• Title/Summary/Keyword: 혼합방법론

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1-β-D-Arabinofuranosyl-cytosine Induces Chromosomal Breaks in vitro (In vitro에서 1-β-D-arabinofuranosyl-cytosine의 염색체 파열 유도)

  • Jeon, In-sang
    • Clinical and Experimental Pediatrics
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    • v.46 no.12
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    • pp.1186-1193
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    • 2003
  • Purpose : Fragile sites are points on chromosomes which tend to break non-randomly when exposed to specific chemical agents or conditions of tissue culture. The chromosomal break induced by the antineoplastic drug, 1-${\beta}$-D-arabinofuranosyl-cytosine(Ara-c), was investigated to study the laboratory conditions in which the incidence of chromosomal break could be enhanced. Besides, the fragile sites induced by Ara-C were investigated and compared to the already known locations of the specific chromosomal alterations observed in specific neoplasms. Methods : T-lymphocytes from theree normal males and three females were cultured for 48 hours. Cells from each individual were exposed to the Ara-C for an additional 24 hours. After the caffeine was added during the last six hours culture, the metaphase chromosomes were prepared following the conventional method. A site was considered fragile if it was found to break two or more per 100 chromosomal breaks in more than four of six individuals tested. Results : Ara-C induced 252.1 chromosomal breaks per 100 mitotic cells and this result was significantly higher than that of the control, which induced 25.2 breaks(P<0.05). The incidence of the chromosomal break by Ara-C was higher, if cultured in the MEM-FA, which has no folic acid, than in the RPMI 1640 which contains enough folic acid(P<0.05). The most common break site by Ara-C was 3p14.2(FRA3B). There were 20 fragile sites induced by Ara-C. Among these 20 fragile sites, seven coincided with the locations of the mapped oncogenes, JUN, SKI, REL, N-MYC, FHIT, MET, ETS-1, and FOS. Conclusion : S phase specific chemotherapeutic agent, Ara-C, induced the expression of the chromosomal fragile sites effectively using the T-lymphocyte in vitro. Some of the fragile sites by Ara-C highly coincided with the oncogenes and neoplasm specific chromosome breakpoints. In this regard, the fragile sites reported here could provide the unknown neoplasm related chromosomal alternation points.

Nucleotide Sequence Analysis of the Hemagglutinin-Neuraminidase Gene of Urabe AM-9 Strain (Urabe AM-9 볼거리 백신주의 Hemagglutinin-Neuraminidase 유전자 염기서열 분석)

  • Lee, Joo Yeon;Kim, Jee Hee;Lee, Jin Soo;Park, Ji Ho;Sohn, Young Mo
    • Pediatric Infection and Vaccine
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    • v.7 no.1
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    • pp.83-93
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    • 2000
  • Purpose : Urabe AM-9 strain was known to be associated with increased aseptic meningitis. The reason for high incidence of vaccine-associated meningitis was known that nucleotide(nt) substituted form G to A at position 1081 of the hemagglutinin-neuraminidase(HN) gene and therefore, glutamic acid changed to lysine at amino acid 335. We assessed by comparing nt sequence of the HN gene form Urabe AM-9 strain with wild strain and documented the correlation between nt substitution and vaccine-associated meningitis. Methods : Two lots of Urabe AM-9 vaccine distributed in Korea and mumps wild strains isolated from 1998 through 1999 were analysed. Analysis was made by nt sequencing following amplification of HN gene by RT-PCR. Results : Nucleotide substitution at position 343, 1476, 1570 was not found in both Urabe AM-9 vaccines and wild strains. But analysis of vaccine strains and wild strains isolated from patients revealed substitution from G to A at nt 1081 of the HN gene. Therefore, it encodes lysine instead of glutamic acid at amino acid 335. There was no mixture from of G and A at nt 1081. Nt at 1470 of one lot of Urabe AM-9 vaccines changed from C to A after Vero cell passage. Nt at 1727 of vaccines and wild strains was substituted A to G, so it encodes glycine instead of aspartic acid. Conclusion : Nucleotide analysis of HN gene revealed that nt 1081 of Urabe AM-9 vaccines and wild strains had wild type AAA($Lys^{335}$) instead of variant type GAA($Glu^{335}$). The results of this study suggest that there was a probability of vaccine-associated meningitis due to Urabe AM-9 in Korea before. But incidence of actual side effect was not evaluated because there was no reporting system in Korea.

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Kinetics and Mechanism of Alkalie Hydrolysis of Cinnamonitrile (II) (Cinnamonitrile의 알카리 가수분해(加水分解) 반응(反應)메카니즘 (II))

  • Sung, Nack Do;Chung, Woo Jin;Kwon, Ki Sung;Park, Byung Kwan
    • Korean Journal of Agricultural Science
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    • v.10 no.2
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    • pp.354-364
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    • 1983
  • Confonmation of (Z)-cinnamonitrile have been studied by molecular orbital theoretically using extended Huckel theory(EHT) and CNDO/2 molecular orbital calculation methods. The results indicate that the stability of conformation is(Z)-gauch>(Z)-planar. The rate constants for alkalie hydrolysis of cinnamonitrile at pH 7.0-14.0 range have been determined by ultra-violet spectrophotometry in 50% methanol at $25^{\circ}C$ and the following rate equation which can be applied over wide pH range was obtained; $${\therefore}k=({\frac{1.41{\times}10^{-14}+1.21{\times}10^7/[H_3O^+]}{2.65{\times}10^{-7}+1.64/[H_3O^+]})+9.14{\times}10^9/[H_3O^+]$$ The rate equation reveals that, at pH 7.0-10.0, the reaction is initiated by the addition of water molecule to unsaturated cabon-carhon double bond of cinnamonitrile and ${\alpha}C-{\beta}C$ bond scission follow subsequently in neutral and alkalie media. At pH 12.0-14.0, in strong alkalie solution, that so-called Michael type nucleophilic addition that the over-all rate constants is only dependent upon the concentration of hydroxide ion occurs competitively and are very complicated. Hence, the reaction mechanism of alkalie hydrolysis of cinnamonitrile which did not carefully before can be fully explained.

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The Effect of Epigallocatechin-3-gallate on HIF-1 α and VEGF in Human Lung Cancer Cell Line (비소세포폐암주에서 저산소상태에 의해 유발된 HIFa-1 α와 VEGF의 발현증가에 미치는 Epigallocatechin-3-gallate의 억제 효과)

  • Song, Joo Han;Jeon, Eun Joo;Kwak, Hee Won;Lee, Hye Min;Cho, Sung Gun;Kang, Hyung Koo;Park, Sung Woon;Lee, Jae Hee;Lee, Byung Ook;Jung, Jae Woo;Choi, Jae Cheol;Shin, Jong Wook;Kim, Ki Jeong;Kim, Jae-Yeol;Park, In Won;Choi, Byoung Whui
    • Tuberculosis and Respiratory Diseases
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    • v.66 no.3
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    • pp.178-185
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    • 2009
  • Background: Epigallocatechin-3-gallate (EGCG) is the major catechin in green tea, and has shown antiproliferative, antiangiogenic, antimetastatic and cell cycle pertubation activity in various tumor models. Hypoxia can be induced because angiogenesis is insufficient for highly proliferating cancer. Hypoxia-inducible factor-1$\alpha$ (HIF-1$\alpha$) and its downstream target, vascular endothelial growth factor (VEGF), are important for angiogenesis, tumor growth and metastasis. The aim of this study was to determine how hypoxia could cause changes in the cellular phenomena and microenvironment in a non-small cell culture system and to examine the effects of EGCG on a HIF-1$\alpha$ and VEGF in A549 cell line. Methods: A549 cells, a non-small cell lung cancer cell line, were cultured with DMEM and 10% fetal bovine serum. A decrease in oxygen tension was induced using a hypoxia microchamber and a $CO_2-N_2$ gas mixture. Gas analysis and a MTT assay were performed. The A549 cells were treated with EGCG (0, 12.5, 25, 50 ${\mu}mol/L$), and then examined by real-time-PCR analysis of HIF-1$\alpha$, VEGF, and $\beta$-actin mRNA. Results: Hypoxia reduced the proliferation of A549 cells from normoxic conditions. EGCG inhibited HIF-1$\alpha$ transcription in A549 cells in a dose-dependent manner. Compared to HIF-1$\alpha$, VEGF was not inhibited by EGCG. Conclusion: HIF-1$\alpha$ can be inhibited by EGCG. This suggests that targeting HIF-1$\alpha$ with a EGCG treatment may have therapeutic potential in non-small cell lung cancers.

Fermented Extracts of Korean Mistletoe with Lactobacillus (FKM-110) Stimulate Macrophage and Inhibit Tumor Metastasis (유산균으로 발효된 한국산 겨우살이 추출물의 Macrophage 자극에 의한 면역학적 활성화와 종양전이 억제효과)

  • Yoon, Taek-Joon;Yoo, Yung-Choon;Kang, Tae-Bong;Lee, Kwan-Hee;Kwak, Jin-Hwan;Baek, Young-Jin;Huh, Chul-Sung;Kim, Jong-Bae
    • Korean Journal of Food Science and Technology
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    • v.31 no.3
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    • pp.838-847
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    • 1999
  • Based on the results that the extract of Korean mistletoe (KM-110) has immunological and anti-tumor activities and its main component is lectin called KML-U, this study was carried out to investigate the immunostimulatory and anti-tumor activities of FKM-110, fermented KM-110 with lactobacillus, as a basic study for the development of functional food with anti-tumor activity. The amount of lectin after fermentation determined by ELISA was varied with the fermentation time and kinds of lactobacillus. Cytotoxic effects of FKM-110 on the various tumor cells was significant and dependent on the concentration of KML-U and the kinds of lactobacillus. FKM-110 stimulated macrophage and resulted in the secretion of some cytokines such as IL-1 and $IFN-{\gamma}$, but this effect was not correlated with the concentration of lectin. FKM-110 fermented with Marshall Lactobacillus casei showed the most potent antitumor activity in experimental and spontaneous metastasis models. When yoghurt produced with KM-110, Marshall Lactobacillus casei and skim milk was administered orally to mouse, the metastasis of tumor cells was significantly inhibited.

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Prognostic Significance of Cyclin D1 Overexpression in Non-Small Cell Lung Cancer (Cyclin D1의 발현이 비소세포폐암의 예후에 미치는 영향)

  • Yang, Seok-Chul;Shin, Dong-Ho;Park, Sung-Soo;Lee, Jung-Hee;Keum, Joo-Seob;Kong, Gu;Lee, Jung-Dal
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.4
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    • pp.776-784
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    • 1998
  • Background: The cyclin D1 gene is one of the most frequently amplified chromosomal regions(11q13) in human carcinomas. In laryngeal and head and neck carcinomas, its overexpression has been shown to be associated with advanced local invasion and presence of lymph node metastases. Cyclin D1 may therefore playa key role in cell growth regulation and tumorigenesis. Lung cancer is a worldwide problem and in many contries it is the most lethal malignancy. As relapse is frequent after resection of early stage non-small cell lung cancer, there is an urgent need to define prognostic factors. Purpose: This study was undertaken to evaluate the prognostic value of the cyclin D1, that is one the G1 cyclins which control cell cycle progression by allowing G1 to S phase transition, on the patients in radically resected non-small cell lung cancer. Method: Total 81 cases of formalin-fixed paraffin-embedded blocks from resected primary non-small cell lung cancer from January 1, 1983 to July 31, 1995 at Hanyang University Hospital were available for both clinical follow-up and immunohistochemical staining using monoclonal antibodies for cyclin D1. Results : The histologic classification of the tumor was based on WHO criteria, and the specimens included 45 squamous cell carcinomas, 25 adenocarcinomas and 11 large cell carcinomas. Cyclin D1 overexpression was noted in 26 cases of 81 cases tested (30.9%). Cyclin D1 expression was not significantly associated with cell types of the tumor, pathological staging and the size of the tumor. But cyclin D1 overexpression was significantly correlated with positive lymph node metastasis(p=0.035). The mean survival duration was $22.76{\pm}3.50$ months in cyclin D1 positive group and $45.38{\pm}5.64$ months in eyclin D1 negative group. There was a nearly significant difference in overall survival between cyclin D1 positive and negative groups(p=0.0515) in radically resected non-small cell lung cancer. Conclusion: Based on this study, cyelin D1 overexpression appears an important poor prognostic indicator in non-small cell lung cancer and may have diagnostic and prognostic importance in the treatment of resectable non-small cell lung cancer.

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Clinical Feature and Dietary Pattern of Infantile Constipation Under Two Years of Age (2세 이하 영유아 변비의 임상 양상과 식이 방식)

  • Kim, Hye-Jin;Moon, Jin Soo;Hwang, Jong Hee;Jang, Hyun Oh;Nam, Seung Yeun;Kim, Dong Wook;Lee, Chong Guk
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.9 no.1
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    • pp.31-38
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    • 2006
  • Purpose: Infantile constipation is one of the most common problems in pediatric gastrointestinal outpatient clinic. We planed to show the clinical feature of infantile constipation and explore the possible relationship between diets and symptoms of constipation. Methods: We analyzed the medical records and telephone questionnaire about infants under 2 years of age with constipation, who visited outpatient clinic of Department of Pediatrics, Ilsan Paik Hospital during the time from March 2002 to February 2005. Data including the symptoms and signs of constipation, diet history, and past-medical history were analyzed. Results: Total 96 infants, 40 male and 56 female, were enrolled in this study. The mean age was 9 months. Twenty-three infants were exclusively breast milk fed (BMF), 20 infants were cow's milk fed (CMF) and 53 infants had a history of mixed feeding with cow's milk and breast milk (MMF). Patients showed painful defecation (95.8%), abdominal distension (53.1%), palpable rectal stool (35.1%), hard stool (30.2%), blood-tinged stool (29.2%) and anal fissure (16.7%). Patients with exclusive cow milk feeding had prominent clinical features, such as anal fissure (p=0.03), hematochezia (p=0.04) and palpable rectal stool (p=0.025). Patients who had a history of larger intake of liquid food had a tendency to get anal skin tag (p<0.05). Conclusion: Exclusive breast milk feeding seemed important to avoid constipation with clinical significance. To educate caregivers in appropriate way of the weaning food may help the infants with constipation.

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The Effect of Surfactant on Neutrophil Apoptosis in Lipopolysaccharide Induced Acute Lung Injury in Rat (기관내 내독소 투여로 유도한 백서의 급성 폐손상 모델에서 surfactant가 호중구의 아포토시스에 미치는 영향)

  • Yoo, Ji-Hoon;Lee, Byoung-Jun;Jeong, Do-Young;Lee, Sang-Hoon;Shin, Jong-Wook;Kim, Jae-Yeol;Park, In-Won;Choi, Byoung-Whui
    • Tuberculosis and Respiratory Diseases
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    • v.53 no.4
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    • pp.409-419
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    • 2002
  • Background : The therapeutic effects of surfactant on acute lung injury derive not only from its recruiting action on collapsed alveoli but also from its anti-inflammatory effects. Pro-apoptotic action on alveolar neutrophils represents one of the important anti-inflammatory mechanisms of surfactant. In the present study, we evaluated the effects of sufactant on the apoptosis of human peripheral and rat alveolar neutrophils. Methods : In the (Ed- the article is not definitely needed but it helps to separate the two prepositions 'in') in vitro study, human neutrophils were collected from healthy volunteers. An equal number of neutrophils ($1{\times}10^6$) (Ed-confirm) was treated with LPS (10, 100, 1000ng/ml), surfactant (10, 100, $1000{\mu}g/ml$), or a combination of LPS (1000ng/ml) and surfactant (10, 100, $1000{\mu}g/ml$). After incubation for 24 hours, the apoptosis of neutrophils was evaluated by Annexin V method. In the in vivo study, induction of acute lung injury in SD rats by intra-tracheal instillation of LPS (5mg/kg) was followed by intra-tracheal administration of either surfactant (30mg/kg) or normal saline (5ml/kg). Tenty-four hours after LPS instillation, alveolar neutrophils were collected and the apoptotic rate was evaluated by Annexin V method. In addition, changes of the respiratory mechanics of rats (respiratory rate, tidal volume, and airway resistance) were evaluated with one chamber body plethysmography before, and 23 hours after, LPS instillation. Results : in the in vitro study, LPS treatment decreased the apoptosis of human peripheral blood neutrophils (control: $47.4{\pm}5.0%$, LPS 10ng/ml; $30.6{\pm}10.8%$, LPS 100ng/ml; $27.5{\pm}9.5%$, LPS 1000ng/ml; $24.4{\pm}7.7%$). The combination of low to moderate doses of surfactant with LPS promoted apoptosis (LPS 1000ng/ml + Surf $10{\mu}g/ml$; $36.6{\pm}11.3%$, LPS 1000ng/ml +Surf $100{\mu}g/ml$; $41.3{\pm}11.2%$). The high dose of surfactant ($1000{\mu}g/ml$) decreased apoptosis ($24.4{\pm}7.7%$) and augmented the anti-apoptotic effect of LPS (LPS 1000ng/ml + Surf $1000P{\mu}g/ml$; $19.8{\pm}5.4%$). In the in vivo study, the apoptotic rate of alveolar neutrophils of surfactant-treated rats was higher than that of normal saline-treated rats ($6.03{\pm}3.36%$ vs. $2.95{\pm}0.58%$). The airway resistance (represented by Penh) of surfactant-treated rats was lower than that of normal saline-treated rats at 23 hours after LPS injury ($2.64{\pm}0.69$ vs. $4.51{\pm}2.24$, p<0.05). Conclusion : Surfactant promotes the apoptosis of human peripheral blood and rat alveolar neutrophils. Pro-apoptotic action on neutrophils represents one of the important anti-inflammatory mechanisms of surfactant.

Research for Space Activities of Korea Air Force - Political and Legal Perspective (우리나라 공군의 우주력 건설을 위한 정책적.법적고찰)

  • Shin, Sung-Hwan
    • The Korean Journal of Air & Space Law and Policy
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    • v.18
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    • pp.135-183
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    • 2003
  • Aerospace force is a determining factor in a modem war. The combat field is expanding to space. Thus, the legitimacy of establishing aerospace force is no longer an debating issue, but "how should we establish aerospace force" has become an issue to the military. The standard limiting on the military use of space should be non-aggressive use as asserted by the U.S., rather than non-military use as asserted by the former Soviet Union. The former Soviet Union's argument is not even strongly supported by the current Russia government, and realistically is hard to be applied. Thus, the multi-purpose satellite used for military surveillance or a commercial satellite employed for military communication are allowed under the U.S. principle of peaceful use of space. In this regard, Air Force may be free to develop a military surveillance satellite and a communication satellite with civilian research institute. Although MTCR, entered into with the U.S., restricts the development of space-launching vehicle for the export purpose, the development of space-launching vehicle by the Korea Air Force or Korea Aerospace Research Institute is beyond the scope of application of MTCR, and Air Force may just operate a satellite in the orbit for the military purpose. The primary task for multi-purpose satellite is a remote sensing; SAR sensor with high resolution is mainly employed for military use. Therefore, a system that enables Air Force, the Korea Aerospace Research Institute, and Agency for Defense Development to conduct joint-research and development should be instituted. U.S. Air Force has dismantled its own space-launching vehicle step by step, and, instead, has increased using private space launching vehicle. In addition, Military communication has been operated separately from civil communication services or broadcasting services due to the special circumstances unique to the military setting. However, joint-operation of communication facility by the military and civil users is preferred because this reduces financial burden resulting from separate operation of military satellite. During the Gulf War, U.S. armed forces employed commercial satellites for its military communication. Korea's participation in space technology research is a little bit behind in time, considering its economic scale. In terms of budget, Korea is to spend 5 trillion won for 15 years for the space activities. However, Japan has 2 trillion won annul budget for the same activities. Because the development of space industry during initial fostering period does not apply to profit-making business, government supports are inevitable. All space development programs of other foreign countries are entirely supported by each government, and, only recently, private industry started participating in limited area such as a communication satellite and broadcasting satellite, Particularly, Korea's space industry is in an infant stage, which largely demands government supports. Government support should be in the form of investment or financial contribution, rather than in the form of loan or borrowing. Compared to other advanced countries in space industry, Korea needs more budget and professional research staff. Naturally, for the efficient and systemic space development and for the prevention of overlapping and distraction of power, it is necessary to enact space-related statutes, which would provide dear vision for the Korea space development. Furthermore, the fact that a variety of departments are running their own space development program requires a centralized and single space-industry development system. Prior to discussing how to coordinate or integrate space programs between Agency for Defense Development and the Korea Aerospace Research Institute, it is a prerequisite to establish, namely, "Space Operations Center"in the Air Force, which would determine policy and strategy in operating space forces. For the establishment of "Space Operations Center," policy determinations by the Ministry of National Defense and the Joint Chief of Staff are required. Especially, space surveillance system through using a military surveillance satellite and communication satellite, which would lay foundation for independent defense, shall be established with reference to Japan's space force plan. In order to resolve issues related to MTCR, Air Force would use space-launching vehicle of the Korea Aerospace Research Institute. Moreover, defense budge should be appropriated for using multi-purpose satellite and communication satellite. The Ministry of National Defense needs to appropriate 2.5 trillion won budget for space operations, which amounts to Japan's surveillance satellite operating budges.

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