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Development of screening systems for modulators on phospholipase-mediated signal transduction

  • Lee, Young-Han-;Min, Do-Sik;Kim, Jae-Ho-;Suh, Pann-Ghill;Ryu, Sung-Ho
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1994.04a
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    • pp.186-186
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    • 1994
  • Many agonists have been known to activate the hydrolysis of membrane phospholipids through the bindings with corresponding receptors on the various cells. Diacylglycerol and inositol 1,4,5-trisphosphate(IP3) generated by the action of phosphoinositide-specific phospholipase C (PI-PLC) are well known second messengers for the activation of protein kinase C and the mobilization of Ca2+ in many cells. Three types of PI-PLC isozyme (${\alpha}$,${\gamma}$, and $\delta$) and several subtrpes for each type have been identified from mammalian sources by purification of enzymes and cloning of their cDNAs. Each type PI-PLC isozyme is coupled to different receptors and mediators, for example, ${\beta}$-types are coupled to the seven-transmembrane-receptors via Gq family of G-proteins and ${\beta}$-types directly to the receptor tyrosine kinases. Specific modulators for the signaling pathway through each type of PI-PLC should be very useful as potential potential candidates for lend substances in developing novel drugs. To establish the sensitive and convenient screening systems for searching modulators on PI-PLC mediated signaling, two kinds of approaches have been tried. (1) Establishment of in vitro assay condition for each type of PI-PLC isozyme: Overexpression by using vaccinia virus and purification of each isozyme was carried out for the preparation of large amounts of enaymes. Optimum and sensitive assay condition for the measurements of PI-ELC activities were established. (2) Development of the cell lines in which each type of PI-PLC is permanently overexpressed: A fibroblast cell line (3T3${\gamma}$1-7) in which PI-PLC-${\gamma}$1 was overexpressed by using pZip-neo expression vector was developed and used for the measurement of PDGF-induced IP3 formation. The responses for IP3 formed in 3T3${\gamma}$1-7 cells by the treatment of PDGF is 8 times more sensitive than those in control cells. 3T3${\gamma}$l-7 cell is useful for the screening of the inhibitors on the PDGF-induced cellular responses from large number of samples in a small volume(50 ${\mu}$l) and short time(5-15 min). Using these systems, we screened hundreds of herb-extracts for the inhibition of PDGF-induced IP3 formation and selected several extracts that showed the inhibition as the candidates for isolation and characterization of active substances. The determination of the acting point of selected extracts or fractions in the PDGF signaling pathway has been analyzing.

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Association Between GSTM1 Polymorphism and Nasopharyngeal Cancer Susceptibility: a Meta-analysis

  • Sun, Zhen-Feng;Zhang, Jia;Xu, Hong-Ming;Wang, Guo-Liang;Dong, Pin
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.11
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    • pp.5817-5821
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    • 2012
  • Background/Aims: Glutathione S-transferase M1 (GSTM1) is a multifunctional enzyme that plays a critical role in the detoxification of varieties of carcinogenic metabolites. Many studies have been conducted to investigate the association between GSTM1 polymorphism and nasopharyngeal cancer (NPC) risk, but the findings among those studies are inconsistent. To assess this relationship more precisely, we performed a meta-analysis of all available studies on the subject. Methods: Case-control studies were identified by searching Pubmed, Embase, ISI Web of Science, and Wanfang databases through September 6, 2012. We used the pooled odds ratio (OR) with its corresponding 95% confidence interval (95%CI) to evaluate the association of GSTM1 polymorphism with NPC susceptibility. Subgroup analyses by pathological types, sex and smoking status were performed to further identify the association. Results: Overall, 11 published studies with 1,513 cases and 2,802 controls were finally included into this meta-analysis according to the inclusion criteria. Meta-analysis of total studies showed that the null genotype of GSTM1 was significantly associated with increased risk of NPC, when comparing with the non-null genotype (OR=1.51, 95%CI=1.33-1.72, POR<0.001). The association was still statistically significant in subgroup analysis of patients with nasopharyngeal squamous cell carcinoma (OR=1.73, 95%CI=1.24-2.42, POR=0.001). Males with the null genotype of GSTM1 were more likely to subject to NPC than females. In addition, the association between the null genotype of GSTM1 and NPC risk was strongest in individuals with exposure to smoking. Sensitivity analysis by sequential omission of any individual studies one at a time further demonstrated the significant association. Conclusions: The findings suggest that the null genotype of GSTM1 is a risk factor for NPC, and there is a gene-smoking interaction in this association.

Design of Adaptive User Interface(AUI) for Bus Information Terminal (Bus Information Terminal(BIT)를 위한 Adaptive User Interface(AUI) 설계)

  • Nam, Doo-Hee
    • The Journal of the Institute of Internet, Broadcasting and Communication
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    • v.11 no.2
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    • pp.89-94
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    • 2011
  • Today, the utilization of communication devices is being increased including information terminals, cell phones, handheld personal digital assistants (PDA) caused by the development of information and communication technology. The development of information and services is speeding up, whereas most communication devices have provided a inefficient hierarchical menu and sequential searching structure. In this study, the Adaptive User Interface is applied to the Bus Information Terminal(BIT) which is one of communication equipment installed in the bus stop. It will be based on analysis of unspecified individuals' preferences and user's directly personalization in the BIT prototype. We expect the results of this study to be possible to provide users with efficient and convenient information acquisition and contribute to the development of public transport use by improving the accessibility and usability of BIT.

Review for Clinical Studies of Oriental Medicine on the Treatment of Ovarian Hyperstimulation Syndrome (난소과자극증후군의 치료에 관한 한의 임상 연구 고찰)

  • Ku, Su-Jeong;Hwang, Deok-Sang;Lee, Jin-Moo;Lee, Chang-Hoon;Jang, Jun-Bock
    • The Journal of Korean Obstetrics and Gynecology
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    • v.33 no.3
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    • pp.60-79
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    • 2020
  • Objectives: This review plans to assess the efficacy and effectiveness of oriental medicine for the treatment of Ovarian Hyperstimulation Syndrome (OHSS) through literature research and overview. Methods: Database searching was conducted to identify relevant randomized controlled trials (RCTs) on oriental medicine for the treatment of Ovarian Hyperstimulation Syndrome. Studies were searched from Journal of Korean Obstetrics and Gynecology, Korean Medical Database, Korean studies Information Service System, China National Knowledge Infrastructure, Cochrane library, PubMed and EmBase up to 21st May, 2020. Results: Seventeen studies were finally selected. Fifteen studies intervened with oral Chinese herb medicine, two studies intervened with acupuncture and moxibustion. Nine studies concluded that intervention with oriental medicine significantly relieved OHSS symptoms. Three studies reporting ovary diameter, four studies reporting abdominal circumference and other four studies reporting pelvic effusion showed significant reduction compared to control groups. Six studies showed significantly shorter duration for hospitalization in intervention groups. Only one study showed significantly higher pregnancy rate. Factors related with vascular permeability and blood cell coagulation were significantly lowered in intervention groups in general. Conclusions: From seventeen studies, oriental medicine relieved OHSS symptoms and showed treatment effectiveness. Further strictly designed studies and long-term observed studies are needed to establish evidences.

A Study on the Behavior of Old People in Outdoor (노인의 외부 생활행태 연구)

  • Chang, Young-Hee
    • Journal of The Korean Digital Architecture Interior Association
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    • v.1 no.1
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    • pp.59-66
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    • 2001
  • The purpose of this study is to improve productivity of architectural space planning(A.S.P,) by computer system and to optimize ASP. A searching algorithm is the best way to slave optimized A.S.P. Because architectural design is too many various site situations and client's demands to specify the general solving methods. This method seek the best design case in all possibility and to be modeled as this; Seongbukgu's case that is city structure former times negative by in facilities utilization of the near street limit. But, case of Gangnamgu and Songpagu is thought that environment and utilization etc. of area life of old people are affinity with quality of life environment of old people when see that is using various area facilities using electric railway and a bus etc. actively. It is looked by the other that individual's special quality uses area facilities according to life partner's existence and nonexistence and family composition and existence and nonexistence of profession and distinction of sex. Show difference of external behavior according to public garden and market and supermarket and welfare facilities etc.'s location in dwelling environment of area and relation about facilities of area has been formed and old people and dwelling environment of area can know that is that do interaction. Environment that access about facilities may have to be easy, and can live that communicating closely with area's inhabitantses may have to consist so that old people may can run various external life.. Notions of the evaluated value is an profit(+) and expense(-) that decide design intention. To adapt real planning, 1. A raster type space cell has logical site informations. 2. To be evaluate various factor. 3. To reflect operator's design mind, they should add an extra weight on evaluated value.

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Nano-scale Proteomics Approach Using Two-dimensional Fibrin Zymography Combined with Fluorescent SYPRO Ruby Dye

  • Choi, Nack-Shick;Yoo, Ki-Hyun;Yoon, Kab-Seog;Maeng, Pil-Jae;Kim, Seung-Ho
    • BMB Reports
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    • v.37 no.3
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    • pp.298-303
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    • 2004
  • In general, a SYPRO Ruby dye is well known as a sensitive fluorescence-based method for detecting proteins by one-or two-dimensional SDS-PAGE (1-DE or 2-DE). Based on the SYPRO Ruby dye system, the combined two-dimensional fibrin zymography (2-D FZ) with SYPRO Ruby staining was newly developed to identify the Bacillus sp. proteases. Namely, complex protein mixtures from Bacillus sp. DJ-4, which were screened from Doen-Jang (Korean traditional fermented food), showed activity on the zymogram gel. The gel spots on the SYPRO Ruby gel, which corresponded to the active spots showing on the 2-D FZ gel, were analyzed by a matrix-assisted laser desorption ionization time of flight (MALDI-TOF) mass spectrometric analysis. Five intracellular fibrinolytic enzymes of Bacillus sp. DJ-4 were detected through 2-D FZ. The gel spots on the SYPRO Ruby dye stained 2-D gel corresponding to 2-D FZ were then analyzed by MALID TOF MS. Three of the five gel spots proved to be quite similar to the ATP-dependent protease, extracellular neutral metalloprotease, and protease of Bacillus subtilis. Also, the extracellular proteases of Bacillus sp. DJ-4 employing this combined system were identified on three gels (e.g., casein, fibrin, and gelatin) and the proteolytic maps were established. This combined system of 2-D zymography and SYPRO Ruby dye should be useful for searching the specific protease from complex protein mixtures of many other sources (e.g., yeast and cancer cell lines).

Trend on the Recycling Technologies for Silicon Sludge by the Patent and Paper Analysis (특허(特許)와 논문(論文)으로 본 실리콘 슬러지의 재활용(再活用) 기술(技術) 동향(動向))

  • Jang, Hee-Dong;Kil, Dae-Sup;Chang, Han-Kwon;Cho, Young-Ju;Cho, Bong-Gyoo
    • Resources Recycling
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    • v.21 no.4
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    • pp.60-68
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    • 2012
  • Silicon wafer for making semiconductor devices and solar cell is used in the semiconductor and solar industry, respectively. Silicon wafer is produced by cutting with silicon ingot and sludge contains silicon occurs from cutting process. Generation of silicon sludge is increasing on developing all industry sectors which have need of semiconductor device. These days it has been widely studied for the recycling technologies of the silicon sludge from view points of economy and efficiency. In this paper, patents and paper on the recycling technologies of the silicon sludge were analyzed. The range of search was limited in the open patents of USA (US), European Union (EU), Japan (JP), Korea (KR) and SCI journals from 1982 to 2011. Patents and journals were collected using key-words searching and filtered by filtering criteria. The trends of the patents and journals was analyzed by the years, countries, companies, and technologies.

Fine Needle Aspiration Cytology of Kimura's Disease of Parotid Gland - Report of A Case Cytologically Failed to Diagnose as Kimura's Disease - (귀밑샘의 기무라병의 세침흡인 세포학적 소견 -세포학적 검사로 예측할 수 없었던 1예 보고-)

  • Kim, Se-Hoon;Kim, Hae-Ryoung;Kim, Sung-Eun;Yang, Woo-Ick;Lee, Kwang-Gil;Hong, Soon-Won
    • The Korean Journal of Cytopathology
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    • v.14 no.2
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    • pp.86-90
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    • 2003
  • Kimura's disease is a chronic inflammatory disorder of unknown cause and is most prevalent among Asians. The cytologic findings of Kimura's disease are significant numbers of eosinophils in a background of lymphoid cells, occasional fragments of collagenous tissue, proliferation oi vessels, and Warthin-Finkeldey polykaryocytes. Among these features, the most important cytologic feature of Kimura's disease is a significant numbers of eosinophils. We experienced a case of Kimura's disease in the parotid gland which we fatted to recognize on cytology due to the apparent paucity of eosinophils. On careful retrograde reviewing of the cytologic findings, a few scattered leukocytes, previously interpreted as polymorphous leukocytes, had bilobed nuclei and coarse green but granular cytoplasm on Papanicolaou preparation. These leukocytes showed obvious orange-red intracyloplasmic granules as in eosionophils on Giemsa stain. The paucity of eosinophils may be due to the thick fibrosis around lymphoid follicles or any technical error during aspiration. Whereas the Warthin-Finkeldey type giant cell is not a sensitive cytologic marker of Kimura's disease, it may be a helpful cytologic feature. To reach a correct cytologic diagnosis of Kimura's disease, It is important to keep in mind that searching for Warthin-Finkeldey type giant cells and evaluation of Giemsa stain for detection of eosinophils would be helpful.

Fermented Unpolished Black Rice (Oryza sativa L.) Inhibits Melanogenesis via ERK, p38, and AKT Phosphorylation in B16F10 Melanoma Cells

  • Sangkaew, Orrarat;Yompakdee, Chulee
    • Journal of Microbiology and Biotechnology
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    • v.30 no.8
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    • pp.1184-1194
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    • 2020
  • Melanin is a major factor that darkens skin color as one of the defense systems to prevent the harmful effects of UV light. However, darkened skin from the localized or systemic accumulation of melanin is viewed in many cultures as an esthetic problem. Consequentially, searching for anti-melanogenic agents from natural sources is very popular worldwide. Previous screening of fermented rice products, obtained from various rice cultivars fermented with different sources of loog-pang (Thai traditional fermentation starter), revealed that the highest ability to reduce the melanin content in B16F10 melanoma cells was from unpolished black rice fermented with a defined starter mixture of microbes isolated from loog-pang E11. The aim of this study was to investigate the mechanism of the fermented unpolished black rice (FUBR) on the inhibition of melanogenesis in B16F10 melanoma cells. The strongest reduction of cellular melanin content was found in the FUBR sap (FUBRS). The melanin reduction activity was consistent with the significant decrease in the intracellular tyrosinase activity. The FUBRS showed no cytotoxic effect to B16F10 melanoma or Hs68 human fibroblast cell lines. It also significantly reduced the transcript and protein expression levels of tyrosinase, tyrosinase-related protein 1 (TYRP-1), TYRP-2, and microphthalmia-associated transcription factor. Furthermore, it induced a significantly increased level of phosphorylated ERK, p38 and Akt signaling pathways, which likely contributed to the negative regulation of melanogenesis. From these results, a model for the mechanism of FUBRS on melanogenesis inhibition was proposed. Moreover, these results strongly suggested that FUBRS possesses anti-melanogenesis activity with high potential for cosmeceutical application as a skin depigmenting agent.

Suppressive effects on the expression of cyclooxygenase-2 and inducible nitric oxide synthase by a natural sesquiterpenoid in lipopolysaccharide-stimulated mouse macrophage cells

  • Min, Hye-Young;Park, Hyen-Joo;Park, Eun-Jung;Lee, Sang-Kook
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2003.11a
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    • pp.101-101
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    • 2003
  • Prostaglandins (PGs) and nitric oxide (NO) produced by inducible cyclooygenase (COX-2) and nitric oxide synthase (iNOS), respectively, have been implicated as important mediators in the process of inflammation and carcinogenesis. On this line, the potential COX-2 or iNOS inhibitors have been considered as anti-inflammatory and cancer chemopreventive agents. In our continuing efforts of searching for novel cancer chemopreventive agents from natural products, we isolated natural sesquiterpenoids as potential COX-2 and iNOS inhibitors in cultured lipopolysaccharide (LPS)-activated mouse macrophage RAW 264.7 cells. Alantolactone, a natural eudesmane-type sesquiterpenoid, exhibited a potent inhibition of COX-2 (IC50 = 0.4 $\mu\textrm{g}$/$m\ell$) and iNOS activity (IC50 = 0.08 $\mu\textrm{g}$/$m\ell$) in the assay system determined by PGE2 and NO accumulation, respectively. The inhibitory potential of alantolactone on the PGE2 and NO production was well coincided with the suppression of COX-2 and iNOS protein and mRNA expression in LPS-induced macrophages. Furthermore, alantolactone inhibited NF-kB but not AP-l binding activity on nuclear extracts evoked by LPS-stimulated macrophage cells, suggesting the possible involvement of NF-kB in the regulation of COX-2 and iNOS expression. In further study with COX-2-expressing human colon HT-29 cells, alantolactone inhibited the cell proliferation, down-regulated COX-2, and inhibited the ERK phosphorylation in the early time. These results suggest that a natural sesquiterpenoid alantolactone might be a potential lead candidate for further developing COX-2 or iNOS inhibitor possessing cancer chemopreventive or anti-inflammatory activity

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