Broad-specificity amino acid racemase, a novel non-antibiotic selectable marker for transgenic plants

Yi‑Chia Kuan,Venkatesan Thiruvengadam,Jia‑Shin Lin,Jia‑Hsin Liu,Tsan‑Jan Chen,Hsin‑Mao Wu,Wen‑Ching Wang,Liang‑Jwu Chen 한국식물생명공학회 2018 Plant biotechnology reports Vol.12 No.1

The broad-specificity amino acid racemase (Bsar) from Pseudomonas putida catalyzes the racemization of various amino acids, offering a flexible and feasible platform to develop a new non-antibiotic selectable marker system for plant transformation. In the present study, we demonstrated that a Bsar variant, Bsar-R174K, that is useful as a selectable marker gene in Arabidopsis and rice that were susceptible to l-lysine and D-alanine. The introduction of wild-type Bsar, Bsar-R174K or Bsar-R174A into E. coli lysine or asparagine auxotrophs was able to rescue the growth of these microorganisms in minimal media supplemented with selectable amino acid enantiomers. The transformation of Arabidopsis with Bsar or Bsar variants based on d-alanine selection revealed that Bsar-R174K had the greatest efficiency (2.40%), superior to kanamycin selectionbased transformation (1.10%). Whereas, l-lysine-based selection exhibited lower efficiency for Bsar-R174K (0.17%). The progenies of selected Bsar-R174K transgenic Arabidopsis revealed normal growth properties. In addition, Bsar-R174K transgenic rice was obtained on l-lysine medium with an efficiency of 0.9%, and the progenies of the transgenic rice revealed morphologically normal phenotypes comparable with their wild-type counterparts. This study presents the first report of broad range amino acid racemase Bsar-R174K as a non-antibiotic selectable marker system applied in transgenic plants.

Synthesis of mesoporous SiO2 xerogel/chitosan mixed-matrix membranes for butanol dehydration

Yi-Feng Lin,Chang-Yu Wu,Ting-Yu Liu,Kun-Yi Andrew Lin,Kuo-Lun Tung,Tsair-Wang Chung 한국공업화학회 2018 Journal of Industrial and Engineering Chemistry Vol.57 No.-

Novel mesoporous SiO2 xerogel/chitosan (CS) mixed-matrix membranes (MMMs) were successfully prepared by incorporating sol–gel synthesized SiO2 xerogels into glutaraldehyde (GA) cross-linked CS membranes. The effects of the SiO2 doping amount and the feed temperature on the performance of the separation of butanol and water was also investigated in this study. The 0.25 wt% SiO2 xerogel/CS MMMs possessed the best pervaporation performance and the largest PSI value of 1102 kg/m2h with a permeate flux of 736 g/m2h (Permeability of 3.25 × 10−5 g m m−2 h −1 Pa−1) and a separation factor of 1498 at a feed temperature of 50 °C. This PSI value is not only comparable but also exceeds that of other membranes reported in the literature. The separation factor and the flux of the SiO2 xerogel/CS MMMs in this study clearly exceed the upper limit of the previously reported MMMs. As a result, the as-prepared novel SiO2 xerogel/CS MMMs showed an improved performance during the butanol and water separation process, which raises the possibility of future pervaporation applications using this novel SiO2 xerogel/CS MMMs.

Decoy receptor 3 is involved in epidermal keratinocyte commitment to terminal differentiation via EGFR and PKC activation

Wu Nan-Lin,Huang Duen-Yi,Hsieh Shie-Liang,Dai Yang-Shia,Lin Wan-Wan 생화학분자생물학회 2022 Experimental and molecular medicine Vol.54 No.-

Decoy receptor 3 (DcR3) is a soluble receptor for Fas ligand, LIGHT and TL1A, but it also exerts effector functions. Previously, we found that DcR3 is upregulated in the serum and lesional skin of patients with psoriasis and is upregulated by EGFR activation in proliferating primary human epidermal keratinocytes. However, the functional role of intracellular DcR3 in keratinocyte differentiation is still incompletely defined. Herein, primary cultured human epidermal keratinocytes were differentiated by phorbol 12-myristate 13-acetate (PMA) treatment, calcium treatment and cell confluence, which are three standard in vitro differentiation models. We found that the constitutive expression of the DcR3 gene and protein was progressively suppressed during terminal differentiation of keratinocytes. These changes were correlated with downregulation of EGFR activation during keratinocyte differentiation. EGFR inhibition by gefitinib further decreased confluence-induced suppression of DcR3 mRNA expression, and, vice versa, knocking down DcR3 expression attenuated EGFR and EGFR ligand expression as well as EGFR activation. Under conditions without a change in cell growth, DcR3 silencing reduced the expression of involucrin and transglutaminase 1 but enhanced the induction of the terminal differentiation markers keratin 10 and loricrin. Of note, DcR3 interacted with PKCα and PKCδ and enhanced PKC activity. In keratinocytes with PKCα and PKCδ silencing, differentiation markers were differentially affected. In conclusion, DcR3 expression in keratinocytes is regulated by EGFR and forms a positive feedback loop to orchestrate constitutive EGFR and PKC activity. During differentiation, DcR3 is downregulated and involved in modulating the pattern of terminal differentiation.

Fluorescence Characteristic Spectra of Domestic Fuel Products through Laser Induced Fluorescence Detection

Wu, Ting-Nien,Chang, Shui-Ping,Tsai, Wen-Hsien,Lin, Cian-Yi Korean Society of Soil and Groundwater Environment 2014 지하수토양환경 Vol.19 No.5

Traditional investigation procedures of soil and groundwater contamination are followed by soil gas sampling, soil sampling, groundwater sampling, establishment of monitoring wells, and groundwater monitoring. It often takes several weeks to obtain the analysis reports, and sometimes, it needs supplemental sampling and analysis to delineate the polluted area. Laser induced fluorescence (LIF) system is designed for the detection of free-phase petroleum pollutants, and it is suitable for on-site real-time site investigation when coupling with a direct push testing tool. Petroleum products always contain polycyclic aromatic hydrocarbon (PAH) compounds possessing fluorescence characteristics that make them detectable through LIF detection. In this study, LIF spectroscopy of 5 major fuel products was conducted to establish the databank of LIF fluorescence characteristic spectra, including gasoline, diesel, jet fuel, marine fuel and low-sulfur fuel. Multivariate statistical tools were also applied to distinguish LIF fluorescence characteristic spectra among the mixtures of selected fuel products. This study successfully demonstrated the feasibility of identifying fuel species based on LIF characteristic fluorescence spectra, also LIF seemed to be uncovered its powerful ability of tracing underground petroleum leakages.

Rhodium-Catalyzed Highly Regioselective C-H Arylation of Imidazo[1,2-a]pyridines with Aryl Halides and Triflates

Yi Liu,Lin He,Guoqiang Yin,Guojie Wu,Yingde Cui 대한화학회 2013 Bulletin of the Korean Chemical Society Vol.34 No.8

A convenient Rh-catalyzed C-H arylation of imidazo[1,2-a]pyridines with a variety of aryl halides or triflates has been reported. This process afforded a range of biaryl compounds in excellent yields and showed high activity and broad scope.

Significance of Expression of Human METCAM/MUC18 in Nasopharyngeal Carcinomas and Metastatic Lesions

Lin, Jin-Ching,Chiang, Cheng-Feng,Wang, Shur-Wern,Wang, Wen-Yi,Kwan, Po-Cheung,Wu, Guang-Jer Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.1

Human METCAM/MUC18, a cell adhesion molecule (CAM) in the immunoglobulin-like gene super family, plays a dual role in the progression of several epithelium cancers; however, its role in the nasopharyngeal carcinoma (NPC) remains unclear. To initiate the study we determined human METCAM/MUC18 expression in tissue samples of normal nasopharynx (NP), NPCs, and metastatic lesions, and in two established NPC cell lines. Immunoblotting analysis was used for the determination in lysates of frozen tissues, and immunohistochemistry (IHC) for expression in formalin-fixed, paraffin-embedded tissue sections of 7 normal nasopharynx specimens, 94 NPC tissue specimens, and 3 metastatic lesions. Human METCAM/MUC18 was expressed in 100% of the normal NP, not expressed in 73% of NPC specimens (or expressed at very low levels in only about 27% of NPC specimens), and expressed again in all of the metastatic lesions. The level of human METCAM/MUC18 expression in NPC tissues was about one fifth of that in the normal NP and metastatic lesions. The low level of human METCAM/MUC18 expression in NPC specimens was confirmed by a weak signal of RT-PCR amplification of the mRNA. Low expression levels of human METCAM/MUC18 in NPC tissues were also reflected in the seven established NPC cell lines. These findings provided the first evidence that diminished expression of human METCAM/MUC18 is an indicator for the emergence of NPC, but increased expression then occurs with metastatic progression, suggesting that huMETCAM/MUC18, perhaps similar to TGF-${\beta}$, may be a tumor suppressor, but a metastasis promoter for NPC.

Prolyl endopeptidase remodels macrophage function as a novel transcriptional coregulator and inhibits fibrosis

Lin Shuang-Zhe,Wu Wei-Jie,Cheng Yu-Qing,Zhang Jian-Bin,Jiang Dai-Xi,Ren Tian-Yi,Ding Wen-Jin,Liu Mingxi,Chen Yuan-Wen,Fan Jian-Gao 생화학분자생물학회 2023 Experimental and molecular medicine Vol.55 No.-

Macrophages are immune cells crucial for host defense and homeostasis maintenance, and their dysregulation is involved in multiple pathological conditions, such as liver fibrosis. The transcriptional regulation in macrophage is indispensable for fine-tuning of macrophage functions, but the details have not been fully elucidated. Prolyl endopeptidase (PREP) is a dipeptidyl peptidase with both proteolytic and non-proteolytic functions. In this study, we found that Prep knockout significantly contributed to transcriptomic alterations in quiescent and M1/M2-polarized bone marrow-derived macrophages (BMDMs), as well as aggravated fibrosis in an experimental nonalcoholic steatohepatitis (NASH) model. Mechanistically, PREP predominantly localized to the macrophage nuclei and functioned as a transcriptional coregulator. Using CUT&Tag and co-immunoprecipitation, we found that PREP was mainly distributed in active cis-regulatory genomic regions and physically interacted with the transcription factor PU.1. Among PREP-regulated downstream genes, genes encoding profibrotic cathepsin B and D were overexpressed in BMDMs and fibrotic liver tissue. Our results indicate that PREP in macrophages functions as a transcriptional coregulator that finely tunes macrophage functions, and plays a protective role against liver fibrosis pathogenesis.

Bevacizumab Regulates Cancer Cell Migration by Activation of STAT3

Wu, Huan-Huan,Zhang, Shuai,Bian, Huan,Li, Xiao-Xu,Wang, Lin,Pu, Yin-Fei,Wang, Yi-Xiang,Guo, Chuan-Bin Asian Pacific Journal of Cancer Prevention 2015 Asian Pacific journal of cancer prevention Vol.16 No.15

There are numerous clinical cases indicating that long-term use of bevacizumab may increase the invasiveness of tumors. However, to date, little is known about underlying molecular mechanisms. Therefore, the purpose of our study was to investigate effects of bevacizumab in four cancer cells lines (WSU-HN6, CAL27, Tca83, and HeLa). It was found to promote migration and invasion in the WSU-HN6 and Tca83 cases, while exerting inhibitory effects in CAL27 and HeLa cells. The signal transducer and activator of transcription (STAT) 3 inhibitors niclosamide and S3I-201 inhibited the STAT3 signal pathway, which is activated by bevacizumab. These inhibitors also substantially blocked bevacizumab-induced migration of WSU-HN6 and Tca83 cells. Bevacizumab upregulated interleukin (IL)-6 and phosphorylated (p)-STAT3 expression time-dependently. Therefore, we propose that bevacizumab has differential effects on the migration of different cancer cell lines and promotes migration via the IL-6/STAT3 signaling pathway.

Gene Expression Profiles of HeLa Cells Impacted by Hepatitis C Virus Non-structural Protein NS4B

( Yi Zheng ),( Lin Bai Ye ),( Jing Liu ),( Wei Jing ),( Khalid A. Timani ),( Xiao Jun Yang ),( Fan Yang ),( Wei Wang ),( Bo Gao ),( Zhen Hui Wu ) 생화학분자생물학회 2005 BMB Reports Vol.38 No.2

By a cDNA array representing 2308 signal transductionrelated genes, we studied the expression profiles of HeLa cells stably transfected by Hepatitis C virus nonstructural protein 4B (HCV-NS4B). The alterations of the expression of four genes were confirmed by real-time quantitative RT PCR; and the aldo-keto reductase family 1, member Cl (AKR1C1) enzyme activity was detected in HCV-NS4B transiently transfected HeLa cells and Huh-7, a human hepatoma cell line. Of the 2,308 genes we examined, 34 were up-regulated and 56 were down-regulated. These 90 genes involved oncogenes, tumor suppressors, cell receptors, complements, adhesions, transcription and translation, cytoskeletion and cellular stress. The expression profiling suggested that multiple regulatory pathways were affected by HCV-NS4B directly or indirectly. And since these genes are related to carcinogenesis, host defense system and cell homeostatic mechanism, we can conclude that HCV-NS4B could play some important roles in the pathogenesis mechanism of HCV.

Electromagnetic Modeling of OLEDs and Its Applications to Advanced OLEDs

Wu, Chung-Chih,Lin, Chun-Liang,Cho, Ting-Yi,Yang, Chih-Jen,Lu, Yin-Jui The Korean Infomation Display Society 2006 Journal of information display Vol.7 No.4

The optical structures and rigorous electromagnetic modeling of OLEDs will be discussed of first and then their applications in analyses and designs of various advanced OLED structures, e.g. microcavity OLEDs, tandem OLEDs and top-emitting OLEDs etc., will be reported.