• 제목/요약/키워드: Bfa1

검색결과 21건 처리시간 0.023초

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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    • 제21권4호
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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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    • 제9권1호
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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

  • ;;;;권오유
    • 대한의생명과학회지
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    • 제13권2호
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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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정수처리공정에서 bisphenol-A의 제거에 관한 연구 (Bisphenol-A Removal in Conventional Water Treatment Systems)

  • 김혜리;이윤진;박선구;남상호
    • 한국환경보건학회지
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    • 제30권1호
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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

  • ;;;;권오유
    • 대한의생명과학회지
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    • 제13권2호
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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
    • 한국가금학회지
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    • 제35권1호
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    • pp.9-13
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    • 2008
  • 양계 산업에서 살모넬라에 의한 질병들 중 가장 중요한 원인체는 S. Gallinarum, S. Purollum, S. Enteritidis로 간주된다. 생화학적 검사에 의한 직접적인 균 분리.동정과 같은 이들 질병에 대한 종래의 진단법은 많은 시간이 소요되며, 특이성 또한 낮다. 본 연구는 3종의 살모넬라균에 의해 야기되는 이들 질병에 대한 빠르고 정확한 진단을 위한 효율적인 진단법에 초점을 두었다. 먼저 종래의 생화학적 검사를 대신하여 새로 고안된 ITSF/ITSR PCR primer를 이용하여 살모넬라균임을 확인하였으며, 증폭된 phase 1 flagellin (fliC) 유전자를 BpmI 또는 BfaI 제한 효소로 처리하여 S. Gallinarum, S. Purollum, S. Enteritidis를 상호 용이하게 감별하였다. 이상의 결과는 3종의 살모넬라균을 효율적으로 진단하기 위해 신속하게 검출하고 감별할 수 있는 유용한 진단법임을 알 수 있었다.

Induction of ER-stress by Heat Shock in the Thyrocytes

  • ;;양영모
    • 대한의생명과학회지
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    • 제12권4호
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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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고 함량 트립토판 생산 GM 벼 개발 및 전사체 분석 (Development of high tryptophan GM rice and its transcriptome analysis)

  • 정유진;;조용구;강권규
    • Journal of Plant Biotechnology
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    • 제42권3호
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    • pp.186-195
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    • 2015
  • Anthranilate synthase (AS)는 트립토판(Trp)과 indole-3-acetic acid, indole alkaloids의 생합성 경로에서 중요한 효소로 작용한다. 트립토판 생합성 상에서 feedback inhibition에 민감하게 반응하는 AS alpha-subunit 관련 OASA2 유전자 영역의 single (F124V) 및 double (S126F/L530D) 점돌연변이로 변형된 유전자의 재조합운반체를 작성하고 이들 유전자들을 Agrobacterium 방법으로 동진벼에 도입하여 형질전환체를 육성하였다. Single 및 double 돌연변이 OsASA2 유전자가 도입된 형질전환 벼 계통들은 nos gene probe를 이용한 TaqMan PCR 방법으로 single copy를 선발하였고, intergenic 계통을 선발하기 위해서 Bfa I 제한효소를 이용하여 RB와 LB 인접서열로부터 IPCR을 통한 FST 분석을 수행하여 4 개의 intergenic 계통을 선발하였다. 도입된 유전자의 발현으로 형질전환 벼는 Trp, IAN 및 IAA가 잎에 가장 많이 축적되었고, 종자의 트립토판 함량도 증가되었다. 후대에서 tryptophan 함량이 높은 S-TG와 D-TG의 두 호모 이벤트 계통을 육성하여 트립토판 함량을 분석한 결과 대조구에 비하여 13~30배 이상 높게 나타났으며, 유리아미노산의 함량도 증가하였다. 이벤트 계통을 이용하여 microarray 분석을 수행한 결과 세포 내 이온 수송, 영양분 공급 등에 영향을 주는 유전자군들이 up-regulation 되었고, 세포 내 기능유전자의 역할을 담당하는 조효소 등이 down-regulation 된 것을 확인 할 수 있었다. 이러한 결과는 선발된 두개의 상동성 이벤트 계통들이 고함량의 유리 트립토판 생산 벼의 육종에 효과적으로 이용될 수 있음을 보여준 결과로 생각된다.

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

  • Baek, Jong-Beom;Chris B. Lyon;Tan, Loon-Seng
    • 한국복합재료학회:학술대회논문집
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    • 한국복합재료학회 2003년도 추계학술발표대회 논문집
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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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말초혈액 유래 간엽전구세포의 골분화 (Osteogenic Differentiation of Circulating Peripheral Blood Derived Mesenchymal Progenitor Cells)

  • 은석찬;김진희;허찬영;백롱민;장학;민경원
    • Archives of Plastic Surgery
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    • 제35권3호
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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.