• Title/Summary/Keyword: Bfa1

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Evidence of an Alternative Route of Cellobiase Secretion in the Presence of Brefeldin A in the Filamentous Fungus Termitomyces clypeatus

  • Banik, Samudra Prosad;Pal, Swagata;Chowdhury, Sudeshna;Ghorai, Shakuntala;Khowala, Suman
    • Journal of Microbiology and Biotechnology
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    • v.21 no.4
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    • pp.412-420
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    • 2011
  • Secretion of cellobiase occurred in a brefeldin A (BFA) uninhibited manner in the filamentous fungus Termitomyces clypeatus. Fluorescence confocal microscopy revealed that application of the drug at a concentration of 50 ${\mu}g$/ml caused arrest of Spitzenkorper assembly at the hyphal tip. This resulted in greater than 30% inhibition of total protein secretion in the culture medium. However, the cellobiase titer increased by 17%, and an additional 13% was localized in the vacuolar fraction en route secretion. The secretory vacuoles formed in the presence of the drug were also found to be bigger (68 nm) than those in the control cultures (40 nm). The enzyme secreted in the presence and absence of BFA revealed a single activity band in both cases in native PAGE and had similar molecular masses (approx. 120 kDa) in SDS-PAGE. The BFA enzyme retained 72% of native glycosylation. It also exhibited a higher stability and retained 98% activity at $50^{\circ}C$, 93.3% activity at pH 9, 63.64% activity in the presence of 1M guanidium hydrochloride, and 50% activity at a glucose concentration of 10 mg/ml in comparison to 68% activity, 75% activity, 36% activity, and 19% activity for the control enzyme, respectively. The observations collectively aimed at the operation of an alternative secretory pathway, distinct from the target of brefeldin A, which bypassed the Golgi apparatus, but still was able to deliver the cargo to the vacuoles for secretion. This can be utilized in selectively enhancing the yield and stability of glycosidases for a successful industrial recipe.

A Case of Bilateral Tension Pneumothorax after the Successful CO2 Laser-assisted Removal of a Bronchial Foreign Body in a Child

  • Mun, In Kwon;Ju, Yeo Rim;Lee, Sang Joon;Woo, Seung Hoon
    • Medical Lasers
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    • v.9 no.1
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    • pp.65-70
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    • 2020
  • Bronchial foreign body aspiration (BFA) is a common but emergent condition in infants and children. Furthermore, it can result in various complications such as atelectasis, pneumonia, bronchiectasis, and pneumothorax. Among these, pneumothorax is a very rare complication. However, it can be fatal without the swift implementation of appropriate treatment. We experienced a case of 16-month-old girl with an aspirated peanut. The foreign body was fixed in her left main bronchus. A CO2 laser was used to safely cut and break the foreign body. Removal was successful after breaking it. But after the process, inflammatory tissue of the tracheal mucosa was ruptured. Bilateral tension pneumothorax followed after the rupture. The patient was treated with bilateral chest tube insertion. Here we present this BFA case with a rare and unexpected complication. We also review the appropriate literature.

Endoplnsmic Reticulum Stress Up-regulates Ferritin Heavy Chain 1 Expression

  • Jin, Cho-Yi;Kwon, Ki-Sang;Yun, Eun-Young;Goo, Tae-Won;Kwon, O-Yu
    • Biomedical Science Letters
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    • v.13 no.2
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    • pp.153-155
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    • 2007
  • Ferritin heavy chain 1 (FTH1) is an ubiquitous and highly conserved protein which plays a major role in iron homeostasis. The expression of FTH1 was specifically enhanced under various condition of endoplasmic reticulum (ER) stresses drugs such as Brefeldin A (BFA), DTT (Dithiothreitol), calcium ionophore A23187 and tunicamycin. We firstly report here that ER-stress induces up-regulated expression of FTH1 in FRTL-5 culture thyrocytes.

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Bisphenol-A Removal in Conventional Water Treatment Systems (정수처리공정에서 bisphenol-A의 제거에 관한 연구)

  • 김혜리;이윤진;박선구;남상호
    • Journal of Environmental Health Sciences
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    • v.30 no.1
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    • pp.59-64
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    • 2004
  • This study was carried out to investigate influencing factors of bisphenol A(BPA) removal characteristic in conventional water treatment systems to be connected with coagulation, sedimentation, filtration and disinfection. The result are summarized as follows; In BPA removal, optimal doses of PAC, alum, ferric chloride were 7.5 mg Al/L, 10.0 mg AI/L, 15.0 mg Fek. PAC was most effective coagulant to remove BPA. In coagulation process, BPA removal efficiency were increased about 2% by adjusting pH of raw water as 6. At temperature rise 1$0^{\circ}C$, BPA removal efficiency were increased 0.94%. but BPA removal efficiency in sand filtration process were under 1 %ie, so that BPA was almost not removed. At free chlorine dose 1, 2 mg/L, the reaction rate constant k in the BPA removal have been calculated to be 0.397, 0.953 min$^{-1}$ . At free chlorine dose 1, 2 mg/1-, degradation reaction of BPA was completed during 10 min and BFA removal efficiencies were 97.66, 99.99% at this time.

Up-regulation of Early Growth Response-1 Expression by Endoplasmic Reticulum Stress

  • Han, Song-Yi;Kwon, Ki-Sang;Yun, Eun-Young;Goo, Tae-Won;Kwon, O-Yu
    • Biomedical Science Letters
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    • v.13 no.2
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    • pp.157-160
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    • 2007
  • Endoplasmic reticulum (ER) plays formation of disulfide bonds and proper folding of secretory proteins. Cellular responses to ER stress enhances the stress-activated kinase pathway and the induces a lot of immediate-early genes. Among of them, the early growth response-1 (Egr-1), a transcription factor, which plays an important role in cell growth, development, differentiation, apoptosis and various types of injury. For that reason, we have tested the expression of Egr-1 against ER stress inducible drugs (tunicamycin, DTT, A23187 and BFA) to understand what kind of aspect occurred by ER stresses.

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Rapid Detection and Discrimination of the Three Salmonella Serotypes, S. Pullorum, S. Gallinarum and S. Enteritidis by PCR-RFLP of ITS and fliC Genes

  • Cha, Se-Yeoun;Jang, Du-Hee;Kim, Sang-Min;Park, Jong-Beom;Jang, Hyung-Kwan
    • Korean Journal of Poultry Science
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    • v.35 no.1
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    • pp.9-13
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    • 2008
  • Salmonella enterica serotype gallinarum biovar Gallinarum or Pullorum and Salmonella enterica serotype Enteritidis are the most important diseases in poultry industry. Transitional diagnosis methods of these diseases such as direct isolation and identification by a biochemical test are time consuming with low specificity. In this study, we have focused on the suitable procedure for the rapid and accurate diagnosis of diseases derived from the three Salmonella strains. We initially confirmed Salmonella species by PCR using a specific ITSF/ITSR primer pair instead of biochemical test, and then the PCR-amplified phase 1 flagellin (fliC) using a specific fliCF/fliCR primer pair was digested with a restriction endonuclease, Bpm I and/or Bfa I, to discriminate among S. Pullorum, S. Gallinarum, and S. Enteritidis. We found that these methods could be applied to field isolates of the three Salmonella strains to detect and to discriminate rapidly for convenient diagnosis.

Induction of ER-stress by Heat Shock in the Thyrocytes

  • Kwon, Ki-Sang;Kwon, O-Yu;Yang, Young-Mo
    • Biomedical Science Letters
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    • v.12 no.4
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    • pp.435-438
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    • 2006
  • In eukaryotes, ER stress induces UPR (unfolded protein response) via IRE1 activation which sends a molecular signal for XBP1 mRNA splicing in the cytosol. During this mRNA splicing, 23 nt removed in which contains PstI site and then resulting XBP1 product is not digested with PstI restriction enzyme. In this study, using this XBP1 mRNA splicing mechanism, the effect of heat shock on thyrocytes is studied, because heat shock response in the thyrocytes needs more study to understand thyroid physiology under alternative environments. ER inducible drugs (tunicamycin, DTT, $Ca^{2+}$ ionopore A23187, BFA) induce ER stress in the thyrocytes. From 3 hours after heat shock, ER stress is induced and which is reversible when heat shock is without. While $Ca^{2+}$ ionopore A23187 is reversible from ER stress by washing out the drug, thapsigagin is irreversible. Other ER inducible drugs are not so sensitive to ER stress repairing. XBP1 mRNA splicing in a cell is very available method to detect ER stress. It needs only a small quantity of total RNA and processing also very easy.

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Development of high tryptophan GM rice and its transcriptome analysis (고 함량 트립토판 생산 GM 벼 개발 및 전사체 분석)

  • Jung, Yu Jin;Nogoy, Franz Marielle;Cho, Yong-Gu;Kang, Kwon Kyoo
    • Journal of Plant Biotechnology
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    • v.42 no.3
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    • pp.186-195
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    • 2015
  • Anthranilate synthase (AS) is a key enzyme in the biosynthesis of tryptophan (Trp), which is the precursor of bioactive metabolites like indole-3-acetic acid and other indole alkaloids. Alpha anthranilate synthase 2 (OsASA2) plays a critical role in the feedback inhibition of tryptophan biosynthesis. In this study, two vectors with single (F124V) and double (S126F/L530D) point mutations of the OsASA2 gene for feedback-insensitive ${\alpha}$ subunit of rice anthranilate synthase were constructed and transformed into wildtype Dongjinbyeo by Agrobacterium-mediated transformation. Transgenic single and double mutant lines were selected as a single copy using TaqMan PCR utilized nos gene probe. To select intergenic lines, the flanking sequence of RB or LB was digested with a BfaI enzyme. Four intergenic lines were selected using a flanking sequence tagged (FST) analysis. Expression in rice (Oryza sativa L.) of the transgenes resulted in the accumulation of tryptophan (Trp), indole-3-acetonitrile (IAN), and indole-3-acetic acid (IAA) in leaves and tryptophan content as a free amino acid in seeds also increased up to 30 times relative to the wildtype. Two homozygous event lines, S-TG1 and D-TG1, were selected for characterization of agronomic traits and metabolite profiling of seeds. Differentially expressed genes (DEGs), related to ion transfer and nutrient supply, were upregulated and DEGs related to co-enzymes that work as functional genes were down regulated. These results suggest that two homozygous event lines may prove effective for the breeding of crops with an increased level of free tryptophan content.

New Hyperbranched Polyimides and Polyamides: Synthesis, Chain-End Functionalizations, Curing Studies, and Some Physical Properties (새로운 Hyperbranchedpolyimidesandpolyamides: 합성, 말단기 변형, 경화 연구, 그리고 물리적 성질)

  • Baek, Jong-Beom;Chris B. Lyon;Tan, Loon-Seng
    • Proceedings of the Korean Society For Composite Materials Conference
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    • 2003.10a
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    • pp.1-2
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    • 2003
  • While aromatic polyimides and polyamides have found widespread use as high performance polymers, the present work addressed the need for organosoluble materials through the use of a hyperbranching scheme. The $AB_2$ monomers were prepared. The $AB_2$ monomers were then polymerized via aromatic fluoride-displacement and Yamazaki reactions to afford the corresponding hydroxyl-terminated hyperbranched polyimides (HT-PAEKI) and amine-terminated hyperbranched polyamides, respectively. HT-FAEKI was then functionalized with allyl and propargyl bromides as well as epichlorohydrin to afford allyl-terminated AT-PAEKI, propargyl-terminated PT-PAEKI, and epoxy (glycidyl)-terminated ET-PAEKI, in that order. All hyperbranched poly(ether-ketone-imide)s were soluble in common organic solvents. AT-PAEKI was blended with a bisphenol-A-based bismaleimide (BFA-BMI) in various weight ratios. Thermal, rheological, and mechanical properties of these blend systems were evaluated. Two characteristic hyperbranched polyamides, which the one has para-electron donating groups to the surface amine groups and the other has para-electron withdrawing groups to the surface amine groups, were selected to compare BMI curing behaviors. The electron rich polymer displayed ordinary Michael addition type exothermic reaction, while electron deficient polymer did display unusual curing behaviors. Based on analytical data, the later system provided the strong evidences to support room temperature curing of BMI by reactive intermediates instead of reactive primary amine groups on the macromolecule surface.

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Osteogenic Differentiation of Circulating Peripheral Blood Derived Mesenchymal Progenitor Cells (말초혈액 유래 간엽전구세포의 골분화)

  • Eun, Seok Chan;Kim, Jin Hee;Heo, Chan Yeong;Baek, Rong Min;Chang, Hak;Minn, Kyung Won
    • Archives of Plastic Surgery
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    • v.35 no.3
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    • pp.229-234
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    • 2008
  • Purpose: There are some reports presenting that peripheral blood contain circulating hematopoietic cells as well as, in significantly smaller quantities, mesenchymal stem cells. The purposes of this study is to isolate and characterize circulating mesenchymal progenitor cells with osteogenic potential from human peripheral blood. Methods: Human buffycoat containing mononuclear cells was harvested from peripheral blood of normal persons and isolated using a density gradient centrifugation and serially subcultured in osteogenic media for 1-4 weeks. The proliferation capability, phase-contrast microscopy, transmission electron microscopy, immunophenotype FACS analysis, Alizarin red staining and RT-PCR assays for osteogenic differentiation potential were performed. Results: The phenotype of cultured cells changed from small round or cuboidal cells at passage 1 into large spindle-shaped fibroblastic morphology cells at passage 4. Surface marker expressed CD14, but did not express CD34, CD80, CD83. Strong positive staining was observed for Alizarin reds in osteogenic medium on day 14, Using RT-PCR, the mRNA levels of bone- specific genes, such as ALP, c-bfa-1 and osteocalcin were detected. Conclusion: A new subset of peripheral blood derived progenitor cells described here has the ability to proliferate and differentiate into osteogenic cell lineages in vitro, and to be candidate for regenerative therapy.