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Stereoselective Bioreduction of Ethyl 3-Oxo-3-(2-Thienyl) Propanoate Using the Short-Chain Dehydrogenase/Reductase ChKRED12

  • Ren, Zhi-Qiang;Liu, Yan;Pei, Xiao-Qiong;Wu, Zhong-Liu
    • Journal of Microbiology and Biotechnology
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    • v.29 no.11
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    • pp.1769-1776
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    • 2019
  • Ethyl (S)-3-hydroxy-3-(2-thienyl) propanoate ((S)-HEES) acts as a key chiral intermediate for the blockbuster antidepressant drug duloxetine, which can be achieved via the stereoselective bioreduction of ethyl 3-oxo-3-(2-thienyl) propanoate (KEES) that contains a 3-oxoacyl structure. The sequences of the short-chain dehydrogenase/reductases from Chryseobacterium sp. CA49 were analyzed, and the putative 3-oxoacyl-acyl-carrier-protein reductase, ChKRED12, was able to stereoselectively catalyze the NADPH-dependent reduction to produce (S)-HEES. The reductase activity of ChKRED12 towards other substrates with 3-oxoacyl structure were confirmed with excellent stereoselectivity (>99% enantiomeric excess) in most cases. When coupled with a cofactor recycling system using glucose dehydrogenase, the ChKRED12 was able to catalyze the complete conversion of 100 g/l KEES within 12 h, yielding the enantiopure product with >99% ee, showing a remarkable potential to produce (S)-HEES.

Violetonostoc minutum gen. et sp. nov. (Nostocales, Cyanobacteria) from a rocky substrate in China

  • Cai, Fangfang;Peng, Xin;Li, Renhui
    • ALGAE
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    • v.35 no.1
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    • pp.1-15
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    • 2020
  • Two strains isolated from a subtropical region in China, were morphologically identified as a Nostoc-like species, but its taxonomic identity was unknown. In this study, these two strains were taxonomically and phylogenetically characterized based on polyphasic approach combining morphological and genetic characteristics. Though both were virtually indistinguishable from Nostoc in field and cultured material, these two strains were phylogenetically distinct from Nostoc based on 16S rRNA phylogeny. The 16S-23S internal transcribed spacer rRNA secondary structure of these strains showed the unique pattern of D1-D1', Box-B, and V3 helix, which distinguished them from other Nostoc-like heterocytous genera. A unique cluster separated from Nostoc sensu stricto supports the establishment of Violetonostoc gen. nov. with the type species as Violetonostoc minutum sp. nov.

Bird's Eye View Semantic Segmentation based on Improved Transformer for Automatic Annotation

  • Tianjiao Liang;Weiguo Pan;Hong Bao;Xinyue Fan;Han Li
    • KSII Transactions on Internet and Information Systems (TIIS)
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    • v.17 no.8
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    • pp.1996-2015
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    • 2023
  • High-definition (HD) maps can provide precise road information that enables an autonomous driving system to effectively navigate a vehicle. Recent research has focused on leveraging semantic segmentation to achieve automatic annotation of HD maps. However, the existing methods suffer from low recognition accuracy in automatic driving scenarios, leading to inefficient annotation processes. In this paper, we propose a novel semantic segmentation method for automatic HD map annotation. Our approach introduces a new encoder, known as the convolutional transformer hybrid encoder, to enhance the model's feature extraction capabilities. Additionally, we propose a multi-level fusion module that enables the model to aggregate different levels of detail and semantic information. Furthermore, we present a novel decoupled boundary joint decoder to improve the model's ability to handle the boundary between categories. To evaluate our method, we conducted experiments using the Bird's Eye View point cloud images dataset and Cityscapes dataset. Comparative analysis against stateof-the-art methods demonstrates that our model achieves the highest performance. Specifically, our model achieves an mIoU of 56.26%, surpassing the results of SegFormer with an mIoU of 1.47%. This innovative promises to significantly enhance the efficiency of HD map automatic annotation.

Overexpression of afsR and Optimization of Metal Chloride to Improve Lomofungin Production in Streptomyces lomondensis S015

  • Wang, Wei;Wang, Huasheng;Hu, Hongbo;Peng, Huasong;Zhang, Xuehong
    • Journal of Microbiology and Biotechnology
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    • v.25 no.5
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    • pp.672-680
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    • 2015
  • As a global regulatory gene in Streptomyces, afsR can activate the biosynthesis of many secondary metabolites. The effect of afsR on the biosynthesis of a phenazine metabolite, lomofungin, was studied in Streptomyces lomondensis S015. There was a 2.5-fold increase of lomofungin production in the afsR-overexpressing strain of S. lomondensis S015 N1 compared with the wild-type strain. Meanwhile, the transcription levels of afsR and two important genes involved in the biosynthesis of lomofungin (i.e., phzC and phzE) were significantly upregulated in S. lomondensis S015 N1. The optimization of metal chlorides was investigated to further increase the production of lomofungin in the afsR-overexpressing strain. The addition of different metal chlorides to S. lomondensis S015 N1 cultivations showed that CaCl2, FeCl2, and MnCl2 led to an increase in lomofungin biosynthesis. The optimum concentrations of these metal chlorides were obtained using response surface methodology. CaCl2 (0.04 mM), FeCl2 (0.33 mM), and MnCl2 (0.38 mM) gave a maximum lomofungin production titer of 318.0 ± 10.7 mg/l, which was a 4.1-fold increase compared with that of S. lomondensis S015 N1 without the addition of a metal chloride. This work demonstrates that the biosynthesis of phenazine metabolites can be induced by afsR. The results also indicate that metal chlorides addition might be a simple and useful strategy for improving the production of other phenazine metabolites in Streptomyces.

Hydrogen's influence on reduced activation ferritic/martensitic steels' elastic properties: density functional theory combined with experiment

  • Zhu, Sinan;Zhang, Chi;Yang, Zhigang;Wang, Chenchong
    • Nuclear Engineering and Technology
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    • v.49 no.8
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    • pp.1748-1751
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    • 2017
  • Reduced activation ferritic/martensitic (RAFM) steels are widely applied as structural materials in the nuclear industry. To investigate hydrogen's effect on RAFM steels' elastic properties and the mechanism of that effect, a procedure of first principles simulation combined with experiment was designed. Density functional theory models were established to simulate RAFM steels' elastic status before and after hydrogen's insertion. Also, experiment was designed to measure the Young's modulus of RAFM steel samples with and without hydrogen charging. Both simulation and experiment showed that the solubility of hydrogen in RAFM steels would decrease the Young's modulus. The effect of hydrogen on RAFM steels' Young's modulus was more significant in water-quenched steels than it was in tempering steels. This indicated that defects inside martensite, considered to be hydrogen traps, could decrease the cohesive energy of the matrix and lead to a decrease of the Young's modulus after hydrogen insertion.

A design of Giga-bit security module using Fully pipe-lined CTR-AES (Full-pipelined CTR-AES를 이용한 Giga-bit 보안모듈 설계)

  • Vinh, T.Q.;Park, Ju-Hyun;Kim, Young-Chul;Kim, Kwang-Ok
    • Journal of the Korea Institute of Information and Communication Engineering
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    • v.12 no.6
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    • pp.1026-1031
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    • 2008
  • Nowdays, homes and small businesses rely more and more PON(Passive Optical Networks) for financial transactions, private communications and even telemedicine. Thus, encryption for these data transactions is very essential due to the multicast nature of the PON In this parer, we presented our implementation of a counter mode AES based on Virtex4 FPGA. Our design exploits three advanced features; 1) Composite field arithmetic SubByte, 2) efficient MixColumn transformation 3) and on-the-fly key-scheduling for fully pipelined architecture. By pipeling the composite field implementation of the S-box, the area cost is reduced to average 17 percent. By designing the on-the-fly key-scheduling, we implemented an efficient key-expander module which is specialized for a pipelined architecture.

Asiatic Acid Protects Dopaminergic Neurons from Neuroinflammation by Suppressing Mitochondrial ROS Production

  • Chen, Dong;Zhang, Xiao-Ya;Sun, Jing;Cong, Qi-Jie;Chen, Wei-Xiong;Ahsan, Hafiz Muhammad;Gao, Jing;Qian, Jin-Jun
    • Biomolecules & Therapeutics
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    • v.27 no.5
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    • pp.442-449
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    • 2019
  • This study sought to evaluate the effects of Asiatic acid in LPS-induced BV2 microglia cells and 1-methyl-4-phenyl-pyridine ($MPP^+$)-induced SH-SY5Y cells, to investigate the potential anti-inflammatory mechanisms of Asiatic acid in Parkinson's disease (PD). SH-SY5Y cells were induced using $MPP^+$ to establish as an in vitro model of PD, so that the effects of Asiatic acid on dopaminergic neurons could be examined. The NLRP3 inflammasome was activated in BV2 microglia cells to explore potential mechanisms for the neuroprotective effects of Asiatic acid. We showed that Asiatic acid reduced intracellular production of mitochondrial reactive oxygen species and altered the mitochondrial membrane potential to regulate mitochondrial dysfunction, and suppressed the NLRP3 inflammasome in microglia cells. We additionally found that treatment with Asiatic acid directly improved SH-SY5Y cell viability and mitochondrial dysfunction induced by $MPP^+$. These data demonstrate that Asiatic acid both inhibits the activation of the NLRP3 inflammasome by downregulating mitochondrial reactive oxygen species directly to protect dopaminergic neurons from, and improves mitochondrial dysfunction in SH-SY5Y cells, which were established as a model of Parkinson's disease. Our finding reveals that Asiatic acid protects dopaminergic neurons from neuroinflammation by suppressing NLRP3 inflammasome activation in microglia cells as well as protecting dopaminergic neurons directly. This suggests a promising clinical use of Asiatic acid for PD therapy.

${\alpha}$-Cyperone Alleviates Lung Cell Injury Caused by Staphylococcus aureus via Attenuation of ${\alpha}$-Hemolysin Expression

  • Luo, M.;Qiu, J.;Zhang, Y.;Wang, J.;Dong, J.;Li, H.;Leng, B.;Zhang, Q.;Dai, X.;Niu, X.;Zhao, S.;Deng, X.
    • Journal of Microbiology and Biotechnology
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    • v.22 no.8
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    • pp.1170-1176
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    • 2012
  • In this study, we aimed to evaluate the effect of ${\alpha}$-cyperone on S. aureus. We used a hemolysin test to examine the hemolytic activity in supernatants of S. aureus cultured with increasing concentrations of ${\alpha}$-cyperone. In addition, we evaluated the production of ${\alpha}$-hemolysin (Hla) by Western blotting. Real-time RT-PCR was performed to test the expression of hla (the gene encoding Hla) and agr (accessory gene regulator). Furthermore, we investigated the protective effect of ${\alpha}$-cyperone on Hla-induced injury of A549 lung cells by live/dead and cytotoxicity assays. We showed that in the presence of subinhibitory concentrations of ${\alpha}$-cyperone, Hla production was markedly inhibited. Moreover, ${\alpha}$-cyperone protected lung cells from Hla-induced injury. These findings indicate that ${\alpha}$-cyperone is a promising inhibitor of Hla production by S. aureus and protects lung cells from this bacterium. Thus, ${\alpha}$-cyperone may provide the basis for a new strategy to combat S. aureus pneumonia.

Diversity and Functions of Endophytic Fungi Associated with Roots and Leaves of Stipa purpurea in an Alpine Steppe at Qinghai-Tibet Plateau

  • Yang, Xiaoyan;Jin, Hui;Xu, Lihong;Cui, Haiyan;Xin, Aiyi;Liu, Haoyue;Qin, Bo
    • Journal of Microbiology and Biotechnology
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    • v.30 no.7
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    • pp.1027-1036
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    • 2020
  • Stipa purpurea is a unique and dominant herbaceous plant species in the alpine steppe and meadows on the Qinghai-Tibet Plateau (QTP). In this work, we analyzed the composition and diversity of the culturable endophytic fungi in S. purpurea according to morphological and molecular identification. Then, we investigated the bioactivities of these fungi against plant pathogenic fungi and 1-aminocyclopropane-1-carboxylate deaminase (ACCD) deaminase activities. A total of 323 fungal isolates were first isolated from S. purpurea, and 33 fungal taxa were identified by internal transcribed spacer primers and grouped into Ascomycota. The diversity of endophytic fungi in S. purpurea was significantly higher in roots as compared to leaves. In addition, more than 40% of the endophytic fungi carried the gene encoding for the ACCD gene. The antibiosis assay demonstrated that 29, 35, 28, 37 and 34 isolates (43.9, 53.1, 42.4, 56.1, and 51.5%) were antagonistic to five plant pathogenic fungi, respectively. Our study provided the first assessment of the diversity of culture-depending endophytic fungi of S. purpurea, demonstrated the potential application of ACCD activity and antifungal activities with potential benefits to the host plant, and contributed to high biomass production and adaptation of S. purpurea to an adverse environment.