• Title/Summary/Keyword: Co-regulator

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Hypertonicity Down-regulates the $1{\alpha},25(OH)_2$ Vitamin $D_3$-induced Osteoclastogenesis Via the Modulation of RANKL Expression in Osteoblast

  • Jeong, Hyun-Joo;Yushun, Tian;Kim, Bo-Hye;Nam, Mi-Young;Lee, Hyun-A;Yoo, Yun-Jung;Seo, Jeong-Taeg;Shin, Dong-Min;Ohk, Seung-Ho;Lee, Syng-Ill
    • International Journal of Oral Biology
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    • v.30 no.1
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    • pp.23-30
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    • 2005
  • Bone remodeling is a process controlled by the action of two major bone cells; the bone forming osteoblast and the bone resorbing osteoclast. In the process of osteoclastogenesis, stromal cells and osteoblast produce RANKL, OPG, and M-CSF, which in turn regulate the osteoclastogenesis. During the bone resorption by activated osteoclasts, extracellular $Ca^{2+}/{PO_4}^{2-}$ concentration and degraded organic materials goes up, providing the hypertonic microenvironment. In this study, we tested the effects of hypertonicity due to the degraded organic materials on osteoclastogenesis in co-culture system. It was examined the cellular response of osteoblastic cell in terms of osteoclastogenesis by applying the sucrose, and mannitol, as a substitute of degraded organic materials to co-culture system. Apart from the sucrose, mannitol, and NaCl was tested to be compared to the effect of organic osmotic particles. The addition of sucrose and mannitol (25, 50, 100, 150, or 200 mM) to co-culture medium inhibited the number of tartrate-resistant acid phosphatase (TRAP) positive multinucleated cells induced by 10 nM $1{\alpha},25(OH)_2vitaminD_3$ ($1{\alpha},25(OH)_2D_3$). However, NaCl did exert harmful effect upon the cells in this co-culture system, which is attributed to DNA damage in high concentration of NaCl. To further investigate the mechanism by which hypertonicity inhibits $1{\alpha},25(OH)_2D_3$-induced osteoclastogenesis, the mRNA expressions of receptor activator of nuclear factor (NF)-kB ligand (RANKL) and osteoprotegerin (OPG) were monitored by RT-PCR. In the presence of sucrose (50 mM), RANKL mRNA expression was decreased in a dose-dependent manner, while the change in OPG and M-CSF mRNA were not occurred in significantly. The RANKL mRNA expression was inhibited for 48 hours in the presence of sucrose (50 mM), but such a decrement recovered after 72 hours. However, there were no considerable changes in the expression of OPG and M-CSF mRNA. Conclusively, these findings strongly suggest that hypertonic stress down-regulates $1{\alpha},25(OH)_2D_3$-induced osteoclastogenesis via RANKL signal pathway in osteoblastic cell, and may playa pivotal role as a regulator that modulates osteoclastogenesis.

Inhibition of poly 3-hydroxybutyrate (PHB) synthesis by phaR deletion in Methylobacterium extorquens AM1 (메탄올자화균 Methylobacterium extorquens AM1의 phaR 유전자 결실을 통한 poly 3-hydroxybutyrate (PHB) 생합성 억제)

  • Kim, Yujin;Lee, Kwanghyun;Kim, Hyeonsoo;Cho, Sukhyeong;Lee, Jinwon
    • Korean Chemical Engineering Research
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    • v.55 no.3
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    • pp.363-368
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    • 2017
  • Methylotrophy is able to use reduced one-carbon compound, such as methanol and methylamine, as a sole carbon source. Methylobacterium extorquens AM1 is the most extensively studied methylotroph utilizing serine-isocitrate lyase cycle. Because the Poly 3-hydroxybutyrate (PHB) synthesis pathway in M. extorquens AM1 is likely to interlink with EMCP (ethylmalonyl-CoA pathway), glyoxylate, and TCA cycles, regulation of PHB production is needed to produce EMCP-derived acid or TCA acids. To adjust carbon flux to PHB production, PhaR, which seems to have function of regulator of PHB synthesis and acetyl-CoA flux, was knocked out in M. extorquens AM1 by using markerless gene deletion methods. As a result, PHB granules were remarkably reduced in the knockout strain ${\Delta}phaR$ compared to parental strain. Although lag phase was extended for 12h, ${\Delta}phaR$ showed similar cell growth and methanol consumption rate compared to wild type.

Characterization of the scr Gene Cluster Involved! in Sucrose Utilization in Bifidobacterium longum (Bifidobacterium longum의 Sucrose 대사 관련 scr 유전자군의 특성 규명)

  • 권태연;이종훈
    • Microbiology and Biotechnology Letters
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    • v.32 no.3
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    • pp.199-205
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    • 2004
  • The nucleotide sequence of 8.6-kb EcoRI fragment containing sucrose phosphorylase gene isolated from Bifidobacterium longum SJ32 was determined. It was found that the fragment contained five open reading frames including the gene cluster for sucrose utilization such as a sucrose phosphorylase (ScrP), a sucrose transporter (ScrT), and a GalR-LacI-type transcriptional regulator (ScrR) identified by amino acid homology. Each gene showed over 94% amino acid homology among various B. longum strains. Genomic organization of the gene cluster is the same as those of other strains of B. longum but different from that of B. lactis. In spite of high homology of each gene among B. longum strains, the difference of flanking sequences of the gene cluster between strains SJ32 and NCC2705 insinuates the horizontal transfer of scrPTR between B. longum strains. The increase of sucrose phosphorylase activity in heterologous E. coli system by the co-expression of scrT with scrP against the single expression of scrP was measured. It seems to be the result of sucrose uptake increment by scrT in the host and is an indirect evidence that scrT is the gene for sucrose transport. The existence of multiple sucrose uptake systems in B. longum is supposed from the findings of several genes besides scrPTR involved in sucrose uptake in the genome of B. longum NCC2705.

Effect of seaweed extracts(GA14), a plant growth regulator, on growth and yield of two rice cultivars (식물생장조절제 Seaweed extracts(GA14)의 수도 품종간 생육 및 수량에 미치는 효과)

  • Song, Jae-Young;Kim, Yong-Il;Park, Bo-Young;Jung, Jae-Young;Choi, Hyun-Gu;Jung, Jong-Tae;Lee, Hee-Bong
    • Korean Journal of Agricultural Science
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    • v.31 no.1
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    • pp.1-8
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    • 2004
  • This study was carried out to identify the effects of seaweed extracts(GA14) on growth of two rice cultivars, Junambyo and Donganbyo. Seedling qualities of two cultivars were better in all items including heading dates at early stage treated at seedling plus 2-3 leaf stages than at single treatment of seedling. Ripening ratio of Junambyo in paddy field was increased 0.3% by seaweed extracts(GA14) treatment, but that of Donganbyo decreased 0.5%. 1,000 grain weight of Junambyo and Donanbyo by seaweed extracts(GA14) treatment was two to four grams higher and the yield of two cultivars was also higher by three to four percent. Appearance characters of two rice cultivars was high in head, while damaged, chalky and crack rate were low at seaweed extracts(GA14) treatment. Protein, moisture and amylose characteristics related to table quality of Junambyo were not different by seaweed extracts treatment, but table values was high in only treatment. Donganbyo was also similar to Junambyo, but table quality was slightly high at non-treatment.

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Transformation of Populus tremuloides Using Agrobacterium rhizogenes (Agrobacterium rhizogenes를 이용한 Populus tremuloides의 형질전환)

  • So, In-Sup;U, Zang-Kual;Ko, Young-Hwan;Lee, Sun-Joo;Hackett, Wesley P.;Riu, Key-Zung
    • Applied Biological Chemistry
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    • v.38 no.2
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    • pp.123-128
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    • 1995
  • Several factors affecting on transformation efficiency were studied to establish a Agrobacterium rhizogenes mediated system for the transformation of Populus species, and We could obtaine tansgenic plantlets expressing the introduced gene. Leaf sections were more sensitive than stem sections to kanamycin and thought to be better material for transformant screening. The bacterial density did not affect on the efficiency of transformation over the range of $4{\times}10^5{\sim}7{\times}10^9\;cfu$. The optimum period for co-cultivation was one day or shorter. Both of the optimum concentrations of cefotaxime and ampicillin in the medium were $250\;{\mu}g/ml$ for elimination of bacteria from the inoculated leaf sections. The addition of acetosyringone in the bacterial culture medium increased transformation rate, and the highest rate was obtained at $50\;{\mu}M$ of acetosyringone. The transformed galls could be selectively induced and gown on the growth regulator-free medium or on the medium containing $100\;{\mu}g/ml$ or higher contrition of kanamycin. The roots were induced from the galls incited by A. rhizogenes within 3 weeks on the growth regulator-free medium as well as on the medium containing growth regulators. The plantlets were regenerated from the galls cultured for 6 weeks on the medium containing 0.05 mg/ml of NAA and 0.5 mg/ml of BA. The expressions of the introduced opine gene in the transformed galls and plantlets were confirmed by the analysis of agropine and mannopine.

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A Design Study of Signal Processor for Small Tracking Radar (소형 추적 레이더를 위한 신호처리기 설계 기술 연구)

  • Choi, Jinkyu;Park, Changhyun;Kim, Younjin;Kim, Hongrak;Kwon, Junbeom;Kim, Gwang-Hee
    • The Journal of the Institute of Internet, Broadcasting and Communication
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    • v.20 no.5
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    • pp.71-77
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    • 2020
  • Recently, the tracking radar has confirmed the necessity of developing a small tracking radar that can be operated without various restrictions in various environments. In addition, the performance of a small tracking radar requires equal to or higher than the existing tracking radar. Such a small tracking radar can be implemented through miniaturization and low power of existing tracking radar. In this paper, the role and function of a signal processor for a small tracking radar are defined and we proposed a method to increase the efficiency of power consumption and miniaturization by minimizing the use of devices required to implement a signal processor for a small tracking radar. Used as a method for miniaturization, a device processor such as DDC and communication controller was implemented in an FPGA to design a signal processor for a small tracking radar. In addition, a low-power signal processor was designed by a power supply using a highly efficient switching regulator. Finally, the signal processor was verified by the performance test of the signal processor for the small tracking radar implemented, the Doppler tracking test using the signal processor on the small tracking radar, and the distance tracking test.

Quality Characteristics, Carbon Dioxide, and Ethylene Production of Asparagus (Asparagus officinalis L.) Treated with 1-Methylcyclopropene and 2-Chloroethylphosphonic Acid during Storage (아스파라거스에서 1-MCP와 CEPA 처리에 따른 CO2 및 에틸렌 발생과 품질특성)

  • Lee, Jung-Soo
    • Horticultural Science & Technology
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    • v.33 no.5
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    • pp.675-686
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    • 2015
  • Asparagus (Asparagus officinalis L.) needs proper post-harvest treatment to prolong its storage life. This study investigated the effect of 1-methylcyclopropene (1-MCP) on the quality and storage life of asparagus. Fresh-harvested asparagus was treated with 1-MCP ($1mg{\cdot}L^{-1}$), CEPA($10mg{\cdot}L^{-1}$), and 1-MCP($1mg{\cdot}L^{-1}$) + CEPA($10mg{\cdot}L^{-1}$) and compared with an untreated control. The carbon dioxide ($CO_2$) production, ethylene production, and morphological characteristics of the preserved asparagus were observed. The flow-system and the static-type measurement methods for ethylene and $CO_2$ production (respiration rate) were used. Weight loss, respiration rate, degree of freshness, and ethylene production were monitored during storage at $7^{\circ}C$. The results further showed that CEPA (2-chloroethylphosphonic acid) treatment had greater effects on $CO_2$ and ethylene production than using the 1-MCP process. The asparagus treated with CEPA or 1-MCP + CEPA had significantly increased the ethylene production rate compared to the control or using only 1-MCP during storage. There were no evident changes in the respiration rate of asparagus under 1-MCP treatment as compared with the control. Using the flow-system, slight differences in the rates of $CO_2$ and ethylene production were noted as compared to using the static type. Findings showed that in using the flow-system, asparagus manifested clearer results as compared with the static type. Weight loss in asparagus was significantly lower in control and 1-MCP treated samples than in those treated with CEPA. Likewise, the $CO_2$ and ethylene production of the CEPA treated samples significantly increased. The 1-MCP treatment reduced the effects of CEPA on weight loss, soluble solids content, and osmolality. The effect was not observed with exogenous ethylene as CEPA treatment had no visible effect as compared to the untreated group. Thus, 1-MCP treatment of asparagus could slightly reduce damage to the quality of asparagus during its distribution where ethylene gas is produced. Therefore, this study suggests that 1-MCP treatment can reduce the damage induced by ethylene gas on asparagus in poor distribution environments.

High Expression of HIF-1α, BNIP3 and PI3KC3: Hypoxia-Induced Autophagy Predicts Cholangiocarcinoma Survival and Metastasis

  • Thongchot, Suyanee;Yongvanit, Puangrat;Loilome, Watcharin;Seubwai, Wanchana;Phunicom, Kutcharin;Tassaneeyakul, Wichittra;Pairojkul, Chawalit;Promkotra, Wisuttiphong;Techasen, Anchalee;Namwat, Nisana
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.14
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    • pp.5873-5878
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    • 2014
  • Hypoxia and autophagy are known to facilitate tumor progression. We here aimed to investigate the role of hypoxia-associated autophagy in cholangiocarcinoma (CCA) survival and metastasis. Immunostaining of hypoxic-responsive proteins (HIF-$1{\alpha}$ and BNIP3) and a key regulator of autophagy (PI3KC3) were examined in CCA tissues and their expression levels were compared with clinicopathological parameters. A hypoxia mimicking condition ($CoCl_2$ treatment) was also tested regarding CCA cell functions. Our results showed that HIF-$1{\alpha}$ (66%), BNIP3 (44%) and PI3KC3 (46%) showed strong staining in human CCA tissues. Positive expression of HIF-$1{\alpha}$ (p=0.033), BNIP3 (p=0.040) and PI3KC3 (p=0.037) was significantly correlated with lymph node metastasis. HIF-$1{\alpha}$ was well associated with BNIP3 (r=0.3, p<0.01) and PI3KC3 (r=0.2, p<0.01). The survival rates of patients who were positive with HIF-$1{\alpha}$ (p=0.047) or co-expressed HIF-$1{\alpha}$ and BNIP3 (p=0.032) or HIF-$1{\alpha}$ and PI3KC3 (p=0.043) were significantly greater than in the negative groups. CCA cells treated with $CoCl_2$ showed an increase in HIF-$1{\alpha}$, BNIP3, PI3KC3 and LC3-II, with increased cell migration and pFAK levels. These data suggest that hypoxia associated autophagy enhances CCA metastasis, resulting in a poor prognosis of CCA.

The Verification Of Green Soil Material Characteristics For Slope Protection (사면 보호를 위한 녹생토 재료 특성 검증)

  • Lee, Byung-Jae;Heo, Hyung-Seok;Noh, Jae-Ho;Jang, Young-Il
    • Asia-pacific Journal of Multimedia Services Convergent with Art, Humanities, and Sociology
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    • v.7 no.6
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    • pp.681-692
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    • 2017
  • In recent years, large-scale construction projects such as road pavement construction and new city construction have been carried out nationwide with by the expansion of social overhead facilities and base on the economic development planning, resulting in a rapid increase in artificial slope damage. The existing vegetation-based re-installation method of the slope surface greening method reveals various problems such as lack of bonding force, drying, and lack of organic matter. In this study, research was carried out using vegetation-based material and environmentally friendly soil additives, were are used in combination with natural humus, Bark compost, coco peat, and vermiculite. Uniaxial compressive strength was measured according to the mixing ratio of soil additives and the strength was analyzed. Experiments were carried out on the characteristics of the soil material to gauge the slope protection properties by using the soil compaction test method wherein the soil and the soil additive materials are mixed in relation to the soil height, the number of compaction, the compaction method (layer) and the curing condition. As a result of the experiment, excellent strength performance was demonstrated in soil additives using gypsum cement, and it satisfied vegetation growth standards by using performance enhancer and pH regulator. It was confirmed that the strength increases with the mixing of soil and soil additive, and the stability of slope protection can be improved.

In Vitro Bioassay for Transforming Growth Factor-$\beta$ Using XTT Method

  • Kim, Mi-Sung;Ahn, Seong-Min;Moon, Aree
    • Archives of Pharmacal Research
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    • v.25 no.6
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    • pp.903-909
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    • 2002
  • Research in the cytokine field has grown exponentially in recent years, and the validity of such studies relies heavily on the appropriate measurement of levels of cytokines in various biological samples. Transforming growth factor (TGF)-$\beta$, a hormonally active polypeptide found in normal and transformed tissue, is a potent regulator of cell growth and differentiation. The most widely used bioassay for TGF-$\beta$ is the inhibition of the proliferation of mink lung epithelial cells. Though detection of [$^3$H]thymidine incorporation is more sensitive than the MTT assay, it presents some disadvantages due to the safety and disposal problems associated with radioisotopes. In this study, we attempted to ascertain the experimental conditions which could be used for measuring the in vitro biological activity of TGF-$\beta$ in a safer and more sensitive way compared with the currently available methods. We compared the commonly used method, the MTT assay, to the XTT assay using different parameters including cell number, incubation time and the wave length used for detecting the product. We examined the anti-proliferative activities of TGF-$\beta$ in three different cell lines: Mv-1-Lu mink lung epithelial cells, MCF10A human breast epithelial cells and H-ras-transformed MCF10A cells. Herein, we present an experimental protocol which provides the most sensitive method of quantifying the biological activity of TGF-$\beta$, with a detection limit of as low as 10 pg/ml: Mv-1-Lu or H-ras MCF10A cells ($1{\times}10^5/well$) were incubated with TGF-$\beta$ at $37^{\circ}C$ in a humidified $CO_2$ incubator for 24 hr followed by XTT treatment and determination of absorbance at 450 or 490 nm. Our results may contribute to the establishment of an in vitro bioassay system, which could be used for the satisfactory quantitation of TGF-$\beta$.