• Title/Summary/Keyword: BL52

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The expression of Foxp3 protein by retroviral vector-mediated gene transfer of Foxp3 in C57BL/6 mice (C57BL/6 마우스에서 Retroviral 벡터를 이용한 Foxp3 유전자의 도입에 의한 Foxp3 단백의 발현 양상)

  • Hwang, Insun;Ha, Danbee;Bing, So Jin;Jeon, Kyong-Leek;Ahn, Ginnae;Kim, Dae Seung;Cho, Jinhee;Lim, Jaehak;Im, Sin-Hyeog;Hwang, Kyu-Kye;Jee, Youngheun
    • Korean Journal of Veterinary Research
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    • v.52 no.3
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    • pp.183-191
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    • 2012
  • The maintenance of peripheral immune tolerance and prevention of chronic inflammation and autoimmune disease require $CD4^{+}CD25^{+}$ T cells (regulatory T cells). The transcription factor Foxp3 is essential for the development of functional, regulatory T cells, which plays a prominent role in self-tolerance. Retroviral vectors can confer high level of gene transfer and transgene expression in a variety of cell types. Here we observed that following retroviral vector-mediated gene transfer of Foxp3, transductional Foxp3 expression was increased in the liver, lung, brain, heart, muscle, spinal cord, kidney and spleen. One day after vector administration, high levels of transgene and gene expression were observed in liver and lung. At 2 days after injection, transductional Foxp3 expression level was increased in brain, heart, muscle and spinal cord, but kidney and spleen exhibited a consistent low level. This finding was inconsistent with the increase in both $CD4^{+}CD25^{+}$ T cell and $CD4^{+}Foxp3^{+}$ T cell frequencies observed in peripheral immune cells by fluorescence-activated cell-sorting (FACS) analysis. Retroviral vector-mediated gene transfer of Foxp3 did not lead to increased numbers of $CD4^{+}CD25^{+}$ T cell and $CD4^{+}Foxp3^{+}$ T cell. These results demonstrate the level and duration of transductional Foxp3 gene expression in various tissues. A better understanding of Foxp3 regulation can be useful in dissecting the cause of regulatory T cells dysfunction in several autoimmune diseases and raise the possibility of enhancing suppressive functions of regulatory T cells for therapeutic purposes.

Fast Coding Mode Decision for MPEG-4 AVC|H.264 Scalable Extension (MPEG-4 AVC|H.264 Scalable Extension을 위한 고속 모드 결정 방법)

  • Lim, Sun-Hee;Yang, Jung-Youp;Jeon, Byeung-Woo
    • Journal of the Institute of Electronics Engineers of Korea SP
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    • v.45 no.6
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    • pp.95-107
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    • 2008
  • In this paper, we propose a fast mode decision method for temporal and spatial scalability to reduce computational complexity of mode decision that used to be computationally one of the most intensive processes of the MPEG-4 AVC|H.264 SE(Scalable Extension) encoding. For temporal scalability, we propose an early skip method and MHM(mode history map) method. The early skip method confines macroblock modes of backward and forward frames within selected a few candidates. The MHM method utilizes stored information of frames inside a GOP of lower levels for the decision of MHM at higher level. For the spatial scalability, we propose the method that uses a candidate mode according to the MHM method and adds the BL_mode as candidates. The proposed scheme reduces the number of candidate modes to reduce computational complexity in mode decision. The proposed scheme reduces total encoding time by about 52% for temporal scalability and 47% for spatial scalability without significant loss of RD performance.

Characteristics of Seed Germination and Seedling Growth of Native Hydrangea serrata for. acuminata (자생 산수국의 종자 발아와 유묘 생육 특성)

  • Lee, Seung Youn;Kim, Kwang Jin;Lee, Jeong Sik
    • FLOWER RESEARCH JOURNAL
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    • v.16 no.2
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    • pp.134-142
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    • 2008
  • This work aims to obtain basic information for seed propagation of Hydrangea serrata for. acuminata. The germination percentage of the seeds taken on 15 November, 30 December, and 23 January was $90.0{\pm}4.16%$, $84.4{\pm}5.52%$, and $88.9{\pm}2.40%$, respectively. This suggest that seeds of Hydrangea serrata for. acuminata are non-dormant seeds. The optimum temperature for germination was $25^{\circ}C$ and light was necessary. Most of the growth parameters (shoot and leaf length, stem diameter, root length, no. of roots, T/R ratio, and fresh and dry wts.) were significantly greater at $25/20^{\circ}C$ and $25^{\circ}C$ than at the other temperatures. Low T/R ratio at relatively cool temperatures (15 and $20^{\circ}C$) was caused by suppressed top growth. In light quality treatment, red light (RL) significantly enhanced stem elongation. The greatest photosynthetic pigments (total chl, chl a/b, and carotenoid) were observed in seedlings grown in blue light (BL), followed by seedlings grown in RL+BL. When blue light was added, higher pigment contents were found. Effect of plug cell size (50, 72, 128, 162 and 200 cells) on the growth of seedlings was investigated. The highest top growth was observed in seedlings grown in 50 cell trays, followed by seedlings grown in 72, 128, 162, and 200 cell trays. However, there was no significant differences between 162 and 200 cell trays. Especially, smaller size leaves were observed in seedlings grown in smaller cell trays (lower volume and high plant density).

Cloning and Expression of a Alkaline Protease from Bacillus clausii I-52 (Bacillus clausii I-52로부터 alkaline protease 유전자의 클로닝 및 발현)

  • Joo, Han-Seung;Choi, Jang Won
    • Journal of agriculture & life science
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    • v.45 no.6
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    • pp.201-212
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    • 2011
  • The alkaline protease gene was cloned from a halo-tolerant alkalophilic Bacillus clausii I-52 isolated from the heavily polluted tidal mud flat of West Sea in Inchon Korea, which produced a strong extracellular alkaline protease (BCAP). Based on the full genome sequence of Bacillus subtilis, PCR primers were designed to allow for the amplification and cloning of the intact pro-BCAP gene including promoter region. The full-length gene consists of 1,143 bp and encodes 381 amino acids, which includes 29 residues of a putative signal peptide and an additional 77-amino-acid propeptide at its N-terminus. The mature BCAP deduced from the nucleotide sequence consists of 275 amino acids with a N-terminal amino acid of Ala, and a relative molecular weight and pI value was 27698.7 Da and 6.3, respectively. The amino acid sequence shares the highest similarity (99%) to the nattokinase precursor from B. subtilis and subtilisin E precursor from B. subtilis BSn5. The substrate specificity indicated that the recombinant BCAP could hydrolyze efficiently the synthetic substrate, N-Suc-Ala-Ala-Pro-Phe-pNA,and did not hydrolyze the substrates with basic amino acids at the P1 site. The recombinant BCAP was strongly inhibited by typical serine protease inhibitor, PMSF, indicating that BCAP is a member of the serine proteases.

High Expression of Water-Soluble Recombinant Antigenic Domains of Toxoplasma gondii Secretory Organelles

  • Yang, Zhaoshou;Ahn, Hye-Jin;Nam, Ho-Woo
    • Parasites, Hosts and Diseases
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    • v.52 no.4
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    • pp.367-376
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    • 2014
  • Recombinant antigenic proteins of Toxoplasma gondii are alternative source of antigens which are easily obtainable for serodiagnosis of toxoplasmosis. In this study, highly antigenic secretory organellar proteins, dense granular GRA2 and GRA3, rhoptrial ROP2, and micronemal MIC2, were analyzed by bioinformatics approach to express as water-soluble forms of antigenic domains. The transmembrane region and disorder tendency of 4 secretory proteins were predicted to clone the genes into pGEX-4T-1 vector. Recombinant plasmids were transformed into BL21 (DE3) pLysS E. coli, and GST fusion proteins were expressed with IPTG. As a result, GST fusion proteins with $GRA2_{25-105}$, $GRA3_{39-138}$, $ROP2_{324-561}$, and $MIC2_{1-284}$ domains had respectively higher value of IgG avidity. The $rGST-GRA2_{25-105}$ and $rGST-GRA3_{39-138}$ were soluble, while $rGST-ROP2_{324-561}$ and $rGST-MIC2_{1-284}$ were not. $GRA2_{31-71}$, intrinsically unstructured domain (IUD) of GRA2, was used as a linker to enhance the solubility. The $rGST-GRA2_{31-71}-ROP2_{324-561}$, a chimeric protein, appeared to be soluble. Moreover, $rGST-GRA2_{31-71}-MIC2_{1-284}$ was also soluble and had higher IgG avidity comparing to $rGST-MIC2_{1-284}$. These 4 highly expressed and water-soluble recombinant antigenic proteins may be promising candidates to improve the serodiagnosis of toxoplasmosis in addition to the major surface antigen of SAG1.

Estimation of Genetic Variance Components of Body Size Measurements in Hanwoo (Korean Cattle) Using a Multivariate Linear Model

  • Lee, Jung-Jae;Kim, Nae-Soo
    • Journal of Animal Science and Technology
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    • v.52 no.3
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    • pp.167-174
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    • 2010
  • The objectives of this study were to quantify the combination values of the principal components and factors calculated using body measurements of Hanwoo (Korean Cattle) and estimate their heritabilities. The technique of multivariate analysis was used to reduce a large number of variables to a smaller number of new variables and characterize cattle according to body shape. The analyses were performed using 1,979 cattle at 12 months of age and 936 cattle at 24 months of age. The data for the analyses was obtained from progeny tests performed on Korean Cattle for 6 years from 2003 to 2008. The phenotypic correlations among these traits were estimated to range from 0.32 to 0.90 at 12 months of age and from 0.21 to 0.82 at 24 months of age. The first principal components (PC1s) indicated a weighed average of overall body measurements, accounting for 99.91% of the total variation for both periods of test. The two first PCs had positive coefficients for all body measurements. The major sources of PC, such as chest girth (CG), body length (BL), rump height (RH), and wither height (WH) were similar for both test periods. The heritabilities for PC1, the first factor score (FS1), and the second factor score (FS2) were estimated by multivariate REML method. The estimated heritabilities for PC1, FS1, and FS2 were 0.33, 0.38, and 0.40, respectively, at 12 months of age and 0.26, 0.76, and 0.58 at 24 months of age. Further studies are needed to determine whether the heritabilities of FS1 and FS2 at 24 months of age were overestimated.

Corn silk extracts did not alter the adiposity and underlying substrate utilization in high-fat diet-induced obese C57BL/6 mice

  • Kang, Eun Young;Kim, Hyun Kyung;Moyo, Knowledge Mudhibadhi;Gang, Gyoungok;Kim, Wooki;Go, Gwang-woong
    • Korean Journal of Food Science and Technology
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    • v.52 no.2
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    • pp.144-148
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    • 2020
  • Corn silk (Okmi-su) was anciently adopted as a material for tea or beverage. Corn silk extracts (CSE) contain bioactive phytochemicals such as phenolic acid, flavonoids, ascorbic acid, tannins, and glycosides. Under the impact of these functional components, CSE has benefits for antioxidation, diuresis, anti-diabetes, and dyslipidemia recovery. Nonetheless, its role in whole-body adiposity was not investigated; therefore, the effects of CSE on obesity were evaluated in high-fat diet-induced obese mice. Mice were assigned to either group (n=12); 1) normal diet (18% kcal from fat), 2) high-fat diet (45% kcal from fat, the control), 3) high-fat diet with CSE (800 mg/kg diet), and 4) high-fat diet with orlistat (500 mg/kg diet, a comparable control for weight loss). Our results showed that body weight, adiposity, and energy expenditure in obese mice were not altered by CSE. Lean body mass tended to decrease by CSE, which can be explained by stimulation of diuresis (p=0.06). In conclusion, our results suggest that dietary consumption of CSE does not influence the adiposity and underlying substrate utilization in high-fat diet-induced obese mice.

Tissue-resident natural killer cells exacerbate tubulointerstitial fibrosis by activating transglutaminase 2 and syndecan-4 in a model of aristolochic acid-induced nephropathy

  • Wee, Yu Mee;Go, Heounjeong;Choi, Monica Young;Jung, Hey Rim;Cho, Yong Mee;Kim, Young Hoon;Han, Duck Jong;Shin, Sung
    • BMB Reports
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    • v.52 no.9
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    • pp.554-559
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    • 2019
  • Despite reports suggesting that tissue-resident natural killer (trNK) cells cause ischemic kidney injury, their contribution to the development of tubulointerstitial fibrosis has not been determined. This study hypothesized that the depletion of trNK cells may ameliorate renal fibrosis by affecting transglutaminase 2/syndecan-4 interactions. Aristolochic acid nephropathy (AAN) was induced in C57BL/6 mice as an experimental model of kidney fibrosis. The mice were treated with anti-asialo GM1 (ASGM1) or anti-NK1.1 antibodies to deplete NK cells. Although both ASGM1 and NK1.1 antibodies suppressed renal $NKp46^+DX5^+$ NK cells, renal $NKp46^+DX5^-$ cells were resistant to suppression by ASGM1 or NK1.1 antibodies during the development of tubulointerstitial fibrosis in the AAN-induced mouse model. Western blot analysis showed that both antibodies increased the expression of fibronectin, transglutaminase 2, and syndecan-4. These findings indicate that trNK cells played an exacerbating role in tubulointerstitial fibrosis by activating transglutaminase 2 and syndecan-4 in the AAN-induced mouse model.

Purification and Characterization of a Recombinant Pea Chloroplastic Fructose-1, 6-bisphosphatase

  • Shin, Eun-Hye;Yoo, Yong-Cheol;Lee, Sang-Won;Hahn, Tae-Ryong
    • Journal of Applied Biological Chemistry
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    • v.44 no.4
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    • pp.167-172
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    • 2001
  • A cDNA fragment encoding the chloroplastic fructose-1, 6-bisphosphatase (FBPase) was cloned via PCR from the cDNA library of pea leaves. The cloned cDNA, about 1.05 kbp without signal sequence, was introduced into a pET-28a vector for expression in E. coli strain BL21(DE3)pLysS. The recombinant FBPase was purified through $Ni^+-NTA$ affinity chromatography and characterized. Molecular mass of the monomer was about 42,000. Enzymatic activity of the purified enzyme as the native pea chloroplastic FBPase was the highest at alkaline pH (pH 9.0). The recombinant enzyme was activated by a reducing agent DTT and was insensitive to AMP. The activation energy (Ea) and Arrehenius frequency factor were 42.67 kcal/mol and $2.65{\times}10^{14}/s$, respectively, slightly higher than those of the native enzyme. $K_M$ and $V_{max}$ were $99.98{\mu}M$ and $52.9{\mu}M/min$, respectively, which were comparable with the native enzyme.

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Angiotensin Converting Enzyme(ACE) Gene Polymorphism in Korean Stroke Patients (한국인(韓國人) 중풍(中風) 환자(患者)의 Angiotensin Converting Enzyme 유전자(遺傳子) 다형성(多形成)에 관(關)한 연구(硏究))

  • Ha, Ji-young;Kim, Chang-hwan;Koh, Hyung-kyun
    • Journal of Acupuncture Research
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    • v.20 no.2
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    • pp.161-172
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    • 2003
  • Objective : This study was designed to investigate the relation between the angiotensin converting enzyme(ACE) gene polymorphism and stroke in the Korean population. Methods : This study was carried out on 58 stroke patients who were hospitalized in the department of acupuncture & moxibustion, college of Oriental Medicine, Kyung-Hee University and 61 healthy control subjects. Blood samples from all subjects were obtaind for DNA extration. The extracted DNA was amplified by polymerase chain reaction(PCR). PCR products were visualized by 2% agarose gel electrophoresis. Results : The sub-genotypes of ACE gene were II homozygotes, ID heterozygotes, DD homozygotes. While the distribution of ACE polymorphism in control subjects was 31%, 51%, 18%, the distribution of it in stoke patients was 33%, 52%, 16%(II, ID, DD). Thus, there was no significant different between the control and stroke groups. Conclusions : we conclude that there is no significant association between ACE gene polymorphism and storke in Korean papulation. However, the findings of this study need to be confirmed in large patients and further studies. Additional epidemiologicallly based studies of the effects and relationship between ACE or other genes and lifesyles with regard to stroke required.

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