• Title/Summary/Keyword: Primary Culture

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Production of a anti-MUC1 monoclonal antibody using a glutathione- S-transferase-MUC1 bacterial fusion protein.

  • Park, Kyu-Hwan;Shin, Chan-Young;You, Byung-Kwon;Ko, Kwang-Ho
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1998.11a
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    • pp.198-198
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    • 1998
  • Muc1 mucin is found in a variety of epithelial tissue and is overexpressed in several epithelial cancer. Recently it is alsol reported that primary Hamster tracheal surface epithelial(HTSE) cells express Muc1 protein and cDNA encoding HTSE muc1 protein has been cloned. Although numerous monoclonal antibodies (mAbs) to human muncins, particularly Muc1 have been produced, no such antibodies to murine Muc1 have been described. We now describe monoclonal antibody, called mAb M1CT, produced to C-terminal region of HTSE Muc1 protein by immunising mice with a glutathion-s-transferase linked fusion protein. In this study, using this antibody(mAb M1CT) we investigated the effect of RA on the expression of Muc1 in HTSE cells. Retinoic acid(RA) plays an essential role in maintaining normal differentiation of tracheal epithelial cells. With RA-deficiency tracheocytes undergo squamous metaplasia, an abnormal differentiation that can be reversed by RA. We had primary culture of HTSE cells under different concentrations of RA. Culture was maintained until the direction of differentiation was determined. Then Western blot analysis with mAb M1CT was performed with the cell lysates from the culture. The expression of Muc1 protein was decreased in dose-dependent manner as the concentration of retinoic acid was decreased. Our result indicates that the expression of Muc1 protein is coordinately regulated with airway mucous cell differentiation by RA pathway. And the antibody, mAb M1CT, produced in this study should provide useful tool to study the expression of Muc1 mucin in differentiation process or disease.

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Inhibition of $IL-1{\beta}$ and IL-6 in Osteoblast-Like Cell by Isoflavones Extracted from Sophorae fructus

  • Joo, Seong-Soo;Kang, Hee-Cheol;Choi, Min-Won;Choi, Young-Wook;Lee, Do-ik
    • Archives of Pharmacal Research
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    • v.26 no.12
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    • pp.1029-1035
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    • 2003
  • Osteoporosis is recognized as one of the major hormonal deficiency diseases, especially in menopausal women and the elderly. When estrogen is reduced in the body, local factors such as IL-1 $\beta$ and IL-6, which are known to be related with bone resorption, are increased and promote osteoclastogenesis, which is responsible for bone resorption. In the present study, we investigated whether glucosidic isoflavones (Isocal, PIII) extracted from Sophorae fructus affect the proliferation of osteoblasts and prevent osteoclastogenesis in vitro by attenuating upstream cytokines such as IL-1$\beta$ and IL-6 in a human osteoblastic cell line (MG-63) and in a primary osteoblastic culture from SD rat femurs. Interestingly, IL-1$\beta$ and IL-6 mRNA were significantly suppressed in osteoblast-like cells treated with 17$\beta$-estradiol (E2) and PIII when compared to positive control (SDB), and this suppression was more effective at $10^{-8}$% than at the highest concentration of $10^{-4}$%. In addition, these were confirmed in protein levels using ELISA assay. In the cell line, the cells showed that E2 was the most effective in osteoblastic proliferation over the whole range of concentration ($10^{-4}%-10^{-12}$%), even though PIII also showed the second greatest effectiveness at $10^{-8}$%. Nitric oxide (NO) was significantly (p<0.05) upregulated in PIII and E2 over the concentration range $10^{-6}% to 10^{-8}$% when compared to SDB, without showing any dose dependency. In bone marrow primary culture, we found by TRAP assay that PIII effectively suppressed osteoclastogenesis next to E2 in comparison with SDB and culture media (control). In conclusion, these results suggest that local bone-resorbing cytokines can be regulated by PIII at lower concentrations and that, therefore, PIII may preferentially induce anti-osteoporosis response by attenuating osteoclastic differentiation and by upregulating NO.

Effects of Glucagon and Insulin on Glutathione Homeostasis: Role of Cellular Signaling Pathways and Glutathione Transport System (Glucagon과 insulin이 glutathione 항상성에 미치는 영향: 세포신호전달체계 및 glutathione transport system의 역할)

  • Kim, Bong-Hee;Oh, Jung-Min;Yun, Kang-Uk;Kim, Chung-Hyeon;Kim, Sang-Kyum
    • Environmental Analysis Health and Toxicology
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    • v.22 no.3
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    • pp.227-233
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    • 2007
  • It has been reported that hepatic glutathione (GSH) levels are decreased in diabetic patients, and glucagon increases hepatic efflux of GSH into blood. The signaling pathways responsible for mediating the glucagon effects on GSH efflux, however, are unknown. The signaling pathways involved in the regulation of GSH efflux in response to glucagon and insulin were examined in primary cultured rat hepatocytes. The GSH concentrations in the culture medium were markedly increased by the addition of glucagon, although cellular GSH levels are significantly decreased by glucagon. Insulin was also increased the GSH concentrations in the culture medium, but which is reflected in elevations of both cellular GSH and protein. Treatment of cells with 8-bromo-cAMP or dibutyryl-cAMP also resulted in elevation of the GSH concentrations in the culture medium. Pretreatment with H89, a selective inhibitor of protein kinase A, before glucagon addition markedly attenuated the glucagon effect. These results suggest that glucagon changes GSH homeostasis via elevation of GSH efflux, which may be responsible for decrease in hepatic GSH levels observed in diabetic condition. Furthermore, the present study implicates cAMP and protein kinase A in mediating the effect of glucagon on GSH efflux in primary cultured rat hepatocytes.

Multiple shoot induction and plant regeneration from axillary buds of Magnolia 'Vulcan'

  • Kim, Tae-Dong;Kim, Ji-Ah;Lee, Na-Nyum;Choi, Chang-Ho
    • Journal of Plant Biotechnology
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    • v.47 no.1
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    • pp.40-45
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    • 2020
  • An efficient protocol for multiple shoot induction and plant regeneration from axillary bud culture of Magnolia 'Vulcan' was developed in the present study. Primary shoots were obtained from axillary bud explants cultured on Murashige and Skoog (MS) medium containing 1.0 mg/L 6-benzylaminopurine (BA). To induce multiple shoots effectively, primary shoot tips were cultured on MS medium supplemented with different concentrations of BA and zeatin at 0, 0.2, 0.5, and 1.0 mg/L. Of these treatments, the MS medium with 0.5 mg/L BA resulted in the highest number of shoots per explant with an average value of 5.9, and it produced the greatest shoot height at 4.8 cm after 12 weeks of culturing. In the rooting of in vitro produced shoots, the greatest percentage of explants forming roots (91.3%), number of roots per explant (9.7), and root length (2.8 cm) were obtained in half-strength MS medium supplemented with 6.0 mg/L indole-3-butyric acid (IBA). Regenerated plantlets were successfully acclimatized and hardened off inside the culture room with 87.5% survival rate. Plants were transferred to a greenhouse with a 97.2% survival rate. The highly efficient shoot multiplication and plant regeneration system reported herein can be used for large-scale clonal propagation of valuable Magnolia species or cultivars.

A Survey of the Perception of the Superior Factors to of Korean Traditional Foods by College Students with Food Related Majors (외식조리식품관련 전공 대학생들의 전통음식 우수요인에 관한 인식조사)

  • Kang, Jae-Hee;Kim, Ji-Eung
    • Journal of the Korean Society of Food Culture
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    • v.24 no.2
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    • pp.155-163
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    • 2009
  • This study was conducted to examine the superior factors and the perception of Korean traditional foods by college students studying in food related industries. The subject group consisted of 65.0% of the students majoring in food service & culinary arts, 12.9% of the students majoring in food & nutrition, and 7.3% of the students majoring in baking & pastry. The nutritional value was evaluated as the top superior factor of Korean traditional foods by the study subjects. In addition, 92.4% of reported that they had eaten Korean traditional foods in the past, 76.8% of whom responded that the reason for having Korean traditional foods was its time-honored tradition. 'Cooking for oneself' was considered to be the most popular way (78.2%) of obtaining Korean traditional foods, while the traditional market (58.6%) was the most popular place to purchase the foods. 'Not having enough time' (47.2%) was the primary reason for not having Korean traditional foods, although 72.9% of the subjects reported that they wanted to learn about Korean foods. 'Standardization of taste, nutritional value, and recipes' was found to be the most important factor (41.3%) required to increase the consumption of Korean traditional foods. Additionally, 56.1% of the subjects responded that they feel there is a need for modernization of the cuisine to meet the taste of the general public. However, 61.4% of the subjects responded that the succession of traditional dietary culture was the primary reason for developing traditional cuisine, which indicates that there is a bright future for Korean traditional foods.

The Evaluation of Historical and Cultural Authenticity of the Three Major Cultural Areas Projects - Targeting Korean culture theme park in Yeongju-Zone and World confucian scholar culture park in Bonghwa-Zone - (3대문화권 사업 계획의 역사문화적 진정성 평가 - 한국문화테마파크 영주지구와 세계유교선비문화공원 봉화지구를 대상으로 -)

  • Jung, Kyoung-Ah;So, Hyun-Su
    • Journal of Korean Society of Rural Planning
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    • v.22 no.1
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    • pp.25-35
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    • 2016
  • This study targets Korean culture theme park in Yeongju-Zone and World confucian scholar culture park in Bonghwa-Zone among the Three Major Cultural Areas Projects supervised by Ministry of Culture, Sports and Tourism. The study analyzes the Historical and Cultural Authenticity of each project on the basis of the reports and the report materials for meeting with related organizations. The results of the study are drawn as follows. First, through the theoretical consideration, this study drew the types of authenticity: originality, identity, specificity and visibility, which are evaluation items on the Historical and Cultural Authenticity. Second, Bonghwa-Zone succeeded in acquiring originality with tangible cultural properties but Yeongju-Zone chose a project site without it. Third, with originality, Bonghwa-zone was evaluated as having resources and concept with high traditional culture connectivity and fulfilled identity. It led the feature showing the high affinity between originality and identity. Fourth, compared to the projects of Andong-Zone in the Three Major Cultural Areas Projects, these two projects failed to acquire the distinctions since the primary and the secondary influence area and major facilities & programs coincided with those of the projects of Andong-Zone. Fifth, compared to Bonghwa-Zone, Yeongju-Zone realized visibility faithfully by the conceptual flexibility of "Korean Culture" and a large-scale development. Sixth, in terms of the Historical and Cultural Authenticity of project plan, it is evaluated that Yeongju-Zone and Bonghwa-Zone only fulfilled visibility and specificity respectively.

The Case Study of Lactobacillus mixture culture fluid on Atopic dermatitis (아토피피부염에 대한 Lactobacillus 혼합배양액의 임상 증례)

  • Jo, Eul-Hwa;Kim, Tae-Keun;Hong, Su-Jung;Jung, Do-Yean;Hwang, Seng-Yean;Ahn, Seong-Hun
    • The Journal of Korean Medicine
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    • v.36 no.3
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    • pp.135-143
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    • 2015
  • Objectives: Recently lactic acid formulation was known as the adjuvant therapy on atopic dermatitis(AD) symptoms because of little side effects. In this study, it was studied that Lactobacillus mixed culture appliment was effective on not on AD symptoms. Methods: The case-photos 30-40 times of AD symptoms from case were observed with literatures. The case-photos were acquired with environmental AD dermatitis experience program which is conducted in SUNCHANG RESERCH INSTITUTE OF HEALTH AND LONGEVITY from 2014.01 to 2014.08. Experience applicants were limited the oral administration and chemical external administration. Results: Lactobacillus mixed culture appliment was effective because of the mitigation or disappearance of AD primary symptoms like pruritus, erythema, edema, effusion, dry skin, scaly, scab etc. AD symptom degree was dependent on lactobacillus mixed culture appliment times, and classified as Reaction Period (RAP), Reduction Period (RDP), Efection Period (EP), Reproduction Period (RPP) on a single mixed culture appliment time. And AD symptoms which were appeared in RPP were classified as Rebound Period (RBP), Effection Period (EP), Subclinical Period (SCP). Conclusions: Lactobacillus mixed culture appliment is considered to be useful for AD symptoms and is estimated to be effective by different mechanism with oral administration or steroid ointment application. Lactobacillus mixed culture appliment is expected to induce a synergistic effects on AD symptom reliefs with the other oral administration.

Effects of Feeder Cells on the Primary Culture of Ovarian Cell Populations from Adult Japanese Medaka (Oryzias latipes)

  • Ryu, Jun Hyung;Gong, Seung Pyo
    • Journal of Animal Reproduction and Biotechnology
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    • v.35 no.1
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    • pp.65-72
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    • 2020
  • Fish ovarian germline stem cells (OGSCs) that have the abilities to self-renew and differentiate into functional gametes can be used in various researches and applications. A main issue to be solved for effective utilization of fish OGSCs is the development of their stable in vitro culture condition, but only few researches about fish OGSC culture have been reported so far. In this study, in order to find the clues to develop the culture condition for OGSCs from Japanese medaka (Oryzias latipes), we tried to establish somatic cell lines as a candidate for the feeder cells and evaluated its supporting effects on the culture of ovarian cell populations from O. latipes. As the results, the somatic cell lines could be established only from the embryonic tissues among three tissues derived from embryos, fins and ovaries. Three embryonic cell lines were tested as a feeder cell for the culture of ovarian cell population and all three cell lines induced cell aggregation formation of the cultured ovarian cells whereas the feeder-free condition did not. Furthermore, a significant cellular proliferation was observed in the ovarian cells cultured on two of three cell lines. As a trial to increase the capacity of the cell lines as a feeder cell that supports the proliferation of the cultured ovarian cells, we subsequently established a stable line that expresses the foreign O. latipes fibroblast growth factor 2 (FGF2) from an embryonic cell line and evaluated its effectiveness as a feeder cell. The ovarian cells cultured on FGF2 expressing feeder cells still formed cell aggregates but did not show a significant increase in cellular proliferation compared to those cultured on non-transformed feeder cells. The results from this study will provide the fundamental information for in vitro culture of medaka OGSCs.

Looking at Organizational Socialization from the Developmental Network Perspective

  • Chang, Jihyun;Kim, Taesung
    • The Journal of Asian Finance, Economics and Business
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    • v.5 no.3
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    • pp.195-206
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    • 2018
  • Paying close attention to those new to an organization, whether fresh or experienced, whose primary interest is in (re)socialization, the current study intends to (1) further the concept of mentoring from a bilateral relationship to a community and culture fostered by developmental networks, (2) propose an integrated conceptual framework for organizational socialization, and (3) suggest implications for practice and future research. This study reviews, analyzes, and integrates research assets and subsequently re-conceptualizes the aggregate information as valid propositions and a conceptual framework. The findings include (1) 11 propositions regarding the relationships among network characteristics (embeddedness, diversity), developmental functions (career support, psychosocial support, and role modeling), and socialization outcomes (learning and attitudinal outcomes); and (2) an integrated conceptual framework that depicts a comprehensive mechanism through which developmental networks conduce to organizational socialization of newcomers. Implications are that developmental networking must be an individual's fundamental competency and an essential part of organizational onboarding processes, and imperative for both members' career development and innovative organizational culture. By integrating research assets on the developmental phenomenon into conceptualizations, this study furthers the concept of mentoring to organizational culture and stimulates a substantive discourse for theory-building towards organizational socialization from the developmental network perspective.

Zinc Increases Chemotactic Activity of Porcine Peripheral Blood Polymorphonuclear Cells

  • Seo, Dong-Hee;Kang, Byeong-Teck;Kang, Ji-Houn;Yang, Mhan-Pyo
    • Journal of Veterinary Clinics
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    • v.35 no.5
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    • pp.195-199
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    • 2018
  • Zinc is necessary for normal functions in the immune system. The objective of the study is to examine the effect of zinc on the chemotactic activity of porcine peripheral blood polymorphonuclear cells (PMNs). A modified Boyden chamber was used to determine the directional migration distance of PMNs. Various concentrations of zinc showed no chemotactic activity to PMNs. However, culture supernatant from peripheral blood mononuclear cells (PBMCs) treated with zinc remarkably increased the chemotactic activity of PMNs when compared with culture supernatant from PBMCs treated without zinc. Culture supernatant from PBMCs treated without zinc also increased the migration distance of PMNs relative to vehicle control (medium alone). Increasing effect in chemotactic activity of PMNs by culture supernatant from PBMCs treated with zinc was inhibited by treatment of porcine anti-interleukin (IL)-8 polyclonal antibody (pAb). This effect was not affected by heat treatment ($4-85^{\circ}C$). This corresponded with heat stable physical characteristics of IL-8. These results suggest that zinc can upregulate the chemotaxis of PMNs, which is primary mediated by IL-8 chemotactic factor released from PBMCs treated with zinc.