• Title/Summary/Keyword: Efficient Production

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Analysis of Transcriptional Activity and Estrogen Responsiveness of Regulatory Elements in Chicken Ovalbumin Promoter (닭 오브알부민 프로모터의 길이에 따른 유전자 발현 활성 및 에스트로겐 반응성 분석)

  • Yang, Hyeon;Kim, Kyung-Woon;Kim, Jeom Sun;Woo, Jae-Seok;Lee, Hwi-Cheul;Choi, Hoonsung;Jung, Sun Keun;Sureshkumar, Shanmugam;Lee, Haesun;Oh, Keon Bong;Byun, Sung June
    • Korean Journal of Poultry Science
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    • v.46 no.1
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    • pp.17-24
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    • 2019
  • Chickens have been considered as well-defined animal bioreactor. The optimized ovalbumin promoter is essential for recombinant protein production in transgenic chicken. Here we try to compare the activity and identify the effect of estrogen on ovalbumin promoter according to each promoter length with estrogen response element (ERE) existence. We cloned two (2.8 and 5.5 kb) ovalbumin promoters that the 5.5 kb contained the ERE but the 2.8 kb did not, and these two promoters were cloned to pGL4.11 vector. Additionally, we constructed another pGL4.11 vector containing of the 4.4 kb (with ERE) ovalbumin promoter deleted with 1 kb between ERE region and the 2.8 kb promoter. For reporter assay, HeLa, MES-SA, LMH/2A, and cEF cells were transfected with all the pGL4.11 vectors. The comparative analysis showed that the mutated 4.4 kb promoter has more potent activity than the 2.8 and 5.5 kb promoters in HeLa, MES-SA, and LMH/2A cells. However, there is no significant difference in cEFs. Also, these cells transfected with the mutated 4.4 kb promoter were treated with the $17{\beta}$-estradiol (0~3,000 nM) and HeLa, MES-SA, and LMH/2A cells showed estrogen responsibilities, but cEFs did not. Besides, the mutated 4.4 kb promoter has still higher activity than the 2.8 and 5.5 kb promoter, and there is no transcriptional induction effect in 2.8 kb promoter at 500 nM estrogen that is blood concentration of laying hens. Hence our study strongly suggested that the mutated 4.4 kb promoter is considered as one of the most efficient length for generating transgenic chicken.

In vitro mass propagation and acclimatization of Haworthia truncata (하월시아 옥선(Haworthia truncata)의 기내 대량 증식 및 순화 조건 구명)

  • Kim, Youn Hee;Lee, Gee Young;Kim, Hye Hyeong;Lee, Jae Hong;Jung, Jae Hong;Lee, Sang Deok
    • Journal of Plant Biotechnology
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    • v.46 no.2
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    • pp.127-135
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    • 2019
  • The purpose of this study was to investigate suitable parts for callus induction and optimal concentrations of growth regulators, contained in the medium affecting shoot and rooting for in vitro mass production of Haworthia truncata. Leaves and flower bud showed 100% callus formation rate at NAA $1{\sim}2mgL^{-1}$ treatment, and NAA $1mgL^{-1}$ + TDZ $2mgL^{-1}$ treatment. The flower stalk showed 75% callus formation rate, at NAA $2mgL^{-1}$ + TDZ $2mgL^{-1}$ treatment in H. truncata. While the rate of callus formation was high in leaves and flower bud, leaves were the most efficient in obtaining most culture parts. Shoot induction rate from callus was highest, at NAA $0.1mgL^{-1}$ treatment in H. truncata. Additionally, the number of shoots formation was 66.3 shoots high, in NAA $1mgL^{-1}$ + BA $0.1mgL^{-1}$ treatment in H. truncata. In the case of acclimatization of regenerated plant, growth characteristics did not show significant difference (95%) shading with respect to the different ratio of substrate mixture, and it was determined that would be appropriate, considering plant height and appearance preference of H. truncata. It was established that optimization of culture condition, was responsible for mass propagation in vitro cultures of H. truncata.

Growth and Useful Component of Angelica gigas Nakai under High Temperature Stress (고온 스트레스에 따른 참당귀의 생육 및 유용성분 특성)

  • Jeong, Dae Hui;Kim, Ki Yoon;Park, Sung Hyuk;Jung, Chung Ryul;Jeon, Kwon Seok;Park, Hong Woo
    • Korean Journal of Plant Resources
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    • v.34 no.4
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    • pp.287-296
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    • 2021
  • Recently, the pace of global climate change has tremendously increased, causing extreme damage to crop production. Here, we aimed to examine the growth characteristics and useful components of Angelica gigas under extreme heat stress, providing fundamental data for its efficient cultivation. Plants were exposed to various experimental temperatures (28℃, 34℃, and 40℃), and their growth characteristics and content of useful components were analyzed. At the experimental site, the ambient and soil temperature were 19.38℃ and 21.34℃, ambient and soil humidity were 81.3 % and 0.18 m3/m3, solar radiation was 162.05 W/m2. Moreover, the soil was sandy-clay-loam (pH 6.65), with 2.66% organic matter, 868.52 mg/kg soil available phosphate, and 0.14% nitrogen. Values of most growth characteristics, including the survival rate (85%), plant height (38.66cm), and fresh and dry weight (41.3 g and 14.24 g), were the highest at 28℃. Although the highest content of useful components was observed at 34℃ (3.24%), there were no significant differences across temperatures. Growth characteristics varied across temperatures due to detrimental effects of heat stress, such as accelerated tissue aging, reduced photosynthesis, and delay of growth. Similar content of useful components across temperatures may be due to poor accumulation of anabolic products caused by impaired growth at extremely high temperatures.

Considerations of Countermeasure Tasks in the Fields of Forest and Forestry in Korea through Case Study on "The Nagoya Protocol (Access to Genetic Resources and Benefit Sharing)" ("유전자원의 접근과 이익공유(ABS)" 사례연구를 통한 국내 산림·임업분야 대응과제 고찰)

  • Lee, Gwan Gyu;Kim, Jun Soon;Jung, Haw young
    • Journal of Korean Society of Forest Science
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    • v.100 no.3
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    • pp.522-534
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    • 2011
  • The aim of this study is to draw forth the tasks for establishing the right of native biology in Korea through the case study on 'Access on genetic resources and Benefit Sharing'. For this purpose, this study decided on its research subject by selecting Hoodia, on which ABS treaty was made the most recently, through the examination of the representative ABS precedents on plant species. This study analyzed the process background of ABS on Hoodia, and compared & analyzed the ABS procedures of 'Bonn Guidelines' adopted by the 6th Conference of the Parties of the Convention on Biological Diversity in 2002 and Hoodia case. Together with the ABS major issues in common drawn as a result of this analysis, and "Nagoya Protocol" adopted by the 10th Conference of the Parties of the Convention on Biological Diversity, this study intended to shed a light on the impending tasks which Korea faces at present and its role relationship. The research results are as follows: 1. It is required that species habitats should be divided based on biological classification and its subsequent community should be established with the development of infrastructure such as a community's independent production, management and monitoring of bio-species. 2. There needs to be a designation of ABS National Focal Point for sharing of ABS-related general information, boosting of implementation of the relevant convention. 3. There needs to be the establishment of ABS convention system consequent on legislative, administrative, political procedures, and designation of the Competent National Authorities for the provision of the format of Prior Informed Consent (PIC) and Mutually Agreed Terms (MAT) and their contents assessment and confirmation. 4. There should be the establishment of integrated management system of ABS-related research and development of forest biological resources and its relevant research projects. 5. There should be information development through the distribution of responsibility and role between the ministries and offices concerned according to bio-resources, and there needs to be efforts in aiming for opening a working group of academic-industrial institutions for developing a mutually interchangeable system. 6. It's required that the efficient access between industrial circles and the people should be promoted by setting up ABS support center of biological resources in ministry and office's charge. 7. There should be a selection of a national supervisory organization for securement of the right of a local community and monitoring of ABS convention implementation, and a countermeasure system for preventing outflow of forest bioresources. Conclusively, it's judged that it will be possible to inquire into the countermeasures for the establishment of the native forest biology dominion through such research results.

Evaluation of the Potential of Nitrogen Plasma to Cosmetics (질소 플라즈마의 화장품 가능성 평가)

  • Lee, So Min;Jung, So Young;Brito, Sofia;Heo, Hyojin;Cha, Byungsun;Lei, Lei;Lee, Sang Hun;Lee, Mi-Gi;Bin, Bum-Ho;Kwak, Byeong-Mun
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.48 no.3
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    • pp.189-196
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    • 2022
  • Plasma refers to an ionized gas that is often referred to as "the fourth phase of matter", following solid, liquid, and gas. Plasma has traditionally been utilized for industrial applications such as welding and neon signs, but its promise in biomedical fields such as cancer treatment and dermatology has lately been recognized. Indeed, due to its beneficial effects in promoting collagen production, improving skin tone, and eliminating harmful bacteria in the skin, plasma treatment constitutes an important target for dermatological research. In this study, a plasma device for cosmetic manufacturing based on nitrogen, the main component of the atmosphere, was designed and assembled. Moreover, nitric oxide (NO) was selected since is easier to follow and evaluate than other nitrogen plasma active species, and its contents were measured to perform a quantitative and qualitative evaluation of plasma. First, an injection method, using different proximities labeled "sinking" and "non sinking" treatments, was performed to test the most efficient plasma treatment method. As a result, it was observed that the formulation obtained by a non sinking treatment was more effective. Furthermore, toner and ampoule were selected as cosmetics formulations, and the characteristics of the formulation and changes in the injected plasma state were observed. In both formulations, the successful injection of NO plasma was 2 times higher in toner formulation than ampoule formulation, and it gradually decreased with time, having dissipated after a week. It was confirmed that the nitrogen plasma used did not affect the stability of the toner and ampoule formulations at low temperature (4 ℃), room temperature (25 ℃), and high temperature (37 ℃ and 50 ℃) conditions. The results of this study demonstrate the potential of plasma cosmetics and highlight the importance of securing the stability of the injected plasma.

A Study on Policy-making, Leadership and Improvement of Professionalism for Audiovisual Archives Management in Korea (국내 시청각 기록관리 정책 리더십 및 전문성 제고 방안 연구)

  • Choi, Hyo jin
    • The Korean Journal of Archival Studies
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    • no.72
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    • pp.91-163
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    • 2022
  • The focus of this paper lies on the fact that the 'management' and 'utilization' of audiovisual archives are still not specialized in both the public and the private sectors. The use of online video platforms including 'YouTube' has became common. Accordingly the production and collection of high-definition and high-capacity audiovisual archives has been rapidly increasing. However, it also emphasizes that there are no references or principles in the current Public Records Act and its enforcement rules, public standards, and guidelines. This paper ultimately examines the provisions that are related to audiovisual archives of the current Public Records Act, which needed to be revised and enacted due to the lack of an audiovisual archives management manual of national institutions, public broadcasters, and organizations can refer to. In addition, this study tries to find out what kind of systems and guidelines are used in audiovisual archives management. This paper examines the current state of standardization of audiovisual records of the National Archives. It also analyses the systems and the guidelines methodically for efficient audiovisual record management in the public records management sector. It suggests the new direction of relevant public standards and guidelines through this research. Futhermore, it measures to activate the audiovisual management policy-making functions of the National Archives. The necessity of establishing a Public Audiovisual Archives as an organization was also reviewed in this paper. The Public Audiovisual Archives will collect Public Audio and Videos systematically and comprehensively through the legal deposit system. And it will be operated by the management and the utilization system so that it can be used for public as a collective memory. Finally, it will takes a charge of a professional role in audiovisual record management field, such as technology standardization to safeguard and protect the copyrights through this process.

A review of the mass-mortalities of sea-cage farm fishes (해상 가두리양식장 양식어류의 대량폐사에 대하여)

  • Han, Jido;Lee, Deok-Chan
    • Journal of fish pathology
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    • v.35 no.1
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    • pp.1-25
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    • 2022
  • The aquaculture industry has developed rapidly over the last three decades and is an important industry that supplies over 15% of humans' animal protein intake; therefore, there is a need to increase production to meet the continuous demand. The fish cage farms on the southern coast (Kyengsangnam-do and Jeollanam-do) of Korea are critical resources in aquaculture because they account for approximately 90% of the national total fish cage farms by water area ratio. However, the current aquaculture environment is being gradually affected by climate change, which is a global issue, and its effects are expected to intensify in the future. Therefore, it is urgently imperative to accurately evaluate the effects of climate change on South Korean aquaculture industries and to develop social and national strategies to minimize damage to the fishing industry. The damage to fish farmed in cage farms on the southern coast is increasing annually and the leading causes are high and low water temperature and red tides, which are directly or indirectly related to climate change. At present, global warming can provide opportunities for aquaculture industrialization of fish or other novel species, with economic implications. However, despite such opportunities, the influx of new species can also cause problems such as ecological disturbances, increase in the reproduction frequency of microalgae such as red tide, increase in disease incidence, and occurrence and periods of high water temperatures in summer. The scale of farmed fish mortality is increasing due to the complex effects of these factors. Increased damages due to fish mortality not only have severe economic impacts on the aquaculture industry, but the social costs of responding to the damage and follow-up measures also increase. various active responses can reduce the mortality damage in fish farms such as improving the management skills in aquaculture, improved species breeding, efficient food management, disease prevention, proactive responses, and system-wide improvements. This review article analyzes the large-scale mortality cases occurring in fish cage farms on the southern coast of Korea and proposes measures to mitigate mortality and enhance responses to such scenarios.

Perceptional Change of a New Product, DMB Phone

  • Kim, Ju-Young;Ko, Deok-Im
    • Journal of Global Scholars of Marketing Science
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    • v.18 no.3
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    • pp.59-88
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    • 2008
  • Digital Convergence means integration between industry, technology, and contents, and in marketing, it usually comes with creation of new types of product and service under the base of digital technology as digitalization progress in electro-communication industries including telecommunication, home appliance, and computer industries. One can see digital convergence not only in instruments such as PC, AV appliances, cellular phone, but also in contents, network, service that are required in production, modification, distribution, re-production of information. Convergence in contents started around 1990. Convergence in network and service begins as broadcasting and telecommunication integrates and DMB(digital multimedia broadcasting), born in May, 2005 is the symbolic icon in this trend. There are some positive and negative expectations about DMB. The reason why two opposite expectations exist is that DMB does not come out from customer's need but from technology development. Therefore, customers might have hard time to interpret the real meaning of DMB. Time is quite critical to a high tech product, like DMB because another product with same function from different technology can replace the existing product within short period of time. If DMB does not positioning well to customer's mind quickly, another products like Wibro, IPTV, or HSPDA could replace it before it even spreads out. Therefore, positioning strategy is critical for success of DMB product. To make correct positioning strategy, one needs to understand how consumer interprets DMB and how consumer's interpretation can be changed via communication strategy. In this study, we try to investigate how consumer perceives a new product, like DMB and how AD strategy change consumer's perception. More specifically, the paper segment consumers into sub-groups based on their DMB perceptions and compare their characteristics in order to understand how they perceive DMB. And, expose them different printed ADs that have messages guiding consumer think DMB in specific ways, either cellular phone or personal TV. Research Question 1: Segment consumers according to perceptions about DMB and compare characteristics of segmentations. Research Question 2: Compare perceptions about DMB after AD that induces categorization of DMB in direction for each segment. If one understand and predict a direction in which consumer perceive a new product, firm can select target customers easily. We segment consumers according to their perception and analyze characteristics in order to find some variables that can influence perceptions, like prior experience, usage, or habit. And then, marketing people can use this variables to identify target customers and predict their perceptions. If one knows how customer's perception is changed via AD message, communication strategy could be constructed properly. Specially, information from segmented customers helps to develop efficient AD strategy for segment who has prior perception. Research framework consists of two measurements and one treatment, O1 X O2. First observation is for collecting information about consumer's perception and their characteristics. Based on first observation, the paper segment consumers into two groups, one group perceives DMB similar to Cellular phone and the other group perceives DMB similar to TV. And compare characteristics of two segments in order to find reason why they perceive DMB differently. Next, we expose two kinds of AD to subjects. One AD describes DMB as Cellular phone and the other Ad describes DMB as personal TV. When two ADs are exposed to subjects, consumers don't know their prior perception of DMB, in other words, which subject belongs 'similar-to-Cellular phone' segment or 'similar-to-TV' segment? However, we analyze the AD's effect differently for each segment. In research design, final observation is for investigating AD effect. Perception before AD is compared with perception after AD. Comparisons are made for each segment and for each AD. For the segment who perceives DMB similar to TV, AD that describes DMB as cellular phone could change the prior perception. And AD that describes DMB as personal TV, could enforce the prior perception. For data collection, subjects are selected from undergraduate students because they have basic knowledge about most digital equipments and have open attitude about a new product and media. Total number of subjects is 240. In order to measure perception about DMB, we use indirect measurement, comparison with other similar digital products. To select similar digital products, we pre-survey students and then finally select PDA, Car-TV, Cellular Phone, MP3 player, TV, and PSP. Quasi experiment is done at several classes under instructor's allowance. After brief introduction, prior knowledge, awareness, and usage about DMB as well as other digital instruments is asked and their similarities and perceived characteristics are measured. And then, two kinds of manipulated color-printed AD are distributed and similarities and perceived characteristics for DMB are re-measured. Finally purchase intension, AD attitude, manipulation check, and demographic variables are asked. Subjects are given small gift for participation. Stimuli are color-printed advertising. Their actual size is A4 and made after several pre-test from AD professionals and students. As results, consumers are segmented into two subgroups based on their perceptions of DMB. Similarity measure between DMB and cellular phone and similarity measure between DMB and TV are used to classify consumers. If subject whose first measure is less than the second measure, she is classified into segment A and segment A is characterized as they perceive DMB like TV. Otherwise, they are classified as segment B, who perceives DMB like cellular phone. Discriminant analysis on these groups with their characteristics of usage and attitude shows that Segment A knows much about DMB and uses a lot of digital instrument. Segment B, who thinks DMB as cellular phone doesn't know well about DMB and not familiar with other digital instruments. So, consumers with higher knowledge perceive DMB similar to TV because launching DMB advertising lead consumer think DMB as TV. Consumers with less interest on digital products don't know well about DMB AD and then think DMB as cellular phone. In order to investigate perceptions of DMB as well as other digital instruments, we apply Proxscal analysis, Multidimensional Scaling technique at SPSS statistical package. At first step, subjects are presented 21 pairs of 7 digital instruments and evaluate similarity judgments on 7 point scale. And for each segment, their similarity judgments are averaged and similarity matrix is made. Secondly, Proxscal analysis of segment A and B are done. At third stage, get similarity judgment between DMB and other digital instruments after AD exposure. Lastly, similarity judgments of group A-1, A-2, B-1, and B-2 are named as 'after DMB' and put them into matrix made at the first stage. Then apply Proxscal analysis on these matrixes and check the positional difference of DMB and after DMB. The results show that map of segment A, who perceives DMB similar as TV, shows that DMB position closer to TV than to Cellular phone as expected. Map of segment B, who perceive DMB similar as cellular phone shows that DMB position closer to Cellular phone than to TV as expected. Stress value and R-square is acceptable. And, change results after stimuli, manipulated Advertising show that AD makes DMB perception bent toward Cellular phone when Cellular phone-like AD is exposed, and that DMB positioning move towards Car-TV which is more personalized one when TV-like AD is exposed. It is true for both segment, A and B, consistently. Furthermore, the paper apply correspondence analysis to the same data and find almost the same results. The paper answers two main research questions. The first one is that perception about a new product is made mainly from prior experience. And the second one is that AD is effective in changing and enforcing perception. In addition to above, we extend perception change to purchase intention. Purchase intention is high when AD enforces original perception. AD that shows DMB like TV makes worst intention. This paper has limitations and issues to be pursed in near future. Methodologically, current methodology can't provide statistical test on the perceptual change, since classical MDS models, like Proxscal and correspondence analysis are not probability models. So, a new probability MDS model for testing hypothesis about configuration needs to be developed. Next, advertising message needs to be developed more rigorously from theoretical and managerial perspective. Also experimental procedure could be improved for more realistic data collection. For example, web-based experiment and real product stimuli and multimedia presentation could be employed. Or, one can display products together in simulated shop. In addition, demand and social desirability threats of internal validity could influence on the results. In order to handle the threats, results of the model-intended advertising and other "pseudo" advertising could be compared. Furthermore, one can try various level of innovativeness in order to check whether it make any different results (cf. Moon 2006). In addition, if one can create hypothetical product that is really innovative and new for research, it helps to make a vacant impression status and then to study how to form impression in more rigorous way.

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Analysis of 5-aza-2'-deoxycytidine-induced Gene Expression in Lung Cancer Cell Lines (폐암 세포주에서 5-aza-2'-deoxycytidine 처치에 의해 발현되는 암항원 유전자 분석)

  • 김창수;이해영;김종인;장희경;박종욱;조성래
    • Journal of Chest Surgery
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    • v.37 no.12
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    • pp.967-977
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    • 2004
  • Background: DNA methylation is one of the important gene expression mechanisms of the cell. When cytosine of CpG dinucleotide in promotor is hypomethylated, expression of some genes that is controlled by this promoter is altered. In this study, the author investigated the effect of DNA demethylating agent, 5-aza-2'-deoxycytidine (ADC), on the expressions of cancer antigen genes, MHC and B7 in 4 lung cancer cell lines, NCIH1703, NCIH522, MRC-5, and A549. Material and Method: After treatment of cell lines, NCIH1703, NCIH522, MRC-5 and A549 with ADC (1 uM) for 48 hours, RT-PCR was performed by using the primers of MAGE, GAGE, NY-ESO-1, PSMA, CEA, and SCC antigen gene. In order to find the optimal ADC treatment condition for induction of cancer antigen, we studied the effect of ADC treatment time and dose on the cancer antigen gene expression. To know the effect of ADC on the expression of MHC or B7 and cell growth, cells were treated with 1 uM of ADC for 72 hours for FACS analysis or cells were treated with 0.2, 1 or 5 uM of ADC for 96 hours for cell counting. Result: After treatment of ADC (1 uM) for 48 hours, the expressions of MAGE, GAGE, NY-ESO-1, and PSMA genes increased in some cell lines. Among 6 MAGE isotypes tested, and gene expression of MAGE-1, -2, -3, -4 and -6 could be induced by ADC treatment. However, CEA gene expression did not change and SCC gene expression was decreased by ADC treatment. Gene expression was generally induced 24 - 28 hours after ADC treatment and expression of MAGE, GAGE, and NY-ESO-1 was maintained at least 14 days after ADC ADC teatment, and expression of MAGE, GAGE, and NY-ESO-1 was maintained at least 14 days after ADC teatment in ADC-Free medium. Most gene expression could be induced at 0.2 uM of ADC, but gene expression increased dependently on ADC treatment dose. The expression of MHC and B7 was not increased by ADC treatment in all four cell lines, and the growth rate of 4 cell lines decreased significantly with the increase of ADC concentrations. Conclusion: Treatment of lung cancer cell lines with ADC increases the gene expression MAGE, GAGE and NY-ESO-1 that are capable of induction of cytotoxic T lymphocyte response. We suggest that treatment with 1 uM of ADC for 48 hours and then culturing in ADC-free medium is optimal condition for induction of cancer antigen. However, ADC has no effect on MHC and B7 induction, additional modification for increase of expression of MHC, B7 and cytokine will be needed for production of efficient cancer cell vaccine.

Effects of Activation Regimens of Recipient Cytoplasm, Culture Condition of Donor Embryos and Size of Blastomeres on Development of Reconstituted Bovine Embryos (수핵 난자의 활성화 방법과 공핵 수정란의 배양체계 및 할구의 크기가 소 핵이식 수정란의 발달에 미치는 영향)

  • 심보웅;조성근;이효종;박충생;최상용
    • Korean Journal of Animal Reproduction
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    • v.22 no.4
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    • pp.425-435
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    • 1998
  • To improve the efficiency of nuclear transplantation in bovine, in this study the development in vitro of nuclear transferred (NT) embryos was compared by different activation regimens of the enucleated oocytes. The effect of developmental stage and culture system of donor nuclei on fusion and development in vitro of NT embryos were also evaluated. Oocytes were collected from Hanwoo ovaries obtained from slaughterhouse and matured in Ham's F-10 supplemented with hormones. After 20~22 h maturation, the oocytes were vortexed to be free from cumulus cells and subsequently their nucleus and the first polar body were removed. Enucleated oocytes were divided into 3 groups for activation; the oocytes of group I were activated with ionomycin for 5 min and subsequently incubated in 6-dimetylarninopurine (DMAP) for 4 h, Those of group II were treated with DMAP for 4 h at 39 h after onset of in vitro maturation (IVM) and those of group III were kept in room temperature ($25^{\circ}C$) for 3 h at 39 h after onset of IVM. After in vitro fertilization (IVF) the embryos for muclear donor were cultured either by group culture (20 embryos /50 ${mu}ell$ drop) or individually (1 embryo /50 ${mu}ell$ drop) for 4 day and 5 day. At day 4 and 5 after IVF, blastomeres were separated in calcium-magnesium free medium, and then classified into small (day 5: $\leq$ 38 ${\mu}{\textrm}{m}$, day 4: $\leq$ 46 ${\mu}{\textrm}{m}$) and large (day 5 : $\geq$ 38 ${\mu}{\textrm}{m}$, day 4 ; $\geq$ 46 ${\mu}{\textrm}{m}$). The separated blastomeres were replaced into enucleated and activated recipient cytoplasm. The blastomere-oocyte complexes were fused by electrically. The NT embryos were cultured in TCM-199 containing 10% FCS in 39$^{\circ}C$, 5% $CO_2$ incubator for 7 day. The results obtained were summarized as follows; There were no differences in fusion and development to blastocyst between groups as group I (68%, 10%), group II (75%, 14%) and group III (73%, 9%), respectively. However, the cell number in blastocyst of NT embryos in group III were significantly fewer than in the other groups (P<0.05). No differences in fusion and development to blastocyst were found between individual or group cultured and between small or large blastomeres of day 4 and day 5 donor embryos. From these results, it was concluded that the combination of ionomycin and DMAP, or treatment of DMAP at 39 h after onset of IVM were useful for the efficient of production of NT bovine embryos, and the individual cultured embryos could be simply used as donor nuclei for NT bovine embryo.

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