Growth Inhibition by Connexin26 Expression in Cultured Rodent Tumor Cells

Hae-Jung Lee,In-Kyung Lee,Kyung-Hwan Seul,Seung-Keun Rhee 한국분자세포생물학회 2002 Molecules and cells Vol.14 No.1

The Connexin (Cx) gene family acts as a tumor sup-pressor. However, it is unclear whether the tumor-suppressing activity acquired by Cx gene transfection is mainly due to the recovery of the gap junction-mediated intercellular communication (GJIC) or to other unknown mechanisms. In order to elucidate the mechanism of the Cx-induced tumor-suppressing ac-tivity, we transfected Cx26 cDNA into a rodent mam-mary tumor cell-line (BICR-M1Rk) in which Cx43 had been normally expressed and a typical pattern of GJIC had been observed. The exogenous Cx26 was mainly localized on the nuclear envelope, whereas most of the endogenous Cx43 resided at the plasma membrane of the transfected BICR-M1Rk. Consistent with the localization of Cx26, GJIC was not increased upon the transfection of Cx26 when it was assessed by a scrape-loading dye transfer technique. A conven-tional [3H]-thymidine incorporation study showed that the growth rate of the Cx26-transfected cells was sig-nificantly decreased (70%), compared to that of the control BICR-M1Rk. Therefore, our results clearly demonstrate that the exogenously expressed Cx26 in the BICR-M1Rk cancer cell-line exerts an anti-proliferate activity in a GJIC-independent manner.

Ginseng Saponin as an Antagonist for Gap Junctional Channels

Rhee, Seung-Keun The Korean Society of Ginseng 2006 Journal of Ginseng Research Vol.30 No.2

Gap junctional channels, allowing rapid intercellular communication and synchronization of coupled cell activities, play crucial roles in many signaling processes, including a variety of cell activities. Consequently, a modulation of the gap junctional intercellular communication (GJIC) should be a potential pharmacological target. In the present, the GJIC of a epithelial-derived rat mammary cells (BICR-M1Rk) was assessed in the presence of ginseng saponin, by using an established method of scrape-loading dye transfer assay. The transfer of Lucifer yellow (diameter: 1.2 nm) among the neighboring BICR-M1Rk cells, in which connexin43 (Cx43) is a major gap junction channel-forming protein, was significantly retarded at a concentration of $10{\mu}g/ml$ ginseng saponin. By using both methods of RT-PCR and Western blotting, it was demonstrated that ginseng saponin modulated neither the mRNA synthesis of Cx43 nor the translational process of Cx43. This ginseng saponin-induced modification of GJIC was a similar phenomenon observed under the $\beta$-glycyrrhetinic acid treatment, a well-known gap junction channel blocker. Taken together, it is reasonable to conclude that the ginseng saponin inhibits GJIC only by modulating the gating property of gap junction channels.

Inhibitory Effect of Bisphenol A on Gap Junctional Intercellular Communication in an Epithelial Cell line of Rat Mammary Tissue

Lee, In-Kyoung,Rhee, Seung-Keun 대한약학회 2007 Archives of Pharmacal Research Vol.30 No.3

An endocrine disruptor, bisphenol-A (BPA), has been reported to have several short-term actions in various cells and tissues. However, the mechanisms of these actions have not been fully elucidated. In order to assess the effect of BPA on the intercellular communication modiated by gap junctions, we conducted the present study in the rat epithelium-derived BICR-M1Rk cell-line, in which connexin 43 (Cx43) is a major gap junction channel-forming protein. The cytotoxicity of BPA toward the cultured cells was evaluated by using both MTT reduction and LDH leakage assay systems. The results showed no appreciable loss in cell viability in the presence of increasing concentrations of BPA (from 0.1 to 3.2 ${\mu}$M) for 1 h incubation. However, most of cell viability was lost when cells were incubated for 24 hr with the same concen-trations of BPA. The BPA acted as an antagonist on gap junction-mediated intercellular communication (GJIC), and the phenomenon was dose-dependent and irreversible. Accord-ing to the data obtained from scrape-leading dye-transfer experiments, three quarters of nor-mal GJIC was reduced by concentration of 0.4 ${\mu}$M BPA for 1 h incubation. To identify there levance of this retardation upon BPA treatment, the GJIC to Cx43 synthesis, the mRNA and protein levels of Cx43 were assessed with RT-PCR and Western-blotting, respectively. The total protein level of Cx43 was almost constant in a wide range of BPA concentrations, as wel las in Cx43 mRNA level. These results suggest that BPA inhibits GJIC through a modulation of the gating of gap junction channels, not through a genomic modulation of Cx43.

Inhibitory Effect of Bisphenol A on Gap Junctional Intercellular Communication in an Epithelial Cell line of Rat Mammary Tissue

In-Kyoung Lee,이승근 대한약학회 2007 Archives of Pharmacal Research Vol.30 No.3

An endocrine disruptor, bisphenol-A (BPA), has been reported to have several short-term actions in various cells and tissues. However, the mechanisms of these actions have not been fully elucidated. In order to assess the effect of BPA on the intercellular communication mediated by gap junctions, we conducted the present study in the rat epithelium-derived BICRM1Rk cell-line, in which connexin 43 (Cx43) is a major gap junction channel-forming protein. The cytotoxicity of BPA toward the cultured cells was evaluated by using both MTT reduction and LDH leakage assay systems. The results showed no appreciable loss in cell viability in the presence of increasing concentrations of BPA (from 0.1 to 3.2 µM) for 1 h incubation. However, most of cell viability was lost when cells were incubated for 24 hr with the same concentrations of BPA. The BPA acted as an antagonist on gap junction-mediated intercellular communication (GJIC), and the phenomenon was dose-dependent and irreversible. According to the data obtained from scrape-loading dye-transfer experiments, three quarters of normal GJIC was reduced by concentration of 0.4 µM BPA for 1 h incubation. To identify the relevance of this retardation upon BPA treatment, the GJIC to Cx43 synthesis, the mRNA and protein levels of Cx43 were assessed with RT-PCR and Western-blotting, respectively. The total protein level of Cx43 was almost constant in a wide range of BPA concentrations, as well as in Cx43 mRNA level. These results suggest that BPA inhibits GJIC through a modulation of the gating of gap junction channels, not through a genomic modulation of Cx43.

Ginseng Saponin as an Antagonist for Gap Junctional Channels

Seung-Keun Rhee 고려인삼학회 2006 Journal of Ginseng Research Vol.30 No.2

Gap junctional channels, allowing rapid intercellular communication and synchronization of coupled cell activities, play crucial roles in many signaling processes, including a variety of cell activities. Consequently, a modulation of the gap junctional intercellular communication (GJIC) should be a potential pharmacological target. In the present, the GJIC of a epithelial-derived rat mammary cells (BICR-M1Rk) was assessed in the presence of ginseng saponin, by using an established method of scrape-loading dye transfer assay. The transfer of Lucifer yellow (diameter: 1.2 ㎚) among the neighboring BICR-M1Rk cells, in which connexin43 (Cx43) is a major gap junction channel-forming protein, was significantly retarded at a concentration of 10 ㎍/㎖ ginseng saponin. By using both methods of RT-PCR and Western blotting, it was demonstrated that ginseng saponin modulated neither the mRNA synthesis of Cx43 nor the translational process of Cx43. This ginseng saponin-induced modification of GJIC was a similar phenomenon observed under the β-glycyrrhetinic acid treatment, a well-known gap junction channel blocker. Taken together, it is reasonable to conclude that the ginseng saponin inhibits GJIC only by modulating the gating property of gap junction channels.

생쥐 난소에서 Preantral Follice의 단순 분리법

김주환,박기상,송해범,전상식,Kim, Ju-Hwan,Park, Kee-Sang,Song, Hai-Bum,Chun, Sang-Sik 대한생식의학회 2000 Clinical and Experimental Reproductive Medicine Vol.27 No.3

Objective: Our present studies were conducted to examine more effective isolating method of preantral follicles from mouse ovaries. Methods: ICR mice (3-6 weeks old) were sacrificed through cervical dislocation and their ovaries were removed and put into watch glasses containing Hams F-10 supplemented with 10% fetal bovine serum (FBS). Preantral follicles were isolated by three different methods; 1) enzymatical method and 2) mincing method, and 3) scraping method. Enzymatical method was carried out as following. Ovaries were bisected with a pair of fine 30G needles. Bisected ovaries were incubated at $37^{\circ}C$ and 5% $CO_2$ incubator in 2-well dish containing Hams F-10 supplemented with collagenase 600 lU/ml and DNAse 20 lU/ml. After 20 min., follicles were isolated by repeated pipetting. Isolated preantral follicles were collected, and the remnant of tissues was placed in incubator and previous procedure was repeated. Mincing method was carried out with a pair of fine 30G needles attached to 1 ml syringes and minced ovary. Scraping method was carried out with a pair of fine 30G needles and scratched to surface of ovary. The differences between isolating methods were analyzed using Student's t-test and Chi-square. Results were considered statistically significant when ${\rho}$ value was less than 0.05. Results: In handling time, mincing or scraping method ($28{\pm}3.42$ min or $16{\pm}1.58$ min) were significantly (p<0.00001) shorter than enzymatical method ($72{\pm}1.69$ min), and scraping method was significantly (p<0.01) shorter than mincing method. Total number of isolated follicles was significantly (p<0.0001) higher in enzymatical method ($49.8{\pm}3.91$) than in mincing or scraping method ($25.3{\pm}2.33$ or $20.5{\pm}1.75$). Isolated follicles in ${\leq}$90${\mu}m$ were significantly (p<0.005) higher in enzymatical method ($15{\pm}1.71$) than in mincing or scraping method ($7.8{\pm}0.98$ or $8.1{\pm}1.31$). In 91~130 ${\mu}m$, isolated follicles were significantly (p<0.0005) higher in enzymatical method ($33{\pm}3.27$) than in mincing or scraping method ($16.3{\pm}1.82$ or $10.7{\pm}1.38$). In ${\geq}$ 131 ${\mu}m$, isolated follicles were not significantly differences between all groups. In equal sizes, the rate of isolated follicles in ${\leq}$ 90 ${\mu}m$ was highest in scraping method (39.6% vs. enzymatical method: 30.1%, p<0.05; mincing method: 30.9%, p=0.11719, NS). Rate of follicles in $91{\sim}130$ ${\mu}m$ was significantly (p<0.05) lower in scraping method (52.7%) than in enzymatical or mincing method (66.3% or 64.5%). Rate of follicles in ${\geq}$131 ${\mu}m$ was highest in scraping method (8.3% vs. enzymatical or scraping method: 3.6%, p<0.05 or 4.6%, p=0.19053, NS). Conclusions: This study suggests that scraping method is simple and useful for isolation of preantral follicles, because this method reduced handling time and recovered enough follicles. The recovered rate of isolated follicles in diameter of 91 ~ 130 ${\mu}m$ was highest in all methods.

뉴스 기사 스크랩의 저작재산권 제한에 대한 검토

신상진 성균관대학교 법학연구원 2020 성균관법학 Vol.32 No.3

뉴스 기사는 인터넷의 등장과 디지털 기술의 발달로 인해 다양한 매체를 통한 신속한 전파가 가능하게 되었으며, 이는 곧 뉴스 기사의 저작재산권 침해 가능성을 증대하는 결과를 초래하였다. 특히 뉴스 기사 ‘스크랩’은 저작재산권 침해가 가장 빈번히 발생하는 행위로서, 법적 보호를 위한 정책적인 고민에 앞서 그 유형에 대한 분석 및 현행 저작권법제 하에서의 저작재산권 제한사유에 대한 실질적인 검토가 필요하다. 뉴스기사 스크랩은 기사의 ‘추출’ 방식에 따라 크게 기사를 직접 이용하는 방식, 기사 링크를 이용하는 방식으로 분류할 수 있다. 또한 추출된 기사의 이용방식에 따라 기사를 책이나 게시판 공간에 직접 모으는 형태, 기사나 링크를 별도의 디지털 매체에 저장하는 형태, 인터넷 기사나 링크를 다른 온라인 공간에 게시하는 형태, 기사 또는 링크를 애초에 이메일과 같은 개인 저장 공간으로 발송하는 형태로 나누어 볼 수 있다. 기사를 직접 이용하는 추출 방식에 의해 기사의 복제권 침해가 발생할 수 있으며, 추출 기사를 다른 온라인 공간에 게시하거나 동일 기사를 다수인에게 이메일로 발송할 경우 전송권 침해가 가능하다. 링크를 이용하는 방식의 경우 기사의 일부를 함께 제공하지 않는 한 링크 자체만으로는 저작재산권 침해가 발생한다고 보기 어렵다. 기사 스크랩은 저작권법상 저작재산권 제한사유인 언론기관의 전재, 사적이용을 위한 복제 등에 해당할 경우 저작재산권 침해가 발생하지 않을 것이며, 특히 위 제한사유 중 공표된 저작물의 인용, 공정이용은 침해 분쟁에 있어 주요한 항변사유로 고려될 수 있다. 저작권법상 공정이용 조항의 도입 이후 공표된 저작물의 인용 조항은 인용저작물과 피인용저작물의 주종관계를 요건으로 하여 제한적으로 적용할 필요가 있는 바, 그렇다면 기사 스크랩 결과물의 경우 인용저작물의 모음에 불과하거나 스크랩 기사를 제하면 나머지 부분으로서 기사에 비해 월등한 가치를 갖는다고 보기 어려운 경우가 많아 위 조항에 의한 저작재산권 제한을 예상하기 어렵다고 본다. 결국, 기사 스크랩에 의한 저작재산권 침해 분쟁에 있어 공정이용 조항의 적용 여부가 다투어질 것인데, 이 때 저작권법상 공정이용 여부 판단의 각 고려요소에 대하여는 LA 타임즈 대 프리리퍼블릭, AP 대 멜트웨어 판결 등 앞선 미국 법원의 판단에서 제시된 기준을 참고할 수 있을 것이다. 특히 시장 대체성과 관련하여서는 기사의 거래현실을 감안하고 기사 이용자의 역할과 스크랩의 이용방식에 따른 특성을 반영한 합리적 판단이 필요하다. These days news articles could be spread quickly through diverse mass media in accordance with the growing use of digital technology and the internet, which resulted in the increased possibility of the infringement of the author’s economic rights of news articles. Especially the infringement could occur most frequently while scraping news articles. Therefore analysis on the pattern of the scraping news articles and the grounds for the limitation on author’s economic rights under the current Copyright act is required. The pattern of scraping news articles could be categorized by the way how to extract news articles into types ; the way to use the article itself directly and the way to use the link of the article. Also the pattern could be categorized by the way how to use the extracted articles into types ; the way to collect the news articles directly in the book or in the bulletin board, the way to store the news articles in the separate digital storage, the way to post the news articles or the link of the articles in the separate website, the way to send the articles to the individual online space such as E-mail. A Right to reproduction would be violated by the way to use the article itself directly. A Right of interactive transmission could be violated by the way posting the news article on the website or sending the articles to the multiple individual through Email. The way to use the article’s link alone is not a violation of the Author’s Economic Rights as long as it does not provide some parts of articles together with the link. The conduct of scraping news articles does not infringe Author’s Economic rights if it falls in the limitations on Author’s Economic rights such as Free Use of Public Works(Article 24-2), Reproduction for private Use (Article 30). Especially the use for Quotation from Works made Public(Article 28) or Fair Use of Works(Article 35-5) could be used as major defense in the copyright infringement litigation. After the introduction of the Article 35-5 ; Fair use of Works in the copyright act, The Article 28; Quotation from Works made Public should be applied restrictively on the condition that there should be master-servant relationship between works made by quotation and the works being quoted. Therefore the application of the Article 28 could hardly be applied in scraping news article because the work products of the news scraping would be the simple collection of the works made by quotation. Also remaining parts except for the scraped article does not have higher value than the scraped article. Accordingly the application of the Fair Use article(Article 35-5) would be the main issue in copyright infringement litigation in relation to the news scraping and when judging the standards for Fair Use article under Copyright act, the case of Los Angeles Times v. Free Republic and the case of Associated Press v. Meltwater would be a good reference. Especially when judging the market substitutability, reasonable approach is required considering the news market situation, news user’s role and the way to use scraped news articles.

거문도 멸치잡이 어구ㆍ어법의 변천과 소멸

곽유석(Kwak Yu-seok) 목포대학교 도서문화연구소 2009 島嶼文化 Vol.0 No.34

멸치잡이가 성했고, 멸치잡이 관련 민속문화가 전해오는 남해 거문도 지역의 멸치잡이 방법을 사적으로 고찰하여 그 변화상을 추적하였다. 또한 멸치잡이가 사라지는 원인에 대해서도 살펴보았다. 거문도에서는 전통적으로 抄網을 써서 멸치를 잡아왔다. 또한 초망의 발전된 형태가 챗배인데, 일제 강점기 전까지는 초망과 챗배가 함께 사용되었다. 거문도는 해양생태계상 갯벌이나 모래밭이 발달해 있지 않고 수심이 깊기 때문에 독살, 죽방, 주목망 등 갯벌생태계에서 멸치를 잡을 수 있는 어구는 발전하지 않았다. 그래서 야간에 횃불로 멸치 떼를 유인해 그물로 떠 잡는 형태로 멸치를 잡아왔고, 1980년대 초반 멸치잡이가 사라질 때까지 이 원리의 방법이 유지 되었다. 일제강점기에 들어와 우리나라에 건착망이 전래되면서 멸치잡이가 대량으로 행해졌다. 그러나 거문도에서는 독자적으로 건착망의 축소 변형인 ‘홀치기’ 어법을 발전시켰다. 거문도 내해는 대형 건착망 선단이 어로작업을 하기에는 좁은 편이다. 또한 대형선단을 꾸리기 위해서는 대규모 자본이 필요하다. 그래서 거문도 사람들은 잡는 원리는 건착망과 같으나 배도 소형이고, 그물 길이도 짧고, 인원도 적게 들며, 장비도 덜 갖춘 형태의 홀치기어법을 고안해 냈다. 해방 후에도 챗배어로와 훌치기어법은 양 산맥을 이루며 거문도에서 멸치를 잡는 수단이었다. 두 어로활동의 차이를 보면, 챗배어로는 전통적으로 해오던 멸치잡이 어법이며, 홀치기는 일제 강점기에 들어왔다. 챗배어로는 1척의 배로 모든 작업을 할 수 있는 반면, 홀치기는 최소한 4척의 배가 동원되어야 한다. 어장도 차이가 난다. 챗배는 거문도 내외해 섬 주위 바다인 반면 홀치기는 거문도 내해에서만 이루어졌다. 거문도에서 멸치잡이 어구ㆍ어법은 홀치기어로, 챗배어로, 들망어로 순으로 1970년대 초반부터 80년대 초반 10여년 사이 사라졌다. 멸치잡이 소멸 원인은 인구 감소ㆍ수익 어종의 고급화 같은 사회경제적 요인, 대형어선과 기술의 발달로 인한 외해에서 어군 차단, 동도방파제 축조로 인한 해양생태계의 변화 등 몇 가지 원인이 복합적으로 작용하였다. This paper is to evaluate anchovy fishing methods in Geomun-do area where historically anchovy fishing has thrived and related folk cultures have been handed down, and to track its changes in times. Also the paper also touches on the reason behind disappearance of anchovy fishing in the region. In Geomun-do, hand scrape net was a general and traditional means for anchovy fishing. Then fixed scraping net in boat, a developed form of hand scrape net, was adopted for the fishing, and both methods were popularly used before the times of Japanese Colonial Era. Considering marine ecosystem around the islands where water is deep with almost no sand beach or mud flat, other types of anchovy fishing method such as stone barrier, bamboo weir or stow net on stakes cannot be utilized. Rather, a method of using torch light to attract a school of anchovy at night and heaving captured anchovies in the nets were generally adopted, and that method kept utilized until anchovy fishing disappeared in the early 1980s. Purse seines method was introduced in Korea during the Japanese Colonial Period enabling anchovy fishing in large scale. In Geomun-do, however, a locally-developed purse net fishing, a smaller form of purse seines method, became more wide-spread due to several factors specific to the region such as relative narrow inland sea of the region for fishing with large vessels and lack of capital to operate those vessel group. In that regard, fishers in Geomun-do borrowed the method and customized it available with smaller vessel, shorter nets, smaller number of fishers and less equipments. Even after liberalization of Korea, fixed scrape net in boat and purse net were still the two major methods for anchovy fishing. When taking a deeper look at the difference between two, major differences are as followed: fixed scrape net is a traditional method for the fishing in the region while purse net is adopted during the Japanese Colonial Period; a single boat is enough to carry out anchovy fishing with fixed scrape net in boat while purse net requires at least four boats; and fixed scrape net fishing covers both inland and open seas around Geomun-do while purse net is only performed on inland sea. Anchovy fishing methods in Geomun-do have disappeared in 10 year times from the early 1970s to the early 80s in the order of purse net, fixed scrape net in boat and lift net. The reasons are many but socioeconomic factors such as population decrease and transition to high value fish, blocked flow of fishes by large-scale fishing vessel and technological development, changes in marine ecosystem from breakwater construction in Dong-do, and several other complicated reasons are considered to be attributable to the disappearance.

괄사요법이 여대생의 신체조성, 혈중지질 및 혈관탄성도에 미치는 영향

강미영(Mi Young Kang),한나리(Na Ri Han),한채정(Chae Jeong Han) 한국인체미용예술학회 2011 한국인체미용예술학회지 Vol.12 No.1

The aim of the study was to investigate the effect of meridian scraping therapy on body composition, lipid profiles and pulse wave velocity in female college students. The subject(n=20) who was female college students participated in this study. They consists of two groups, one is control group(n=10), another is experimental group(meridian scraping massage group(n=10). To the experimental group was given the meridian scraping therapy two times per week for 8 weeks and to the control group was given the swedish massage two times per week for 8 weeks after then measured body weight(body weight, BMI, FAT, FM), lipid profiles(TC, TG, HDL and LDL), pulse wave velocity. The results of the study, repeated ANOVA by using Windows 14.0 SPSS program after 8 weeks intervention. The significant level was set to a=.05. The results are as follows. The body weight, BMI, %FAT and FM were not significant differences in the groups. TG was significantly decreased in the meridian scraping therapy group more than in the control group(p<.01). LDL was significantly decreased in the meridian scraping therapy group more than in the control group(p<.01). Pulse wave velocity was not significant differences in the groups. In conclusion, meridian scraping therapy had positive effects in TG and LDL levels. Analysis of date suggested a conclusion that merdian scraping therapy is effective to health for female college students

괄사요법(刮痧療法)에 대한 고찰(考察)

유진호,고형균,강성길,Yu, Jin-Ho,Koh, Hyung-Kyun,Kang, Sung-Keel 대한침구의학회 2000 대한침구의학회지 Vol.17 No.3

Objectives : Meridian Scraping Therapy(MST: Kwal-Sa: 刮痧) has not well known in Korea. But it's a kind of Traditional Physical Therapy wide spreaded in China and Taiwan. So I want to introduce this new Therapy by summarizing the basic theories and practical manipulations. Conclusions : To analyse the meaning of the character Kwal-Sa, Kwal(刮) means scraping the skin by some tools and Sa(痧) means originally the red spots on skin of measles but in this therapy it's the red spot on skin after the treatment. Meridian Scraping Therapy is based on the meridiology and holism which are the basic theory of Oriental Medicine. The main effects of Meridian Scraping Therapy are draining of pathogenic factors, promoring the flow of ki and blood, tonify yang, balancing the harmony of intemal organs and enhancing the immuno-ability in body. Rhinoceros corpus is common matrial of MST-plate which is the tool of manipulation. And by this plate several manipulation is performed according to the symptom and the locations of pain. The merits of MST are that easily learned by practician, the effects of the treatment are very quickly manifest and no significant side-effects. And additionally this therapy performed not only for treat diseases but prevent it and preserve health.