• Title/Summary/Keyword: Protein Deposition

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Effects of MeOH Extract from Stem Bark of Plantocracy strobilacea on the Metabolism of Amyloid Precursor Protein in Neuroblastoma Cells (화향수(化香樹) 수피(樹皮)의 메탄올 추출물이 신경세포에서 아밀로이드 전구단백질의 대사에 미치는 영향)

  • Jiang, Gui Bao;Leem, Jae Yoon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.27 no.1
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    • pp.57-62
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    • 2013
  • Alzheimer's disease (AD), one of the most common forms of dementia, is characterized pathologically by the presence of intracellular neurofibrillary tangles and deposition of ${\beta}$-amyloid ($A{\beta}$) peptides of 40-42 residues, which are generated by processing of amyloid precursor protein (APP). $A{\beta}$ has been believed to be neurotoxic and now is also considered to have a role on the mechanism of memory dysfunction. Here, we show that MeOH extract from stem bark of Platycarya strobilacea Sieb. et. Zucc. (PSM) affects on the processing of APP from the APPswe over-expressing Neuro2a cell line. We found that PSM may regulate the processing of APP and increase the sAPP${\alpha}$. PSM does not change the protein level of presenilin and nicastrin, however, it reduces the protein expression level of BACE1. In addition, PSM reduces the secretion level of $A{\beta}42$ and $A{\beta}40$ from the cell line without toxicity. We suggest that Platycarya strobilacea may be useful as a herbal medicine to treat Alzheimer's disease.

β-Secretase (BACE1) Purification by Refolding Method and Complex with Hispidin

  • Lim, Ji-Hong;Lee, Bo Ram;Park, Hee Won;Hong, Bum Soo;Lim, Beong Ou;Kim, Young Jun
    • Journal of the Korean Chemical Society
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    • v.58 no.6
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    • pp.553-559
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    • 2014
  • Alzheimer's disease (AD) is a devastating neurodegenerative disease that represents the most common form of dementia among the elderly population. The deposition of aggregated ${\beta}$-amyloid ($A{\beta}$) senile plaques in the human brain is a classic observation in the neuropathology of AD, yet an understanding of the mechanism of their formation remains elusive. $A{\beta}$ is formed through endoproteolysis of the amyloid precursor protein (APP) by ${\beta}$-secretase (BACE1, ${\beta}$-site APP-cleaving enzyme) and ${\gamma}$-secretase. In this study, BACE1 protein was successfully over-expressed, purified, and refolded and utilized in a binding study with hispidin. We developed a simpler refolding method using a urea gradient and size-exclusion gel filtration to purify an active BACE1 protein variant, in larger quantities than that reported previously, and measured the binding affinity of hispidin to the BACE1 protein variant through isothermal titration calorimetry.

Effect of Choice Feeding on Performance, Gastrointestinal Development and Feed Utilization of Broilers

  • Yao, Junhu;Tian, Xiaoyan;Xi, Haibo;Han, Jincheng;Xu, Ming;Wu, Xiaobing
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.1
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    • pp.91-96
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    • 2006
  • The experiment was conducted to study the effect of choice feeding on growth performance, carcass quality, gastrointestinal development and feed utilization of 22-49 days old broilers. One hundred and forty four 22-day-old broilers were randomly allocated to 3 treatments with 4 replicates per treatment and 12 birds per replicate. Three feeding regimes are complete diet (control), ground corn and protein concentrate (treatment I), and soybean meal and balancer (treatment II). Protein concentrate is the residue part of complete diet without corn, and balancer is the residue part of complete diet without soybean meal. Treatment I and II are designed for the broilers to freely choose the two parts of diet. The results showed that: (1) broilers under choice feeding (treatment I and II) had lower performances compared with the control; (2) gastrointestinal development and the efficiency ratios that broilers converted dietary crude protein and lysine to body weight gain were improved in treatment I (p<0.05); (3) there were no significant differences in the apparent metabolizabilities of dietary dry matter, crude protein and gross energy, and deposition ratios of dietary nitrogen and energy, and carcass quality among three feeding regimes (p>0.05).

High glucose induces differentiation and adipogenesis in porcine muscle satellite cells via mTOR

  • Yue, Tao;Yin, Jingdong;Li, Fengna;Li, Defa;Du, Min
    • BMB Reports
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    • v.43 no.2
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    • pp.140-145
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    • 2010
  • The present study investigated whether the mammalian target of rapamycin (mTOR) signal pathway is involved in the regulation of high glucose-induced intramuscular adipogenesis in porcine muscle satellite cells. High glucose (25 mM) dramatically increased intracellular lipid accumulation in cells during the 10-day adipogenic differentiation period. The expressions of CCAAT/enhancer binding protein-$\alpha$ (C/EBP-$\alpha$) and fatty acid synthase (FAS) protein were gradually enhanced during the 10-day duration while mTOR phosphorylation and sterol-regulatory- element-binding protein (SREBP)-1c protein were induced on day 4. Moreover, inhibition of mTOR activity by rapamycin resulted in a reduction of SREBP-1c protein expression and adipogenesis in cells. Collectively, our findings suggest that the adipogenic differentiation of porcine muscle satellite cells and a succeeding extensive adipogenesis, which is triggered by high glucose, is initiated by the mTOR signal pathway through the activation of SREBP-1c protein. This process is previously uncharacterized and suggests a cellular mechanism may be involved in ectopic lipid deposition in skeletal muscle during type 2 diabetes.

Comparative Study on the Structural and Thermodynamic Features of Amyloid-Beta Protein 40 and 42

  • Lim, Sulgi;Ham, Sihyun
    • Proceeding of EDISON Challenge
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    • 2014.03a
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    • pp.237-249
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    • 2014
  • Deposition of amyloid-${\beta}$ ($A{\beta}$) proteins is the conventional pathological hallmark of Alzheimer's disease (AD). The $A{\beta}$ protein formed from the amyloid precursor protein is predominated by the 40 residue protein ($A{\beta}40$) and by the 42 residue protein ($A{\beta}42$). While $A{\beta}40$ and $A{\beta}42$ differ in only two amino acid residues at the C-terminal end, $A{\beta}42$ is much more prone to aggregate and exhibits more neurotoxicity than $A{\beta}40$. Here, we investigate the molecular origin of the difference in the aggregation propensity of these two proteins by performing fully atomistic, explicit-water molecular dynamics simulations. Then, it is followed by the solvation thermodynamic analysis based on the integral-equation theory of liquids. We find that $A{\beta}42$ displays higher tendency to adopt ${\beta}$-sheet conformations than $A{\beta}40$, which would consequently facilitate the conversion to the ${\beta}$-sheet rich fibril structure. Furthermore, the solvation thermodynamic analysis on the simulated protein conformations indicates that $A{\beta}42$ is more hydrophobic than $A{\beta}40$, implying that the surrounding water imparts a larger thermodynamic driving force for the self-assembly of $A{\beta}42$. Taken together, our results provide structural and thermodynamic grounds on why $A{\beta}42$ is more aggregation-prone than $A{\beta}40$ in aqueous environments.

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Comparisons of Adherence Level of Micro-organisms According to Contact Lens Materials and Protein Deposition and Disinfection Efficacy of Multipurpose Solution (콘택트렌즈 재질 및 침착 단백질에 따른 균 흡착 정도와 다목적용액의 살균력 비교)

  • Sung, Hyung Kyung;Kim, So Ra;Park, Mijung
    • Journal of Korean Ophthalmic Optics Society
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    • v.20 no.1
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    • pp.35-42
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    • 2015
  • Purpose: The present study was aimed to compare the difference in adherence level of microorganisms according to contact lens materials and protein deposition and to evaluate disinfection efficacy of multipurpose solution. Methods: The evaluations of micro-organisms' adherence and disinfection efficacy of multi-purpose solution were conducted by employing the Part 2. Regimen Procedure for Disinfecting Regiments in the Disinfection Efficacy Testing under the "FDA Evaluation Criteria & Method". Results: Pseudomonas aeruginosa, Serratia marcescens, Candida albicans except Staphylococcus aureus adhered more on etafilcon A lens and disinfection efficacy of total 4 products investigated was almost perfect except Candida albicans. The 3 micro-organisms except Serratia marcescens adhered more to albumin-predeposited lens. Disinfection efficacy of multi-purpose solution was higher against the micro-organisms adhered to albumin-deposited lens than against the micro-organisms adhered to the lysozyme-deposited lens. Furthermore, disinfection efficacy of multi-purpose solution was different according to types of micro-organisms. Conclusions: It was revealed that the type of micro-organisms, the lens materials and type of absorbed tear protein affected the amount of adhered micro-organisms to contact lens and that adhesion of tear protein could induce the change of disinfection efficacy of multi-purpose solution. It suggest that the hygienic condition of contact lens can vary by these factors influencing on disinfection efficacy and the occurrence of adverse effect can be affected.

Cloning and Expression of the Duck Leptin Gene and the Effect of Leptin on Food Intake and Fatty Deposition in Mice

  • Dai, Han Chuan;Long, Liang Qi;Zhang, Xiao Wei;Zhang, Wei Min;Wu, Xiao Xiong
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.6
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    • pp.850-855
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    • 2007
  • Leptin is the adipocyte-specific product of the obese gene and plays a major role in food intake and energy metabolism. Leptin research was mainly focused on mammalian species, but understanding of leptin and its function in poultry is very poor. In this study, the duck leptin gene was amplified using the reverse transcription-polymerase chain reaction (RT-PCR) from duck liver RNA. The cDNA fragment was inserted into the pET-28a expression vector, and the resulting plasmid was expressed in Escherichia coli BL21 (DE3). Experimental mice were given an intraperitoneal injection of 10 mg/kg leptin dissolved in phosphate buffered saline (PBS), while the control mice were injected with PBS. The effect of leptin on food intake, body weight and fatty deposition in mice was detected. Sequence analysis revealed that duck leptin had a length of 438 nucleotides which encoded a peptide with 146 amino acid residues. The sequence shares highly homology to other animals. The coding sequence of duck leptin was 84 and 86% identical to human and pig leptin nucleotides sequence. Highest identity was with the rat coding sequence (95%). The identity of the amino acid sequence was 84, 82 and 96% respectively compared to that of the human, pig and rat. Results of SDS-PAGE analysis indicated that a fusion protein was specifically expressed in E. coli BL21 (DE3). The purified product was found to be biologically active during tests. Continuous administration of recombinant duck leptin inhibited food intake. Despite the decrease of food intake, leptin significantly induced body weight and fatty deposition. These changes were accompanied by a significant down-secretion of plasma glucose, cholesterol, triglyceride and insulin levels in mice. The observations provide evidence for an inhibitory effect of leptin in the regulation of food intake and for a potential role of duck leptin in the regulation of lipogenesis.

Role of TGF-β1/SMADs signalling pathway in resveratrol-induced reduction of extracellular matrix deposition by dexamethasone-treated human trabecular meshwork cells

  • Amy Suzana Abu Bakar;Norhafiza Razali;Renu Agarwal;Igor Iezhitsa;Maxim A. Perfilev;Pavel M. Vassiliev
    • The Korean Journal of Physiology and Pharmacology
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    • v.28 no.4
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    • pp.345-359
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    • 2024
  • Deposition of extracellular matrix (ECM) in the trabecular meshwork (TM) increases aqueous humour outflow resistance leading to elevation of intraocular pressure (IOP) in primary open-angle glaucoma, which remains the only modifiable risk factor. Resveratrol has been shown to counteract the steroid-induced increase in IOP and increase the TM expression of ECM proteolytic enzymes; however, its effects on the deposition of ECM components by TM and its associated pathways, such as TGF-β-SMAD signalling remain uncertain. This study, therefore, explored the effects of trans-resveratrol on the expression of ECM components, SMAD signalling molecules, plasminogen activator inhibitor-1 and tissue plasminogen activator in dexamethasone-treated human TM cells (HTMCs). We also studied the nature of molecular interaction of trans-resveratrol with SMAD4 domains using ensemble docking. Treatment of HTMCs with 12.5 µM trans-resveratrol downregulated the dexamethasone-induced increase in collagen, fibronectin and α-smooth muscle actin at gene and protein levels through downregulation of TGF-β1, SMAD4, and upregulation of SMAD7. Downregulation of TGF-β1 signalling by trans-resveratrol could be attributed to its effect on the transcriptional activity due to high affinity for the MH2 domain of SMAD4. These effects may contribute to resveratrol's IOP-lowering properties by reducing ECM deposition and enhancing aqueous humour outflow in the TM.

Gene-editing techniques and their applications in livestock and beyond

  • Tae Sub Park
    • Animal Bioscience
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    • v.36 no.2_spc
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    • pp.333-338
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    • 2023
  • Genetic modification enables modification of target genes or genome structure in livestock and experimental animals. These technologies have not only advanced bioscience but also improved agricultural productivity. To introduce a foreign transgene, the piggyBac transposon element/transposase system could be used for production of transgenic animals and specific target protein-expressing animal cells. In addition, the clustered regularly interspaced short palindromic repeat-CRISPR associated protein 9 (CRISPR-Cas9) system have been utilized to generate chickens with knockout of G0/G1 switch gene 2 (G0S2) and myostatin, which are related to lipid deposition and muscle growth, respectively. These experimental chickens could be the invaluable genetic resources to investigate the regulatory pathways and mechanisms of improvement of economic traits such as fat quantity and growth. The gene-edited animals could also be applicable to the livestock industry.

Fluorescence Immunoassy of HDL and LDL Using Protein A LB Film

  • Choi, Jeong-Woo;Park, Jun-Hyo;Lee, Woo-Chang;Oh, Byung-Keun;Min, Jun-Hong;Lee, Won-Hong
    • Journal of Microbiology and Biotechnology
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    • v.11 no.6
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    • pp.979-985
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    • 2001
  • A fluorometric detection technique for HDL (High Density Lipoprotein) and LDL (Low Density Lipoprotein) was developed for application in a fiber-optic immunosensor using a protein A Langmuir-Blodgget (LB) film. For the fluorescence immunoassay, antibodies specific to HDL or LDL were imobilied on the protein A LB film, and a fluorescence amplification method was developed to overcome their weak fluorescence. The deposition of protein A using the LB technique was monitored using a surface pressure-are $({\pi}-A)$ curve, and the antibody immobilization of the protein A LB film was experimentally verified. The immobilized antibody was used to separate only HDL and LDL from a sample, then the fluorescence of he separated HDL or LDL was amplified. The amount of LDL or HDL was measured using the developed fiber optic fluorescence detection system. The optical properties resulting from the reaction of HDL or LDL with o-phtaldialdehyde, detection range, response time, and stability of the immunoassay were all investigated. The respective detection ranges for HDL and LDL were sufficient to diagnose the risk of coronary heart disease. The amplification step increased the sensitivity, while selective separation using the immobilized antibody led to linearity in the sensor signal. The regeneration of the antibody-immobilized substrate could produce a stable and reproducible immunosensor.

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