• Title/Summary/Keyword: In Vitro Development

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Effects of Activin on Testosterone-primed Immature Rainbow Trout Gonadotropin Release in vitro (Testosterone 처리한 미성숙 무지개송어 뇌하수체의 세포배양계에서 생식소자극초르몬 분비에 대한 Activin의 효과)

  • KIM Dae-Jung;HAN Chang-Hee;AIDA Katsumi
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.32 no.2
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    • pp.204-210
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    • 1999
  • The present studies were conducted to evaluate the effects of activin-A on gonadotropins (GTHs) release in testosterone-treated immature rainbow trout Oncorhpynchus mykiss. The administration of testosterone elevated pituitary level of GTH II but not of GTH I. In this study using primary cultures of dispersed pituitary cells in static incubation, dose-dependent increases in GTH II release was observed in the activin-treated group at day 3 of incubation (long-term incubation), but not at day 1 of incubation (short-term incubation). Dopamine, a potent inhibitor of gonadotropin-releasing hormone (GnRH)-stimulated GTH II release in rainbow trout, was only partially effective in decreasing actvin-induced GTH II release. Furthermore, salmon GnRH (sGnRH)-stimulated GTH II release was not potentiated by the pretreatment with activin. However, the control mechanisms of GTH I release by activin and other hormones were not observed in the all tested experiments. The results of these studies support the contention that in contrast with the usual stimulatory effects of activin on GTH release in mammals, activin exerts long-term stimulatory actions on GTH II release in rainbow trout. The control mechanism of GTH I release, however, is a question that remains to be elucidated.

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Protective effect of Korean Red Ginseng against glucocorticoid-induced osteoporosis in vitro and in vivo

  • Kim, Jinhee;Lee, Hyejin;Kang, Ki Sung;Chun, Kwang-Hoon;Hwang, Gwi Seo
    • Journal of Ginseng Research
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    • v.39 no.1
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    • pp.46-53
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    • 2015
  • Background: Glucocorticoids (GCs) are commonly used in many chemotherapeutic protocols and play an important role in the normal regulation of bone remodeling. However, the prolonged use of GCs results in osteoporosis, which is partially due to apoptosis of osteoblasts and osteocytes. In this study, effects of Korean Red Ginseng (KRG) on GC-treated murine osteoblastic MC3T3-E1 cells and a GC-induced osteoporosis mouse model were investigated. Methods: MC3T3-E1 cells were exposed to dexamethasone (Dex) with or without KRG and cell viability was measured by the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. Realtime polymerase chain reaction was performed to evaluate the apoptotic gene expression; osteogenic gene expression and alkaline phosphatase (ALP) activity were also measured. Western blotting was performed to evaluate the mitogen-activated protein kinase (MAPK) proteins. A GC-induced osteoporosis animal model was used for in vivo study. Results and conclusion: The MTT assay revealed that Korean Red Ginseng (KRG) prevents loss of cell viability caused by Dex-induced apoptosis in MC3T3E1 cells. Real-time polymerase chain reaction data showed that groups treated with both Dex and KRG exhibited lower mRNA levels of caspase-3 and -9, whereas the mRNA levels of Bcl2, IAPs, and XIAP increased. Moreover, groups treated with both Dex and KRG demonstrated increased mRNA levels of ALP, RUNX2, and bone morphogenic proteins as well as increased ALP activity in MC3T3-E1 cells, compared to cells treated with Dex only. In addition, KRG increased protein kinase B (AKT) phosphorylation and decreased c-Jun N-terminal kinase (JNK) phosphorylation. Moreover, microcomputed tomography analysis of the femurs showed that GC implantation caused trabecular bone loss. However, a significant reduction of bone loss was observed in the KRG-treated group. These results suggest that the molecular mechanism of KRG in the GC-induced apoptosis may lead to the development of therapeutic strategies to prevent and/or delay osteoporosis.

Practical Propagation Methods for Production of Prothalli and Sporophytes in Deparia pycnosora (Christ) M. Kato

  • Jang, Bo Kook;Park, Kyungtae;Cho, Ju Sung;Lee, Cheol Hee
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.04a
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    • pp.43-43
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    • 2019
  • Deparia pycnosora (Christ) M. Kato is a fern used as ornamental plant. In addition, it is called "Teol-go-sa-ri" in Korean name. The aim of this study was to develop a practical propagation method of D. pycnosora using tissue culture technique. Prothallus obtained from spore germination was the used as experiment materials. The prothalli (300 mg) used in all experiments were sub-cultured for 8-week intervals. The most suitable media for prothallus propagation were identified by culturing 300 mg of prothalli in $1/4{\times}$, $1/2{\times}$, $1{\times}$, $2{\times}$ MS medium and in Knop medium for 8 weeks. Also, the prothalli were cultured by chopping with a scalpel. In addition, sucrose, activated charcoal, and total nitrogen source were added in different concentrations based on the culture medium selected. Cultures were maintained at a temperature of $25{\pm}1^{\circ}C$, light intensity of $30{\times}1.0{\mu}mol-m-2{\cdot}s-1$, and a photoperiod of 16/8 h (light/dark) in in vitro. The results showed that optimum was achieved prothallus fresh weight and development in $1{\times}$ MS medium. When other components were added to the basic $1{\times}$ MS medium, prothallus propagation was maximized in $1{\times}$ MS medium supplemented with 2% sucrose, 0.2% activated charcoal, and 60 mM total nitrogen. To select a suitable soil mixture for sporophyte formation, 1.0 g of prothallus was blended with distilled water, spread on five combinations of different soil substrates (decomposed granite, horticultural substrates, peat moss, and perlite), and cultivated for 12 weeks. The sporophyte cultures were maintained at a temperature of $25{\pm}1^{\circ}C$, light intensity of $43{\pm}2.0{\mu}mol-m-2{\cdot}s-1$, humidity of $84{\pm}1.4%$, and a photoperiod of 16/8 h (light/dark). As a results, horticultural substrate alone, 2:1 (v:v) mixtures of horticultural substrate and perlite, and 2:1 mixtures of horticultural substrate and decomposed granite induced 208.0, 201.3 and 248.8 sporophytes per pot, respectively. Therefore, this result could provide a practical mass propagation method of D. pycnosora

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Use of Chicken Meat and Processing Technologies (가금육의 이용과 가공기술)

  • Ahn, Dong-Uk
    • Proceedings of the Korea Society of Poultry Science Conference
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    • 2003.07b
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    • pp.67-88
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    • 2003
  • The consumption of poultry meat (chicken and turkey) grew the most during the past few decades due to several contributing factors such as low price. product research and development. favorable meat characteristics, responsive to consumer needs, vertical integration and industry consolidation, new processing equipments and technology, and aggressive marketing. The major processing technologies developed and used in chicken processing include forming/restructuring, tumbling, curing, smoking, massaging, injection, marination, emulsifying, breading, battering, shredding, dicing, and individual quick freezing. These processing technologies were applied to various parts of chicken including whole carcass. Product developments using breast, thigh, and mechanically separated chicken meat greatly increased the utilization of poultry meat. Chicken breast became the symbol of healthy food, which made chicken meat as the most frequent menu items in restaurants. However, the use of and product development for dark meat, which includes thigh, drum, and chicken wings were rather limited due to comparatively high fat content in dark meat. Majority of chicken are currently sold as further processed ready-to-cook or ready-to-eat forms. Major quality issues in chicken meat include pink color problems in uncured cooked breast, lipid oxidation and off-flavor, tenderness PSE breast, and food safety. Research and development to ensure the safety and quality of raw and cooked chicken meat using new processing technologies will be the major issues in the future as they are now. Especially, the application of irradiation in raw and cooked chicken meat products will be increased dramatically within next 5 years. The market share of ready-to-eat cooked meat products will be increased. More portion controlled finished products, dark meat products, and organic and ethnic products with various packaging approaches will also be introduced.

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Effect of Plant Growth Regulators on Bulblet Formation and Plant Regeneration in Fritillaria thunbergii Miq. (패모조직배양에서 생장조절 물질이 자구형성 및 식물체 재생에 미치는 영향)

  • Park, Chul-Hyoung;Ryu, Jeom-Ho;Han, Kwang-Soo;Doo, Hong-Soo;Choi, Sun-Young
    • Korean Journal of Medicinal Crop Science
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    • v.4 no.2
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    • pp.119-125
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    • 1996
  • To improve the efficiency of mass propagation in vitro of Fritillaria thunbergii, bulb scale and nodes were cultured in LS medium supplemented with the combination of 2, 4-D and kinetin or NAA and BA. The number and size of bulb, the number of adventitious shoot, the ratio of callus formation, rooting, and the effects of light and dark on the culture, plant regeneration from calli, and the gelling substances were investigated. The combination of 2, 4-D and kinetin in media was more effective than the media of NAA and BA for the bulblet formation. The media supplemented with 2 mg/L 2, 4-D and 1 mg/L kinetin, $1{\sim}2\;mg/L$ 2, 4-D without kinetin and $1{\sim}3\;mg/L$ BAA only were effective in the adventitious shoot development. Callus formation and root formation, respectively were effective in the medium supplemented with 2mg/L 2, 4-D and 1mg/L kinetin. In bulb formation, the medium with 5 mg/L kinetin was effective, and the most of bulbs were formed from the axillary bud of node part. In bulb formation, shoot growth, callus and root formation, the light culture for 16 hours per day was better than that in the dark culture. Bulb was nicely formed in the medium with 0. 2 mg/L 2, 4-D, 1 mg/L kinetin. The medium without hormone was most effective for plant regeneration. The phytagel was more effective than agar in the medium as the gelling agent.

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광어(Paralichtys olivaceus)의 스쿠치카감염증(感染症) -스쿠치카섬모충(纖毛蟲)의 배양성상(培養性状).약제감수성(藥劑感受性).병원성(病源性)-

  • 길수 수;일향진일;오 명주;생도삼내자;목촌교구;삼 립성;야촌철일;회면량남
    • Journal of fish pathology
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    • v.6 no.2
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    • pp.205-218
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    • 1993
  • On the development of hirame(Paratichtys olivaceus) culture, outbreak of scuticociliata infection was reported to cause severe damage in Japan. To establish effective measures for isolation and cultivation of this ciliate, we tried to culture this pathogenic ciliate using medium for bacteria and fish cell lines in vitro. Scuticociliata from the brain tissues of infected fish was aseptically inoculated to CHSE-214 cells cultured in MEM-10 without antibiotic. Scuticociliata grew well and the number of ciliate reached $10^6\;cells/ml$ at temperatures of $15^{\circ}C$ to $20^{\circ}C$ for 10d. The number of ciliate cultured in the cell lines is 10 times higher than the numbers cultured in the liquid medium alone. This ciliata could be cloned by dilution method. Scuticociliata isolated could grow well on 42 different cell lines that were established from marine fish, warm freshwater fish, and salmonids. This ciliate could be preserved in liquid nitrogen for more than 6 months. Subsequently, we observed the optimal temperature and salinity for growth, and tested the sensitivities of this organism to formaldehyde, flagyl(Metronidazole), Ekuteshin(Combination compound of sulfamonometoxin and ormethoprim), and ozonixation. Optimal temperature for growth was $25^{\circ}C$ and salinity was 1.0 to 1.5%. Washed scuticociliata was killed by formaldehyde at the concentration of 50ppm for 10min, but was not completely killed even at a high concentration of 400ppm for 20min in MEM-5. Flagyl and Ekuteshin can inhibit the growth of scuticociliata at the concentration of 1,000 and $100{\mu}g/ml$ in MEM-10, respectively. More than 99% of this scuticociliata could be killed by ozonization at a dose equivalent to $1.0mg/\ell$ oxidant for 30sec in sea water. Isolated scuticociliata showed the pathogenicity to the cultured hirame by artificial infection(I. P. injection, $10^5\;cells$/fish). The number of scuticociliata in the water could be counted by most probable number(MPN) method using tissue culture, and the minimum detectable number was $1.8\;cells/\ell$. The number in the reservoir tank for water supply to the culture tank was 110 cells/l. After cleaning by elimination of the sediments from of the reservoir tank and disinfected with formaldehyde, number of scuticociliata decreased and was counted less than $1.8\;cells/\ell$ and infection rate of cultured hirame was decreased.

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Analysis of Molecular Expression in Adipose Tissue-Derived Mesenchymal Stem Cells : Prospects for Use in the Treatment of Intervertebral Disc Degeneration

  • Jin, Eun-Sun;Min, Joongkee;Jeon, Sang Ryong;Choi, Kyoung Hyo;Jeong, Je Hoon
    • Journal of Korean Neurosurgical Society
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    • v.53 no.4
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    • pp.207-212
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    • 2013
  • Objective : Recent studies have shown encouraging progress toward the use of autogenic and allogenic mesenchymal stem cells (MSCs) to arrest, or even lead to partial regeneration in, intervertebral disc (IVD) degeneration. However, this technology is still in its infancy, and further development is required. The aim of this study was to analyze whether rat adipose-derived mesenchymal stem cells (ADMSC) can differentiate towards IVD-like cells after treatment with transforming growth factor ${\beta}3$ (TGF-${\beta}3$) in vitro. We also performed quantitative analysis of gene expression for ADMSC only, ADMSCs treated with TGF-${\beta}3$, and co-cultured ADMSCs treated with TGF-${\beta}3$. Methods : ADMSCs were sub-cultured to homogeneity and used in fluorocytometry assays for CD11, CD45, and CD90/Thy1. ADMSCs were differentiated in spheroid culture towards the chondrogenic lineage by the presence of TGF-${\beta}3$, dexamethasone, and ascorbate. We also co-cultured pure ADMSCs and nucleus pulposus cells in 24-well plates, and performed immunohistochemical staining, western blotting, and RT-PCR for quantitative analysis of gene expression. Results : Results of fluorocytometry were positive for CD90/Thy1 and negative for CD11 and CD45. TGF-${\beta}3$-mediated induction of ADMSCs led to the expression of the differentiation markers of intervertebral disc-like cells, such as aggrecan, collagen II, and sox-9. Co-cultured ADMSCs treated with TGF-${\beta}3$ showed higher expression of differentiation markers and greater extracellular matrix production compared with ADMSCs treated with TGF-${\beta}3$ alone. Conclusion : ADMSC treated with TGF-${\beta}3$ may be an attractive source for regeneration therapy in degenerative IVD. These findings may also help elucidate the pathologic mechanism of MSC therapy in the degeneration of IVD in vivo.

Effect of explant's position and culture method on shoot proliferation and micro-cuttings for a rare and endangered species, Abeliophyllum distichum Nakai (희귀 및 멸종위기 식물 미선나무(Abeliophyllum distichum Nakai)의 절편위치 및 치상방법에 따른 기내증식 및 미세삽목)

  • Lee, Na Nyum;Kim, Ji-Ah;Kim, Yong-Wook;Choi, Yong Eui;Moon, Heung Kyu
    • Journal of Plant Biotechnology
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    • v.42 no.3
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    • pp.228-234
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    • 2015
  • Using either the apical or axillary bud of the endangered species Abeliophyllum distichum Nakai, we tested the effect of bud position and culture method on shoot proliferation and rooting. In shoot proliferation, the axillary bud explant was more effective than the apical bud and the effect was fostered by BA treatment, whereas no differences were observed in shoot elongation by the explant position. Spontaneous rooting was observed in the MS basal medium and resulted in conspicuous differences in the explant position : more than 80% in apical bud explant and 28% in axillary bud explant was achieved, respectively. The positional effects were also observed in BA pre-treatments: generally vertical culture method appeared to be better in shoot proliferation, growth, and rooting than that of the horizontal culture method regardless of the BA pre-treatment duration. The highest shoot multiplication was achieved through the vertical culture method with axillary bud explant, whereas the best shoot elongation and rooting was obtained using the vertical culture method with the apical bud explant. Apical bud explant was superior to axillary bud explant in ex vitro micro-cuttings and revealed a significant difference in shoot growth and root development. The above results suggest that explant position and culture method influence the efficiency of micropropagation for a rare and endangered plant Abeliophyllum distichum.

Baseline Sensitivity and Monitoring for the Resistance to Benomyl of Fusarium Species Causing Bakanae Disease in Korea (국내 벼 키다리병균의 Benomyl 약제에 대한 감수성 기준 및 저항성 변화)

  • Choi, Hyo-Won;Lee, Yong Hwan;Hong, Sung Kee;Lee, Young Kee;Lee, Jae Guem;Kim, Hyo Won
    • The Korean Journal of Mycology
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    • v.43 no.4
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    • pp.260-266
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    • 2015
  • To examine the changes in resistance to benomyl of Fusarium species causing bakanae disease, Fusarium isolates were collected in Korea, and pathogenicity tests were performed using rice seeds in vitro. Minimum inhibitory concentration (MIC) and effective concentration of 50% ($EC_{50}$) values of isolates were examined using the agar dilution method. High frequency distribution of MIC values to benomyl against isolates collected in 2006~2007 and 2013~2014 years were $1.5625{\sim}3.125{\mu}g/mL$ and more than $25{\mu}g/mL$, respectively. The mean $EC_{50}$ value of isolates to benomyl increased from $1.6397{\mu}g/mL$ in 2006~2007 to $2.4892{\mu}g/mL$ in 2013~2014. Based on MIC and $EC_{50}$ values of isolates, the moderate resistance of benomyl were determined as more than $25{\mu}g/mL$ of MIC and less $2.4{\mu}g/mL$ of $EC_{50}$ value, and resistant isolates to benomyl were determined as more than $2.4{\mu}g/mL$ of $EC_{50}$ value. Compared with the ratio of resistant isolates in 2006~2007, the ratio of resistance isolates in 2013~2014 increased from 12.5% to 36.4%. In addition, multiple resistant isolates to prochloraz as well as benomyl increased to 20.3% in 2013~2014.

Regulation of Matrix Metalloproteinase-1 Expression by the Homeodomain Transcription Factor Caudal in Drosophila Intestine (초파리 장조직에서 Caudal 전사조절인자에 의한 matrix metalloproteinase-1 발현 조절)

  • Lee, Shin-Hae;Hwang, Mi-Sun;Choi, Yoon-Jeong;Kim, Young-Shin;Yoo, Mi-Ae
    • Journal of Life Science
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    • v.22 no.12
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    • pp.1600-1607
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    • 2012
  • The matrix metalloproteinase (MMP) family plays essential roles in physiological processes such as embryonic development, angiogenesis, wound healing, and tissue homeostasis as a consequence of MMPr capacity for breaking down many types of extracellular matrix proteins. Imbalanced regulation of MMP expression can also lead to pathological conditions such as tumor progression. We recently reported that the Drosophila Mmp1 gene is highly expressed in the digestive tract and is required for the maintenance of intestinal homeostasis such as by restriction of uncontrolled intestinal stem cell proliferation. However, the regulatory mechanisms of MMP gene expression in the intestine remain unclear. In this study, we determined that the expression of Mmp1 is regulated by the homeodomain transcription factor Caudal. Experiments using the targeted expression of Caudal under the regulation of Gal4-UAS system indicated that endogenous Caudal is required for the Mmp1 gene expression in the adult Drosophila intestine and that exogenous Caudal induces Mmp1 expression. Transient transfection experiments indicated that Caudal can activate the promoter activity of Mmp1 and that several putative Caudal binding sites in the 5'-flanking region of the Mmp1 gene may be critical to the upregulation by Caudal. Our data suggest that Mmp1 is one of the target genes of Caudal in physiological normal condition and in tumorigenesis.