• Title/Summary/Keyword: Li2CO3

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Soluble Production of CMP-Neu5Ac Synthetase by Co-expression of Chaperone Proteins in Escherichia coli (샤페론 단백질 동시 발현기술을 이용한 수용성 CMP-Neu5Ac Synthetase 생산)

  • Choi, Hwa Young;Li, Ling;Cho, Seung Kee;Lee, Won-Heong;Seo, Jin-Ho;Han, Nam Soo
    • Microbiology and Biotechnology Letters
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    • v.42 no.2
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    • pp.190-193
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    • 2014
  • CMP-Neu5Ac synthetase is a key enzyme for the synthesis of CMP-Neu5Ac, which is an essential precursor of sialylated glycoconjugates. For the soluble expression of the CMP-Neu5Ac synthetase gene (neuA) from Escherichia coli K1, various heat shock proteins were co-expressed in E. coli BL21 (DE3) Star. In order to do this, a pG-KJE8 plasmid, encoding genes for GroEL-ES and DnaK-DnaJ-GrpE, was co-transformed with neuA and was expressed at $20^{\circ}C$ by the addition of 0.01 mM IPTG and 0.005 mg/ml L-arabinose. The co-expression of a variety of heat shock proteins resulted in the remarkably improved production of soluble CMP-Neu5Ac synthetase in E. coli.

Soluble Expression of the Fucosyltransferase Gene from Helicobacter pylori in Escherichia coli by Co-expression of Molecular Chaperones (샤페론단백질동시발현기술을이용하여 Helicobacter pylori 유래의 fucosyltransferase의수용성생산)

  • Lee, A Reum;Li, Ling;Shin, So-Yeon;Moon, Jin Seok;Eom, Hyun-Ju;Han, Nam Soo
    • Microbiology and Biotechnology Letters
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    • v.43 no.3
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    • pp.212-218
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    • 2015
  • Fucosyltransferases (FucTs) catalyze fucosyl transfer from guanosine-diphosphate fucose (GDP-β-L-fucose) to acceptor molecules to form fucosyloligosaccharides with α-glycosidic linkages. However, when FucT genes have been expressed in Escherichia coli, most cases have resulted in the production of inclusion bodies. In this study, to overcome this drawback, molecular chaperones were co-expressed with α1,2-fucosyltransferase (FucT2) in E. coli. For this, the pACYC184 vector, having genes for chaperones such as GroEL, GroES, DnaK, DnaJ, and GrpE, were transformed into E. coli BL21 (DE3) star harboring pHFucT2, including the FucT2 gene from Helicobacter pylori 26695. The results from SDS-PAGE showed that 5 chaperones were successfully expressed and the soluble fraction of FucT2 was also increased. HPLC analysis revealed that the coexpression of chaperone proteins resulted in a 5-fold increase in the total activity of fucosyltransferase in E. coli. In conclusion, the FucT2 expression system developed in this study can be used as a useful tool for the synthesis of fucosyloligosaccharides.

Identification of AMPK activator from twelve pure compounds isolated from Aralia Taibaiensis: implication in antihyperglycemic and hypolipidemic activities

  • Li, Yuwen;Park, Jongsun;Wu, Yin;Cui, Jia;Jia, Na;Xi, Miaomiao;Wen, Aidong
    • The Korean Journal of Physiology and Pharmacology
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    • v.21 no.3
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    • pp.279-286
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    • 2017
  • The root bark extract of Aralia taibaiensis is used traditionally for the treatment of diabetes mellitus in China. The total saponin extracted from Aralia Taibaiensis (sAT) has effective combined antihyperglycemic and hypolipidemic activities in experimental type 2 diabetic rats. However, the active compounds have not yet been fully investigated. In the present study, we examined effects of twelve triterpenoid saponins on AMP-activated protein kinase (AMPK) activation, and found that compound 28-O-${\beta}$-D-glucopyranosyl ester (AT12) significantly increased phosphorylation of AMPK and Acetyl-CoA carboxylase (ACC). AT12 effectively decreased blood glucose, triglyceride (TG), free fatty acid (FFA) and low density lipoprotein-cholesterol (LDL-C) levels in the rat model of type 2 diabetes mellitus (T2DM). The mechanism by which AT12 activated AMPK was subsequently investigated. Intracellular ATP level and oxygen consumption were significantly reduced by AT12 treatment. The findings suggested AT12 was a novel AMPK activator, and could be useful for the treatment of metabolic diseases.

Effect of Exogenous Xylanase Supplementation on the Performance, Net Energy and Gut Microflora of Broiler Chickens Fed Wheat-based Diets

  • Nian, F.;Guo, Y.M.;Ru, Y.J.;Li, F.D.;Peron, A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.3
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    • pp.400-406
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    • 2011
  • An experiment was carried out to assess the effects of xylanase supplementation on the performance, net energy and gut microflora of broilers fed a wheat-based diet. Day-old male broiler chicks were allocated to two dietary treatments. Each treatment was composed of six replicate cages of seven broilers per cage. The diets were wheat-based and offered as mash. The treatments included i) basal diet deficient in metabolizable energy; and ii) basal diet supplemented with a commercial xylanase added at 4,000 U/kg feed. Bird performance, nutrient utilization and gut microbial populations were measured. Heat production and net energy were determined using an auto-control, open circuit respiration calorimetry apparatus. Results showed that exogenous xylanase supplementation improved feed conversion efficiency (p<0.05) and increased diet AME (+4.2%; p<0.05), as well as heat production (HP), net energy for production (NEp), production of $CO_2$, and consumption of $O_2$. The respiratory quotient (RQ) was also increased (p<0.01) by the addition of xylanase. NEp value was increased by 26.1% while daily heat production per kg metabolizable body weight was decreased by 26.2% when the xylanase was added. Xylanase supplementation numerically increased the ileal digestibility of protein and energy by 3 and 6 percentage units respectively (p>0.05). The ileal digestibility of hemicellulose was significantly improved by xylanase addition (p<0.05).

Isopsoralen Induces Differentiation of Prechondrogenic ATDC5 Cells via Activation of MAP Kinases and BMP-2 Signaling Pathways

  • Li, Liang;Eun, Jae-Soon;Nepal, Manoj;Ryu, Jae-Ha;Cho, Hyoung-Kwon;Choi, Bo-Yun;Soh, Yun-Jo
    • Biomolecules & Therapeutics
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    • v.20 no.3
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    • pp.299-305
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    • 2012
  • Endochondral bone formation is the process by which mesenchymal cells condense to become chondrocytes, which ultimately form new bone. The process of chondrogenic differentiation and hypertrophy is critical for bone formation and as such is regulated by many factors. In this study, we aimed to indentify novel factors that regulate chondrogenesis. We investigated the possible role of isopsoralen in induction of chondrogenic differentiation in clonal mouse chondrogenic ATDC5 cells. Isopsoralen treatment stimulated the accumulation of cartilage nodules in a dose-dependent manner. Further, ATDC5 cells treated with isopsoralen were stained more intensely with Alcian blue than control cells, suggesting that isopsoralen increases the synthesis of matrix proteoglycans. Similarly, isopsoralen markedly induced the activation of alkaline phosphatase activity compared with control cells. Isopsoralen enhanced the expressions of chondrogenic marker genes such as collagen II, collagen X, OCN, Smad4 and Sox9 in a time-dependent manner. Furthermore, isopsoralen induced the activation of extracellular signal-regulated kinase (ERK) and p38 MAP kinase, but not that of c-jun N-terminal kinase (JNK). Isopsoralen significantly enhanced the protein expression of BMP-2 in a time-dependent manner. PD98059 and SB 203580, inhibitors of ERK and p38 MAPK, respectively, decreased the number of stained cells treated with isopsoralen. Taken together, these results suggest that isopsoralen mediates a chondromodulating effect by BMP-2 or MAPK signaling pathways, and is therefore a possible therapeutic agent for bone growth disorders.

Computerized bone age estimation system based on China-05 standard

  • Yin, Chuangao;Zhang, Miao;Wang, Chang;Lin, Huihui;Li, Gengwu;Zhu, Lichun;Fei, Weimin;Wang, Xiaoyu
    • Advances in nano research
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    • v.12 no.2
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    • pp.197-212
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    • 2022
  • The purpose of this study is to develop an automatic software system for bone age evaluation and to evaluate its accuracy in testing and feasibility in clinical practice. 20394 left-hand radiographs of healthy children (2-18 years old) were collected from China Skeletal Development Survey data of 1998 and China Skeletal Development Survey data of 2005. Three experienced radiologists and China-05 standard maker jointly evaluate the stages of bone development and the reference bone age was determined by consensus. 1020 from 20394 radiographs were picked randomly as test set and the remaining 19374 radiographs as training set and validation set. Accuracy of the automatic software system for bone age assessment is evaluated in test set and two clinical test sets. Compared with the reference standard, the automatic software system based on RUS-CHN for bone age assessment has a 0.04 years old mean difference, ±0.40 years old in 95% confidence interval by single reading, a 85.6% percentage agreement of ratings, a 93.7% bone age accuracy rate, 0.17 years old of MAD, 0.29 years old of RMS; Compared with the reference standard, the automatic software system based on TW3-C RUS has a 0.04 years old mean difference, a ±0.38 years old in 95% confidence interval by single reading, a 90.9% percentage agreement of ratings, a 93.2% bone age accuracy rate, a 0.16 years of MAD, and a 0.28 years of RMS. Automatic software system, AI-China-05 showed reliably accuracy in bone age estimation and steady determination in different clinical test sets.

A 6-Week Oral Toxicity Study of Oral Cholera Vaccine in Sprague-Dawley Rats

  • Baek, Yeong-Ok;Choi, Seuk-Keun;Shin, Seo-Ho;Koo, Kyo-Hwan;Choi, Ho-Young;Cha, Seung-Bum;Li, Yong-Chun;Yoo, Hyeon-Jeong;Lee, Joo-Young;Kil, Ki-Hyun;Kim, Hak-Soo;Kang, Min-Soo;Kang, Boo-Hyun;Kim, Kap-Ho;Bae, Jin-Sook
    • Toxicological Research
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    • v.28 no.4
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    • pp.225-233
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    • 2012
  • The present study was carried out to examine the toxicity and target organs of oral cholera vaccine (OCV) after repeated oral administration in Sprague-Dawley rats for 6 weeks (3 administrations, once every 2 weeks). OCV is an inactivated oral cholera vaccine that contains Vibrio cholerae and confers protection against cholera caused by V. cholera serogroups O1 (Inaba and Ogawa serotypes) and O139 (strain 4260B). The animals were orally administered either OCV placebo (negative control) or OCV at a dose equivalent to 240 times the anticipated human dose. Throughout the administration period, no significant change was detected in clinical signs, body weight, food or water consumption, urinalysis results, hematological and clinical biochemistry test results, organ weights, necropsy, or histopathological examination results. Minor changes were found in hematological and clinical biochemistry tests; however, these changes were within normal ranges. The above results suggest that oral administration of OCV in rats did not induce any toxicologically meaningful changes, and the target organs could not be determined. This study was conducted in accordance with the guidelines established by Good Laboratory Practice (2009-183, KFDA, December 22, 2009) and the OECD Principles of Good Laboratory Practice (1997).

Effect of Submergence and Air Exposure of the Shoot on Growth, Nutrient Uptake and Photosynthesis in Monochoria vaginalis Presl. (물달개비 경엽(莖葉)의 침수여부(沈水與否)에 따른 생장(生長), 양분흡수(養分吸收) 및 광합성(光合成) 비교(比較))

  • Soh, C.H.;Yang, K.S.;Kwon, Y.W.
    • Korean Journal of Weed Science
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    • v.16 no.1
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    • pp.14-20
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    • 1996
  • Growth, nutrient uptake and photosynthesis as affected by submersion of shoot in pickerel weed (Monochoria vaginalis Presl.) were determined. The shoots of pickerel weeds in hydroponic culture were subjected to the submerged or emerged condition at 3- or 5-leaf stage for 8 or 10 days. Under submerged condition, growth in plant height was enhanced, but leaf number, leaf area, fresh and dry weight were reduced compared to those under the emerged condition. Similar responses in growth to submergence were obtained with the pickerel weeds rooted in the soil. Under submergence, chlorophyll content increased during the first 2 days, but thereafter remarkably decreased at 3-leaf stage and after the first 4 days at 5-leaf stage. Compared to the emerged condition, uptakes of $NH_4\;^+$-N, $NO_3\;^-$-N, $P_2O_5$ and $K^+$ were reduced, but uptakes of $Ca^{++}$ and $Mg^{++}$ increased under the submerged condition. Photosynthetic rate of shoot under water, measured by $CO_2$electrode, showed the maximum by 210 ${\mu}$moles $HCO_3\;^-$/g F.W. at the 8th day after submergence(DAS) at 3-leaf stage and 320 ${\mu}$moles $HCO_3\;^-$/g F. W. at 6 DAS at 5-leaf stage. These results indicate that pickerel weeds grow much better when the shoot is air-exposed and are less tolerable to submergence at 3 leaf-stage than at 5-leaf stage.

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The MDM2 SNP309T>G Polymorphism Increases Bladder Cancer Risk among Caucasians: a Meta-analysis

  • Wang, Huai-Gao;Wu, Qing-Yun;Zhou, Hui;Peng, Xin-Sheng;Shi, Meng-Jie;Li, Jie-Mei;Zhou, Yan-Fang
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.13
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    • pp.5277-5281
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    • 2014
  • Published studies have evaluated associations between the MDM2 SNP309T>G polymorphism and bladder cancer susceptibility. However, these generated inconsistent results. The aim of the present investigation was to quantify the strength of association between MDM2 SNP309T>G polymorphism and bladder cancer risk by conducting a meta-analysis. We searched PubMed and Embase for related studies that had been published in English before April 1, 2014 and associations were assessed by summarizing the odds ratios (ORs) with the corresponding 95% confidence intervals (CIs). Five case-control studies with a total of 972 cases and 1,012 controls were finally identified to be eligible for the meta-analysis. Overall, the results indicated that there was no significant association between the MDM2 SNP309T>G polymorphism and bladder cancer risk (for the allele model G vs. T: OR=1.08, 95% CI 0.85-1.36, p=0.54; for the co-dominant model GG vs. TT: OR=1.20, 95% CI 0.74-1.93, p=0.46; for the dominant model GG+GT vs. TT: OR=0.98, 95% CI 0.80-1.20, p=0.83; for the recessive model GG vs. GT+TT: OR=1.20, 95% CI 0.83-1.74, p=0.33). However, on subgroup analysis by ethnicity, significant associations were found in Caucasians in three models (for the allele model G vs. T: OR=1.41, 95% CI 1.10-1.81, p=0.006; for the co-dominant model GG vs. TT: OR=2.16, 95% CI 1.28-3.63, p=0.004; for the recessive model GG vs. GT+TT: OR=2.06, 95% CI 1.31-3.22, p=0.002). In summary, the present meta-analysis provides evidence that the genotype for the MDM2 SNP309T>G polymorphism may be associated with genetic susceptibility to bladder cancer among Caucasians.

Magnetic Tunnel Junctions with AlN and AlO Barriers

  • Yoon, Tae-Sick;Yoshimura, Satoru;Tsunoda, Masakiyo;Takahashi, Migaku;Park, Bum-Chan;Lee, Young-Woo;Li, Ying;Kim, Chong-Oh
    • Journal of Magnetics
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    • v.9 no.1
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    • pp.17-22
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    • 2004
  • We studied the magnetotransport properties of tunnel junctions with AlO and AlN barriers fabricated using microwave-excited plasma. The plasma nitridation process provided wider controllability than the plasma oxidization for the formation of MTJs with ultra-thin insulating layer, because of the slow nitriding rate of metal Al layers, comparing with the oxidizing rate of them. High tunnel magnetoresistance (TMR) ratios of 49 and 44% with respective resistance-area product $(R{\times}A) of 3 {\times} 10^4 and 6 {\times} 10^3 {\Omega}{\mu}m^2$ were obtained in the Co-Fe/Al-N/Co-Fe MTJs. We conclude that AlN is a hopeful barrier material to realize MTJs with high TMR ratio and low $R{\times}A$ for high performance MRAM cells. In addition, in order to clarify the annealing temperature dependence of TMR, the local transport properties were measured for Ta $50{\AA} /Cu 200 {\AA}/Ta 50 {\AA}/Ni_{76}Fe_{24} 20 {\AA}/Cu 50 {\AA}/Mn_{75}Ir_{25} 100 {\AA}/Co_{71}Fe_{29} 40 {\AA}/Al-O$ junction with $d_{Al}= 8 {\AA} and P_{O2}{\times}t_{0X}/ = 8.4 {\times} 10^4$ at various temperatures. The current histogram statistically calculated from the electrical current image was well in accord with the fitting result considering the Gaussian distribution and Fowler-Nordheim equation. After annealing at $340^{\circ}C$, where the TMR ratio of the corresponding MTJ had the maximum value of 44%, the average barrier height increased to 1.12 eV and its standard deviation decreased to 0.1 eV. The increase of TMR ratio after annealing could be well explained by the enhancement of the average barrier height and the reduction of its fluctuation.