• 제목/요약/키워드: two-step culture process

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생쥐 생식줄기세포의 체외 분리 및 증식 (In Vitro Isolation and Proliferation of Mouse Male Germ-Line Stem Cells)

  • 김수경;김계성
    • 한국수정란이식학회지
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    • 제18권3호
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    • pp.243-248
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    • 2003
  • 1. 생쥐 고환으로부터 얻은 세포를 배양하여 군집을 형성하는 것을 관찰할 수 있었으며, AP, SSEA-1, -3, -4과 Integrin $\alpha$6, $\beta$1 및 Oct4의 발현을 확인하였다. 2. 생쥐 생식줄기세포를 3-5일정도 배양하게 되면, 여러 층으로 이루어진 군집을 이루게 되는데 이는 생쥐 배아줄기세포나 배아생식줄기세포의 형태와 같은 것이었다. 3. 생쥐 생식줄기세포를 체외에서 효과적으로 분리, 배양할 수 있는 조건을 확립하였다.

Screening and Evaluation of Antibacterial Metabolites from Entomopathogenic Fungi

  • Shin, Tae-Young;Ko, Seung-Hyun;Lee, Won-Woo;Bae, Sung-Min;Choi, Jae-Bang;Woo, Soo-Dong
    • International Journal of Industrial Entomology and Biomaterials
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    • 제26권2호
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    • pp.89-94
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    • 2013
  • Entomopathogenic fungal species have been investigated for their potential use as biological control agents owing to their natural role as insect pathogens. These fungi produce a wide range of secondary metabolites with high therapeutic values, such as antibiotics and cytotoxic substances. To evaluate the antibacterial activity of entomopathogenic fungi, 10 isolates from Korean soil were selected and tested for their activity against Escherichia coli by using fungal culture filtrates. Antibacterial activity was assessed using a two-step process: (1) a screening assay for the selection of fungal isolates and (2) a quantitative assay to evaluate the activity of select fungi. Although 4 fungal isolates were selected through the screening assay, only 3 fungal isolates, from Beauveria bassiana and Metarhizium anisopliae, showed high antibacterial activity according to the quantitative assay. The antibacterial activity of selected fungal culture filtrates was stable when exposed to heat and proteolytic enzyme treatments, which indicated that the antibacterial compound is not a protein. These entomopathogenic fungal metabolites might be useful as a source for bacterial control and in the pharmaceutical industry.

The Factors Affecting Unsafe Behaviors of Iranian Workers: A Qualitative Study Based on Grounded Theory

  • Malakoutikhah, Mahdi;Jahangiri, Mehdi;Alimohammadlou, Moslem;Faghihi, Seyed Aliakbar;Kamalinia, Mojtaba
    • Safety and Health at Work
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    • 제12권3호
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    • pp.339-345
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    • 2021
  • Background: Some researchers state that they are not yet able to provide a deep understanding of the underlying causes of unsafe behaviors (UBs). Therefore, the present study was conducted to investigate the attitudes and experiences of Iranian workers of UBs. Methods: This present study was conducted in 35 industries using a semistructured interview based on grounded theory. Forty participants were interviewed, including 13 industrial safety and health experts and 27 workers and supervisors. The analysis of the present study consisted of a three-step coding process including open, axial, and selective coding. Results: The results showed that the factors affecting UBs could be classified into three categories: organizational, individual, and socioeconomic factors. Organizational factors were divided into 6 parts: procedure and environmental conditions, communications, monitoring, organizational safety culture, resource allocation, and human resources. Socioeconomic factors had three subcategories: community safety culture, type of organizational ownership, and economic problems. Finally, the individual factors were classified into two categories of personality traits and individual competence. Conclusion: The results showed that organizational factors were the most categorized, and it is estimated that this factor has a more important role in the UBs. Of course, to better understand the close relationship between these factors and find the weight and importance of each factor, it needs to measure it with multicriteria decision systems.

Expression and Purification of Biologically Active Human Bone Morphogenetic Protein-4 in Recombinant Chinese Hamster Ovary Cells

  • Cha, Minyub;Han, Nara;Pi, Jia;Jeong, Yongsu;Baek, Kwanghee;Yoon, Jaeseung
    • Journal of Microbiology and Biotechnology
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    • 제27권7호
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    • pp.1281-1287
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    • 2017
  • Bone morphogenetic protein-4 (BMP-4) is considered to have therapeutic potential for various diseases, including cancers; however, the high expression of biologically active recombinant human BMP-4 (rhBMP-4) needed for its manufacture for therapeutic purposes has yet to be established. In the current study, we established a recombinant Chinese hamster ovary (rCHO) cell line overexpressing rhBMP-4 as well as a production process using 7.5-l bioreactor (5 L working volume). The expression of the mature rhBMP-4 was significantly enhanced by recombinant furin expression. The combination of a chemically defined medium and a nutrient supplement solution for high expression of rhBMP-4 was selected and used for bioreactor cultures. The 11-day fed-batch cultures of the established rhBMP-4-expressing rCHO cells in the 7.5-L bioreactor produced approximately 32 mg/l of rhBMP-4. The mature rhBMP-4 was purified to homogeneity from the culture supernatant using a two-step chromatographic procedure, resulting in a recovery rate of approximately 55% and a protein purity greater than 95%. The N-terminal amino acid sequences and N-linked glycosylation of the purified rhBMP-4 were confirmed by N-terminal sequencing and de-N-glycosylation analysis, respectively. The mature purified rhBMP-4 has been proved to be functionally active, with an effective dose concentration of $EC_{50}$ of 2.93 ng/ml.

The Two-Component Protease NS2B-NS3 of Dengue Virus Type 2: Cloning, Expression in Escherichia coli and Purification of the NS2B, NS3(pro) and NS2B-NS3 Proteins

  • Champreda, Veerawat;Khumthong, Rabuesak;Subsin, Benchamas;Angsuthanasombat, Chanan;Panyim, Sakol;Katzenmeier, Gerd
    • BMB Reports
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    • 제33권4호
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    • pp.294-299
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    • 2000
  • Proteolytic processing of the dengue virus serotype 2 polyprotein precursor is catalyzed by a host signal peptidase and a virus encoded two-component protease consisting of the nonstructural proteins, NS2B and NS3. We expressed in Escherichia coli the NS2B, NS3(pro) and NS2B-NS3 proteins from the dengue virus type 2 strain 16681 as N-terminal fusions with a hexahistidine affinity tag under the control of the inducible trc promoter. All fusion proteins were purified to >90% purity by detergent extraction of inclusion bodies and a single step metal chelate chromatography. Proteins were refolded on-column and recovered with yields of 0.5, 6.0 and 1.0 mg/l of E. coli culture that was grown to $OD_{600}=1.0$ for NS2B, NS3(pro) and NS2B-NS3, respectively. Purified proteins gave strong signals in Western blots using $Ni^{2+}-nitrilotriacetic$ acid as a probe for the presence of the polyHis tag. During the purification process, $(His)_{6}NS2B-NS3$ was apparently not autoproteolytically cleaved at the NS2B/NS3 site.

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문화간호를 위한 한국인의 민간 돌봄에 대한 연구 : 출생을 중심으로 (Study on Folk Caring in Korea for Cultural Nursing)

  • 고성희;조명옥;최영희;강신표
    • 대한간호학회지
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    • 제20권3호
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    • pp.430-458
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    • 1990
  • Care is a central concept of nursing. Nursing would not exist without caring. Care and quality of life are closely related. Human behavior is a manifestation of culture. We can say that caring and nursing care are expression of culture. The nurse must understand the relationship of culture with care for ensure quality nursing care. But knowledge of cultural factors in nursing is not well developed. Time and in - depth study are needed to find meaningful relationships between culture and care. Nurses recognized the importance of culturally appropriate nursing There are two care systems in culturally based nursing. The folk care system and the professional nursing care system. The folk care system existed long before the professional nursing care system was introduced into this culture. If the discrepancy between these two care systems is great, the client may receive inappropriate nursing care. Culture and subcaltures are diverse and dynamic in nature. Nurses need to know the caring behaviors, patterns, and their meaning in their own culture. In Korea we have taken some first step to study cultural nursing phenomena. It is not our intent necessarily to return to the past and develop a nationalistic of nursing, but to identify the core of traditional caring and relate that to professional nursing care. Our Assumptions are as follows : 1) Care is essential for human growth, well being and survial. 2) 7here are diverse and universal forma, expressions, patterns, and processes of human care that exist transcul - turally. 3) The behaviors and functions of caring differ according to the social structure of each culture. 4) Cultures have folk and professional care values, beliefs, and practices. To promote the quality of nursing care we must understand the folk care value, beliefs, and practices. We undertook this study to understand caring in our traditional culture. The Goals of this study were as follows : 1) To identify patterns in caring behavior, 2) To identify the structural components of caring, and 3) To understand the meaning and some principles of caring. We faised several questions in this study. Who is the care-giver? Who is the care-receipient? Was the woman the major care -giver at any time? What are the patterns in caring behavior? What art the priciples underlying the caring process? We used an interdisciplinary team approach, composed of representatives from nursing and anthropology, to contribute in -depth understanding of caring through a socicaltural perspeetive. A Field study was conducted in Ro-Bong, a small agricultural kinship village. The subjects were nine women and one man aged be or more years of age. Data were collected from january 15 to 21, 1990 through opem-ended in-depth interviews and observations. The interview focused on caring behaviors sorrounding birth, aging, death and child rearing. We analysed these data for meaning, pattern and priciples of caring. In this report we describe caring behaviors surrounding childbirth. The care-givers were primarily mothers- in -low, other women in the family older than the mother - to- be, older neighbor woman, husbands, and mothers of the mother-to- be. The care receivers were the mother-to-be the baby, and the immediate family as a component of kinship. Emerging caring behavior included praying, helping proscribing, giving moral advice(Deug - Dam), showing concern, instructing, protecting, making preparations, showing consideration, touching, trusting, encouraging, giving emotional comfort, being with, worrying about, being patient, preventing problems, showing by an example, looking after bringing up, taking care of postnatal health, streng thening the health condition, entering into another's feelings(empathizing), and sharing food, joy and sorrow The emerging caring component were affection, touching, nurtuing, teaching, praying, comforting, encouraging, sharing. empathizing, self - discipline, protecting, preparing, helping and compassion. Emerging principles of. caring were solidarity, heir- archzeal relationships, sex - role distinction. Caring during birth expresses the valve of life and reflects the valued traditional beliefs that human birth is given by god and a unique unifying family event reaching back to include the ancestors and foreward to later generations. In addition, We found positive and rational foundations for traditionl caring behaviors surrounding birth, these should not be stigmatized as inational or superstitious. The nurse appropriately adopts the rational and positive nature of traditional caring behaviors to promote the quality of nursing care.

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Photoproduction of Hydrogen from Acetate by Rhodopseudomonas: Effect of Culture Conditions and Sequential Dark/Light Fermentation

  • Oh, You-Kwan;Seol, Eun-Hee;Park, Sung-Hoon
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2003년도 생물공학의 동향(XIII)
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    • pp.422-427
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    • 2003
  • Rhodopseudomonas palustris P4 can produce $H_2$ either from CO by water-gas shift reaction or from various sugars by anaerobic fermentation. Fermentative $H_2$ production by P4 is fast, but its yield is relatively low due to the formation of various organic acids. In order to increase $H_2$ production yield from glucose, P4 was investigated for the photo-fermentation of acetate which is a major by-product of fermentative $H_2$ production. Experiments were performed in batch modes using both light-grown and dark-grown cells. When the dark-grown P4 was challenged with light and acetate, $H_2$ was produced with the consumption of acetate after a lag period of 25 h. $H_2$ production was inhibited when a nitrogen source, especially ammonium, is present. When the dark-fermentation broth containing acetate was adopted for photo-fermentation with light-grown cells, $H_2$ production and concomitant acetate consumption occurred without a lag period. The $H_2$ yield was estimated as 2.4 - 2.8 mol $H_2/mol$ acetate and the specific $H_2$ production rate was as 9.8 ml $H_2/g$ cell${\cdot}$h, The fact that a single strain can perform both dark- and light-fermentation gives a great advantage in process development Compared to a one-step dark-fermentation, the combined dark- and light-fermentation can increase the $H_2$ production yield on glucose by two-fold.

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Inhibition of Tumor Growth in a Mouse Xenograft Model by the Humanized Anti-HGF Monoclonal Antibody YYB-101 Produced in a Large-Scale CHO Cell Culture

  • Song, Seong-Won;Lee, Song-Jae;Kim, Chang-Young;Song, Jae-Kyung;Jung, Eui-Jung;Choi, Yong Bock;Min, Sung-Won;Oh, Jong-Won
    • Journal of Microbiology and Biotechnology
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    • 제23권9호
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    • pp.1327-1338
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    • 2013
  • The humanized anti-hepatocyte growth factor (HGF) monoclonal antibody (mAb) YYB-101 is a promising therapeutic candidate for treating various cancers. In this study, we developed a bioprocess for large-scale production of YYB-101 and evaluated its therapeutic potential for tumor treatment using a xenograft mouse model. By screening diverse chemically defined basal media formulations and by assessing the effects of various feed supplements and feeding schedules on cell growth and antibody production, we established an optimal medium and feeding method to produce 757 mg/l of YYB-101 in flask cultures, representing a 7.5-fold increase in titer compared with that obtained under non-optimized conditions. The optimal dissolved oxygen concentration for antibody production was 70% $pO_2$. A pH shift from 7.2 to 7.0, rather than controlled pH of either 7.0 or 7.2, resulted in productivity improvement in 5 L and 200 L bioreactors, yielding 737 and 830 mg/ml of YYB-101, respectively. The YYB-101 mAb highly purified by affinity chromatography using a Protein A column and two-step ion exchange chromatography effectively neutralized HGF in a cell-based assay and showed potent tumor suppression activity in a mouse xenograft model established with human glioblastoma cells.

Typology of ROII Patterns on Cluster Analysis in Korean Enterprises

  • Kim, Young Sun;Kwon, Oh Jun;Kim, Ki Sik;Rhee, Kyung Yong
    • Safety and Health at Work
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    • 제3권4호
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    • pp.278-286
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    • 2012
  • Objectives: Authors investigated the pattern of the rate of occupational injuries and illnesses (ROII) at the level of enterprises in order to build a network for exchange of experience and knowledge, which would contribute to workers' safety and health through safety climate of workplace. Methods: Occupational accidents were analyzed at the manufacturing work site unit. A two step clustering process for the past patterns regarding the ROII from 2001 to 2009 was investigated. The ROII patterns were categorized based on regression analysis and the patterns were further divided according to the subtle changes with Mahalanobis distance and Ward's linkage. Results: The first clustering of ROII through regression analysis showed 5 different functions; 29 work sites of the linear function, 50 sites of the quadratic function, 95 sites of the logarithm function, 62 sites of the exponential function, and 54 sites of the sine function. Fourteen clusters were created in the second clustering. There were 3 clusters in each function categorized in the first clustering except for sine function. Each cluster consisted of the work sites with similar ROII patterns, which had unique characteristics. Conclusion: The five different patterns of ROII suggest that tailored management activities should be applied to every work site. Based on these differences, the authors selected exemplary work sites and built a network to help the work sites to share information on safety climate and accident prevention measures. The causes of different patterns of ROII, building network and evaluation of this management model should be evaluated as future researches.

뉴이미지론의 위상과 두 패러다임 : J. Baudrillard와 J. Lacan을 중심으로 (Two Paradigms of the New Image Theory : J. Baudrillard and J. Lacan)

  • 최광진
    • 조형예술학연구
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    • 제2권
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    • pp.193-221
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    • 2000
  • The postmodern culture since the later 20C breaks downa tradition a relation between the reality and languages or sign images expressing it. It develops in the way to review the meaning on the object's imitation or the representation to have been followed since Plato and represent the new state and concept of expressed things. Also, The visual art leads an change of paradigm by images giving up the visual resemblance or the function of representation and endowing them with the new sense. This essay has a purpose to study an important discussion about this change centered on Baudrillard and Lacan. A sociologist Baudrillard promotes the concept of 'simulation' through detecting the reality and the social and historical state of the image. Studying on the course of this change, he calls the step that the image escapes from the stage to reflect the reality and become the pure imitation by itself simulation. The image in the stage of simulation is called 'hyperreality' because it doesn't have any an indicator or a substitute and happens by models without the original or the reality. So he asserts that art is not to contain some absoluteness or transcendency as the past, but to be as the spectacle with characteristics of meaningless, emptiness, contingency. Lacan dismantles the concept of the absolute Cogito to have become the center of the western ideology, and creates the concept of 'Other'. He concludes also the reality exists but can't be captured, and it's impossible for the thinking subject can reach it. The concept of new image which can be thought as the Symbolic in Lacan is 'Signifier without Signified' since it isn't possible to be the transcendent Signifier fixing the meaning finally in it. His 'Gaze' theory is which to be emitted in other's area determines the subject. Equally Baudrillard and Lacan sets up the new state of the image through the end of representation system As for Baudrillard, art intends to the worthlessness and is nothing but imagination. But in Lacan a picture represents the subject being in process by the dialectic of desire.

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