• Title/Summary/Keyword: depletion activity

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Adequate concentration of B cell leukemia/lymphoma 3 (Bcl3) is required for pluripotency and self-renewal of mouse embryonic stem cells via downregulation of Nanog transcription

  • Kang, Songhwa;Yun, Jisoo;Kim, Da Yeon;Jung, Seok Yun;Kim, Yeon Ju;Park, Ji Hye;Ji, Seung Taek;Jang, Woong Bi;Ha, Jongseong;Kim, Jae Ho;Baek, Sang Hong;Kwon, Sang-Mo
    • BMB Reports
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    • v.51 no.2
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    • pp.92-97
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    • 2018
  • B cell leukemia/lymphoma 3 (Bcl3) plays a pivotal role in immune homeostasis, cellular proliferation, and cell survival, as a co-activator or co-repressor of transcription of the $NF-{\kappa}B$ family. Recently, it was reported that Bcl3 positively regulates pluripotency genes, including Oct4, in mouse embryonic stem cells (mESCs). However, the role of Bcl3 in the maintenance of pluripotency and self-renewal activity is not fully established. Here, we report the dynamic regulation of the proliferation, pluripotency, and self-renewal of mESCs by Bcl3 via an influence on Nanog transcriptional activity. Bcl3 expression is predominantly observed in immature mESCs, but significantly decreased during cell differentiation by LIF depletion and in mESC-derived EBs. Importantly, the knockdown of Bcl3 resulted in the loss of self-renewal ability and decreased cell proliferation. Similarly, the ectopic expression of Bcl3 also resulted in a significant reduction of proliferation, and the self-renewal of mESCs was demonstrated by alkaline phosphatase staining and clonogenic single cell-derived colony assay. We further examined that Bcl3-mediated regulation of Nanog transcriptional activity in mESCs, which indicated that Bcl3 acts as a transcriptional repressor of Nanog expression in mESCs. In conclusion, we demonstrated that a sufficient concentration of Bcl3 in mESCs plays a critical role in the maintenance of pluripotency and the self-renewal of mESCs via the regulation of Nanog transcriptional activity.

Improvement of Peroxidase Productivity by Optimization of Medium Composition and Cell Inoculum Size in Suspension Cultures of Sweet Potato (Ipomoea batatas) (고구마(Ipomoea batatase)현탁배양에서 배지조성 및 세포접종량의 적정화에 의한 Pemxidase생산성 향상)

  • 곽상수;김수경;정경희;유순희;박일현;유장렬
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.2
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    • pp.91-97
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    • 1994
  • To improve the productivity of peroxidase (POD) of cell line SP-47 derived from cell suspension cultures of sweet potato (Ipomoea batatas (L) Lam.cv White Star), we optimized culture conditions including the composition and concentration of plant growth regulators and carbon source, and the cell inoculum size. When one g (fr wt) of cells was inoculated into 50 mL TL medium supplemented with l mg/L 2,4-D and 30g/L sucrose in 300 mL Erlenmeyer flask at 25$^{\circ}C$ in the dark (100rpm), the POD activity per g cell dry wt was maximized to be about 6,800 units after 25 days of subculture, which was about 30 times higher than that of intact roots of horseradish plants grown in the greenhouse, but the cell growth was maximum after 15 days of subculture. The protein content per g cell dry wt maintained almost plateau and after 25 days of subculture decreased as culture Proceeded further whereas the POD specific activity (unit/mg protein) was about two times higher after subculture and continuously increased from 12 days to the end of cultures (40 days). The POD isozyme patterns showed almost the same regardless of cell growth stage, but some acidic isozymes were slightly increased after 25 days of subculture. These results indicate that POD activity in suspension cultures of sweet potato is closely associated with cell growth and stresses derived from cell culture renditions and medium depletion. Due to its high POD activity the SPL47cell line seems to be suitable for the mass production of POD.

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Depletion Effects of Chick Germinal Crescent's Primordial Germ Cells by Heat Activated Busulfan Injection (닭 생식반월의 Busulfan 가온 주입방법에 의한 원시생식세포 제거 효과)

  • Jeong, Dong-Kee
    • Development and Reproduction
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    • v.11 no.3
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    • pp.219-226
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    • 2007
  • This study was conducted to identify optimistic primordial germ cells'(PGCs) migration activity using heat activated busulfan treatment for the increasing germline chimerism. Donar PGCs viability tests of important conditions for useful germ line chimerism indicated approximately $70{\sim}80%$ viability was time dependent. Transplantation experiments of PGCs into recipient embryos after busulfun treatment, showed the treatment group having 23.5% viability. By comparison, the control group showed 4.8% viability. The 96 hour treatment group and the 118 hour treatment group of the cultured PGCs showed high migration activity. Generally, the transplantation method would consider morphological and physiological characteristics before transplantation. In the present study, the effect of busulfan on migration activity showed viability highest at 53.4% after 48-hour incubation time. However, a previous study showed the best condition for transplantation time to be prior to the 48-hour incubation period, when the chicken embryo does not yet have a developed blood vessel system. In conclusion, an important condition for the production of a transgenic chicken is that most donor PGCs migrate into the recipient embryo without any inhibitory factors. The present results suggest, perhaps by using this modified method of transplantation, it can produce a more efficient chimeric germ line, transgenic chicken.

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Peroxidase Activity of Peroxidasin Affects Endothelial Cell Growth (내피 세포 성장에 영향을 미치는 PXDN의 peroxidase 활성)

  • Kyung A Ham;Seong Bin Jo;Min Ju Lee;Young Ae Joe
    • Journal of Life Science
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    • v.33 no.1
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    • pp.8-14
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    • 2023
  • Peroxidasin (PXDN), a multidomain heme peroxidase containing extracellular matrix (ECM) motifs, as well as a catalytic domain, catalyzes the sulfilimine crosslink of collagen IV (Col IV) to reinforce Col IV scaffolds. We previously reported that PXDN is required for endothelial cell (EC) survival and growth signaling through sulfilimine crosslink-dependent matrix assembly. In this study, we examined whether peroxidase activity is required for PXDN function in ECs. First, we constructed a mutant PXDN by point mutation of two highly conserved amino acids, Q823 and D826, which are present in the active site of the peroxidase domain. After isolation of HEK293 clones highly expressing the mutant protein, conditioned medium (CM) was obtained after incubating the cells in serum-free medium for 24 hours and then analyzed by Western blot analysis under nonreducing conditions. The results revealed that the mutant PXDN formed a trimer and that it was cleaved by proprotein convertase-like wild-type (WT) PXDN. However, peroxidase activity was not detected in the CM containing the mutant PXDN, in contrast to that of WT PXDN. In addition, the sulfilimine crosslink ability of the mutant PXDN was lost. Moreover, the CM containing the mutant PXDN failed to promote the growth of PXDN-depleted ECs, unlike the CM containing WT PXDN. These results suggest that the peroxidase activity of PXDN affects EC growth by forming a sulfilimine crosslink.

Dietary Tea Catechin Inclusion Changes Plasma Biochemical Parameters, Hormone Concentrations and Glutathione Redox Status in Goats

  • Zhong, Rongzhen;Xiao, Wenjun;Ren, Guopu;Zhou, Daowei;Tan, Chuanyan;Tan, Zhiliang;Han, Xuefeng;Tang, Shaoxun;Zhou, Chuanshe;Wang, Min
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.12
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    • pp.1681-1689
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    • 2011
  • The beneficial effects of tea catechins (TCs) are related not only to their antioxidant potential but also to the improvement of animal meat quality. In this study, we assessed the effects of dietary TC supplementation on plasma biochemical parameters, hormone responses, and glutathione redox status in goats. Forty Liuyang goats were randomly divided into four equal groups (10 animals/group) that were assigned to four experimental diets with TC supplementation at 4 levels (0, 2,000, 3,000 or 4,000 mg TC/kg DM feed). After a 60-day feeding trial, all goats were slaughtered and sampled. Dietary TC treatment had no significant effect on blood biochemical parameters, however, low-density lipoprotein cholesterol (p<0.001), triglyceride (p<0.01), plasma urea nitrogen (p<0.01), and glucose (p<0.001) decreased and total protein (p<0.01) and albumin (p<0.05) increased with the feeding time extension, and day 20 was the turning point for most of changes. Interactions were found in glutathione (p<0.001) and the ratio of reduced and oxidized glutathione (p<0.05) in whole blood between treatment and feeding time. Oxidized glutathione in blood was reduced (p<0.05) by 2,000 mg TC/kg feed supplementation, and a similar result was observed in longissimus dorsi muscle. Though plasma glutathione peroxidase (p<0.01) and glutathione reductase (p<0.05) activities were affected by treatment and feeding time interactions, and glutathione S-transferases activity increased with feeding day extension, no changed values appeared in longissimus dorsi muscle. In conclusion, dietary TC supplementation affected the concentrations of some blood metabolites and accelerated GSH depletion in the blood of goats. In terms of less high-density lipoprotein cholesterol, the highest insulin and IGF-I concentrations, the highest ratio of reduced and oxidized glutathione in plasma, the dosage of 2,000 mg TC/kg feed might be desirable for growing goats to prevent glutathione depletion and keep normal physiological metabolism.

Beneficial Roles of Azospirillum as Potential Bioinoculant for Eco-Friendly Agriculture (친환경농업을 위한 유용미생물 Azospirillum의 효율적 이용)

  • Gadagi, Ravi;Park, Myoung-Su;Lee, Hyoung-Seok;Seshadri, Sundaram;Chung, Jong-Bae;Sa, Tong-Min
    • Korean Journal of Soil Science and Fertilizer
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    • v.36 no.5
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    • pp.290-303
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    • 2003
  • Modern agriculture has been heavily dependent on chemical fertilizers to meet the food demands of ever increasing population. Progressive depletion of major plant nutrients in soil due to intensive cultivation practices has also necessitated the use of higher dose of chemical fertilizers particularly in soils where the organic matter content is very low. Indiscriminate use of chemical fertilizers and pressure on agriculturists to enhance per area crop yields has led to fast depletion of fossil fuel resources with concomitant increase in the prices of chemical fertilizers and also led to environmental pollution. Hence, the current trend throughout the world is to explore the possibility of using alternate nutrient sources or increasing the efficiency of chemical fertilizers by supplementing them with organic fertilizers and bioinoculants comprising largely microbes like, bacteria, fungi, algae etc to enhance nitrogen and phosphates in the soil thus creating a sustainable agricultural environment. Among the different microbial inoculants or biofertilizers, Azospirillum could be a potential candidate due to its non specific host root colonization. It had the capability to fix $N_2$ in wide pH regimes and even in presence of combined nitrogen. Azospirillum inoculation can increase the crop yield to 10-25% and substitute 25% of recommended doses of nitrogenous fertilizers. Apart from nitrogen fixation, Azospirillum is also involved in the root improvement, the activity which was attributed to an increase in the rate of water and mineral uptake by roots. The ability of Azospirillum to produce phytohormones was reported to enhance the root respiration rate, metabolism and root proliferation. They have also been reported to produce polyhydroxybutyrate, that can be used as a biodegradable thermosplastic. A lot of studies have addressed improvements in enhancing its efficiency to fix nitrogen fixation and hormone production.

Mitochondrial Dysfunction and Cancer (미토콘드리아 기능 이상과 암)

  • Han, Yu-Seon;Jegal, Myeong-Eun;Kim, Yung-Jin
    • Journal of Life Science
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    • v.29 no.9
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    • pp.1034-1046
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    • 2019
  • The mitochondria is the major cellular organelle of energy metabolism for the supply of cellular energy; it also plays an important role in controlling calcium regulation, reactive oxygen species (ROS) production, and apoptosis. Mitochondrial dysfunction causes various diseases, such as neurodegenerative diseases, Lou Gehrig's disease, cardiovascular disease, mental disorders, diabetes, and cancer. Most of the diseases are age-related diseases. In this review, we focus on the roles of mitochondrial dysfunction in cancer. Mitochondrial dysfunction induces carcinogenesis and is found in many cancers. The factors that cause mitochondrial dysfunction differ depending on the types of carcinoma, and those factors could cause cancer malignancy, such as resistance to therapy and metastasis. Mitochondrial dysfunction is caused by a lack of mitochondria, an inability to provide key substances, or a dysfunction in the ATP synthesis machinery. The main factor associated with cancer malignancy is mtDNA depletion. Mitochondrial dysfunction would leads to malignancy through changes in molecular activity or expression, but it is not known in detail which changes lead to cancer malignancy. In order to explore the relationship between mitochondrial dysfunction and cancer malignancy in detail, mitochondria dysfunctional cell lines are constructed using chemical methods such as EtBr treatment or gene editing methods, including shRNA and CRISPR/Cas9. Those mitochondria dysfunctional cell lines are used in the study of various diseases caused by mitochondrial dysfunction, including cancer.

Expression of Invertase in Recombinant Saccharomyces cerebisiae Containing SUC2 Gene (SUC2 Gene을 갖는 재조합 Saccharomyces cerebisiae의 Invertase 발현특성)

  • 정상철;장재권;김인규;변유량
    • Microbiology and Biotechnology Letters
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    • v.17 no.3
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    • pp.263-268
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    • 1989
  • To maximize the performance of recombinant cell fermentation process through optimizing environmental conditions, the production of invertase from recombinant Saccharomyces cerebisiae Containing SUC2 gene was studied as a model. The recombinant cells showed biphasic growth on glucose. Since the promoter of the SUC2 is regulated by the concentration of glucose in the medium, expression of invertase by recombinant yeast began when the glucose concentration decreased in a range of 0.25-0.4 g/L during the batch culture. Plasmid segregation occured frequently during glucose fermentation, and infrequently during ethanol oxidation. A rapid appearance of invertase activity with glucose was observed under nonaerated condition, and the maximum specific invertase activity was about 1.5 times as high as under aerobic condition, In fed batch culture, when n low level of glucose was continuously supplied to the tormentor after the time of glucose depletion during growth phase, specific and total invertase activity increased about 1.7 and 2.9 fold, respectively, in a batch culture.

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Assessment of soil moisture-vegetation-carbon flux relationship for agricultural drought using optical multispectral sensor (다중분광광학센서를 활용한 농업가뭄의 토양수분-식생-이산화탄소 플럭스 관계 분석)

  • Sur, Chanyang;Nam, Won-Hob
    • Journal of Korea Water Resources Association
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    • v.56 no.11
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    • pp.721-728
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    • 2023
  • Agricultural drought is triggered by a depletion of moisture content in the soil, which hinders photosynthesis and thus increases carbon dioxide (CO2) concentrations in the atmosphere. The aim of this study is to analyze the relationship between soil moisture (SM) and vegetation activity toward quantifying CO2 concentration in the atmosphere. To this end, the MODerate resolution imaging spectroradiometer (MODIS), an optical multispectral sensor, was used to evaluate two regions in South Korea for validation. Vegetation activity was analyzed through MOD13A1 vegetation indices products, and MODIS gross primary productivity (GPP) product was used to calculate the CO2 flux based on its relationship with respiration. In the case of SM, it was calculated through the method of applying apparent thermal inertia (ATI) in combination with land surface temperature and albedo. To validate the SM and CO2 flux, flux tower data was used which are the observed measurement values for the extreme drought period of 2014 and 2015 in South Korea. These two variables were analyzed for temporal variation on flux tower data as daily time scale, and the relationship with vegetation index (VI) was synthesized and analyzed on a monthly scale. The highest correlation between SM and VI (correlation coefficient (r) = 0.82) was observed at a time lag of one month, and that between VI and CO2 (r = 0.81) at half month. This regional study suggests a potential capability of MODIS-based SM, VI, and CO2 flux, which can be applied to an assessment of the global view of the agricultural drought by using available satellite remote sensing products.

Korean Red Ginseng extract ameliorates melanogenesis in humans and induces antiphotoaging effects in ultraviolet B-irradiated hairless mice

  • Saba, Evelyn;Kim, Seung-Hyung;Lee, Yuan Yee;Park, Chae-Kyu;Oh, Jae-Wook;Kim, Tae-Hwan;Kim, Hyun-Kyoung;Roh, Seong-Soo;Rhee, Man Hee
    • Journal of Ginseng Research
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    • v.44 no.3
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    • pp.496-505
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    • 2020
  • Background: Panax ginseng is a marvelous herbal remedy for all ailments of body. That may be why it is called Panax, which means "cure for all". Melanin is a pigment that gives color to our skin; however, increased melanin production can lead to tumor formation. Human exposure to ultraviolet B radiation has increased extensively owing to the increased sunlight due to global warming. Consequently, a phenomenon called photoaging has been observed for all skin colors and types. As a result of this phenomenon, a set of enzymes called matrix metalloproteinases, which serve as degradation enzymes for extracellular matrix proteins, mainly collagen, is increased, causing depletion of collagen and resulting in early wrinkle formation. Methods: Therefore, in our study, we used the murine melanoma cell line B16/F10 to study the inhibition of melanogenesis by Korean Red Ginseng (KRG) extract in vitro and HRM-2 hairless mice exposed to artificial ultraviolet B to examine the efficacy of KRG in vivo. We prepared a 3% red ginseng extract cream and evaluated its effects on human skin. Results: Our results demonstrated that KRG induced potent suppression of tyrosinase activity and melanin production in B16/F10 cells; moreover, it reduced the transcription and translation of components involved in the melanin production pathway. In the in vivo experiments, KRG potently suppressed the expression of matrix metalloproteinases, reduced wrinkle formation, and inhibited collagen degradation. On human skin, ginseng cream increased skin resilience and skin moisture and enhanced skin tone. Conclusion: Therefore, we conclude that KRG is an excellent skin whitening and antiaging product.