded but when one is caught in a crisis, the other one claims to be of help. Unlike this, the sisters in Nabiwa Unggungkwi have a bond that cannot be broken. They are half-sisters that bind each other so severely that they hinder each other's growth and they eventually end up disintegrating. Through such analyses, it is shown that issues of human nature are dealt with more acutely by breaking the 'young stepmother' away from convention by placing her in the position of the victim to amplify the conflicting relationship between sisters, unlike in previous pieces. This study was significant in that it looked into how previously repetitive character type changes appeared in full-length novels in conditions that clearly display the writer's determination to leave behind a masterpiece.

Tyrosinase Inhibition-mediated Anti-melanogenic Effects by Catechin Derivatives Extracted from Ulmus parvifolia (참느릅나무에서 추출된 catechin 유도체 화합물의 멜라닌 생성 억제 효과)

  • Taehyeok Hwang;Hyo Jung Lee;Dong-Min Kang;Kyoung Mi Moon;Jae Cheal Yoo;Mi-Jeong Ahn;Dong Kyu Moon;Dong Kyun Woo
    • Journal of Life Science
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    • v.33 no.2
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    • pp.169-175
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    • 2023
  • As a protective defensive mechanism against ultraviolet (UV) light exposure in skin tissue, melanocytes produce the pigment melanin. Tyrosinase plays a key role in melanin production in melanocytes. However, the overproduction of melanin can lead to lesions, such as freckles and dark spots. Thus, it is clinically important to find a modulating molecule to control melanogenesis by regulating tyrosinase expression and/or activity. It is known that catechin, a plant flavonoid, can reduce melano- genesis through the downregulation of tyrosinase expression. Here, we tested whether catechin derivatives isolated from the stem bark of Ulmus parvifolia have an effect on melanin production by regulating tyrosinase in mouse melanoma cells and in vitro mushroom tyrosinase. The catechin derivatives used in this study included C5A, C7A, C7G, and C7X. Treatments using these catechin derivatives reduced melanin production in mouse melanoma B16F10 cells in which melanogenesis was stimulated by α-MSH. Notably, the anti-melanogenic effects of catechin derivatives were similar to those of kojic acid, a well-known anti-melanogenic molecule. Both C5A and C7A directly inhibited the activity of tyrosinase isolated from mushrooms in vitro. Furthermore, our in silico computational simulation showed that these two compounds were expected to bind to the active site of tyrosinase, which is similar to kojic acid. In addition, all four catechin derivatives reduced tyrosinase protein expression. In summary, our results showed that catechin derivatives can reduce melanogenesis by regulating tyrosinase activity or expression. Thus, this study suggests that catechin derivatives isolated from U. parvifolia can be novel modulators of melanin production.

Study of the Sludge Formation Mechanism in Advanced Packaging Process and Prevention Method for the Sludge (어드밴스드 패키징 공정에서 발생할 수 있는 슬러지의 인자 확인 및 형성 방지법의 제안)

  • Jiwon Kim;Suk Jekal;Ha-Yeong Kim;Min Sang Kim;Dong Hyun Kim;Chan-Gyo Kim;Yeon-Ryong Chu;Neunghi Lee;Chang-Min Yoon
    • Journal of the Korea Organic Resources Recycling Association
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    • v.31 no.1
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    • pp.35-45
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    • 2023
  • In this study, the sludge formation in the wastewater drain from the advanced packaging process mechanisms are revealed as well as the key factors, materials, and sludge prevention methods using surfactant. Compared with that of conventional packaging process, advanced packaging process employ similar process to the semiconductor fabrication process, and thus many processes may generate wastewater. In specific, a large amount of wastewater may generate during the carrier wafer bonding, photo, development, and carrier wafer debonding processes. In order to identify the key factors for the formation of sludge during the advanced packaging process, six types of chemicals including bonding glue, HMDS, photoresist (PR), PR developer, debonding cleaner, and water are utilized and mixing evaluation is assessed. As a result, it is confirmed that the black solid sludge is formed, which is originated by the sludge seed formation by hydrolysis/dehydration reaction of HMDS and sludge growth via hydrophobic-hydrophobic binding with sludge seed and PR. For the sludge prevention investigation, three surfactants of CTAB, PEG, and shampoo are mixed with the key materials of sludge, and it is confirmed that the sludge formations are successfully suppressed. The underlying mechanism behind the sludge formation is that the carbon tails of the surfactant bind to PR with hydrophobic-hydrophobic interaction and inhibit the reaction with HMDS-based slurry seeds to prevent the sludge formation. In this regard, it is expected that various problems like clogging in drains and pipes during the advanced packaging process may effectively solve by the injection of surfactants into the drains.

Kinesin Superfamily Protein 5A (KIF5A) Binds to ArfGAP1, ADP-ribosylation Factor GTPase-activating Protein 1 (Kinesin Superfamily Protein 5A (KIF5A)와 ADP-ribosylation Factor GTPase-activating Protein 1 (ArfGAP1)의 결합)

  • Myoung Hun Kim;Se Young Pyo;Eun Joo Chung;Young Joo Jeong;Sung Woo Park;Mi Kyoung Seo;Won Hee Lee;Sang-Hwa Urm;Mooseong Kim;Dae-Hyun Seog
    • Journal of Life Science
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    • v.34 no.5
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    • pp.333-338
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    • 2024
  • Kinesin-1 is a heterotetrameric protein composed of two heavy chains (KHCs, also known as KIF5s) with a motor domain and two light chains (KLCs) without a motor domain. KIF5 has three subtypes, namely, KIF5A, KIF5B, and KIF5C, which share high amino acid homology except in their carboxy (C)-terminal region. KIF5A is responsible for transporting cargo within the cell. The adaptor proteins that bind to the C-terminal region of KIF5A mediate between kinesin-1 and cargo. However, the proteins regulating the intracellular cargo transport of kinesin-1 have not yet been fully identified. In this study, we identified ADP-ribosylation factor GTPase-activating protein 1 (ArfGAP1), which is involved in the intracellular trafficking of lysosomes, as a binding partner of KIF5A. KIF5A binds to the C-terminal region of ArfGAP1, and ArfGAP1 binds to the C-terminal region of KIF5A but does not interact with KIF5B, KIF5C, kinesin light chain 1 (KLC1), or KIF3A. When co-expressed in mammalian cells, ArfGAP1 co-localized with KIF5A and co-immunoprecipitated with KIF5A, KIF5B, and KLC1, but not with KIF3B. These results suggest that kinesin-1 may be regulated by ArfGAP1 in the intracellular transport of cargo.

Twitter Issue Tracking System by Topic Modeling Techniques (토픽 모델링을 이용한 트위터 이슈 트래킹 시스템)

  • Bae, Jung-Hwan;Han, Nam-Gi;Song, Min
    • Journal of Intelligence and Information Systems
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    • v.20 no.2
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    • pp.109-122
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    • 2014
  • People are nowadays creating a tremendous amount of data on Social Network Service (SNS). In particular, the incorporation of SNS into mobile devices has resulted in massive amounts of data generation, thereby greatly influencing society. This is an unmatched phenomenon in history, and now we live in the Age of Big Data. SNS Data is defined as a condition of Big Data where the amount of data (volume), data input and output speeds (velocity), and the variety of data types (variety) are satisfied. If someone intends to discover the trend of an issue in SNS Big Data, this information can be used as a new important source for the creation of new values because this information covers the whole of society. In this study, a Twitter Issue Tracking System (TITS) is designed and established to meet the needs of analyzing SNS Big Data. TITS extracts issues from Twitter texts and visualizes them on the web. The proposed system provides the following four functions: (1) Provide the topic keyword set that corresponds to daily ranking; (2) Visualize the daily time series graph of a topic for the duration of a month; (3) Provide the importance of a topic through a treemap based on the score system and frequency; (4) Visualize the daily time-series graph of keywords by searching the keyword; The present study analyzes the Big Data generated by SNS in real time. SNS Big Data analysis requires various natural language processing techniques, including the removal of stop words, and noun extraction for processing various unrefined forms of unstructured data. In addition, such analysis requires the latest big data technology to process rapidly a large amount of real-time data, such as the Hadoop distributed system or NoSQL, which is an alternative to relational database. We built TITS based on Hadoop to optimize the processing of big data because Hadoop is designed to scale up from single node computing to thousands of machines. Furthermore, we use MongoDB, which is classified as a NoSQL database. In addition, MongoDB is an open source platform, document-oriented database that provides high performance, high availability, and automatic scaling. Unlike existing relational database, there are no schema or tables with MongoDB, and its most important goal is that of data accessibility and data processing performance. In the Age of Big Data, the visualization of Big Data is more attractive to the Big Data community because it helps analysts to examine such data easily and clearly. Therefore, TITS uses the d3.js library as a visualization tool. This library is designed for the purpose of creating Data Driven Documents that bind document object model (DOM) and any data; the interaction between data is easy and useful for managing real-time data stream with smooth animation. In addition, TITS uses a bootstrap made of pre-configured plug-in style sheets and JavaScript libraries to build a web system. The TITS Graphical User Interface (GUI) is designed using these libraries, and it is capable of detecting issues on Twitter in an easy and intuitive manner. The proposed work demonstrates the superiority of our issue detection techniques by matching detected issues with corresponding online news articles. The contributions of the present study are threefold. First, we suggest an alternative approach to real-time big data analysis, which has become an extremely important issue. Second, we apply a topic modeling technique that is used in various research areas, including Library and Information Science (LIS). Based on this, we can confirm the utility of storytelling and time series analysis. Third, we develop a web-based system, and make the system available for the real-time discovery of topics. The present study conducted experiments with nearly 150 million tweets in Korea during March 2013.

Sesquiterpenoids Bioconversion Analysis by Wood Rot Fungi

  • Lee, Su-Yeon;Ryu, Sun-Hwa;Choi, In-Gyu;Kim, Myungkil
    • 한국균학회소식:학술대회논문집
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    • 2016.05a
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    • pp.19-20
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    • 2016
  • Sesquiterpenoids are defined as $C_{15}$ compounds derived from farnesyl pyrophosphate (FPP), and their complex structures are found in the tissue of many diverse plants (Degenhardt et al. 2009). FPP's long chain length and additional double bond enables its conversion to a huge range of mono-, di-, and tri-cyclic structures. A number of cyclic sesquiterpenes with alcohol, aldehyde, and ketone derivatives have key biological and medicinal properties (Fraga 1999). Fungi, such as the wood-rotting Polyporus brumalis, are excellent sources of pharmaceutically interesting natural products such as sesquiterpenoids. In this study, we investigated the biosynthesis of P. brumalis sesquiterpenoids on modified medium. Fungal suspensions of 11 white rot species were inoculated in modified medium containing $C_6H_{12}O_6$, $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ for 20 days. Cultivation was stopped by solvent extraction via separation of the mycelium. The metabolites were identified as follows: propionic acid (1), mevalonic acid lactone (2), ${\beta}$-eudesmane (3), and ${\beta}$-eudesmol (4), respectively (Figure 1). The main peaks of ${\beta}$-eudesmane and ${\beta}$-eudesmol, which were indicative of sesquiterpene structures, were consistently detected for 5, 7, 12, and 15 days These results demonstrated the existence of terpene metabolism in the mycelium of P. brumalis. Polyporus spp. are known to generate flavor components such as methyl 2,4-dihydroxy-3,6-dimethyl benzoate; 2-hydroxy-4-methoxy-6-methyl benzoic acid; 3-hydroxy-5-methyl phenol; and 3-methoxy-2,5-dimethyl phenol in submerged cultures (Hoffmann and Esser 1978). Drimanes of sesquiterpenes were reported as metabolites from P. arcularius and shown to exhibit antimicrobial activity against Gram-positive bacteria such as Staphylococcus aureus (Fleck et al. 1996). The main metabolites of P. brumalis, ${\beta}$-Eudesmol and ${\beta}$-eudesmane, were categorized as eudesmane-type sesquiterpene structures. The eudesmane skeleton could be biosynthesized from FPP-derived IPP, and approximately 1,000 structures have been identified in plants as essential oils. The biosynthesis of eudesmol from P. brumalis may thus be an important tool for the production of useful natural compounds as presumed from its identified potent bioactivity in plants. Essential oils comprising eudesmane-type sesquiterpenoids have been previously and extensively researched (Wu et al. 2006). ${\beta}$-Eudesmol is a well-known and important eudesmane alcohol with an anticholinergic effect in the vascular endothelium (Tsuneki et al. 2005). Additionally, recent studies demonstrated that ${\beta}$-eudesmol acts as a channel blocker for nicotinic acetylcholine receptors at the neuromuscular junction, and it can inhibit angiogenesis in vitro and in vivo by blocking the mitogen-activated protein kinase (MAPK) signaling pathway (Seo et al. 2011). Variation of nutrients was conducted to determine an optimum condition for the biosynthesis of sesquiterpenes by P. brumalis. Genes encoding terpene synthases, which are crucial to the terpene synthesis pathway, generally respond to environmental factors such as pH, temperature, and available nutrients (Hoffmeister and Keller 2007, Yu and Keller 2005). Calvo et al. described the effect of major nutrients, carbon and nitrogen, on the synthesis of secondary metabolites (Calvo et al. 2002). P. brumalis did not prefer to synthesize sesquiterpenes under all growth conditions. Results of differences in metabolites observed in P. brumalis grown in PDB and modified medium highlighted the potential effect inorganic sources such as $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ on sesquiterpene synthesis. ${\beta}$-eudesmol was apparent during cultivation except for when P. brumalis was grown on $MgSO_4$-free medium. These results demonstrated that $MgSO_4$ can specifically control the biosynthesis of ${\beta}$-eudesmol. Magnesium has been reported as a cofactor that binds to sesquiterpene synthase (Agger et al. 2008). Specifically, the $Mg^{2+}$ ions bind to two conserved metal-binding motifs. These metal ions complex to the substrate pyrophosphate, thereby promoting the ionization of the leaving groups of FPP and resulting in the generation of a highly reactive allylic cation. Effect of magnesium source on the sesquiterpene biosynthesis was also identified via analysis of the concentration of total carbohydrates. Our current study offered further insight that fungal sesquiterpene biosynthesis can be controlled by nutrients. To profile the metabolites of P. brumalis, the cultures were extracted based on the growth curve. Despite metabolites produced during mycelia growth, there was difficulty in detecting significant changes in metabolite production, especially those at low concentrations. These compounds may be of interest in understanding their synthetic mechanisms in P. brumalis. The synthesis of terpene compounds began during the growth phase at day 9. Sesquiterpene synthesis occurred after growth was complete. At day 9, drimenol, farnesol, and mevalonic lactone (or mevalonic acid lactone) were identified. Mevalonic acid lactone is the precursor of the mevalonic pathway, and particularly, it is a precursor for a number of biologically important lipids, including cholesterol hormones (Buckley et al. 2002). Farnesol is the precursor of sesquiterpenoids. Drimenol compounds, bi-cyclic-sesquiterpene alcohols, can be synthesized from trans-trans farnesol via cyclization and rearrangement (Polovinka et al. 1994). They have also been identified in the basidiomycota Lentinus lepideus as secondary metabolites. After 12 days in the growth phase, ${\beta}$-elemene caryophyllene, ${\delta}$-cadiene, and eudesmane were detected with ${\beta}$-eudesmol. The data showed the synthesis of sesquiterpene hydrocarbons with bi-cyclic structures. These compounds can be synthesized from FPP by cyclization. Cyclic terpenoids are synthesized through the formation of a carbon skeleton from linear precursors by terpene cyclase, which is followed by chemical modification by oxidation, reduction, methylation, etc. Sesquiterpene cyclase is a key branch-point enzyme that catalyzes the complex intermolecular cyclization of the linear prenyl diphosphate into cyclic hydrocarbons (Toyomasu et al. 2007). After 20 days in stationary phase, the oxygenated structures eudesmol, elemol, and caryophyllene oxide were detected. Thus, after growth, sesquiterpenes were identified. Per these results, we showed that terpene metabolism in wood-rotting fungi occurs in the stationary phase. We also showed that such metabolism can be controlled by magnesium supplementation in the growth medium. In conclusion, we identified P. brumalis as a wood-rotting fungus that can produce sesquiterpenes. To mechanistically understand eudesmane-type sesquiterpene biosynthesis in P. brumalis, further research into the genes regulating the dynamics of such biosynthesis is warranted.

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Activation of NF-${\kappa}B$ in Lung Cancer Cell Lines in Basal and TNF-${\alpha}$ Stimulated States (폐암 세포에서 기저 상태와 TNF-${\alpha}$ 자극 시 NF-${\kappa}B$의 활성화)

  • HwangBo, Bin;Lee, Seung-Hee;Lee, Choon-Taek;Yoo, Chul-Gyu;Han, Sung-Koo;Shim, Young-Soo;Kim, Young-Whan
    • Tuberculosis and Respiratory Diseases
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    • v.52 no.5
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    • pp.485-496
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    • 2002
  • Background : The NF-${\kappa}B$ transcription factors control various biological processes including the immune response, acute phase reaction and cell cycle regulation. NF-${\kappa}B$ complexes are retained in the cytoplasm in the basal state and various stimuli cause a translocation of the NF-${\kappa}B$ complexes into the nucleus where they bind to the ${\kappa}B$ elements and regulate the transcription of the target genes. Recent reports also suggest that NF-${\kappa}B$ proteins are involved in oncogenesis, tumor growth and metastasis. High expression of NF-${\kappa}B$ expression was reported in many cancer cell lines and tissues. The constitutive activation of NF-${\kappa}B$ was also reported in several cancer cell lines supporting its role in cancer development and survival. The anti-apoptotic action of NF-${\kappa}B$ is important for cancer survival. NF-${\kappa}B$ also controls the expression of several proteins that are important for cellular adhesion (ICAM-1, VCAM-1) suggesting a role in cancer metastasis. In lung cancer, high expression levels of the NF-${\kappa}B$ subunit p50 and c-Rel were reported. In fact, high expression does not mean a high activity, and the activation pattern of NF-${\kappa}B$ in lung cancer has not been reported. Materials and Methods : In this study, the NF-${\kappa}B$ nuclear binding activity in the basal and TNF-${\alpha}$ stimulated states were exmined in various lung cancer cell lines and compared with the normal bronchial epithelial cell line. Twelve lung cancer cell lines including the non-small cell and small cell lung cancer cell lines (A549, NCI-H358, NCI-H441, NCI-H552, NCI-H2009, NCI-H460, NCI-H1229, NCI-H1703, NCI-H157, NCI-H187, NCI-H417, NCI-H526) and BEAS-2B bronchial epithelial cell line were used. To evaluate the NF-${\kappa}B$ expression and DNA binding activity, western blot analysis and an electrophoretic mobility shift assay with the nuclear protein extracts. Results : The basal expressions of the p65 and p50 subunits were observed in the BEAS-2B cell line and all lung cancer cell lines except for NCI-H358 and NCI-H460. The expression levels of p65 and p50 were increased 30 minutes after stimulation with TNF-${\alpha}$ in BEAS-2B and in 10 lung cancer cell lines. In the NCI-H358 and NCI-H460 cell lines, p65 expression was not observed in the basal and stimulated states and the two p50 related protein levels were higher after stimulation with TNF-${\alpha}$ These new proteins were smaller than p50 and are thought to be variants of p50. In the basal state, NF-${\kappa}B$ was nearly activated in the BEAS-2B and all lung cancer cell lines. The DNA binding activity of the NF-${\kappa}B$ complexes was markedly higher after stimulation with TNF-${\alpha}$ In the BEAS-2B and all lung cancer cell line except for NCI-H358 and NCI-H460, the activated NF-${\kappa}B$ complex was a p65/p50 heterodimer. In the NCI-H358 and NCI-H460 lung cancer cell lines, the NF-${\kappa}B$ complex was variant of a p50/p50 homodimer. Conclusion : The NF-${\kappa}B$ activation pattern in the lung cancer cell lines and the normal bronchial epithelial cell lines was similar except for the activation of a variant of the p50/p50 homodimer in some lung cancer cell linse.

An Intervention Study on Integration of Family Planning and Maternal/Infant Care Services in Rural Korea (가족계획과 모자보건 통합을 위한 조산원의 투입효과 분석 -서산지역의 개입연구 평가보고-)

  • Bang, Sook;Han, Seung-Hyun;Lee, Chung-Ja;Ahn, Moon-Young;Lee, In-Sook;Kim, Eun-Shil;Kim, Chong-Ho
    • Journal of Preventive Medicine and Public Health
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    • v.20 no.1 s.21
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    • pp.165-203
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    • 1987
  • This project was a service-cum-research effort with a quasi-experimental study design to examine the health benefits of an integrated Family Planning (FP)/Maternal & Child health (MCH) Service approach that provides crucial factors missing in the present on-going programs. The specific objectives were: 1) To test the effectiveness of trained nurse/midwives (MW) assigned as change agents in the Health Sub-Center (HSC) to bring about the changes in the eight FP/MCH indicators, namely; (i)FP/MCH contacts between field workers and their clients (ii) the use of effective FP methods, (iii) the inter-birth interval and/or open interval, (iv) prenatal care by medically qualified personnel, (v) medically supervised deliveries, (vi) the rate of induced abortion, (vii) maternal and infant morbidity, and (viii) preinatal & infant mortality. 2) To measure the integrative linkage (contacts) between MW & HSC workers and between HSC and clients. 3) To examine the organizational or administrative factors influencing integrative linkage between health workers. Study design; The above objectives called for quasi-experimental design setting up a study and control area with and without a midwife. An active intervention program (FP/MCH minimum 'package' program) was conducted for a 2 year period from June 1982-July 1984 in Seosan County and 'before and after' surveys were conducted to measure the change. Service input; This study was undertaken by the Soonchunhyang University in collaboration with WHO. After a baseline survery in 1981, trained nurses/midwives were introduced into two health sub-centers in a rural setting (Seosan county) for a 2 year period from 1982 to 1984. A major service input was the establishment of midwifery services in the existing health delivery system with emphasis on nurse/midwife's role as the link between health workers (nurse aids) and village health workers, and the referral of risk patients to the private physician (OBGY specialist). An evaluation survey was made in August 1984 to assess the effectiveness of this alternative integrated approach in the study areas in comparison with the control area which had normal government services. Method of evaluation; a. In this study, the primary objective was first to examine to what extent the FP/MCH package program brought about changes in the pre-determined eight indicators (outcome and impact measures) and the following relationship was first analyzed; b. Nevertheless, this project did not automatically accept the assumption that if two or more activities were integrated, the results would automatically be better than a non-integrated or categorical program. There is a need to assess the 'integration process' itself within the package program. The process of integration was measured in terms of interactive linkages, or the quantity & quality of contacts between workers & clients and among workers. Intergrative linkages were hypothesized to be influenced by organizational factors at the HSC clinic level including HSC goals, sltrurture, authority, leadership style, resources, and personal characteristics of HSC staff. The extent or degree of integration, as measured by the intensity of integrative linkages, was in turn presumed to influence programme performance. Thus as indicated diagrammatically below, organizational factors constituted the independent variables, integration as the intervening variable and programme performance with respect to family planning and health services as the dependent variable: Concerning organizational factors, however, due to the limited number of HSCs (2 in the study area and 3 in the control area), they were studied by participatory observation of an anthropologist who was independent of the project. In this observation, we examined whether the assumed integration process actually occurred or not. If not, what were the constraints in producing an effective integration process. Summary of Findings; A) Program effects and impact 1. Effects on FP use: During this 2 year action period, FP acceptance increased from 58% in 1981 to 78% in 1984 in both the study and control areas. This increase in both areas was mainly due to the new family planning campaign driven by the Government for the same study period. Therefore, there was no increment of FP acceptance rate due to additional input of MW to the on-going FP program. But in the study area, quality aspects of FP were somewhat improved, having a better continuation rate of IUDs & pills and more use of effective Contraceptive methods in comparison with the control area. 2. Effects of use of MCH services: Between the study and control areas, however, there was a significant difference in maternal and child health care. For example, the coverage of prenatal care was increased from 53% for 1981 birth cohort to 75% for 1984 birth cohort in the study area. In the control area, the same increased from 41% (1981) to 65% (1984). It is noteworthy that almost two thirds of the recent birth cohort received prenatal care even in the control area, indicating that there is a growing demand of MCH care as the size of family norm becomes smaller 3. There has been a substantive increase in delivery care by medical professions in the study area, with an annual increase rate of 10% due to midwives input in the study areas. The project had about two times greater effect on postnatal care (68% vs. 33%) at delivery care(45.2% vs. 26.1%). 4. The study area had better reproductive efficiency (wanted pregancies with FP practice & healthy live births survived by one year old) than the control area, especially among women under 30 (14.1% vs. 9.6%). The proportion of women who preferred the 1st trimester for their first prenatal care rose significantly in the study area as compared to the control area (24% vs 13%). B) Effects on Interactive Linkage 1. This project made a contribution in making several useful steps in the direction of service integration, namely; i) The health workers have become familiar with procedures on how to work together with each other (especially with a midwife) in carrying out their work in FP/MCH and, ii) The health workers have gotten a feeling of the usefulness of family health records (statistical integration) in identifying targets in their own work and their usefulness in caring for family health. 2. On the other hand, because of a lack of required organizational factors, complete linkage was not obtained as the project intended. i) In regards to the government health worker's activities in terms of home visiting there was not much difference between the study & control areas though the MW did more home visiting than Government health workers. ii) In assessing the service performance of MW & health workers, the midwives balanced their workload between 40% FP, 40% MCH & 20% other activities (mainly immunization). However, $85{\sim}90%$ of the services provided by the health workers were other than FP/MCH, mainly for immunizations such as the encephalitis campaign. In the control area, a similar pattern was observed. Over 75% of their service was other than FP/MCH. Therefore, the pattern shows the health workers are a long way from becoming multipurpose workers even though the government is pushing in this direction. 3. Villagers were much more likely to visit the health sub-center clinic in the study area than in the control area (58% vs.31%) and for more combined care (45% vs.23%). C) Organization factors (admistrative integrative issues) 1. When MW (new workers with higher qualification) were introduced to HSC, it was noted that there were conflicts between the existing HSC workers (Nurse aids with less qualification than MW) and the MW for the beginning period of the project. The cause of the conflict was studied by an anthropologist and it was pointed out that these functional integration problems stemmed from the structural inadequacies of the health subcenter organization as indicated below; i) There is still no general consensus about the objectives and goals of the project between the project staff and the existing health workers. ii) There is no formal linkage between the responsibility of each member's job in the health sub-center. iii) There is still little chance for midwives to play a catalytic role or to establish communicative networks between workers in order to link various knowledge and skills to provide better FP/MCH services in the health sub-center. 2. Based on the above findings the project recommended to the County Chief (who has power to control the administrative staff and the technical staff in his county) the following ; i) In order to solve the conflicts between the individual roles and functions in performing health care activities, there must be goals agreed upon by both. ii) The health sub·center must function as an autonomous organization to undertake the integration health project. In order to do that, it is necessary to support administrative considerations, and to establish a communication system for supervision and to control of the health sub-centers. iii) The administrative organization, tentatively, must be organized to bind the health worker's midwive's and director's jobs by an organic relationship in order to achieve the integrative system under the leadership of health sub-center director. After submitting this observation report, there has been better understanding from frequent meetings & communication between HW/MW in FP/MCH work as the program developed. Lessons learned from the Seosan Project (on issues of FP/MCH integration in Korea); 1) A majority or about 80% of the couples are now practicing FP. As indicated by the study, there is a growing demand from clients for the health system to provide more MCH services than FP in order to maintain the achieved small size of family through FP practice. It is fortunate to see that the government is now formulating a MCH policy for the year 2,000 and revising MCH laws and regulations to emphasize more MCH care for achieving a small size family through family planning practice. 2) Goal consensus in FP/MCH shouBd be made among the health workers It administrators, especially to emphasize the need of care of 'wanted' child. But there is a long way to go to realize the 'real' integration of FP into MCH in Korea, unless there is a structural integration FP/MCH because a categorical FP is still first priority to reduce the rate of population growth for economic reasons but not yet for health/welfare reasons in practice. 3) There should be more financial allocation: (i) a midwife should be made available to help to promote the MCH program and coordinate services, (in) there should be a health sub·center director who can provide leadership training for managing the integrated program. There is a need for 'organizational support', if the decision of integration is made to obtain benefit from both FP & MCH. In other words, costs should be paid equally to both FP/MCH. The integration slogan itself, without the commitment of paying such costs, is powerless to advocate it. 4) Need of management training for middle level health personnel is more acute as the Government has already constructed 90 MCH centers attached to the County Health Center but without adequate manpower, facilities, and guidelines for integrating the work of both FP and MCH. 5) The local government still considers these MCH centers only as delivery centers to take care only of those visiting maternity cases. The MCH center should be a center for the managment of all pregnancies occurring in the community and the promotion of FP with a systematic and effective linkage of resources available in the county such as i.e. Village Health Worker, Community Health Practitioner, Health Sub-center Physicians & Health workers, Doctors and Midwives in MCH center, OBGY Specialists in clinics & hospitals as practiced by the Seosan project at primary health care level.

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