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Duong Bich Thuy,Duong Minh Cuong,Campbell James,Nguyen Van Minh Hoang,Nguyen Huu Hien,Bui Thi Bich Hanh,Nguyen Van Vinh Chau,McLaws Mary-Louise 대한감염학회 2021 Infection and Chemotherapy Vol.53 No.3
Little is known about antibiotic-resistant Gram-negative bacteria (GNB) intestinal carriage among healthcare workers (HCWs) in Vietnam. All HCWs at a tertiary intensive care units were asked to undertake weekly rectal swabs. Among 40 participants, 65% (26/40) carried extended spectrum β-lactamases (ESBL)/AmpC β-lactamase-producing Escherichia coli. Two HCWs colonized with ESBL/AmpC β-lactamase-producing Klebsiella pneumoniae. One HCW colonized with Acinetobacter baumannii. No one carried Pseudomonas spp.. A quarter (10/40) of HCWs were identified as persistent and frequent carriers. There is an urgent need to screen antibiotic-resistant GNB among HCWs and improve HCWs’ hand hygiene compliance to reduce the transmission of antibiotic-resistant GNB in the hospital.
Soluble Expression and Purification of Bioactive Interleukin 33 in E. coli
Bich Hang Do,박상수,Grace G. Kwon,NGUYEN MINH TAN,강효정,송정아,유지원,Anh Ngoc Nguyen,장재평,장미희,이선주,소성준,심성락,진종화,이경진,Mark J Osborn,최한 한국생물공학회 2017 Biotechnology and Bioprocess Engineering Vol.22 No.3
Interleukin-33 (IL-33) is one of the important alarmins of the immune system and possesses dual functions as an anti- or pro-inflammatory molecule. The production of this cytokine in E. coli is hampered by the insoluble expression in the cytoplasm, resulting in inclusion body formation. In this study, the expression of IL-33 was optimized by fusing the N-terminus of IL-33 with several solubilizing tags that act as chaperones for proper protein folding: maltose binding protein (MBP), b´a´ domain of protein disulfide isomerase (PDIb´a´) and glutathione Stransferase (GST). The expression of the fusion proteins was stimulated by 0.5 mM IPTG at different temperatures, 37, 30, 25, and 18oC. As a result, IL-33 was expressed highly and in soluble form in the cytoplasm of E. coli when fused with MBP or PDIb´a´ tags in the presence of 0.5 mM IPTG at 25 or 30oC. We describe a simple purification procedure of IL-33 from the PDIb´a´-IL-33 construct using immobilized metal affinity chromatography (IMACs) with supplementary of tobacco etch virus (TEV) protease for tag removal. The high bioactivity of purified IL-33 on the proliferation and activation of macrophages was confirmed by MTT and nitrite releasing assays using RAW 264.7 These data show an improved method for producing high grade and yield IL-33.
Bich Ngoc Vu,Jae Yong Yoo,Ki Seong Ko,Young Eun Lee,Ha Na Choi,Yoo Na Lee,Mi Hui Jang,Kyun Oh Lee 한국당과학회 2022 한국당과학회 학술대회 Vol.2022 No.07
To cope with multiple stresses caused by various biotic and abiotic stimuli and consequently increased accumulation of misfolded proteins, eukaryotic cells activate a series of adaptive mechanisms that are known as the unfolded protein response (UPR). In response to cellular stresses, phosphorylation of the α subunit of eukaryotic initiation factor 2 (eIF2α) attenuates general protein synthesis. However, it also favors the selective translation of mRNAs with upstream open reading frames (uORFs) such as ATF4, a key regulator of the integrated stress response (ISR), in mammalian cells. The ISR is an evolutionarily conserved translational and transcriptional program initiated upon phosphorylation of eIF2α. However, no studies to date have been conducted on the ISR mediated by phosphorylation of eIF2α, and key regulators responsible for the ISR have not been identified in plants. In this study, we performed in silico analysis to investigate functional orthologs of the ATF4, and confirmed that some genes belong to bZIP family containing uORFs in the 5′UTR are potential candidates. However, it is not well understood whether they are involved in plant ISR mediated by phosphorylation of eIF2α. During prolonged ER stress, transcription of these genes increased by 2-3 fold. Arabidopsis plants overexpressing showed a more sensitive phenotype than Col-0 and knockout mutant plants under tunicamycin-induced ER stress condition, while plants overexpressing with uORF did not show any difference. In addition, the expression level of bZIP GFP fusion protein without uORF is increased under ER stress conditions compare to normal conditions. These results suggest that uORF-containing bZIP genes may have a role in plant ISR. * Corresponding
Serum Amyloid A1: A Biomarker for Neutrophilic Airway Inflammation in Adult Asthmatic Patients
Bich Tra Cao Thi,Quoc Quang Luu,최영우,Yang Eun-Mi,Trinh Hoang Kim Tu,신유섭,박해심 대한천식알레르기학회 2022 Allergy, Asthma & Immunology Research Vol.14 No.1
Purpose: We evaluated the role of serum amyloid A1 (SAA1) in the pathogenesis of airway inflammation according to the phenotype of asthma. Methods: One hundred twenty-two asthmatic patients and 60 healthy control subjects (HCs) were enrolled to measure SAA1 levels. The production of SAA1 from airway epithelial cells (AECs) and its effects on macrophages and neutrophils were investigated in vitro and in vivo. Results: The SAA1 levels were significantly higher in sera of asthmatic patients than in those of HCs (P = 0.014); among asthmatics, patients with neutrophilic asthma (NA) showed significantly higher SAA1 levels than those with non-NA (P < 0.001). In vitro, polyinosinic:polycytidylic acid (Poly I-C) treatment markedly enhanced the production of SAA1 from AECs, which was further augmented by neutrophils; SAA1 could induce the production of interleukin (IL)-6, IL-8, and S100 calcium-binding protein A9 from AECs. Additionally, SAA1 activated neutrophils and macrophages isolated from peripheral blood of asthmatics, releasing neutrophil extracellular traps (NETs) and secreting proinflammatory cytokines presenting M1 phenotype, respectively. In ovalbumin-induced asthma mice, Poly I-C treatment significantly increased SAA1 levels as well as IL-17A/interferon-gamma/IL-33 levels in bronchoalveolar lavage fluid (BALF), leading to airway hyperresponsiveness and inflammation. The highest levels of SAA1 and neutrophilia were noted in the BALF and sera of the NA mouse model, followed by the mixed granulocytic asthma (MA) model. Especially, SAA1 induced IL-17/retinoic acid receptor-related orphan receptor γt expression from activated CD4+ T lymphocytes in asthmatic mice. Conclusions: The results show that SAA1 could induce neutrophilic airway inflammation by activating neutrophils along with NET formation, M1 macrophages, and Th2/Th17 predominant cells, contributing to the phenotype of NA or MA.
Presidential Turnover and Discontinuity in the Philippines’ China Policy
Bich T. Tran 경남대학교 극동문제연구소 2019 ASIAN PERSPECTIVE Vol.43 No.4
Philippines-China relations have experienced dramatic turns, from a “golden age” to “Asia’s most toxic bilateral relationship,” followed by a new “spring time,” as described by the media. Why does the Philippines’ China policy oscillate so much? This article looks at the role of the president in the Philippines’ foreign policymaking by examining the country’s China policy under the leadership of three Philippines’ presidents: Gloria Macapagal Arroyo, Benigno Aquino III, and Rodrigo Duterte. The findings show that although individual presidents have opportunities to promote their own preferences in dealing with China, they are not free from constraints. Despite showing great oscillations in rhetoric, the actual policies undertaken by each administration have never completely shifted to one or another extreme of the Sino-American spectrum: balancing against China using the alliance with the United States, or bandwagoning with China at the cost of losing the alliance with the United States.
( Bich Hang Do ),( Minh Tan Nguyen ),( Jung-a Song ),( Sangsu Park ),( Jiwon Yoo ),( Jaepyeong Jang ),( Sunju Lee ),( Seoungjun So ),( Yejin Yoon ),( Inki Kim ),( Kyungjin Lee ),( Yeon Jin Jang ),( Ha 한국미생물생명공학회(구 한국산업미생물학회) 2017 Journal of microbiology and biotechnology Vol.27 No.12
Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is considered as an antitumor agent owing to its ability to induce apoptosis of cancer cells without imparting toxicity toward most normal cells. TRAIL is produced in poor yield because of its insoluble expression in the cytoplasm of E. coli. In this study, we achieved soluble expression of TRAIL by fusing maltose-binding protein (MBP), b’a’ domain of protein disulfide isomerase (PDIb’a’), or protein disulfide isomerase at the N-terminus of TRAIL. The TRAIL was purified using subsequent immobilized metal affinity chromatography and amylose-binding chromatography, with the tag removal using tobacco etch virus protease. Approximately 4.5 mg of pure TRAIL was produced from 125 ml flask culture with a purification yield of 71.6%. The endotoxin level of the final product was 0.4 EU/μg, as measured by the Limulus amebocyte lysate endotoxin assay. The purified TRAIL was validated and shown to cause apoptosis of HeLa cells with an EC<sub>50</sub> and Hill coefficient of 0.6 ± 0.03 nM and 2.41 ± 0.15, respectively. The high level of apoptosis in HeLa cells following administration of purified TRAIL indicates the significance and novelty of this method for producing high-grade and high-yield TRAIL.