• Title/Summary/Keyword: HE-4 protein

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Improving the Surface Functionality of Curdlan by Conjugation with Unfolding Protein through Naturally Occurring Maillard Reaction

  • Nakamura, Soichiro;Ogawa, Masahiro;Saeki, Hiroki;Saito, Masayoshi;Miyasaka, Satoko;Hata, Junya;Adachi, Naoko;Hwang, Jae-Kwan
    • Preventive Nutrition and Food Science
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    • v.5 no.4
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    • pp.200-204
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    • 2000
  • Protein conjugation of curdlan belonging to $\beta$-1, 3-glucan was carried out to improve it surface functionalities. The glucan was mixed with phosvitin, {TEX}$$\alpha$_{s}${/TEX}-casein, lysozyme or ovalbumin, respectively. The mixture was freeze-dried, and he resulting powder was incubated at 6$0^{\circ}C$ and 79% relative humidity for 12 days in order to generate a controlled Maillard reaction between curdlan and proteins. conjugation with unfolding proteins, i.e., phosvitin and {TEX}$$\alpha$_{s}${/TEX}-casein, drastically increased the solubility of the glucan, whereas lysozyme and ovalbmin did not. The solubility in water of curdlan was 3.44% for the phosvitin conjugate and 1.09% for the {TEX}$$\alpha$_{s}${/TEX}-casein conjugate. SDS-slab polyacrylamide gel electrophoresis showed that curdlan was solubilized due to covalent binding with phosvitin. Emulsifying properties of curdlan were substantially improved by the conjugation with phosvitin and {TEX}$$\alpha$_{s}${/TEX}-casein. Emulsion stability of the curdlan-phosvitin conjugate was about 2.9 times greater than that of the curdlan-phosvitin mixture.

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RNAi-based Knockdown of Multidrug Resistance-associated Protein 1 is Sufficient to Reverse Multidrug Resistance of Human Lung Cells

  • Shao, Shu-Li;Cui, Ting-Ting;Zhao, Wei;Zhang, Wei-Wei;Xie, Zhen-Li;Wang, Chang-He;Jia, Hong-Shuang;Liu, Qian
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.24
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    • pp.10597-10601
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    • 2015
  • Up-regulation of multidrug resistance-associated protein 1 (MRP1) is regarded as one of the main causes for multidrug resistance (MDR) of tumor cells, leading to failure of chemotherapy-based treatment for a multitude of cancers. However, whether silencing the overexpressed MRP1 is sufficient to reverse MDR has yet to be validated. This study demonstrated that RNAi-based knockdown of MRP1 reversed the increased efflux ability and MDR efficiently. Two different short haipin RNAs (shRNAs) targeting MRP1 were designed and inserted into pSilence-2.1-neo. The shRNA recombinant plasmids were transfected into cis-dichlorodiamineplatinum-resistant A549 lung (A549/DDP) cells, and then shRNA expressing cell clones were collected and maintained. Real time PCR and immunofluorescence staining for MRP1 revealed a high silent efficiency of these two shRNAs. Functionally, shRNA-expressing cells showed increased rhodamine 123 retention in A549/DDP cells, indicating reduced efflux ability of tumor cells in the absence of MRP1. Consistently, MRP1-silent cells exhibited decreased resistance to 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) and DDP, suggesting reversal of MDR in these tumor cells. Specifically, MRP1 knockdown increased the DDP-induced apoptosis of A549/DDP cells by increased trapping of their cell cycling in the G2 stage. Taken together, this study demonstrated that RNAi-based silencing of MRP1 is sufficient to reverse MDR in tumor cells, shedding light on possible novel clinical treatment of cancers.

Preliminary Proteomic Analysis of Thiobacillus ferrooxidans Growing on Elemental Sulphur and Fe2+ Separately

  • He, Zhi-guo;Hu, Yue-Hua;Zhong, Hui;Hu, Wei-Xin;Xu, Jin
    • BMB Reports
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    • v.38 no.3
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    • pp.307-313
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    • 2005
  • Thiobacillus ferrooxidans is one of the most important bacterium used in bioleaching, and can utilize $Fe^{2+}$ or sulphide as energy source. Growth curves for Thiobacillus ferrooxidans have been tested, which show lag, logarithmic, stationary and aging phases as seen in other bacteria. The logarithmic phases were from 10 to 32 hours for Thiobacillus ferrooxidans cultivated with $Fe^{2+}$ and from 4 to 12 days for Thiobacillus ferrooxidans cultivated with elemental sulphur. Differences of protein patterns of Thiobacillus ferrooxidans growing on elemental sulphur and $Fe^{2+}$ separately were investigated after cultivation at $30^{\circ}C$ by the analysis of two-dimensional gel electrophoresis (2-DE), matrix-assisted laser desorption/ ionization (MALDI)-Mass spectrometry and ESI-MS/MS. From the 7 identified protein spots, 11 spots were found more abundant when growing on elemental sulphur. By contrast 6 protein spots were found decreased at elemental cultivation condition. Among the proteins identified, cytochrome C have been previously identified as necessary elements of electron-transfering pathway for Thiobacillus ferrooxidans to oxidize $Fe^{2+}$; ATP synthase alpha chain and beta are expressed increased when Thiobacillus ferrooxidans cultivated with $Fe^{2+}$ as energy source. ATP synthase Beta chain is the catalytic subunit, and ATP synthase alpha chain is a regulatory subunit. The function of ATPase produces ATP from ADP in the presence of a proton gradient across the membrane.

Functional analysis of a homologue of the FLORICAULA/LEAFY gene in litchi (Litchi chinensis Sonn.) revealing its significance in early flowering process

  • Ding, Feng;Zhang, Shuwei;Chen, Houbin;Peng, Hongxiang;Lu, Jiang;He, Xinhua;Pan, Jiechun
    • Genes and Genomics
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    • v.40 no.12
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    • pp.1259-1267
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    • 2018
  • Litchi (Litchi chinensis Sonn.) is an important subtropical fruit crop with high commercial value due to its high nutritional values and favorable tastes. However, irregular bearing attributed to unstable flowering is a major ongoing problem for litchi producers. Previous studies indicate that low-temperature is a key factor in litchi floral induction. In order to reveal the genetic and molecular mechanisms underlying the reproductive process in litchi, we had analyzed the transcriptome of buds before and after low-temperature induction using RNA-seq technology. A key flower bud differentiation associated gene, a homologue of FLORICAULA/LEAFY, was identified and named LcLFY (GenBank Accession No. KF008435). The cDNA sequence of LcLFY encodes a putative protein of 388 amino acids. To gain insight into the role of LcLFY, the temporal expression level of this gene was measured by real-time RT-PCR. LcLFY was highly expressed in flower buds and its expression correlated with the floral developmental stage. Heterologous expression of LcLFY in transgenic tobacco plants induced precocious flowering. Meantime, we investigated the sub-cellular localization of LcLFY. The LcLFY-Green fluorescent protein (GFP) fusion protein was found in the nucleus. The results suggest that LcLFY plays a pivotal role as a transcription factor in controlling the transition to flowering and in the development of floral organs in litchi.

Discovery of Cyclin-dependent Kinase Inhibitor, CR229, Using Structure-based Drug Screening

  • Kim, Min-Kyoung;Min, Jae-Ki;Choi, Bu-Young;Lim, Hae-Young;Cho, Youl-Hee;Lee, Chul-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.17 no.10
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    • pp.1712-1716
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    • 2007
  • To generate new scaffold candidates as highly selective and potent cyelin-dependent kinase (CDK) inhibitors, structure-based drug screening was performed utilizing 3D pharmacophore conformations of known potent inhibitors. As a result, CR229 (6-bromo-2,3,4,9-tetrahydro-carbolin-1-one) was generated as the hit-compound. A computational docking study using the X-ray crystallographic structure of CDK2 in complex with CR229 was evaluated. This predicted binding mode study of CR229 with CDK2 demonstrated that CR229 interacted effectively with the Leu83 and Glu81 residues in the ATP-binding pocket of CDK2 for the possible hydrogen bond formation. Furthermore, biochemical studies on inhibitory effects of CR229 on various kinases in the human cervical cancer HeLa cells demonstrated that CR229 was a potent inhibitor of CDK2 ($IC_{50}:\;3\;{\mu}M$), CDKI ($IC_{50}:\;4.9\;{\mu}M$), and CDK4 ($IC_{50}:\;3\;{\mu}M$), yet had much less inhibitory effect ($IC_{50}:>20\;{\mu}M$) on other kinases, such as casein kinase 2-${\alpha}1$ (CK2-${\alpha}1$), protein kinase A (PKA), and protein kinase C (PKC). Accordingly, these data demonstrate that CR229 is a potent CDK inhibitor with anticancer efficacy.

Chronic cold stress-induced myocardial injury: effects on oxidative stress, inflammation and pyroptosis

  • Hongming Lv;Yvxi He;Jingjing Wu; Li Zhen ;Yvwei Zheng
    • Journal of Veterinary Science
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    • v.24 no.1
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    • pp.2.1-2.14
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    • 2023
  • Background: Hypothermia is a crucial environmental factor that elevates the risk of cardiovascular disease, but the underlying effect is unclear. Objectives: This study examined the role of cold stress (CS) in cardiac injury and its underlying mechanisms. Methods: In this study, a chronic CS-induced myocardial injury model was used; mice were subjected to chronic CS (4℃) for three hours per day for three weeks. Results: CS could result in myocardial injury by inducing the levels of heat shock proteins 70 (HSP70), enhancing the generation of creatine phosphokinase-isoenzyme (CKMB) and malondialdehyde (MDA), increasing the contents of tumor necrosis factor-α (TNF-α), high mobility group box 1 (HMGB1) interleukin1b (IL-1β), IL-18, IL-6, and triggering the depletion of superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH). Multiple signaling pathways were activated by cold exposure, including pyroptosis-associated NOD-like receptor 3 (NLRP3)-regulated caspase-1-dependent/Gasdermin D (GSDMD), inflammation-related toll-like receptor 4 (TLR4)/myeloid differentiation factor 88 (MyD88)-mediated nuclear factor kappa B (NF-κB), and mitogen-activated protein kinase (MAPK), as well as oxidative stressinvolved thioredoxin-1/thioredoxin-interacting protein (Txnip) signaling pathways, which play a pivotal role in myocardial injury resulting from hypothermia. Conclusions: These findings provide new insights into the increased risk of cardiovascular disease at extremely low temperatures.

The Second-order Scattering of the Interaction of Pd Nanoparticles with Protein and Its Analytical Application

  • Guo, Xiaoyan;He, Baolin;Sun, Chuntao;Zhao, Yanxi;Huang, Tao;Liew, Kongyong;Liu, Hanfan
    • Bulletin of the Korean Chemical Society
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    • v.28 no.10
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    • pp.1746-1750
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    • 2007
  • The second-order scattering (SOS) phenomenon of the interaction of Pd nanoparticles with protein was reported and a simple, sensitive, palladium nanoparticle-based assay for trace amount of protein with SOS technique was developed. The SOS intensities were significantly enhanced due to the interaction of Pd nanoparticles with bovine serum albumin (BSA) or human serum albumin (HSA) at pH 3.5 or 4.0, respectively. The maximum SOS peak appeared at 260/520 nm (λex/λem). The optimal experiment conditions, affecting factors and the influence of some coexisting substances were checked. The SOS intensity increased proportionally with the increase of Pd concentration below 3.0 × 10?5 mol·L?1, while declined gradually above 4.0 × 10?5 mol·L?1. BSA within the range of 0.01-2.6 μg·mL?1 and HSA of 0.01-1.7 μg·mL?1 can be detected with this method and the detection limits were 2.3 and 11.2 ng·mL?1, respectively. The method was successfully applied to the quantitative detection of total protein content in human serum samples with the maximum relative standard deviation (RSD) lower than 2.6% and the recoveries over the range of 99.5-100.5%.

Effects of Dietary Attitudes on the Nutritional Status of Nurses in Kyungnam Province

  • Bae, Ji-Hyun;Youn, Hyun Sook;Choi, Yun Young
    • Nutritional Sciences
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    • v.7 no.4
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    • pp.247-253
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    • 2004
  • This study has investigated the effect of dietary attitudes on nutritional status of nurses living in Kyungnam province. A total of 249 muses working at hospitals in the Kyungnam area participated in this study. The general characteristics and dietary attitudes of the subjects were surveyed using a self-administered questionnaire, and nutrient intakes were examined using one day 24-hour recall method from November 14 to December 20, in 2001. The data of nutrient intakes were analyzed by the Computer Aided Nutritional Analysis Program and then the diet quality was estimated using the Index of Nutritional Quality (INQ). The subjects were classified as the high score group (17.3 %), medium score group (54.2%), and low score group (28.5%) based on dietary attitude score. The results were as follows: Average age of subjects was 27.1 years old, average nursingexperience was 5.7 years, and 70.7% of the subjects graduated from a junior college. The rate of shift work (45.4%) and non-shift work (54.6%) were similar. The average score on dietary attitudes in thehigh score group was 78.8, and those in he medium score group and in the low score group were 58.6 and 40.8 out of 100 points respectively. The average intake of energy (p < 0.001), carbohydrate (p < 0.001), protein (p < 0.05), animal protein (p < 0.01), animal fat (p < 0.05), vitamin $B_1$ (p < 0.01), vitamin $B_2$ (p < 0.0l), niacin (p < 0.001), Ca (p < 0.001), Fe (p < 0.001), animal Fe (p < 0.01), P (p < 0.001), and crude fiber (p < 0.00l), were significantly higher in the high score group than those in the medium score group and low score group. In the high score group, the average intake of protein, vitamin $B_1$, niacin, vitamin C and P were above 125% of the Korean RDA, and in medium score group, those of vitamin C and P, and in low score group, that of vitamin C were above 125% of the Korean RDA. In the high score group, nutrient of intake below 75% of the Korean RDA was Ca, and those in medium score group were vitamin $B_2$, Ca, and Fe, and those in low score group were energy, vitamin $B_2$, Ca and Fe. The intake ratio of Ca and P showed an unbalanceof 1:2, and that of animal protein/plant protein, and that of animal fat/plant fat were high (1.45-1.64) in all group;. The dietary attitude had a positive correlation (p < 0.00l) with nutrition intake except vitamin A and vitamin C.

Effect of the Various Recovery Diets on the Body Composition of Protein-depleted Rat (각종 회복식이가 무단백식이를 섭취한 흰쥐의 체구성 성분에 미치는 영향)

  • Choi, Young-Ran;Suh, Dong-Hee;Kim, Mi-Kyung;Kim, Sook-He
    • Journal of Nutrition and Health
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    • v.10 no.1
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    • pp.1-9
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    • 1977
  • The changes of the total body composition, internal organs, skeletal muscles, and epididymal fat pad in rats fed protein depletion diet and 4 different recovery diets were examined. Seventy-eight male Sprague-Dawley rats weighing $212{\pm}2$ gr were used, and the results were as follows : A) After the 2 weeks of protein depletion, body weight decreased about 20% from the initial weight. It was mainey due to the total body lipid reduction. Among various organ weights, liver and spleen reduced $35{\sim}58%$, kidney and heart reduced $18{\sim}30%$, and muscles reduced $2{\sim}13%$, while brain, epididymal fat pad were not changed significantly. In regarding protein and lipid contents of these tissues, protein in liver, lipid in muscle, and both in spleen were markedly reduced. B) With the 2 weeks of feeding recovery diets, the increases of body weight were different among 4 Groups. High-fat group gained at the highest level (67%), and high-CHO group the lowest (30%). Total body composition (%) cf the standard and high-protein groups recovered to the level of 0 day protein depletion, while protein in the high-fat group and water in the high-CHO group decreased, and fat in these 2 groups increased. Weights of organs and muscles of the high-protein and high-fat groups were similar to the standard group and tllose of the high-CHO group were lower than the standard group. Composition of organs and muscle in the high-protein group was similar to the standard group, while the N contents of the high-fat and high-CHO groups were lower an the lipid content of the high-fat group was higher than the standard group. The weight and lipid content of epididymal fat pad were the highest in the high-fat group.

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Fine mapping of rice bacterial leaf blight resistance loci to major Korean races of Xoo (Xanthomonas oryzae)

  • Lee, Myung-Chul;Choi, Yu-Mi;Lee, Sukyeung;Yoon, Hyemyeong;Oh, Sejong
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.10a
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    • pp.73-73
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    • 2018
  • Bacterial leaf blight(BLB), caused by X. oryzae pv. oryzae(Xoo), is one of the most destructive diseases of rice due to its high epidemic potential. Understanding BLB resistance at a genetic level is important to further improve the rice breeding that provides one of the best approaches to control BLB disease. In the present investigation, a collection of 192 accessions was used in the genome-wide association study (GWAS) for BLB resistance loci against four Korean races of Xoo that were represented by the prevailing BLB isolates under Xoo differential system. A total of 192 accessions of rice germplasm were selected on the basis of the bioassay using four isolated races of Xoo such as K1, K2, K3 and K3a. The selected accessions was used to prepare 384-plex genotyping by sequencing (GBS) libraries and Illumina HiSeq 2000 paired- end read was used for GBS sequencing. GWAS was conducted using T ASSEL 5.0. The T ASSEL program uses a mixed linear model (MLM). T he results of the bioassay using a selected set of 192 accessions showed that a large number of accessions (93.75%) were resistant to K1 race, while the least number of accessions (34.37%) resisted K3a race. For races K2 and K3, the resistant germplasm proportion remained between 66.67 to 70.83%. T he genotypic data produced SNP matrix for a total of 293,379 SNPs. After imputation the missing data was removed, which exhibited 34,724 SNPs for association analysis. GWAS results showed strong signals of association at a threshold of [-log10(P-value)] more than5 (K1 and K2) and more than4 (K3 and K3a) for nine of the 39 SNPs, which are plausible candidate loci of resistance genes. T hese SNP loci were positioned on rice chromosome 2, 9, and 11 for K1 and K2 races, whereas on chromosome 4, 6, 11, and 12 for K3 and K3a races. The significant loci detected have also been illustrated, NBS-LRR type disease resistance protein, SNARE domain containing protein, Histone deacetylase 19, NADP-dependent oxidoreductase, and other expressed and unknown proteins. Our results provide a better understanding of the distribution of genetic variation of BLB resistance to Korean pathogen races and breeding of resistant rice.

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