Recent Advances on Sorting Methods of High-Throughput Droplet-Based Microfluidics in Enzyme Directed Evolution

Droplet-based microfluidics has been widely applied in enzyme directed evolution (DE), in either cell or cell-free system, due to its low cost and high throughput. As the isolation principles are based on the labeled or label-free characteristics in the droplets, sorting method contributes mostly to the efficiency of the whole system. Fluorescence-activated droplet sorting (FADS) is the mostly applied labeled method but faces challenges of target enzyme scope. Label-free sorting methods show potential to greatly broaden the microfluidic application range. Here, we review the developments of droplet sorting methods through a comprehensive literature survey, including labeled detections [FADS and absorbance-activated droplet sorting (AADS)] and label-free detections [electrochemical-based droplet sorting (ECDS), mass-activated droplet sorting (MADS), Raman-activated droplet sorting (RADS), and nuclear magnetic resonance-based droplet sorting (NMR-DS)]. We highlight recent cases in the last 5 years in which novel enzymes or highly efficient variants are generated by microfluidic DE. In addition, the advantages and challenges of different sorting methods are briefly discussed to provide an outlook for future applications in enzyme DE

Research Progress on Extraction, Functional Properties and Modification Approach of Quinoa Protein

Quinoa is considered as a pseudo-cereal and recommended by FAO as the most suitable "whole food" for human consumption. With the rapid development of the health industry, the demand for quinoa products is increasing. According to the FAO standards, quinoa protein has a balanced amino acid composition, containing nine essential amino acids. It is also rich in lysine and methionine compared with other grains and legumes. Therefore, more and more studies have focused on the nutritional value and functional properties of quinoa protein. In this study, the composition, nutritional value and extraction methods of quinoa protein are reviewed, and the research progress in the functional properties such as emulsification, foaming, gelation and digestion properties of quinoa protein are also discussed. In addition, due to the enhanced functional properties of quinoa protein after modification, the effects of physical and biological modification on the functional properties of quinoa protein are also summarized in this paper. Quinoa protein, as a high-quality protein, can be deeply explored and applied in the processing of plant proteins, promoting the industrialization of quinoa protein

High sensitivity flexible Lamb-wave humidity sensors with a graphene oxide sensing layer

This paper reports high performance flexible Lamb wave humidity sensors with a graphene oxide sensing layer. The devices were fabricated on piezoelectric ZnO thin films deposited on flexible polyimide substrates. Two resonant peaks, namely the zero order antisymmetric (A0) and symmetric (S0) mode Lamb waves, were observed and fitted well with the theoretical analysis and modelling. With graphene oxide microflakes as the sensing layer, the sensing performance of both wave modes was investigated. The humidity sensitivity of the A0 mode is 145.83 ppm per %RH (at humidity 85%RH), higher than that of S0 mode of 89.35 ppm per %RH. For the first time, we have demonstrated that the flexible humidity sensors work as usual without noticeable deterioration in performance even under severe bending conditions up to 1500 με. Also the sensors showed an excellent stability upon repeated bending for thousand times. All the results demonstrated that the Lamb wave flexible humidity sensors have a great potential for application in flexible electronics

High mobility group box 1 promotes radioresistance in esophageal squamous cell carcinoma cell lines by modulating autophagy

Resistance to radiotherapy results in relapse and treatment failure in locally advanced esophageal squamous cell carcinoma (ESCC). High mobility group box 1 (HMGB1) is reported to be associated with the radioresistance in bladder and breast cancer. However, the role of HMGB1 in the radiotherapy response in ESCC has not been fully elucidated. Here, we investigated the role of HMGB1 to radioresistance in ESCC clinical samples and cell lines. We found that HMGB1 expression was associated with tumor recurrence after postoperative radiotherapy in locally advanced ESCC patients. HMGB1 knockdown in ESCC cells resulted in increased radiosensitivity both in vitro and in vivo. Autophagy level was found depressed in HMGB1 inhibition cells and activation of autophagy brought back cell's radioresistance. Our results demonstrate that HMGB1 activate autophagy and consequently promote radioresistance. HMGB1 may be used as a predictor of poor response to radiotherapy in ESCC patients. Our finding also highlights the importance of the utility of HMGB1 in ESCC radiosensitization.Peer reviewe

Triboelectric effect based instantaneous self-powered wireless sensing with self-determined identity

Sensors are the foundation of modern Internet of Things, artificial intelligent, smart manufacturing etc, but most of them require power to operate without spontaneous unique identifiable function. Herein we propose a novel instantaneous force-driven self-powered self-identified wireless sensor based on triboelectric effect to meet the huge demand of true self-powered wireless sensors. The device consists of a microswitch controlled triboelectric nanogenerator (TENG) in parallel with a capacitor-inductor oscillating circuit, and a wireless transmitter. The system is fully powered by the output of the TENG to generate a resonant frequency containing sensing and device identity information, which is then coupled to the transmitter for realizing a long-range wireless communication. The device, with the multiple functions of energy harvesting, sensing, identity generation and wireless signal transmission, is a standalone device, which responds to each trigger without losing sensing information. It eliminates the requirement of electric components for traditional wireless communication, such as rectification circuit, energy storage units, microprocessor, wireless communication chip, etc. Thus, we developed a true self-powered identifiable wireless sensor with great potential for widespread applications

Controlling gene expression with deep generative design of regulatory DNA

Design of de novo synthetic regulatory DNA is a promising avenue to control gene expression in biotechnology and medicine. Using mutagenesis typically requires screening sizable random DNA libraries, which limits the designs to span merely a short section of the promoter and restricts their control of gene expression. Here, we prototype a deep learning strategy based on generative adversarial networks (GAN) by learning directly from genomic and transcriptomic data. Our ExpressionGAN can traverse the entire regulatory sequence-expression landscape in a gene-specific manner, generating regulatory DNA with prespecified target mRNA levels spanning the whole gene regulatory structure including coding and adjacent non-coding regions. Despite high sequence divergence from natural DNA, in vivo measurements show that 57% of the highly-expressed synthetic sequences surpass the expression levels of highly-expressed natural controls. This demonstrates the applicability and relevance of deep generative design to expand our knowledge and control of gene expression regulation in any desired organism, condition or tissue

The Therapeutic Effect of Adipose-Derived Mesenchymal Stem Cells for Radiation-Induced Bladder Injury

This study was designed to investigate the protective effect of adipose derived mesenchymal stem cells (AdMSCs) against radiation-induced bladder injury (RIBI). Female rats were divided into 4 groups: (a) controls, consisting of nontreated rats; (b) radiation-treated rats; (c) radiation-treated rats receiving AdMSCs; and (d) radiation-treated rats receiving AdMSCs conditioned medium. AdMSCs or AdMSCs conditioned medium was injected into the muscular layer of bladder 24 h after radiation. Twelve weeks after radiation, urinary bladder tissue was collected for histological assessment and enzyme-linked immunosorbent assay (ELISA) after metabolic cage investigation. At the 1 w, 4 w, and 8 w time points following cells injection, 3 randomly selected rats in RC group and AdMSCs group were sacrificed to track injected AdMSCs. Metabolic cage investigation revealed that AdMSCs showed protective effect for radiation-induced bladder dysfunction. The histological and ELISA results indicated that the fibrosis and inflammation within the bladder were ameliorated by AdMSCs. AdMSCs conditioned medium showed similar effects in preventing radiation-induced bladder dysfunction. In addition, histological data indicated a time-dependent decrease in the number of AdMSCs in the bladder following injection. AdMSCs prevented radiation induced bladder dysfunction and histological changes. Paracrine effect might be involved in the protective effects of AdMSCs for RIBI

Identification of m5C-related lncRNAs signature to predict prognosis and therapeutic responses in esophageal squamous cell carcinoma patients

Abstract Esophageal squamous cell carcinoma (ESCC) has a dismal prognosis because of atypical early symptoms and heterogeneous therapeutic responses. 5-methylcytosine (m5C) modification plays an important role in the onset and development of many tumors and is widespread in long non-coding RNA (lncRNA) transcripts. However, the functions of m5C and lncRNAs in ESCC have not been completely elucidated. Herein, this study aimed to explore the role of m5C-related lncRNAs in ESCC. The RNA-seq transcriptome profiles and clinical information were downloaded from the TCGA-ESCC database. Pearson analysis was used to identify m5C-related lncRNAs. Then we established the m5C-related lncRNAs prognostic signature (m5C-LPS) using univariate Cox and least absolute shrinkage and selection operator (LASSO) regression analysis. Then, the prognostic value of m5C-LPS was evaluated internally and externally using the TCGA-ESCC and GSE53622 databases through multiple methods. We also detected the expression of these lncRNAs in ESCC cell lines and patient tissues. Fluorescence in situ hybridization (FISH) was used to detect the prognostic value of specific lncRNA. In addition, clinical parameters, immune status, genomic variants, oncogenic pathways, enrichment pathways, and therapeutic response features associated with m5C-LPS were explored using bioinformatics methods. We constructed and validated a prognostic signature based on 9 m5C-related lncRNAs (AC002091.2, AC009275.1, CAHM, LINC02057.1, AC0006329.1, AC037459.3, AC064807.1, ATP2B1-AS1, and UBAC2-AS1). The quantitative real-time polymerase chain reaction (qRT-PCR) revealed that most lncRNAs were upregulated in ESCC cell lines and patient tissues. And AC002091.2 was validated to have significant prognostic value in ESCC patients. A composite nomogram was generated to facilitate clinical practice by integrating this signature with the N stage. Besides, patients in the low-risk group were characterized by good clinical outcomes, favorable immune status, and low oncogenic alteration. Function enrichment analysis indicated that the risk score was associated with mRNA splicing, ncRNA processing, and DNA damage repair response. At the same time, we found significant differences in the responses to chemoradiotherapy between the two groups, proving the value of m5C-LPS in treatment decision-making in ESCC. This study established a novel prognostic signature based on 9 m5C-related lncRNAs, which is a promising biomarker for predicting clinical outcomes and therapeutic response in ESCC

The amino acid sequence determines protein abundance through its conformational stability and reduced synthesis cost

Data and models used in the study

Synthesis of Metal-Organic Frameworks through Enzymatically Recycled Polyethylene Terephthalate

Polyethylene terephthalate (PET) as one of the most produced plastics contributes to global waste pollution. Upcycling PET into value-added products therefore is of environmental and economic interest. Terephthalic acid (TPA), the monomer of PET, is a common linker for metal–organic framework (MOF) synthesis; thus, PET-to-MOF upcycling raises much research attention. However, conventional PET-to-MOF upcycling often requires PET depolymerization with strong acids or bases and high temperatures, which can lead to environmental and energy penalties. As an alternative, PETase offers a sustainable approach to depolymerizing PET under mesophilic and mild pH conditions. Here we report UiO-66, MOF-5, and MIL-101 syntheses using enzymatically recycled TPA as linkers. The enzymatically recycled TPA demonstrated low impurity, and the obtained MOFs possessed comparable crystallinity, thermal stability, and surface area. These results reveal the feasibility of MOF synthesis by using enzymatically recycled PET