• Title/Summary/Keyword: cell identity

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Molecular Cloning, Characterization and Expression Analysis of an ILF2 Homologue from Tetraodon nigroviridis

  • Wang, Hui-Ju;Shao, Jian-Zhong;Xiang, Li-Xin;Shen, Jia
    • BMB Reports
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    • v.39 no.6
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    • pp.686-695
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    • 2006
  • Interleukin-2 enhancer binding factor 2 (ILF2) was reported to regulate transcription of interleukin-2 (IL-2), a central cytokine in the regulation of T-cell responses. This property of ILF2 was well characterized in human and mammals, but little is known in bony fish. In this paper, an ILF2 homologue was cloned and well characterized from Tetraodon nigrovirid is for the further investigation of the function of ILF2 in bony fish. The full-length Tetraodon ILF2 cDNA was 1380 bp in size and contained an open reading frame (ORF) of 1164 bp that translates into a 387 amino-acid peptide with a molecular weight of 42.9 kDa, a 5' untranslated region (UTR) of 57 bp, and a 3' UTR of 159 bp containing a poly A tail. The deduced peptide of Tetraodon ILF2 shared an overall identity of 58%~93% with other known ILF2 sequences, and contained two N-glycosylation sites, two N-myristoylation sites, one RGD cell attachment sequence, six protein kinase C phosphorylation sites, one amino-terminal RGG-rich single-stranded RNA-binding domain, and a DZF zinc-finger nucleic acid binding domain, most of which were highly conserved through species compared. Constitutive expression of Tetraodon ILF2 was observed in all tissues examined, including gill, gut, head kidney, spleen, liver, brain and heart. The highest expression was detected in heart, followed by liver, head kidney and brain. Stimulation with LPS did not significantly alter the expression of Tetraodon ILF2. Gene organization analysis showed that the Tetraodon ILF2 gene have fifteen exons, one more than other known ILF2 genes in human and mouse. Genes up- and down-stream from the Tetraodon ILF2 were Rpa12, Peroxin-11b, Smad4, Snapap and Txnip homologue, which were different from that in human and mouse.

Mechanism of Corni Fructus Induced Vasorelaxation in Rabbit Carotid Artery (산수유의 혈관이완효과 기전에 대한 연구)

  • Kim, Hyung Jun;Park, Sun Young;Kim, Tae Yeon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.30 no.2
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    • pp.101-108
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    • 2016
  • This study is conducted to investigate vasorelaxant effect of Corni Fructus(CF) on rabbit carotid artery. To determine vasorelaxant effect of CF on rabbit carotid artery, arterial sections with intact or removed endothelium were used in this organ bath study. After being contracted by phenylephrine(PE), arterial sections were treated with CF extract in a dose-dependent manner. To identity its mechanism, the contracted arterial sections by PE were pretreated with indomethacin(IM), tetraethylammonium chloride(TEA), Nω-nitro-L-arginine(L-NNA) or methylene blue(MB) and 1.0 ㎎/㎖ CF extract. We also studied to confirm the effect on influx of extracellular calcium chloride(Ca2+) of the CF extract in rabbit carotid artery. To measure the cytotoxicity of the CF extract, cell viability of human umbilical vein endothelial cell(HUVEC) was measured by MTT assay. Generation of nitric oxide(NO) was also measured by Griess reagent. The arterial sections with intact endothelium were relaxed significantly by CF extract, but this effect was inhibited in the arterial sections with damaged endothelium. The vasorelaxant effect was inhibited significantly when arterial sections were pretreated with IM, TEA, L-NNA, MB. In Ca2+-free krebs solution, increasing of arterial contraction by Ca2+ was also inhibited by CF significantly. The treatment of CF extract increased NO concentration in HUVEC. This study suggested that the vasorelaxant effect of CF extract would be related with endothelium derived relaxing factor(EDRF) such as NO, prostacyclin(PGI2), endothelium derived hyperpolarization factor(EDHF).

Analysis of the relationship between the end weight trait and the gene ADGRL2 in purebred landrace pigs using a Genome-wide association study

  • Kang, Ho-Chan;Kim, Hee-Sung;Lee, Jae-Bong;Yoo, Chae-Kung;Choi, Tae-Jeong;Lim, Hyun-Tae
    • Korean Journal of Agricultural Science
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    • v.45 no.2
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    • pp.238-247
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    • 2018
  • The overall consumption of meat is increasing as the level of national income increases. The end weight is a trait closely associated with dressed meat. Genome-wide association study (GWAS) is an effective method of analyzing genetic variation and gene identification associated with a number of natural alternative traits because it can detect variations. So this paper did a GWAS analysis to identity the location on the genome related to the end weight in purebred landrace pigs and to explore the relevant candidate gene. This study identified a significant single nucleotide poly morphism (SNP) marker in chromosome 6 (ASGA0029422, $p=1.22{\times}10^{-6}$). Adhesion G protein-coupled receptor L2 (ADGRL2) was found to be the candidate gene at the identified SNP marker location. ADGRL2 genes have been found to be associated with cell development in relation to the external and internal environment of a cell. In addition, genotype and statistical analyses were done on nine variations on the exon of ADGRL2. The results show that the SNP marker (ASGA0029422, $p=1.32{\times}10^{-6}$) was significant, but the significance of the nine variations on the ADGRL2 exon was not verified. However, by performing further experiments and functional studies on other SNPs showing possible genetic ADGRL-Exon mutations, objects with high associations of high-end weights can be selected.

Molecular Identification of Vaginal Lactobacillus spp. Isolated from Korean Women

  • CHANG, CHUNG EUN;SYLVIA I. PAVLOVA;LIN TAO;EUN-KI KIM;SEUNG CHUL KIM;HYUN SHIK YUN;JAE-SEONG SO
    • Journal of Microbiology and Biotechnology
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    • v.12 no.2
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    • pp.312-317
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    • 2002
  • Indigenous lactobacilli were isolated from vaginas of Korean women for possible use in ecological treatment of bacterial vaginosis. Vaginal swab samples were obtained from a gynecological clinic and streaked on Rogosa SL agar plates to select the most predominant lactobacilli in each sample. The preliminary identification of the isolates as lactobacilli was based on microscopic observation of Gram-positive rod-shaped cell morphology. The initial characterization was performed on 108 isolates in terms of their cell surface hydrophobicity (CSH), antimicrobial activity, and hydrogen peroxide (H₂O₂) production capability, and 10 isolates were then selected for further molecular identification. For a rapid procedure to identify lactobacilli, polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analyses of the l6S rRNA genes were applied. The 10 selected lactobacilli and 9 different reference strains of Lactobacillus spp. were characterized by PCR-RFLP where the amplified l6S rDNA was digested with 7 different restriction endonucleases prior to analysis. DNA sequencing of the 16S rRNA gene of one particular isolate, KLB 46, that had been identified as L. crispatus by the PCR-RFLP analysis, further confirmed its identity as L. crispatus.

Anti-inflammatory Activity on LPS-stimulated in vitro RAW 264.7 Cells and in vivo Zebrafish of Heterosigma akshiwo

  • Kim, Junseong;Choi, Youn Kyung;Lee, Ji-Hyeok;Kim, Seo-Young;Kim, Hyun-Soo;Jeon, You-Jin;Heo, Soo-Jin
    • Journal of Chitin and Chitosan
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    • v.22 no.3
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    • pp.185-193
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    • 2017
  • Red tide Heterosigma akashiwo (H. akashiwo), a microscopic alga of the class Raphidophyceae, causes extensive damage to all marine ecosystems. It is essential to reduce the damage to marine ecosystems for them to be used as a resource. In this study, we used organic solvent fractionation to obtain an ethyl acetate-methanol extract from H. akashiwo (HAEM80) and then evaluated its anti-inflammatory activity in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells and a zebrafish model. HAME80 markedly inhibited the production of nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$). It also down-regulated the protein expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) and decreased the secretion of interleukin-$1{\beta}$ ($IL-1{\beta}$) in LPS-stimulated RAW 264.7 cells. HAME80 reduced yolk edema and improved the survival rate of LPS-stimulated zebrafish embryos; in addition, the extract significantly reduced the production of ROS and NO and attenuated cell death in this model. Gas chromatography-mass spectrometry (GC-MS) of the extract was used to confirm the identity of peaks 1-20. Taken together, our data suggest that H. akashiwo is a beneficial anti-inflammatory agent.

Antagonistic Potentiality of Actinomycete-Derived Extract with Anti-Biofilm, Antioxidant, and Cytotoxic Capabilities as a Natural Combating Strategy for Multidrug-Resistant ESKAPE Pathogens

  • Mohamed H. El-Sayed;Fahdah A. Alshammari;Mohammed H. Sharaf
    • Journal of Microbiology and Biotechnology
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    • v.33 no.1
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    • pp.61-74
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    • 2023
  • The global increase in multidrug-resistant (MDR) bacteria has inspired researchers to develop new strategies to overcome this problem. In this study, 23 morphologically different, soil-isolated actinomycete cultures were screened for their antibacterial ability against MDR isolates of ESKAPE pathogens. Among them, isolate BOGE18 exhibited a broad antibacterial spectrum, so it was selected and identified based on cultural, morphological, physiological, and biochemical characteristics. Chemotaxonomic analysis was also performed together with nucleotide sequencing of the 16S rRNA gene, which showed this strain to have identity with Streptomyces lienomycini. The ethyl acetate extract of the cell-free filtrate (CFF) of strain BOGE18 was evaluated for its antibacterial spectrum, and the minimum inhibitory concentration (MIC) ranged from 62.5 to 250 ㎍/ml. The recorded results from the in vitro anti-biofilm microtiter assay and confocal laser scanning microscopy (CLSM) of sub-MIC concentrations revealed a significant reduction in biofilm formation in a concentration-dependent manner. The extract also displayed significant scavenging activity, reaching 91.61 ± 4.1% and 85.06 ± 3.14% of 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), respectively. A promising cytotoxic ability against breast (MCF-7) and hepatocellular (HePG2) cancer cell lines was obtained from the extract with IC50 values of 47.15 ± 13.10 and 122.69 ± 9.12 ㎍/ml, respectively. Moreover, based on gas chromatography-mass spectrometry (GC-MS) analysis, nine known compounds were detected in the BOGE18 extract, suggesting their contribution to the multitude of biological activities recorded in this study. Overall, Streptomyces lienomycini BOGE18-derived extract is a good candidate for use in a natural combating strategy to prevent bacterial infection, especially by MDR pathogens.

Rhizospheric-Derived Nocardiopsis alba BH35 as an Effective Biocontrol Agent Actinobacterium with Antifungal and Plant Growth-Promoting Effects: In Vitro Studies

  • Mohamed H. El-Sayed;Abd El-Nasser A. Kobisi;Islam A. Elsehemy;Mohamed A. El-Sakhawy
    • Journal of Microbiology and Biotechnology
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    • v.33 no.5
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    • pp.607-620
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    • 2023
  • The biocontrol approach using beneficial microorganisms to control crop diseases is becoming an essential alternative to chemical fungicides. Therefore, new and efficient biocontrol agents (BCA) are needed. In this study, a rhizospheric actinomycete isolate showed unique and promising antagonistic activity against three of the most common phytopathogenic fungi, Fusarium oxysporum MH105, Rhizoctonia solani To18, and Alternaria brassicicola CBS107. Identification of the antagonistic strain, which was performed according to spore morphology and cell wall chemotype, suggested that it belongs to the Nocardiopsaceae. Furthermore, cultural, physiological, and biochemical characteristics, together with phylogenetic analysis of the 16S rRNA gene (OP869859.1), indicated the identity of this strain to Nocardiopsis alba. The cell-free filtrate (CFF) of the strain was evaluated for its antifungal potency, and the resultant inhibition zone diameters ranged from 17.0 ± 0.92 to 19.5 ± 0.28 mm for the tested fungal species. Additionally, the CFF was evaluated in vitro to control Fusarium wilt disease in Vicia faba using the spraying method under greenhouse conditions, and the results showed marked differences in virulence between the control and treatment plants, indicating the biocontrol efficacy of this actinomycete. A promising plant-growth promoting (PGP) ability in seed germination and seedling growth of V. faba was also recorded in vitro for the CFF, which displayed PGP traits of phosphate solubilization (48 mg/100 ml) as well as production of indole acetic acid (34 ㎍/ml) and ammonia (20 ㎍/ml). This study provided scientific validation that the new rhizobacterium Nocardiopsis alba strain BH35 could be further utilized in bioformulation and possesses biocontrol and plant growth-promoting capabilities.

TGF-$\beta$3 Selectively Induces Mouse IgA and IgG2b isotype (TGF-$\beta$3는 마우스 IgA, IgG2b 항체의 선택적 유도작용)

  • 이은경;박석래;전계택;김평현;이세원;최의열
    • Korean Journal of Microbiology
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    • v.35 no.2
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    • pp.164-168
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    • 1999
  • TGF-$\beta$3 is among five TGF-$\beta$ isolorms and shows 80% sequence identity to TGF-$\beta$I, a prototype of TGF--$\beta$. It has been reported that TGF-$\beta$I, particularly in the presence of IL-2 or L-5, increases the pmduction of IgA and IgG2b isoiypes by LPS-actwated murine B cells. We examined the effect of TGF-P3 on Ig synlhesis by B cells from different lymphoid origins. IgA induction by TGP-$\beta$3 was mardnal in LPS-activated spleen B cell culture, while 1gA production was markedly enhanced in the culture shulated with TGF-$\beta$P3 and L-5. In addition, number of IgA secreting cells was increased by TGF-$\beta$P3. Under the same conditions, TGP-$\beta$3 alone was enough to increase IgG2b production but IgM and 1gGl. Sirmlar patiem of IgA and IgGZb enbancement by TGF-$\beta$3 and L-5 was observed in the cullures of mesenteric lymph node B cells. Thus, overall effect of TGF-$\beta$3 on Ig synthesis was quite similar to that of TGF-$\beta$I. Nonetheless, it remains to be underslood whether TGF-$\beta$3 is an important modulator in B cell differentiation since regulation of TGF-$\beta$3 expression is considered to differ from that of TGF-$\beta$I

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Growth and Physiological Properties of Wild Type and Mutants of Halomonas subglaciescola DH-l in Saline Environment

  • Ryu, Hye Jeong;Jeong, Yoo Jung;Park, Doo Hyun
    • Journal of Microbiology
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    • v.42 no.3
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    • pp.174-180
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    • 2004
  • A halophilic bacterium was isolated from fermented seafood. The 16S rDNA sequence identity between the isolate and Halomonas subglaciescola AJ306801 was above 95%. The isolate that did not grow in the condition without NaCl or in the condition with other sodium (Na$\^$+/) or chloride ions (Cl$\^$-/) instead of NaCl was named H. subglaciescola DH-l. Two mutants capable of growing without NaCl were obtained by random mutagenesis, of which their total soluble protein profiles were compared with those of the wild type by two-dimensional electrophoresis. The external compatible solutes (betaine and choline) and cell extract of the wild type did not function as osmoprotectants, and these parameters within the mutants did not enhance their growth in the saline environment. In the proton translocation test, rapid acidification of the reactant was not detected for the wild type, but it was detected for the mutant in the condition without NaCl. From these results, we derived the hypothesis that NaCl may be absolutely required for the energy metabolism of H. subglaciescola DH-l but not for its osmoregulation, and the mutants may have another modified proton translocation system that is independent of NaCl, except for those mutants with an NaCl-dependent system.

Cloning and Characterization of UV-B Inducible Chalcone Synthase from Grape Cell Suspension Culture System and Its Expression Compared with Stilbene Synthase

  • Song, Won-Yong;In, Jun-Gyo;Lim, Yong-Pyo;Park, Kwan-Sam
    • Journal of Photoscience
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    • v.7 no.2
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    • pp.53-58
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    • 2000
  • We performed the cloning of a chalcone synthase (CHS) gene, the key enzyme in the anthocyanin biosynthesis, from the cDNA library constructed with grape suspension cells irradiated UV-B. The PCR fragment was used to cloning the CHS gene. One CHS cDNA clone containing an open reading frame and a partial stilbene synthase (STS)cDNA, the stilbene-type phytoalexin, were isolated. The CHS cDNA clone (VCHS) showed 87% sequence homology with VvCHS (V.vinifea) and 72.3% identity with VSTSY(V.vinifea). its amino acid sequences were longer than any other CHS genes as 454 residues. Two genes were weakly expressed in white light irradiated cells, but highly induced in UV-B irradiated condition during 32 hours. Interestingly, the STS was quickly and abundantly expressed from 2 hours when supplemented with jasmonic acid (JA) and the maximum expression was observed at 4 hours and then gradually decreased. But, the additional UV-B or white light quickly degraded the STS expression than only JA treated grape suspension cells. The CHS also was rapidly induced with JA and the synergistical effect was observed at the addigional light treatment of UV-B or white light. These results are indicated that CHS and STS have different response mechanisms against the environmental stresses.

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