Access tunnel engineering to optimize the catalytic cycle of carbohydrate hydrolases with buried active site

The active site of many enzymes is buried inside the protein core and is connected with the surrounding solvent by access tunnels. An emerging approach to optimize these enzymes properties is the engineering of structural features governing the exchange of ligands between the active sites and bulk solvent. However, it is still challenging to redesign the access tunnels of enzymes catalyzing biopolymers like carbohydrate hydrolases because of the extremely complicated substrate structure. In this study, structure-guided saturated mutagenesis was performed to reconstruct all three access tunnels of xylanase S7-xyl from Bacillus halodurans S7, which results in a mutant 254-RL1 with 3.4-fold increase in specific activity. Structural comparison and kinetic analysis revealed that products egress is the rate-limiting step in the catalytic cycle of S7-xyl. The products release tunnel in S7-xyl was experimentally validated, and not the tunnel radius but the length determining the products release efficiency. Application assessment showed that relieving the inhibition of reducing sugars on mutant 254-RL1 could accelerate the hydrolysis efficiency of cellulase on different pretreated lignocellulose materials, representing a good candidate in enzyme cocktails for lignocellulose biodegradation. In addition, the same strategy was successfully utilized to improve the specific activities of three other xylanases with buried active site, suggesting the general application of tunnel engineering to optimize carbohydrate hydrolases with buried active site

Retrieval Oriented Masking Pre-training Language Model for Dense Passage Retrieval

Pre-trained language model (PTM) has been shown to yield powerful text representations for dense passage retrieval task. The Masked Language Modeling (MLM) is a major sub-task of the pre-training process. However, we found that the conventional random masking strategy tend to select a large number of tokens that have limited effect on the passage retrieval task (e,g. stop-words and punctuation). By noticing the term importance weight can provide valuable information for passage retrieval, we hereby propose alternative retrieval oriented masking (dubbed as ROM) strategy where more important tokens will have a higher probability of being masked out, to capture this straightforward yet essential information to facilitate the language model pre-training process. Notably, the proposed new token masking method will not change the architecture and learning objective of original PTM. Our experiments verify that the proposed ROM enables term importance information to help language model pre-training thus achieving better performance on multiple passage retrieval benchmarks.Comment: Search LM part of the "AliceMind SLM + HLAR" method in MS MARCO Passage Ranking Leaderboard Submissio

Fansubbing in China:Technology-facilitated activism in translation

This paper seeks to explore the socio-political tensions between freedom and constraints in the Chinese fansubbing networks. It approaches the development of fansubbing in China as a process of technology democratisation with the potential to liberate ordinary citizens from authoritarian and commercial imperatives, enabling them to contest official state domination. The paper draws on the strategies adopted by fansubbing groups to organise their working practices and interactive social activities with a view to engaging target audiences. Both facets complement each other and bring to the fore the ‘gamified’ system of fansubbing networks. Gamification enables ordinary citizens to translate, distribute and consume foreign audiovisual products in a strategic move that pits collective activism against government dominance.</jats:p

Unsupervised Boundary-Aware Language Model Pretraining for Chinese Sequence Labeling

Boundary information is critical for various Chinese language processing tasks, such as word segmentation, part-of-speech tagging, and named entity recognition. Previous studies usually resorted to the use of a high-quality external lexicon, where lexicon items can offer explicit boundary information. However, to ensure the quality of the lexicon, great human effort is always necessary, which has been generally ignored. In this work, we suggest unsupervised statistical boundary information instead, and propose an architecture to encode the information directly into pre-trained language models, resulting in Boundary-Aware BERT (BABERT). We apply BABERT for feature induction of Chinese sequence labeling tasks. Experimental results on ten benchmarks of Chinese sequence labeling demonstrate that BABERT can provide consistent improvements on all datasets. In addition, our method can complement previous supervised lexicon exploration, where further improvements can be achieved when integrated with external lexicon information.Comment: 12 pages, 2 figures, 7 tables, EMNLP 202

Ideological Manipulation of Controversial Information:The Unusual Case of the Chinese-subtitled Version of House of Cards

In recent years, owing to the spread of cultural and economic globalisation, Western values and ideologies have become increasingly accessible via media products to the general public in the People’s Republic of China (Mainland China, or China). Although translation enables the exchange of socio-cultural values and ideas across different languages and cultures, it is susceptible to manipulation in terms of political and ideological control. This paper examines interventions by Chinese authorities in the online distribution of foreign audio-visual products. Such interventions are undertaken to secure broadcasting and distribution rights of the government which also encompass more conventional media from other countries, such as television, in promoting mainstream ideological values and political agendas.  Censorship institutions supervise the selection of specific media products to be imported and translated for the Chinese market not only to ensure ease of comprehension but also for political correctness, with a view to avoid any clashes of cultures and ideologies. In the last decade, Chinese audiences have the rise of new media facilitating the wide-spread online viewing of foreign TV and thereby changing the dynamic of access and dissemination of foreign entertainment media in China. We argue that censorship institutions in China opt to adopt alternative strategies, or in many cases more relaxed criteria, in appraising foreign audiovisual products, in order to cater the demands of the new generation of Chinese audiences in those officially supervised media rather than suppressing their needs outright. We will dwell into the Chinese-subtitled version of the second season of the American political drama House of Cards (Netflix 2014), as released by Sohu, the largest online franchise in China. We consider the outcome of subtitling "honest" in terms of the exposure of sensitive information and controversial cultural and political allusions to the reality in contemporary China. By analysing possible reasons behind such “honest” exposure, we argue that an attempt at ideological manipulation has been made in this subtitled version, serving rather than subverting the political objectives of the Chinese government.

Ex Vivo Assay of Electrical Stimulation to Rat Sciatic Nerves: Cell Behaviors and Growth Factor Expression

Neurite outgrowth and axon regeneration are known to benefit from electrical stimulation. However, how neuritis and their surroundings react to electrical field is difficult to replicate by monolayer cell culture. In this work freshly harvested rat sciatic nerves were cultured and exposed to two types of electrical field, after which time the nerve tissues were immunohistologically stained and the expression of neurotrophic factors and cytokines were evaluated. ELISA assay was used to confirm the production of specific proteins. All cell populations survived the 48 h culture with little necrosis. Electrical stimulation was found to accelerate Wallerian degeneration and help Schwann cells to switch into migratory phenotype. Inductive electrical stimulation was shown to upregulate the secretion of multiple neurotrophic factors. Cellular distribution in nerve tissue was altered upon the application of an electrical field. This work thus presents an ex vivo model to study denervated axon in well controlled electrical field, bridging monolayer cell culture and animal experiment. It also demonstrated the critical role of electrical field distribution in regulating cellular activities

Magnolin Protects against Contrast-Induced Nephropathy in Rats via Antioxidation and Antiapoptosis

BACKGROUND: Magnolin is the major active ingredient of the herb Magnolia fargesii which has anti-inflammatory and antioxidative effects. Oxidative stress and apoptosis are involved in the pathogenesis of contrast-induced nephropathy (CIN). We hypothesize that Magnolin could protect against CIN through antioxidative and antiapoptotic properties. METHODS: To test whether Magnolin could attenuate CIN, oxidative stress and apoptosis, in vivo and in vitro, we utilized a rat model of ioversol-induced CIN and a cell model of oxidative stress in which HK2 cells were treated with H2O2. Rats were assigned to 4 groups (n = 6 per group): control group, ioversol group (ioversol-induced CIN), vehicle group (CIN rats pretreated with vehicle), and Magnolin group (CIN rats pretreated with 1 mg/kg Magnolin). RESULTS: The results showed that magnolin ameliorated the renal tubular necrosis, apoptosis, and the deterioration of renal function (P < 0.05). Furthermore, Magnolin reduced the renal oxidative stress, suppressed caspase-3 activity, and increased Bcl-2 expression in vivo and in vitro. CONCLUSION: Magnolin might protect CIN in rats through antioxidation and antiapoptosis

Effect of molecular distillation on the anti-inflammatory activity and neurotoxicity of Asarum essential oil

Asarum essential oil (AEO) has been shown to have good pharmacological activities for the anti-inflammatory and analgesic effects, but increasing the dose may cause toxicity. Therefore, we studied the toxic and pharmacodynamic components of AEO by molecular distillation (MD). Anti-inflammatory activity was assessed using RAW264.7 cells. Neurotoxicity was assessed in PC12 cells and the overall toxicity of AEO was evaluated in the mouse acute toxicity assay. The results showed that AEO is primarily composed of safrole, methyl eugenol, and 3,5-dimethoxytoluene. After MD, three fractions were obtained and contained different proportions of volatile compounds relative to the original oil. The heavy fraction had high concentrations of safrole and methyl eugenol, while the light fraction contained high concentrations of α-pinene and β- pinene. The original oil and all three fractions exhibited anti-inflammatory effects, but the light fraction demonstrated more excellent anti-inflammatory activity than the other fractions. Asarum virgin oil and MD products are all neurotoxic. The exposure of PC12 cells to high concentrations of AEO resulted in abnormal nuclei, an increased number of apoptotic cells, increased ROS formation, and decreased SOD levels. Moreover, the results of acute toxicity tests in mice revealed that the light fractions were less toxic than virgin oils and other fractions. In summary, the data suggest that the MD technology enables the enrichment and separation of essential oil components and contributes to the selection of safe concentrations of AEO

Screening for Anti-Inflammation Quality Markers of Lianhua Qingwen Capsule Based on Network Pharmacology, UPLC, and Biological Activity

Influenza is a common respiratory infectious disease. In China, Lianhua Qingwen capsule (LHQWC), a drug with significant clinical efficacy and few side effects, is commonly used to treat influenza. However, the composition of LHQWC is complicated, and currently used quality control methods cannot ensure its consistency. In this study, combined with its clinical efficacy, the targets of LHQWC were screened using network pharmacology. Then, anti-inflammation quality markers of LHQWC were screened and judged by combined chemical with biological evaluation. Cyclooxygenase-2 (COX-2) was identified as one of the main targets of the anti-inflammatory activity of LHQWC. The rate of inhibition of COX-2 by different batches of LHQWC was determined. Furthermore, seven components of LHQWC were identified. The potential quality markers were screened by spectral-effect relationship. As a result, chlorogenic acid, isochlorogenic acid B, and isochlorogenic acid C were identified and confirmed as anti-inflammatory quality markers of LHQWC. We hope that these findings provide a scientific basis for the accurate quality control of LHQWC and serve as a reference for the quality control of other drugs