• Title/Summary/Keyword: culture-dependent

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Effect of MLN8237, a Novel Aurora A Kinase Inhibitor, on the Spontaneous Fragmentation of Ovulated Mouse Oocytes

  • Park, Ji-Hun;Choi, Tae-Saeng
    • Reproductive and Developmental Biology
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    • v.35 no.4
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    • pp.499-502
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    • 2011
  • Aurora A kinase is a mitotic serine/threonine kinase whose proposed functions include the maturation of centrosomes, G2/M transition, alignment of chromosomes at metaphase, and cytokinesis. In this study, we investigated the effect of MLN8237, an aurora A kinase inhibitor, on the postovulatory aging of oocytes based on the frequency of oocyte fragmentation, cdk1 kinase activity, and cyclin B degradation. The fragmentation of ovulated oocytes during prolonged culture was inhibited by treatment with MLN8237 in a concentration-dependent manner. The frequency of fragmented oocytes was significantly lower in oocytes treated with 2 ${\mu}M$ MLN8237 (13%) than in control oocytes (64%) after two days of culture. Most of the control (non-fragmented) oocytes (91%) were activated after two days of culture. In comparison, only 22% of the MLN8237-treated oocytes were activated; the rest of the oocytes (78%) were still in metaphase with an abnormal spindle and dispersed chromosomes. Next, cdk1 activity and the level of cyclin B were examined. The level of cyclin B and cdk1 activity in MLN8237-treated oocytes were nearly equal to those in control oocytes. Our results indicate that MLN8237 inhibited the fragmentation of ovulated oocytes during prolonged culture, although it blocked the spontaneous decrease in activity of cdk1 and degradation of cyclin B. This mechanism of inhibition is different from that in oocytes treated with nocodazole, which have high levels of cdk1 activity and cyclin B.

Antifungal Activity of Serratia marcescens Culture Extracts against Phytopathogenic Fungi: Possibility for the Chitinases Role

  • Cho, Moo-Je;Lee, Sang-Yeol;Gal, Sang-Wan;Hwang, Jae-Ryoung;Yoon, Hae-Won
    • Journal of Microbiology and Biotechnology
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    • v.2 no.3
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    • pp.209-214
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    • 1992
  • Serratia marcescens co-cultured with various phytopathogenic fungi, including Rhizopus stolonifer, Helminthosporium allii, Pyricularia oryzae, Fusarium oxysporium and Collectothricom cassiicola, in an LB- agar medium containing 1.5% swollen chitin, significantly inhibitied fungal growth. Fungal hyphae grew rapidly outward from the culture dish center, but the hyphal extensions of the pathogenic fungi were significantly inhibited in a perimetric contact area with S. marcescens. This was especially evident in pathogenic fungi which have a high chitin content in their cell walls. The extracellular chitinase activities of S. marcescens were increased seven fold by the addition of 1.5% swollen chitin to the LB-broth, compared to chitinase activities in a culture medium without chitin. The type of induction was dependent on the various forms of chitin used. When the culture supernatant of S. marcescens or the chitinases of Streptomyces griceus purchased from Sigma Chemical Co., were incubated with the mycelium of F. oxysporium, the mycelium gradually burst as cultivation time progressed and completely lysed after incubation for 2 days. On the other hand, E. coli extract did not hydrolyze the F. oxysporium mycelium at all. These data showed that the chitinolytic activities of S. marcescens play important roles in the biochemical control of phytopathogenic fungi.

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Effect of Co-Culture with Mammalian Spermatozoa on In Vitro Maturation of Porcine Cumulus-Enclosed Germinal Vesicle Oocytes

  • Kim Byung Ki
    • Reproductive and Developmental Biology
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    • v.28 no.4
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    • pp.235-240
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    • 2004
  • In vitro maturation of denuded porcine immature oocytes can be enhanced by co-incubation with spermatozoa even before fertilization. This study was to determine whether the addition of spermatozoa into the culture medium could influence the nuclear maturation of porcine cumulus-enclosed germinal vesicle (GV) oocytes. Cumulus-oocyte complexes (COCs) were collected from follicles of 3- to 5-mm diameter. Porcine COCs were cultured in tissue culture medium containing spermatozoa. After 48 h culture, oocytes were examined for evidence of GV breakdown, metaphase I, anaphase-telophase I, and metaphase II. The proportion of oocytes reaching at metaphase II was significantly (P < 0.05) increased in the oocytes cultured in media containing spermatozoa compared to those in media without spermatozoa (52.3% vs 12.5%). No difference in the percentage of metaphase II was observed among the different periods of spermatozoa exposure and among the spermatozoa from different species. The proportion of oocytes reaching metaphase II was significantly different between high and low concentrations of spermatozoa. The present study suggests that manunalian spermatozoa contain a substance(s) that improves nuclear in vitro maturation of porcine cumulus-enclosed GV oocytes. Enhancing effect of spermatozoa for in vitro maturation of oocytes is a highly dose-dependent.

The Effects of Premarital Adult Child's Perceived Self-Differentiation and Dynamics of Emotional System between Parent and Child on Trait Anxiety (미혼성인자녀가 지각하는 자아분화와 부모-자녀간 정서체계 역동성이 특성불안에 미치는 영향)

  • Kwon Mi-Ae;Kim Phil-Sook;Park Ji-Hyun;Kim Tae-Hyun
    • Journal of the Korean Home Economics Association
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    • v.43 no.11 s.213
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    • pp.31-48
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    • 2005
  • The purpose of this study was to explore the direct and indirect effects of premarital adult child's perceived self-differentiation and parent-child dynamics of emotional system on trait anxiety. The subjects of the study numbered 277, and correlation and path analysis was used with SPSS 10.0. The major findings were as follows: 1. It was shown that self-differentiation, over involvement and perceived criticism of premarital adult children were correlated with trait anxiety. And, among demographic variables, education, income, and marital status of the subjects' parents were related with trait anxiety. 2. Parent-child differentiation, over involvement, perceived criticism, and sex had direct effects on premarital adult child's self-differentiation. The variables that had direct effects on trait anxiety, which was the final dependent variable were self-differentiation, parent-child differentiation, perceived criticism, and income while over involvement an indirect effect on it.

Effect of Epididymal Fluid on In Vitro Maturation and Subsequent Sperm Penetration in Porcine Follicular Oocytes

  • Kim, Byung-Ki
    • Reproductive and Developmental Biology
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    • v.33 no.3
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    • pp.125-131
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    • 2009
  • The aim of this study is to investigate the effect of porcine epididymal fluid (pEF) on in vitro-maturation and subsequent fertilization of porcine follicular oocytes. Porcine cumulus-oocytes complexes retrieved from antral follicles were cultured in tissue culture medium (TCM)-l99 supplemented with pEF of different concentrations. At 48 h after culture, development of oocytes to germinal vesicle (GV) breakdown, metaphase I, anaphase-telophase I, and metaphase II were examined Significant (p<0.05) increase in the proportion of oocytes developed to MII stage was observed in oocytes cultured in pEF-containing TCM-l99 than in oocytes cultured in pEF-free TCM-199 (46.2% vs 16.7%), which was a dose-dependent manner. Subsequently, the proportion of monospermic fertilization were significantly (p<0.05) increased in oocytes cultured in the TCM supplemented with pEF than those cultured in pEF-free TCM-199 (51.0% vs 24.1%). In the second series of experiment, the percentage of MII oocytes was significantly (p<0.05) increased after exposure of oocytes to pEF during the first 22 h period of culture than after exposure of oocytes to pEF during the next 24 h of culture, while no significant difference in the percentage of monospermy was observed. The results of this study demonstrate that pEF contains at least enhancing component(s) for nuclear maturation.

The Effects of Ginseng Saponin on Prolactin Secretion in Rats (인삼(人蔘) Saponin이 Prolactin 분비(分泌)에 미치는 효과(效果))

  • Baik, Duck-Woo;Lee, Soe-Yun;Chi, Hyung-Joon
    • Korean Journal of Pharmacognosy
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    • v.9 no.1
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    • pp.33-39
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    • 1978
  • The present study is involved with the prolactin secretion from anterior pituitary gland by ginseng saponin since it was handled down by tradition that ginseng might influence the milk secretion when it was given to nursing mother. To investigate the effect of saponin on the prolactin production or release from the anterior pituitary gland, cell culture study and whole animal studies were carried out. For the cell culture study, enzymatically dispersed anterior pituitary cells of rat anterior pituitary gland in HEPES buffers containing trypsin were used. Ginseng saponin was added to the culture media and the amount of prolactin produced in the cell culture media was determined by radloimmunoassay(RIA) technique. Dose-dependent increases of prolactin with ginseng saponin were observed, whereas, no change was observed without ginseng treatment. For the whole animal study, normal and castrated rats which previously cannulated into the heart via the right juglar vein were used. The prolactin concentration in plasma were determined by using the technique of RIA. In normal rats, prolactin concentration in plasma were elevated dramatically after 1 hour of ginseng saponin administration, whereas, instantaneous increases were observed in castrated rats. For prolactin assay by RIA, NIAMDD Rat Prolactin Kit and NIAMDD Rat Prolactin RP-1 were used as standard. The results indicate that ginseng saponins increase the release of prolactin from the anterior pituitary gland and production of prolactin from the cell in rats.

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A Study on the Differences of Kimchi Consumption according to Household Characteristics (가구 특성에 따른 김치 소비량 차이에 관한 연구)

  • Park, Sung Hoon
    • Journal of the Korean Society of Food Culture
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    • v.34 no.2
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    • pp.159-167
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    • 2019
  • The purpose of this study is to provide basic information to improve understanding of contemporary kimchi culture in Korea. Many Koreans are now purchasing kimchi at markets, while the proportion of self-preparation is gradually decreasing. This commodification tendency of kimchi is considered to be associated with changes in consumer's behavior and attitude. In this study, a linear regression and a logistic regression model were used to identify relationships between kimchi consumption behavior and household characteristics. The results showed that the probability of kimjang activity was positively related with family size, possession of a kimchi refrigerator, self-preparation practice, and the intensity of sharing behavior. I also found that kimchi consumption volume per capita of 'purchasing' household was greater than that of 'self-preparing ' or 'sharing-dependent' households, and that the number of family members was inversely related with kimchi consumption volume per capita. The inverse relationship between family size and kimchi consumption volume per capita is considered to be contrary to the widespread thoughts in Korea, which have been developed while experiencing kimchi preparation and consumption in traditional extended families. I think that the relationship comes from differences in menu varieties, which appear to vary with family size. This issue will be investigated in subsequent studies.

The Effect of Lactobacillus Mixture Culture Fluid Extracts on Atopic Dermatitis Chemokine Expression of in HaCaT Cells (HaCaT 세포에서 Lactobacillus 혼합배양액 추출물이 아토피관련 케모카인 발현에 미치는 효과)

  • Hong, Soo-Jeong;Lee, Won-Jae;Jo, Eul-Hwa;Ahn, Seong-Hun
    • Korean Journal of Acupuncture
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    • v.34 no.2
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    • pp.82-87
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    • 2017
  • Objectives : Recently the case of lactobacillus mixture culture fluid appliment was reported. In this study, anti-inflamation effects and anti-allergy effects were studied by stimulus of lactobacillus mixture culture fluid extracts in HaCaT cells. Methods : The atopic dermatitis were induced by TNF-${\alpha}$ and interferon-${\gamma}$ in HaCaT cells. TARC/CCL17, MDC/CCL22, RANTES/CCL5 and ROS production were investigated to explain anti-inflamation and allergy effects of lactobacillus mixture culture fluid with cell-enzyme-linked Immunosorbent assay in 450 nm, 485 nm, 535 nm with spectro-fluorometer. Results : The extracts of lactobacillus mixture culture fluid were decreased TARC/CCL17, MDC/CCL22, RANTES/CCL5 expressions and ROS production with a concentration dependent manner. Conclusions : The effects mechanism of Lactobacillus mixed culture fluid for atopic dermatitis symptoms were considered to be explain anti-inflamation and allergy effects via control of cytokine, chemokine and ROS production, and the fluid could be applied in skin cells directly. But classified AD symptom degrees reported in clinical case before as Reaction Period, Reduction Period, Effect Period, Reproduction Period and Rebound Period could not be explained. Further study will be expected.

Effects of Chitosan on Human Gingival Fibroblasts in Vitro (키토산이 치은섬유아세포에 미치는 영향)

  • Kim, Ok-Su;Chung, Hyun-Ju
    • Journal of Periodontal and Implant Science
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    • v.32 no.1
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    • pp.235-247
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    • 2002
  • The aim of this study was to evaluate the effects of chitosan coating on the attachment, proliferation, functional and morphological change of human gingival fibroblasts. Primary culture of human gingival fibroblasts were grown in Dulbecco's modified Eagle's medium with 10% fetal bovine serum and 1% antibiotics. In experimental group, cells were inoculated in the multiwell plates coated with chitosan in concentration of 0.02, 0.2, and 2 mg/ml. Cell counting and MTT assay were done after 0.5, 1.5, 3, 6 and 24 hours of incubation to evaluate the cell attachment, and then after 2 and 7 days of culture to evaluate the cell proliferation. The alkaline phosphatase activity was measured after 4 and 7 days of culture and the ability to produce mineralized nodules was evaluated after 21 days of culture. The results were as follows : The morphology of cells on the chitosan-coated well was round or spheric. Round cells were aggregated since 6 hours of culture and showed nodule-like appearance after 24 hours of culture and did not achieved confluency at 7 days. The attachment of gingival fibroblasts was inhibited by chitosan coating with a tendency of dose dependent pattern. But, cellular activity of unit cell was higher than control. The proliferation of gingival fibroblasts was inhibited by chitosan coating at 2 mg/ml(P<0.01), while the cell proliferation at 0.02, 0.2 $mg/m{\ell}$ was comparable to the control well. Total alkaline phosphatase activity was inhibited by chitosan coating and decreased in the course of time. While ALP activity of unit cell was the highest at 2mg/ml after 4 days of culture. Finally, gingival fibroblasts produced the mineralized nodule at 2 mg/ml. In summary, the attachment, proliferation, and alkaline phosphatase activity of gingival fibroblasts were influenced differently by the concentration of coated chitosan. From this study, it could be used as the matrix of tissue engineering for gingiva without inhibition on proliferation of gingival fibroblasts using chitosan at the optimal concentration (0.02mg/ml).

Optimization of Human Thrombopoietin Production in Insert Cells Using Baculovirus Expression System (베큘로 바이러스 발현 시스템에 의한 곤충세포에서의 인간 트롬보포이에틴 생산 최적화)

  • 고여욱;손미영;박상규;안혜경;박승국;박명환;양재명
    • KSBB Journal
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    • v.13 no.2
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    • pp.181-186
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    • 1998
  • In order to obtain high-level production of recombinant human thrombopoietin (rhTPO) in insect cell line, HTI-TN-5B1-4 (TN5), conditions for optimal rhTPO expression such as multiplicity of infection (MOI), the cell density at infection, harvesting time and type of culture method as well as growth media were determined. When TN5 cells were cultured as anchorage-dependent state in 60-mm dish, cell density $2\times^6$ cells,MOI of 10 and Garvesting the culture media at 72 hr post-infection wrere the cinditions for highest rh TPO production. High production of rhTPO was also achieved by using EXPRESS FIVE serum free media rather than SF900II serum free media-1. Anchorage-dependent TN5 cells were adapted as a suspension culture when they were grown in the presence of heparin. TN5 cells were successfully cultured at 0.2 L scale in suspension culture without having aggregation. When TN5 cells were cultured as suspension state, cell density of $0.6\times10^6$ cells/mL, MOI of 1 and harvesting the culture media at 72 hr post-infection, gave the highest yield of rhTPO.

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