• Title/Summary/Keyword: E protein

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Enrichment and verification of differentially expressed miRNAs in bursa of Fabricius in two breeds of duck

  • Luo, Jun;Liu, Junying;Liu, Hehe;Zhang, Tao;Wang, Jiwen;He, Hua;Han, Chunchun
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.7
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    • pp.920-929
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    • 2017
  • Objective: The bursa of Fabricius (BF) is a central humoral immune organ belonging specifically to avians. Recent studies had suggested that miRNAs were active regulators involved in the immune processes. This study was to investigate the possible differences of the BF at miRNA level between two genetically disparate duck breeds. Methods: Using Illumina next-generation sequencing, the miRNAs libraries of ducks were established. Results: The results showed that there were 66 differentially expressed miRNAs and 28 novel miRNAs in bursa. A set of abundant miRNAs (i.e., let-7, miR-146a-5p, miR-21-5p, miR-17~92) which are involved in immunity and disease were detected and the predicted target genes of the novel miRNAs were associated with duck high anti-adversity ability. By gene ontology analysis and enriching KEGG pathway, the targets of differential expressed miRNAs were mainly involved in immunity and disease, supporting that there were differences in the BF immune functions between the two duck breeds. In addition, the metabolic pathway had the maximum enriched target genes and some enriched pathways that were related to cell cycle, protein synthesis, cell proliferation and apoptosis. It indicted that the difference of metabolism may be one of the reasons leading the immune difference between the BF of two duck breeds. Conclusion: This data lists the main differences in the BF at miRNAs level between two genetically disparate duck breeds and lays a foundation to carry out molecular assisted breeding of poultry in the future.

Inflammatory mediator regulation of the Zizyphus jujube leaf fractions in the LPS-stimulated Raw264.7 mouse machrophage (LPS로 염증이 유도된 Raw 264.7 대식세포에서 대추(Zizyphus jujube) 잎 분획물의 염증매개물질 억제)

  • Kim, Ye Jin;Son, Dae-Yeul
    • Food Science and Preservation
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    • v.21 no.1
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    • pp.114-120
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    • 2014
  • Zizyphus jujube leaf fractions (ZLFs) showed no cytotoxic effects of up to $100{\mu}g/mL$, while the anti-inflammatory effects of ZLFs were analyzed by checking the productions of nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$), cyclooxygenase-2 (COX-2), and inflammatory cytokines TNF-${\alpha}$, IL-$1{\beta}$, and IL-6 in the lipopolysaccharide (LPS)-stimulated Raw264.7 macrophage up to the concentration of $100{\mu}g/mL$. ZLFs ($100{\mu}g/mL$) demonstrated a strong anti-inflammatory activity that reduced 61~85% of NO and 71~100% of $PGE_2$ production in the LPS-stimulated Raw264.7 macrophage. Even the low ZLFs concentration of $1{\mu}g/mL$ have reduced NO and $PGE_2$ production by 34~64%. Expressions of COX-2 protein were also effectively inhibited by the ZLFs. Furthermore, the TNF-${\alpha}$, IL-$1{\beta}$, and IL-6 production were significantly suppressed through the treatment of ZLFs at concentrations of 1, 10, and $100{\mu}g/mL$. In the order of the Zizyphus jujube leaf water fraction (ZLWF) < buthanol fraction (ZLBF) < ethyl acetate fraction (ZLEF) showed anti-inflammatory activity. In particular, the ethyl acetate fraction ZLEF at $100{\mu}g/mL$ showed an excellent anti-inflammatory activity by reducing the production of NO, $PGE_2$, COX-2, and inflammatory cytokines (TNF-${\alpha}$, IL-$1{\beta}$, IL-6) in the level of Raw264.7 macrophage without LPS-stimulation or even better. The results of our study suggest the potential of ZLEF for use as an excellent ant-inflammatory inhibiting mediator and may be used as a therapeutic approach to various inflammatory diseases.

Agastache rugosa Leaf Extract Inhibits the iNOS Expression in ROS 17/2.8 Cells Activated with TNF-$\alpha$ and IL-$\beta$

  • Oh Hwa Min;Kang Young Jin;Kim Sun Hee;Lee Young Soo;Park Min Kyu;Heo Ja Myung;Sun Jin Ji;Kim Hyo Jung;Kang Eun Sil;Kim Hye Jung;Sea Han Geuk;Lee Jae Heun;YunChoi Hye Sook
    • Archives of Pharmacal Research
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    • v.28 no.3
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    • pp.305-310
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    • 2005
  • It has been suggested that nitric oxide (NO) derived from inducible nitric oxide synthase (iNOS) may act as a mediator of cytokine-induced effects on bone turn-over. NO is also recognized as an important factor in bone remodeling, i.e., participating in osteoblast apoptosis in an arthritic joint. The components of Agastache rugosa are known to have many pharmacological activities. In the present study, we investigated the effects of Agastache rugosa leaf extract (ELAR) on NO production and the iNOS expression in ROS 17/2.8 cells activated by a mixture of inflammatory cytokines including TNF-$alpha$ and IL-1$\beta$. A preincubation with ELAR significantly and concentration-dependently reduced the expression of iNOS protein in ROS 17/2.8 cells activated with the cytokine mixture. Consequently, the NO production was also significantly reduced by ELAR with an IC$_{50}$ of 0.75 mg/mL. The inhibitory mechanism of iNOS induction by ELAR prevented the activation and translocation of NF-$\kappa$B (p65) to the nucleus from the cytosol fraction. Furthermore, ELAR concentration-dependently reduced the cellular toxicity induced by sodium nitroprusside, an NO-donor. These results suggest that ELAR may be beneficial in NO-mediated inflammatory conditions such as osteoporosis.

Chemical Components and Anti-oxidant Activities of Black Currant (블랙 커런트의 화학성분 및 항산화 활성)

  • Jeong, Chang-Ho;Jang, Chi-Weon;Lee, Koo-Yul;Kim, Il-Hun;Shim, Ki-Hwan
    • Food Science and Preservation
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    • v.19 no.2
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    • pp.263-270
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    • 2012
  • The chemical components and anti-oxidant activities of black currant were investigated. The pH, soluble solid and total acidity values were 3.36, 15.11 $^{\circ}Brix$, and 1.65%, respectively. The Hunter L, a, and b values were 18.20, 5.13, and 1.08, respectively. The proximate compositions were as follows; moisture, 77.64%; nitrogen free extract, 17.41%; crude fiber, 3.08%; crude protein, 1.28%; crude ash, 0.31%; and crude lipid, 0.28%, respectively. The mineral elements were K (177.36 mg/100 g), P (54.74 mg/100 g), and Ca (26.45 mg/100 g). The free sugar components were glucose (7.71%) and fructose (5.88%). The amino acid contents of the black currant were very rich in glutamic acid (105.73 mg/100 g) and deficient in cystine (5.29 mg/100 g). The ascorbic acid and total phenolic contents were 112.19 mg/100 g and 34.48 mg GAE/g, respectively. The ABTS and DPPH radical scavenging activity levels were 99.48% and 89.03% at the 10 and 1.25 mg/mL concentrations. The reducing power and FRAP of the black currant were dose-dependent. Thus, black currant can be an effective source of functional food substances, i.e., natural anti-oxidants.

Effects of Soybean Powder and $Chungkukjang$ Powder on Quality Characteristics of Bread (대두분과 청국장분 첨가가 제빵특성에 미치는 영향)

  • Ko, Eun-Sook;Lee, Won-Jong
    • Food Science and Preservation
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    • v.19 no.2
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    • pp.223-228
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    • 2012
  • The effects of soybean powder and $chungkukjang$ powder on the quality characteristics of bread were investigated. The nutritional elements of the soybean powder, $chungkukjang$ powder and wheat flour differed remarkably. The average crude protein, ash, crude lipid, and fiber of soybean powder and $chungkukjang$ powder were approximately 3, 15, 8 and 5 times higher than those of the wheat flour, respectively. The properties of the dough to which soybean powder and $chungkukjang$ powder were added, i.e., its water absorption (%), development time (min) and stability (min) were measured with a farinograph. The water absorption, and development time of the farinogram increased, but its stability decreased when soybean powder and $chungkukjang$ powder were added to white wheat flour. As long as the mixture rate of soybean powder and $chungkukjang$ powder increased, the loaf volume of the bread declined, but the weight significantly increased. The L value decreased, but the a and b values increased in bread crumb that was made from soybean powder and $chungkukjang$ powder. The white wheat flour to which 10% soybean powder and 5% $chungkukjang$ powder were added had the highest scores in flavor, color, texture, taste, and overall acceptability in the sensory evaluation.

In Silico Analysis of Gene Function and Transcriptional Regulators Associated with Endoplasmic Recticulum (ER) Stress (Endoplasmic recticulum stress와 관련된 유전자기능과 전사조절인자의 In silico 분석)

  • Kim, Tae-Min;Yeo, Ji-Young;Park, Chan-Sun;Rhee, Moon-Soo;Jung, Myeong-Ho
    • Journal of Life Science
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    • v.19 no.8
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    • pp.1159-1163
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    • 2009
  • It has been postulated that endoplasmic (ER) stress is involved in the development of several diseases. However, the detailed molecular mechanisms have not been fully understood. Therefore, we characterized a genetic network of genes induced by ER stress using cDNA microarray and gene set expression coherence analysis (GSECA), and identified gene function as well as several transcription regulators associated with ER stress. We analyzed time-dependent gene expression profiles in thapsigargin-treated Sk-Hep1 using an oligonucleotide expression chip, and then selected functional gene sets with significantly high expression coherence which was processed into functional clusters according to the expression similarities. The functions related to sugar binding, lysosome, ribosomal protein, ER lumen, and ER to golgi transport increased, whereas the functions with mRNA processing, DNA replication, DNA repair, cell cycle, electron transport chain and helicase activity decreased. Furthermore, functional clusters were investigated for the enrichment of regulatory motifs using GSECA, and several transcriptional regulators associated with regulation of ER-induced gene expression were found.

Subacute Oral Toxicity Evaluation of Freeze-Dried Powder of Locusta migratoria

  • Kwak, Kyu-Won;Kim, Sun Young;An, Kyu Sup;Kim, Yong-Soon;Park, Kwanho;Kim, Eunsun;Hwang, Jae Sam;Kim, Mi-Ae;Ryu, Hyeon Yeol;Yoon, Hyung Joo
    • Food Science of Animal Resources
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    • v.40 no.5
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    • pp.795-812
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    • 2020
  • Novel food sources have enormous potential as nutritional supplements. For instance, edible insects are considered as an alternative food source due to their higher protein content; moreover, they are economically efficient reproducers and have high in nutritional value. In this study, we investigated the toxicity of the freeze-dried powder of Locusta migratoria (fdLM), known to contain rich proteins as well as fatty acids. The objective of the present study was to evaluate the subacute toxicity of fdLM in male and female Sprague-Dawley (SD) rats. The SD rats were divided into four groups based on the dosage of fdLM administered: dosage of 0 (vehicle control), 750, 1,500, and 3,000 mg/kg/day were administered for 28 days. Toxicological assessments including observations on food consumption, body and organ weights, clinical signs, mortality, ophthalmologic tests, urinalyses, hematologic tests, clinical chemistry tests, gross findings, and histopathology tests were performed. Clinical signs, urinalyses, hematology, serum biochemistry tests, and organ weight examinations revealed no fdLM-related toxicity. The no-observed-adverse-effect level for fdLM was higher than 3,000 mg/kg/day in rats of both sexes; therefore, fdLM, in conclusion, can be considered safe as an edible alternative human and animal food source material.

Purification of Pig Muscle Stem Cells Using Magnetic-Activated Cell Sorting (MACS) Based on the Expression of Cluster of Differentiation 29 (CD29)

  • Choi, Kwang-Hwan;Kim, Minsu;Yoon, Ji Won;Jeong, Jinsol;Ryu, Minkyung;Jo, Cheorun;Lee, Chang-Kyu
    • Food Science of Animal Resources
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    • v.40 no.5
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    • pp.852-859
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    • 2020
  • The muscle stem cells of domestic animals are of interest to researchers in the food and biotechnology industries for the production of cultured meat. For producing cultured meat, it is crucial for muscle stem cells to be efficiently isolated and stably maintained in vitro on a large scale. In the present study, we aimed to optimize the method for the enrichment of pig muscle stem cells using a magnetic-activated cell sorting (MACS) system. Pig muscle stem cells were collected from the biceps femoris muscles of 14 d-old pigs of three breeds [Landrace×Yorkshire×Duroc (LYD), Berkshire, and Korean native pigs] and cultured in skeletal muscle growth medium-2 (SkGM-2) supplemented with epidermal growth factor (EGF), dexamethasone, and a p38 inhibitor (SB203580). Approximately 30% of total cultured cells were nonmyogenic cells in the absence of purification in our system, as determined by immunostaining for cluster of differentiation 56 (CD56) and CD29, which are known markers of muscle stem cells. Interestingly, following MACS isolation using the CD29 antibody, the proportion of CD56+/CD29+ muscle stem cells was significantly increased (91.5±2.40%), and the proportion of CD56 single-positive nonmyogenic cells was dramatically decreased. Furthermore, we verified that this method worked well for purifying muscle stem cells in the three pig breeds. Accordingly, we found that CD29 is a valuable candidate among the various marker genes for the isolation of pig muscle stem cells and developed a simple sorting method based on a single antibody to this protein.

Characterization of Endoglucanase (F-I-III) Purified from Trichoderma sp. C-4 (새로운 섬유소분해 균주 Trichoderma sp. C-4에서 분리한 Endoglucanase (F-I-III)에 대한 연구)

  • Sul Ok Ju;Chung Dae Kyun;Han In Seob;Jeong Choon Soo
    • Korean Journal of Microbiology
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    • v.41 no.1
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    • pp.81-86
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    • 2005
  • One of the endoglucanases, F-I-III, was purified from the culture filtrate of T. sp. C-4 through procedures including chromatography on Sephacryl S-200, DEAE-Sepharose A-50, and Chromatofocusing on Mono-P (FPLC). The molecular weight of the enzyme was determined to be about 56,000 Da by SDS-PAGE, and pI of 4.9 by analytical isoelectric focusing. F-I-III showed the highest enzyme activity at $55^{\circ}C$, and the pH optimum of the enzyme was 5.0. There was no loss of activity when the enzyme was incubated at $50^{\circ}C$ for 24 hours. The specific activity of the enzyme F-I-III toward the CMC was 315.4 U/mg. The Km value for $PNPG_2$ of F-I-III was 2.69 mM. N-terminal sequence of F-I-III was analyzed to be QPGTSTPEVHPKKLTTYK. It showed $95\%$ of homology to that of EGI from T. reesei. The presence of some metal ions (1 mM) had only a little effect on CMCase activity. The treatment of the reducing agents resulted in the increase of endoglucanase activity.

Molecular Cloning and Characterization of a Large Subunit of Salmonella typhimurium Glutamate Synthase (GOGAT) Gene in Escherichia coli

  • Chung Tae-Wook;Lee Dong-Ick;Kim Dong-Soo;Jin Un-Ho;Park Chun;Kim Jong-Guk;Kim Min-Gon;Ha Sang-Do;Kim Keun-Sung;Lee Kyu-Ho;Kim Kwang-Yup;Chung Duck-Hwa;Kim Cheorl-Ho
    • Journal of Microbiology
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    • v.44 no.3
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    • pp.301-310
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    • 2006
  • Two pathways of ammonium assimilation and glutamate biosynthesis have been identified in microorganisms. One pathway involves the NADP-linked glutamate dehydrogenase, which catalyzes the amination of 2-oxoglutarate to form glutamate. An alternative pathway involves the combined activities of glutamine synthetase, which aminates glutamate to form glutamine, and glutamate synthase, which transfers the amide group of glutamine to 2-oxoglutarate to yield two molecules of glutamate. We have cloned the large subunit of the glutamate synthase (GOGAT) from Salmonella typhimurium by screening the expression of GOGAT and complementing the gene in E. coli GOGAT large subunit-deficient mutants. Three positive clones (named pUC19C12, pUC19C13 and pUC19C15) contained identical Sau3AI fragments, as determined by restriction mapping and Southern hybridization, and expressed GOGAT efficiently and constitutively using its own promoter in the heterologous host. The coding region expressed in Escherichia coli was about 170 kDa on SDS-PAGE. This gene spans 4,732 bases, contains an open reading frame of 4,458 nucleotides, and encodes a mature protein of 1,486 amino acid residues (Mr =166,208). The EMN-binding domain of GOGAT contains 12 glycine residues, and the 3Fe-4S cluster has 3 cysteine residues. The comparison of the translated amino acid sequence of the Salmonella GOGAT with sequences from other bacteria such as Escherichia coli, Salmonella enterica, Shigella flexneri, Yersinia pestis, Vibrio vulnificus and Pseudomonas aeruginosa shows sequence identity between 87 and 95%.