• Title/Summary/Keyword: primed

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Improved Procedure for Large-scale Isolation of Mitochondrial DNA from Mammalian Tissues

  • Hong, Sung-Soo;Lee, Chung-Choo
    • Animal cells and systems
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    • v.3 no.1
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    • pp.73-78
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    • 1999
  • Although there are several methods for the preparation of mitochondrial DNA (mtDNA) from mammalian tissues, most are relatively long ultracentrifugation or manipulations by a small-scale method. We escribed a rapid method for large-scale extraction of mtDNA from human placental and horse liver tissues. The method is based on the preparation and homogenization of tissues, urification of crude mitochondria by differential centrifugations and isolation of mtDNA by alkaline Iysis. It was improved from Pre-existing methods by replacing some steps with simpler ones and discarding many others. This method gives a high yield of pure mtDNA(approximately 1-5mg from one placenta; ca. 400-600 g wet weight), depending on its sources (fresh tissue gave better results than frozen one). The resulting mtDNA indicated that this method can yield mtDNA in sufficient purity and quantity to identify the direct restriction analysis on agarose gel, random-primed labeling as a probe, and end labeling. Therefore, the method is ideal for obtaining good mtDNA samples to conduct routine restriction fragment length polymorphism (RFLP) analyses of natural populations for genetic studies.

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Production and Characteriuation of Monoclonal Antibodies against Human Interferon-$\alpha$ (인터페론 알파에 대한 단세포 군항체의 제조 및 특성)

  • Park, Kyung-Hee;Lee, Ihn-Sook
    • The Korean Journal of Zoology
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    • v.35 no.1
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    • pp.1-7
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    • 1992
  • Seven monoclonal antibodies were produced by fusing splenocytes from Balb/C mouse immunized with partially purified human interferon-a (HUIFN-a) with NSO plasmacytoma cells. aery were identified as five IgG class (432.22: IgG2b/n, 460.52: IgG2b/a , 548.46: IgG2a/n , 573.10: IgG2b/h , 625.12: IgG2b/n ), one IgA class (460.50: IgA/n ) and one IsM class (465.27: IgA/n ), and all of them revealed highly sensitive to HUIFN- a IgG class monoclonal antibodies have pts ranged from 8.2 to 8.6. Ascites fluids produced from primed Balb/c mice and were purified through column chromatography. The cytopathic effect (CPE) inhibition assay to examine neutralization of HuIFU-a by IgG class monoclonal antibodies, gave that MAbs 460.52, 548.46, 573.10 can neutralize HUIFU- a arith varying degrees except 432.22. Therefore, it is deduced that these various monoclonal antibodies may recognize the distinct epitopes on HUIFN-a.

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Effects of Quercus dentata and Quercus acutissima Extracts on the Activations of Macrophage RAW264.7 Primed with $IFN-{\gamma}$ (곡피(?皮)와 상목피(橡木皮) 추출물이 대식세포 RAW264.7 활성화에 미치는 영향)

  • Jo, Jin-Hee;Seong, Nak-Sull;Lee, Young-Jong
    • The Korea Journal of Herbology
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    • v.21 no.1
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    • pp.89-100
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    • 2006
  • Objectives : The effects of methanol extracts from the cortex of Quercus dentata Thunb. and Quercus acutissima Carruth, on the activation of macrophage were examined. Methods : Methanol extracts of Quercus dentata (QD) and Quercus acutissima (QA) were applied to cell line RAW264,7 (macrophage), and their effects were examined. Results : 1. Extracts from QD and QA had no specific influence on the cell growth. 2. Extracts from QD and QA did not activate macrophage independently, but the addition of $IFN-{\gamma}$ facilitated the generation of macrophage's nitric oxide(NO). 3. QD and QA extracts increased the manifestation of iNOS gene, when macrophage was activated by $IFN-{\gamma}$. 4. QD and QA extracts increased the manifestation of $TNF-{\alpha}$ in macrophage, which took 2 hours. 5. QD and QA extracts increased the generation of $PGE_2$ in macrophage. Conclusion : QD and QA activate macrophage in the presence of $IFN-{\gamma}$. After activation is primarily facilated by $IFN-{\gamma}$, it works on macrophage secondarily for the manifestation of iNOS gene and for the generation of $TNF-{\alpha}$.

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Effect of Priming and Preimbibition on Germination of Zoysiagrass Seeds (Priming과 침지처리가 들잔디의 종자발아에 미치는 영향)

  • 김태일;구자형
    • Asian Journal of Turfgrass Science
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    • v.8 no.1
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    • pp.1-11
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    • 1994
  • Studies were conducted to evaluate the effect of priming and preimbibition on gerraination of unscarified and KOH-scarified zoysiagrass (Zoysia japonica Steud.) seeds, respectively. Germi-nation percentages at 35˚C of seeds primed in $KNO_3$ and Ca($NO_3$)$_2$ were significantly higher than that of untreated seeds. The optimum conditions for priming unscarified seeds were in aerated $KNO_3$ solution at O4 M for 4 days at 20˚C. No additional benefit of $GA_3$ on increasing germination was not found in seed priming with solution of $KNO_3$ + $GA_3$ or in seeds preimbibed with $GA_3$ after priming. Scarified seeds preimbibed with distilled water(DW) for 4 days at 20˚C germinated higher and with fewer days to 50% of total germination(T 50) than untreated seeds. Preimbibition with $GA_3$ solution at 50 ppm for 4 days at 5, 10, 15 and 20˚C enabled up to 97% of seeds to germinate and reduced days to T 50 about 1.1 to 1.2 days at 35˚C Especially, seeds preimbibed with $GA^2$˚$_3_2_1$$\times$ had greatly higher germination and lower T 50 than untreated or DW-preimbibibed seeds at 30˚C, which is inadequate to germination in general. Preimbibed seed performance was unchanged after storage at room temperature for 50 days. Seeds preimbibed with GA $_3$ had higher rate of seedling emergence and faster growth of shoot than untreated seeds after sowing at 30˚C in growing medium in growth chamber.

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Platelet Shape Changes and Cytoskeleton Dynamics as Novel Therapeutic Targets for Anti-Thrombotic Drugs

  • Shin, Eun-Kyung;Park, Hanseul;Noh, Ji-Yoon;Lim, Kyung-Min;Chung, Jin-Ho
    • Biomolecules & Therapeutics
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    • v.25 no.3
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    • pp.223-230
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    • 2017
  • Platelets play an essential role in hemostasis through aggregation and adhesion to vascular injury sites but their unnecessary activation can often lead to thrombotic diseases. Upon exposure to physical or biochemical stimuli, remarkable platelet shape changes precede aggregation or adhesion. Platelets shape changes facilitate the formation and adhesion of platelet aggregates, but are readily reversible in contrast to the irrevocable characteristics of aggregation and adhesion. In this dynamic phenomenon, complex molecular signaling pathways and a host of diverse cytoskeleton proteins are involved. Platelet shape change is easily primed by diverse pro-thrombotic xenobiotics and stimuli, and its inhibition can modulate thrombosis, which can ultimately contribute to the development or prevention of thrombotic diseases. In this review, we discussed the current knowledge on the mechanisms of platelet shape change and also pathological implications and therapeutic opportunities for regulating the related cytoskeleton dynamics.

THE ANTICARIOGENIC EFFECT OF F IN PRIMER, BONDING AGENT AND COMPOSITE RESIN IN THE CAVOSURFACE ENAMEDL AREA (Primer Bonding agent, 복합레진 내의 볼소의 법랑질에 대한 항우식 작용)

  • Park, Sung-Ho
    • Restorative Dentistry and Endodontics
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    • v.21 no.2
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    • pp.635-641
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    • 1996
  • This study was designed to evaluate the anti cariogenic effect of F in primers, bonding agents, composite resins or glass ionomer cements in enamel. Twenty-five extracted teeth were selected and a cavity was prepared on either the buccal or the lingual surface of each tooth. After pumicing and etching, the samples were divided into 5 groups. In group A, the samples were primed, bonded and filled with ART bond and Brilliant Enamel (Coltene, Switzerland). Group B composed of Optibond and Herculited XRV (Kerr, USA), group C composed of Syntac and Tetric(Vivadent, Lichtenstein), and group D composed of Scotch-bond Multipurpose and Z 100 (3M, YSA). In group E, the samples were filled with glass ionomer cement (Fuji II LC, Japan), All surfaces except the 2mm beyond the cavosurface margin of the sample were protected, and samples were then put into an acid buffer for 3 days to develop the initial caries. The samples were then sectioned through the filling body into thin wafers and then examined with a polarizing microscope under water imbibition. The fluoride in primer, bonding agent, or composite filling material did not prevent the initial caries in the enamel area adjacent to the filling body whereas the fluoride in the glass ionomer did prevent the initial stage caries.

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Recession and YOLO: The Influence of Negative Perception of Economic Situation on Present-Biased Preference (경기 불황과 욜로(YOLO): 지각된 부정적 경제 상황이 소비자의 현재에 편향된 선호에 미치는 영향)

  • Jung, Bohee;Jeong, Hyewook
    • Journal of Digital Convergence
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    • v.19 no.11
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    • pp.135-144
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    • 2021
  • This study investigates the underlying mechanism of YOLO in millennial consumer, especially the influence of perceived economic recession on the present-biased preference. In addition, it was attempted to expand the implicit theory by proposing the individual's entity belief as a mediator for the effect of perceived economic situation on consumers' present-biased seeking behavior. In three experimental studies, undergraduate students who both highly primed and measured negative economic situation showed more favorable attitudes towards present-biased persuasive message and related products. The results of this research provides practical implication for marketers especially in the current situation experiencing economic slowdown due to low economic growth and COVID 19.

Negative evidence on the transgenerational inheritance of defense priming in Arabidopsis thaliana

  • Yun, Se-Hun;Noh, Bosl;Noh, Yoo-Sun
    • BMB Reports
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    • v.55 no.7
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    • pp.342-347
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    • 2022
  • Defense priming allows plants to enhance their immune responses to subsequent pathogen challenges. Recent reports suggested that acquired resistances in parental generation can be inherited into descendants. Although epigenetic mechanisms are plausible tools enabling the transmission of information or phenotypic traits induced by environmental cues across generations, the mechanism for the transgenerational inheritance of defense priming in plants has yet to be elucidated. With the initial aim to elucidate an epigenetic mechanism for the defense priming in plants, we reassessed the transgenerational inheritance of plant defense, however, could not observe any evidence supporting it. By using the same dipping method with previous reports, Arabidopsis was exposed repeatedly to Pseudomonas syringae pv tomato DC3000 (Pst DC3000) during vegetative or reproductive stages. Irrespective of the developmental stages of parental plants that received pathogen infection, the descendants did not exhibit primed resistance phenotypes, defense marker gene (PR1) expression, or elevated histone acetylation within PR1 chromatin. In assays using the pressure-infiltration method for infection, we obtained the same results as above. Thus, our results suggest that the previous observations on the transgenerational inheritance of defense priming in plants should be more extensively and carefully reassessed.

SARS-CoV-2-Specific T Cell Responses in Patients with COVID-19 and Unexposed Individuals

  • Min-Seok Rha;A Reum Kim;Eui-Cheol Shin
    • IMMUNE NETWORK
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    • v.21 no.1
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    • pp.2.1-2.11
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    • 2021
  • Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection causes coronavirus disease 2019 (COVID-19), an ongoing pandemic disease. In the current review, we describe SARS-CoV-2-specific CD4+ and CD8+ T-cell responses in acute and convalescent COVID-19 patients. We also discuss the relationships between COVID-19 severity and SARS-CoV-2-specific T-cell responses and summarize recent reports regarding SARS-CoV-2-reactive T cells in SARS-CoV-2-unexposed individuals. These T cells may be cross-reactive cells primed by previous infection with human common-cold coronaviruses. Finally, we outline SARS-CoV-2-specific T-cell responses in the context of vaccination. A better understanding of SARS-CoV-2-specific T-cell responses is needed to develop effective vaccines and therapeutics.

Population analysis of the toxic dinoflagellate genus Alexandrium by novel molecular markers

  • Kim, Choong-jae;Kim, Sook-Yang;Kim, Kui-Young;Kang, Young-Sil;Kim, Hak-Gyoon;Kim, Chang-Hoon
    • Proceedings of the Korean Aquaculture Society Conference
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    • 2003.10a
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    • pp.134-135
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    • 2003
  • The geographic expansion of the toxic dinoflagellates genus Alexandrium has been shown to be world wide ranging. The members of the genus Alexandrium ocnstituted of 20-30 species did not show substantial differences in their morphology, which is mostly referred in the 'tamarensis species complex', except some species. Though rDNA sequences variations are very few and pseudogene types are so diverse that it is difficult to use them as the specific markers. In this study, we outlined Korean and Japanese A, tamarense and A. catenella regional isolates by phylogenetic analysis inferred from no cutting alignments of LSU rDNA D1-D2 and SSU rDNA sequences to group these regional isolates. The results were compared to RFLP patterns of PCR products targeted chloroplast DNA. Lastly screening of highly repeated microsatellite DNA which is frequently used for population analysis in eukaryotes was conducted. A. catenella regional strains identified by the sequencing of rDNA D1-D2 domain were divided into at least 3 groups of type E, CMC and Chinese type, divergence root may not be deep comparing with that of A. tamarense whose pseudogenes are very variable. Results of RFLP pattern and the phylogeny of the unknown gene targeting chloroplast showed that Korean and Japanese A. catenella regional isolates were divided into 3 types: Korean, Japanese and the third CMC types. Population-specific PCR amplification with Japanese A. catenella type-specific PCR primers was useful method for population analysis of A. catenella. Various types of satellite sequences such as 5 nucleotides repeats were obtained from A. tamarense and A. catenella. The 5 nucleotides repeats were primed at the both 3'and 5' ends, and these repeats were prominent as longer repeated motifs. This repeated DNA was intercalated as internal sequences containing various types subrepeats. It is expected that these satellite DNA would be a useful molecular population marker through detail comparison among Alexandrium regional isolates to trace their transferring pathway and to prevent their human-associated their regional extents.

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