• Title/Summary/Keyword: high-level expression

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Characteristics of Oncolytic Adenovirus Replication and Gene Expression in Hypoxic Condition

  • Kim, Hong-Sung
    • Biomedical Science Letters
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    • v.17 no.3
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    • pp.185-190
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    • 2011
  • Adenovirus type 5 (Ad5) vectors have been used for gene transfer to a wide variety of cell types in vivo and in vitro. The advantages of adenovirus vectors include the high titer of virus readily obtained in large scale preparations, their ability to transduce dividing and non dividing cells, and the high level of transgene expression. Since adenovirus vectors do not integrate in host cell DNA, there is a lack of insertional mutagenesis. However, many human tumor cells lack expression of the adenovirus 5 receptors and contain areas of hypoxia. In order to identify the pattern of replication and gene expression of oncolytic adenovirus in hypoxic condition, multiple different fiber modified Ads (Ad5F/S11, Ad5F/S35, Ad5F/K7, Ad5F/K21, and Ad5F/RGD) was compared. The replication of all fiber modified adenovirus was inhibited in hypoxic condition in HEK 293 cells, but gene expression has variety on different tumor cell lines and the level of coxackievirus and adenovirus receptor (CAR) expression. These data suggest that CAR expression pattern and hypoxic condition of tumor are considered for optimal oncolytic adenovirus application.

Effects of High Fat Diet on Serum Leptin and Insulin Level and Brown Adipose Tissue UCP 1 Expression in Rats (흰쥐에서 고지방식이가 혈중 렙틴 및 인슐린과 갈색지방조직의 UCP 1 발현에 미치는 영향)

  • 홍경희;강순아;김소혜;조여원
    • Journal of Nutrition and Health
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    • v.34 no.8
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    • pp.865-871
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    • 2001
  • The adipose tissue hormone leptin has been proposed to be involved in the regulation of flood intake and energy expenditure via thermogenesis by uncoupling protein(UCP) in brown adipose tissue(BAT). The objective of the study was to examine the effects of high fat diet on the serum leptin levels, BAT UCPl expression and the body fat mass in rats after weaning. During experimental period of 12 weeks, 4 male Sprague-Dawley rats were killed for the baseline experiment at 4 weeks of age while the remaining rats were fed the two different diets: the control diet AIN-76A(n = 20), high fat(beef tallow) diet(n = 20) ad libitum, which provided 11.7% or 40% of calories as fat, respectively. At 16 weeks of age, the increase in the food efficiency ratio(FER) was related to fat mass in rats on high fat diet. Serum leptin level was increased by age and dietary high fat. There was no difference in serum insulin level between groups until 10 weeks of age, but rats fed high fat diet for 12 weeks showed hyperinsulinemia. The amount of body fat pads was increased significantly in high fat group compared to normal diet group. Visceral fat mass affected acutely by high fat diet, as a result, it was higher in rats fed high fat diet for 2 weeks than normal diet. At 16 weeks of age, BAT and visceral fat mass were significantly high in high fat group. Also, the serum leptin levels reflected the amount of body fat mass. BAT UCPI mRNA expression increased with age and dietary high fat. This study demonstrates that dietary high fat increased serum leptin levels, BAT UCPI expression and body fat mass. Futhermore, in rats fed high fat diets, the increases in leptin and UCPI expression counteracts only in part the excess adiposity and obesity.

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Effects of Dietary Levan on Adiposity, Serum Leptin and UCP Expression in Obese Rats Fed High Fat Diet (고지방 식이로 유도된 비만쥐에서 식이 레반이 체지방 형성 및 혈청 렙틴과 UCP 발현에 미치는 영향)

  • 강순아;홍경희;김소혜;장기효;김철호;조여원
    • Journal of Nutrition and Health
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    • v.35 no.9
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    • pp.903-911
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    • 2002
  • The effects of dietary levan, high-molecular-weight $\beta$-2,6-linked fructose polymer, on adiposity, serum leptin and UCP expression in rats fed high fat diet were studied. The adipose tissue hormone, leptin has been proposed to be involved in the regulation of food intake and energy expenditure. Uncoupling protein (UCP), a mitochondrial protein that uncouples the respiratory chain from oxidative phosphorylation, generates heat instead of ATP, thereby increase energy expenditure. To determine whether the dietary levan may have the anti-obesity effect, 4 wk old Sprague Dawley male rats fed high fat diet for 6 wks to induce obesity, and subsequently fed one of three diets for further 6 wks: 1) high fat (40% of calories) diet without levan 2) with 3% (w/w) levan 3) with 5% levan. For the comparison, control group fed AIN-76A diet. Visceral and peritoneal fat masses were lower in high fat diet with levan groups compared to high fat diet group. The effect of levan was dose-dependent. Adipocyte size was significantly reduced in the levan diet groups compared to the no levan diet group. Serum cholesterol level was not affected by levan containing diet, while the serum HDL cholesterol level was higher in leven diet groups. In addition, serum triglyceride level was markedly reduced by levan containing diet, thus lower than that of control group. Serum leptin was reduced by levan containing diet and lower in 5% levan group compared to 3% levan group (p < 0.001), as a result, serum leptin and insulin levels of 5% levan group were reduced to level of control group. Futhermore, the serum leptin level reflected the adiposity. The expression of UCP 1, and UCP 2 in brown adipose tissue was up-regulated by levan containing diet. In conclusions, levan containing diet reduced adiposity and serum triglyceride but increased UCP expression in the obese rats fed high fat diet. (Korean J Nutrition 35(9) : 903~911, 2002)

Expressional Modulation of Aquaporin 1 and 9 in the Rat Epididymis by an Anabolic-Androgenic Steroid, Nandrolone Decanoate

  • Lee, Ki-Ho
    • Development and Reproduction
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    • v.25 no.4
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    • pp.245-255
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    • 2021
  • The spermatozoa become mature in the epididymis which is divided into initial segment and caput, corpus, and cauda epididymis. The water movement across the epididymal epithelium is important for creating luminal microenvironment for sperm maturation. Aquaporins (Aqps) are water channel proteins, and expression of Aqps is regulated by androgens. The current research was focused to examine expressional regulation of Aqp1 and Aqp9 by an androgenic-anabolic steroid, nandrolone decanoate (ND). The ND at the low dose (2 mg/kg body weight/week) or high dose (10 mg) was subcutaneously administrated into male rats for 2 or 12 weeks. Transcript levels of Aqp1 and Aqp9 were determined by quantitative real-time polymerase chain reaction (PCR) analyses. In the initial segment, level of Aqp1 was decreased with 12 week-treatment, while Aqp9 level was decreased by the high dose treatment for 12 weeks. In the caput epididymis, Aqp9 expression was decreased by the low dose treatment. The 2 week-treatment resulted in an increase of Aqp1 level but a decrease of Aqp9 expression in the corpus epididymis. In the corpus epididymis, the 12 week-treatment at the low dose caused the reduction of Aqp1 and Aqp9 levels, but the high dose treatment resulted in an increase of Aqp1 expression and a decrease of Aqp9 level. In the cauda epididymis, Aqp1 expression was decreased by 2 and 12 week-treatments, while increases of Aqp9 levels was detected with the high dose treatment for 2 weeks and with 12 week-treatment. These findings indicate differential regulation of Aqp1 and Aqp9 expression among epididymal segments by ND.

Survey of the heterogeneous gene expression in olive flounder muscle using the luciferase reporter gene system

  • Hong, Suhee;Lee, Sang-Jun
    • Journal of fish pathology
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    • v.16 no.3
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    • pp.147-152
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    • 2003
  • The CMV promoter driven luciferase reporter gene coding plasmid (pcDNA-luc) was constructed and used as a model for DNA immunization study. Expression of the recombinant luciferase protein was confirmed in vitro in RTG-2 cell line before using in vivo study in olive flounder. In dose response study, the maximum expression of the luciferase gene was found in the group injected with 10-15μg of plasmid DNA. The kinetic study showed that the luciferase gene expression was reached at the maximum level at one day after injection and slightly decreased after then but significantly high level of expression was sustained until the conducted experiment of 7 days. In the study of tissue distribution of gene expression, it was found that luciferase gene was expressed at the significant level in immune organs such as gill and spleen, located far from the injected site, suggesting the systemic distribution of the intramuscularly injected DNA in olive flounder.

High-level Production of Recombinant Human IFN-$\alpha2a$ with Co-expression of $tRNA^{Arg(AFF/AGA)}$ in High-cell-density Cultures of Escherichia coli

  • Shin, Chul-Soo;Hong, Min-Seon;Shin, Hang-Chel;Lee, Jeewon
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.6 no.4
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    • pp.301-305
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    • 2001
  • The co-expression of the arg U gene in a double-vector expression system of recombi-nant Escherichia coli BL22(DE3)[pET-IEN2a+pAC-argU] significantly enhanced the production level of reconminant human interferon -$\alpha$2a(rhIFN-$\alpha$2a) in high cell density cultures, compared to a recombinant E. coli culture containing only the single expression vector, pET-IEN2a. The dry cell mass concentration increased to almost 100 g/L, and more than 4 g/L of rhIFN-$\alpha$2a was accumu-lated in the culture broth. Evidently, the synthesis of rhIFN-$\alpha$2a was strongly dependent on the pre-induction growtih rate and more efficient at a higher specific growth rate. The additional sup-ply of tRN $A^{Arg(AGG/AGA)}$ enhanced the expression level of the rhIFN-$\alpha$2a gene in the early stage of the post-induction phase, yet thereafter the specific production rate of rhIFN-$\alpha$2a rapidly de-creased due to severe segregational instability of plasmid vector pET-IEN2a. It would appear that the plasmid instability with only occurred to pET-IEN2a in the double vector system, was re-lated to the effect of translational stress due to the over expression of rhIFN-$\alpha$2a.

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Prognostic Value of Phosphorylated mTOR/RPS6KB1 in Non-small Cell Lung Cancer

  • Zhang, Yong;Ni, Huan-Juan;Cheng, De-Yun
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.6
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    • pp.3725-3728
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    • 2013
  • Background: The mammalian target of rapamycin (mTOR) /RPS6KB1 activation has recently been implicated in tumour development, but its role in lung cancer remains unclear. The aim of this study was to explore the role of mTOR/RPS6KB1 signaling pathway in non-small-cell lung cancer (NSCLC). Methods: Immunohistochemistry was performed to assess the expression of phosphorylated mammalian target of rapamycin (p-mTOR) and its downstream ribosomal phosphorylated RPS6KB1 (p-RPS6KB1) in NSCLC patients. We also analyzed p-mTOR/p-RPS6KB1 protein expression in 45 fresh NSCLC tissues using Western blotting. Results: The expression level of p-mTOR and p-RPS6KB1 was significantly higher in NSCLC tumor specimens than that in adjacent noncancerous normal lung tissues (P<0.01). p-mTOR expression correlated with p-RPS6KB1. Furthermore, high expression level of p-mTOR or p-RPS6KB1 in NSCLC was associated with a shorter overall survival (both P<0.01). Multivariate analysis indicated high level of p-mTOR expression was an independent prognostic factor (HR=2.642, 95%CI 1.157-4.904, p=0.002). Conclusions: p-mTOR and p-RPS6KB1 could be useful prognostic markers for NSCLC.

Association between expression levels and growth trait-related SNPs located in promoters of the MC4R and MSTN genes in Spinibarbus hollandi

  • Yang, Yang;Lan, Zhaojun;Shu, Hu;Zhou, Huiqiang;Jiang, Xiaolu;Hou, Liping;Gu, Pinghua
    • Genes and Genomics
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    • v.40 no.11
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    • pp.1119-1125
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    • 2018
  • Melanocortin 4 receptor: (MC4R) and Myostatin (MSTN) are two important growth trait-related genes in animals. In this study, we showed that two SNPs, MC4R-719A>G and MSTN-519C>T, found in the promoters of the MC4R and MSTN genes, respectively, are both associated with growth traits in Spinibarbus hollandi. Furthermore, we observed that there were significant associations between the expression levels of the MC4R and MSTN genes and these two growth trait-related SNPs. The expression level of MC4R gene in brain was lower in GG genotype fish with extremely high growth performance than that in AA genotype fish with extremely low growth performance. Expression level of the MSTN gene in muscle was lower in TT genotype fish with extremely high growth performance than that in CC and CT genotype fish with lower growth performance. The results indicated that these SNPs located in the promoters of MC4R and MSTN are associated with growth-related traits through modification of gene expression levels. The MSTN and MC4R SNPs may have useful application in effective marker-assisted selection aimed to increase output in S. hollandi.

Expression of the Pokemon Proto-oncogene in Nasopharyngeal Carcinoma Cell Lines and Tissues

  • Jiao, Wei;Liu, Fei;Tang, Feng-Zhu;Lan, Jiao;Xiao, Rui-Ping;Chen, Xing-Zhou;Ye, Hui-Lan;Cai, Yong-Lin
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.11
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    • pp.6315-6319
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    • 2013
  • To study the differentiated expression of the proto-oncogene Pokemon in nasopharyngeal carcinoma (NPC) cell lines and tissues, mRNA and protein expression levels of CNE1, CNE2, CNE3 and C666-1 were detected separately by reverse transcription polymerase chain reaction (RT-PCR), real-time PCR and Western-blotting. The immortalized nasopharyngeal epithelial cell line NP69 was used as a control. The Pokemon protein expression level in biopsy specimens from chronic rhinitis patients and undifferentiated non keratinizing NPC patients was determined by Western-blotting and arranged from high to low: C666-1>CNE1>CNE2> CNE3>NP69. The Pokemon mRNA expression level was also arranged from high to low: CNE1>CNE2>NP69>C666-1>CNE3. Pokemon expression of NP69 and C666-1 obviously varied from mRNA to protein. The Pokemon protein level of NPC biopsy specimens was obviously higher than in chronic rhinitis. The data suggest that high Pokemon protein expression is closely associated with undifferentiated non-keratinizing NPC and may provide useful information for NPC molecular target therapy.

A Study for the Optimal Expression Level Definition of BIM Model in Construction Phase (시공단계 BIM모델의 적정표현수준 정의를 위한 기초 연구)

  • Noh, Hae-ra;Kim, Ji-yun;Lee, Da-un;Yun, Seok-heon
    • Korean Journal of Computational Design and Engineering
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    • v.21 no.4
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    • pp.378-388
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    • 2016
  • As the needs for the mega-sized, high-rise and irregular shape of building increase, domestic application of BIM in construction industry is increasing in recent years. In this circumstance, interests in BIM modeling and modeling detail level have been also increased. BIM that is based on a three-dimensions, requires a specific and precise building expressions. And model expression level required for each phase such as design, construction, and maintenance of the construction project could be different as its needs. Because that it is not clear to define the criteria for the expression level of construction BIM model, It is difficult to decide how detail and what to model is appropriate in the practical application of BIM model. It could be the reason why the productivity in construction project became lower and lower. In this study we consider the way how to improve the productivity of BIM and analyze the appropriate representation level of construction BIM model through the experts' survey. The results of study could suggest the basis for an appropriate expression level of construction BIM model.