Nanobody-based VSR7 tracing shows clathrin-dependent TGN to Golgi recycling.

Receptor-mediated transport of soluble proteins is nature's key to empowering eukaryotic cells to access a plethora of macromolecules, either by direct accumulation or as products from resulting biochemical pathways. The transport efficiency of these mechanisms results from the receptor's capability to capture, transport, and release ligands on the one hand and the cycling ability that allows for performing multiple rounds of ligand transport on the other. However, the plant VACUOLAR SORTING RECEPTOR (VSR) protein family is diverse, and their ligand-specificity and bidirectional trafficking routes and transport mechanisms remain highly controversial. Here we employ nanobody-epitope interaction-based molecular tools to assess the function of the VSR 7 in vivo. We demonstrate the specificity of the VSR7 for sequence-specific vacuolar sorting signals, and we trace its anterograde transport and retrograde recycling route. VSR7 localizes at the cis-Golgi apparatus at steady state conditions and transports ligands downstream to release them in the trans-Golgi network/early endosome (TGN/EE) before undergoing clathrin-dependent recycling from the TGN/EE back to the cis-Golgi.

A combination of TEXTCNN model and Bayesian classifier for microblog sentiment analysis.

More and more individuals are paying attention to the research on the emotional information found in micro-blog comments. TEXTCNN is growing rapidly in the short text space. However, because the training model of TEXTCNN model itself is not very extensible and interpretable, it is difficult to quantify and evaluate the relative importance of features and themselves. At the same time, word embedding can't solve the problem of polysemy at one time. This research suggests a microblog sentiment analysis method based on TEXTCNN and Bayes that addresses this flaw. First, the word embedding vector is obtained by word2vec tool, and based on the word vector, the ELMo word vector integrating contextual features and different semantic features is generated by ELMo model. Second, the local features of ELMo word vector are extracted from multiple angles by using the convolution layer and pooling layer of TEXTCNN model. Finally, the training task of emotion data classification is completed by combining Bayes classifier. On the Stanford Sentiment Classification Corpus data set SST (Stanford Sentiment Classification Corpus Data bank), the experimental findings demonstrate that the model in this paper is compared with TEXTCNN, LSTM, and LSTM-TEXTCNN models. The Accuracy, Precision, Recall, and F1-score of the experimental results of this research have all greatly increased. Their values are respectively 0.9813, 0.9821, 0.9804 and 0.9812, which are superior to other comparison models and can be effectively used for emotional accurate analysis and identification of events in microblog emotion analysis.

High strength and biodegradable dielectric film with synergistic alignment of chitosan nanofibrous networks and BNNSs.

The development of biodegradable and robust dielectric capacitors with high breakdown strength and energy density are indispensable. Herein, the high strength chitosan/edge hydroxylated boron nitride nanosheets (BNNSs-OH) dielectric film was fabricated via combining the dual chemically-physically crosslinking and the drafting orientation strategy, which could induced BNNSs-OH and chitosan crosslinked network alignment within the film via covalent and hydrogen bonding interaction, leading to the comprehensive reinforcement of tensile strength from 126 to 240 MPa, the Eb from 448 to 584 MV m-1, the in-plane thermal conductivity from 1.46 to 5.95 W m-1 K-1 and energy storage density from 7.22 to 13.71 J cm-1, superior than the comprehensive evaluation of the reported polymer dielectrics. The dielectric film could be completely degraded in soil in 90 days, which opened a new path for the development of next-generation environment-friendly dielectrics with excellent mechanical and dielectric properties.

Discovery and Mechanistic Investigation of Piperazinone Phenylalanine Derivatives with Terminal Indole or Benzene Ring as Novel HIV-1 Capsid Modulators.

HIV-1 capsid (CA) performs multiple roles in the viral life cycle and is a promising target for antiviral development. In this work, we describe the design, synthesis, assessment of antiviral activity, and mechanistic investigation of 20 piperazinone phenylalanine derivatives with a terminal indole or benzene ring. Among them, F2-7f exhibited moderate anti-HIV-1 activity with an EC50 value of 5.89 µM, which was slightly weaker than the lead compound PF74 (EC50 = 0.75 µM). Interestingly, several compounds showed a preference for HIV-2 inhibitory activity, represented by 7f with an HIV-2 EC50 value of 4.52 µM and nearly 5-fold increased potency over anti-HIV-1 (EC50 = 21.81 µM), equivalent to PF74 (EC50 = 4.16 µM). Furthermore, F2-7f preferred to bind to the CA hexamer rather than to the monomer, similar to PF74, according to surface plasmon resonance results. Molecular dynamics simulation indicated that F2-7f and PF74 bound at the same site. Additionally, we computationally analyzed the ADMET properties for 7f and F2-7f. Based on this analysis, 7f and F2-7f were predicted to have improved drug-like properties and metabolic stability over PF74, and no toxicities were predicted based on the chemotype of 7f and F2-7f. Finally, the experimental metabolic stability results of F2-7f in human liver microsomes and human plasma moderately correlated with our computational prediction. Our findings show that F2-7f is a promising small molecule targeting the HIV-1 CA protein with considerable development potential.

Design, Synthesis, and Mechanistic Study of 2-Pyridone-Bearing Phenylalanine Derivatives as Novel HIV Capsid Modulators.

The AIDS pandemic is still of importance. HIV-1 and HIV-2 are the causative agents of this pandemic, and in the absence of a viable vaccine, drugs are continually required to provide quality of life for infected patients. The HIV capsid (CA) protein performs critical functions in the life cycle of HIV-1 and HIV-2, is broadly conserved across major strains and subtypes, and is underexploited. Therefore, it has become a therapeutic target of interest. Here, we report a novel series of 2-pyridone-bearing phenylalanine derivatives as HIV capsid modulators. Compound FTC-2 is the most potent anti-HIV-1 compound in the new series of compounds, with acceptable cytotoxicity in MT-4 cells (selectivity index HIV-1 > 49.57; HIV-2 > 17.08). However, compound TD-1a has the lowest EC50 in the anti-HIV-2 assays (EC50 = 4.86 ± 1.71 µM; CC50= 86.54 ± 29.24 µM). A water solubility test found that TD-1a showed a moderately increased water solubility compared with PF74, while the water solubility of FTC-2 was improved hundreds of times. Furthermore, we use molecular simulation studies to provide insight into the molecular contacts between the new compounds and HIV CA. We also computationally predict drug-like properties and metabolic stability for FTC-2 and TD-1a. Based on this analysis, TD-1a is predicted to have improved drug-like properties and metabolic stability over PF74. This study increases the repertoire of CA modulators and has important implications for developing anti-HIV agents with novel mechanisms, especially those that inhibit the often overlooked HIV-2.

Environmental regulation, market demand, and green innovation: spatial perspective evidence from China.

This paper used a dynamic spatial panel model to empirically analyze the effects of environmental regulation, market demand, and the associated spatial spillovers on regional green innovation in China, from which the following was found. (1) The environmental regulations had a positive "U-shaped" effect on local green innovation and a positive neighborhood spillover effect, and market demand had a significant positive effect on local green innovation and a "ripple effect." (2) The mechanism analysis found that the environmental regulations tended to inhibit regional green innovation input through a "cost compliance" effect, and market demand had a stronger incentive effect on innovation input than the environmental regulations, thus promoting the improvement of green innovation level. (3) The environmental regulation and market demand effects on green innovation had obvious spatial and temporal heterogeneity. The results of this study could help to promote regional green innovation by formulating reasonable environmental policies and stimulating the vitality of green technology market.

The circRNA_102911/miR-129-5p/SOX6 axis is involved with T lymphocyte immune function in elderly patients with laparoscopic left hepatectomy for hepatolithiasis.

The aim of the present study was to investigate the impact of laparoscopic left hepatectomy (LLH) for hepatolithiasis on the T lymphocyte immune changes of elderly patients and to analyze underlying mechanisms of action behind these changes. A total of 164 patients who underwent LLH due to left-sided hepatolithiasis were recruited. In terms of T lymphocyte immune changes, it was found that firstly, the basic quantity of peripheral lymphocytes in the elderly group was significantly lower than that in a younger preoperative group. Secondly, after surgical trauma, the immune function of T lymphocytes had a significant decline and lasted longer when compared with younger patients, which was reflected by the perioperative changes in the T lymphocyte proliferative ability, levels of IL-2 secreted by T lymphocytes and the percentage of CD3+/CD4+ T lymphocytes in the peripheral blood. Circular RNA (circRNA) 102911 (102911) was upregulated and microRNA (miR)-129-5p was downregulated in CD3+/CD4+ T lymphocytes from elderly patients with LLH for hepatolithiasis. Furthermore, the overexpression of 102911 inhibited the proliferation of CD3+/CD4+ T lymphocytes as well as promoting cell apoptosis, with the opposite effects being observed on knockdown of 102911. miR-129-5p is involved in the proliferation and apoptosis of CD3+/CD4+ T lymphocytes and may be a promising target of 102911. Moreover, SOX6 is a downstream molecule of miR-129-5p. Immune function and number of T lymphocytes decreased significantly after surgical trauma compared to younger patients, and this decline lasted longer in older patients treated with LLH for hepatolithiasis. The 102911/miR-129-5p/SOX6 axis was found to be involved in T lymphocytes immune function, which provided a novel insight for the treatment of elderly patients with hepatolithiasis.

An AGAMOUS-like factor is associated with the origin of two domesticated varieties in Cymbidium sinense (Orchidaceae).

Cymbidium has been artificially domesticated for centuries in Asia, which produced numerous cultivated varieties. Flowers with stamenoid tepals or those with multiple tepals have been found in different species of Cymbidium; however, the molecular basis controlling the formation of these phenotypes is still largely unknown. Previous work demonstrated that AGAMOUS/AG lineage MADS genes function in floral meristem determinacy as well as in reproductive organs development in both dicots and monocots, indicating a possible relationship with the origin of two flower varieties in Cymbidium. Here, we characterized and analyzed two AG lineage paralogues, CsAG1 and CsAG2, from Cymbidium sinense, both of which were highly expressed in the gynostemium column of a standard C. sinense. Interestingly, we detected ectopic expression of CsAG1 rather than CsAG2 in all floral organs of a stamenoid-tepal variety and significant down-regulation of CsAG1 in a variety with multiple tepals. Over-expression of CsAG1 in wild type Arabidopsis resulted in petal-to-stamen homeotic conversion, suggesting a conserved C-function of CsAG1 in the development of Cymbidium flower. Altogether, our results supported a hypothesis that disruption of a single AG-like factor would be associated with the formation of two domesticated varieties in C. sinense.

Genetic Diversity and Genome-Wide Association Study of Major Ear Quantitative Traits Using High-Density SNPs in Maize.

Kernel and ear traits are key components of grain yield in maize (Zea mays L.). Investigation of these traits would help to develop high-yield varieties in maize. Genome-wide association study (GWAS) uses the linkage disequilibrium (LD) in the whole genome to determine the genes affecting certain phenotype. In this study, five ear traits (kernel length and width, ear length and diameter, cob diameter) were investigated across multi-environments for 2 years. Combining with the genotype obtained from Maize SNP50 chip, genetic diversity and association mapping in a set of 292 inbred lines were performed. Results showed that maize lines were clustered into seven subgroups and a total of 20 SNPs were found to be associated with ear traits significantly (P < 3.95E-05). The candidate genes identified by GWAS mainly encoded ubiquitin-activation enzymes (GRMZM2G015287), carotenoid cleavage dioxygenase (GRMZM2G446858), MYB-CC type transfactor, and phosphate starvation response protein 3, and they were associated with kernel length (KL) and ear diameter (ED), respectively. Moreover, two novel genes corresponding to RNA processing and fructose metabolism were found. Further, the SNPs detected by GWAS were confirmed by meta-QTL analysis. These genes and SNPs identified in the study would offer essential information for yield-related genes clone and breeding program in maize.

Transcriptome-Wide Analysis Reveals the Origin of Peloria in Chinese Cymbidium (Cymbidium sinense).

An orchid flower exhibits a zygomorphic corolla with a well-differentiated labellum. In Cymbidium sinense, many varieties with peloric or pseudopeloric flowers have been bred during centuries of domestication. However, little is known about the molecular basis controlling orchid floral zygomorphy and the origin of these varieties. Here, we studied the floral morphogenesis of C. sinense and transcriptome-wide enriched differentially expressed genes among different varieties. The floral zygomorphy of C. sinense is established in the early developmental process. Out of 27 MIKCC-MADS factors, we found two homeotic MADS genes whose expression was down-regulated in peloric varieties but up-regulated in pseudopeloric varieties. CsAP3-2 expressed in the inner floral organs co-operates with a labellum-specific factor CsAGL6-2, asymmetrically promoting the differentiation of inner tepals. Interestingly, we detected exon deletions on CsAP3-2 in peloric varieties, indicating that loss of B-function results in the origin of peloria. Additional petaloid structures developed when we ectopically expressed these genes in Arabidopsis, suggesting their roles in floral morphogenesis. These findings indicate that the interplay among MADS factors would be crucial for orchid floral zygomorphy, and mutations in these factors may have maintained during artificial selection.

Enhanced removal of uranium(VI) by nanoscale zerovalent iron supported on Na-bentonite and an investigation of mechanism.

The reductive removal of U(VI) by nanoscale zerovalent iron (NZVI) was enhanced by using Na(+)-saturated bentonite (Na-bent) as the support, and the mechanism for the enhanced removal were investigated comprehensively. Under the same experimental conditions, NZVI supported on the negatively charged Na-bent showed much higher removal efficiency (99.2%) of cationic U(VI) than either bare NZVI (48.3%) or NZVI supported on the positively charged bentonite (Al-bent) did. Subsequent experimental investigations revealed the unique roles of bentonite on enhancing the reactivity and reusability of NZVI. First, Na-bent can buffer the pH in reaction media, besides preventing NZVI from aggregation. Second, Na-bent promoted the mass transfer of U(VI) from solution to NZVI surface, leading to the enhanced removal efficiency. Third, the bentonite may transfer some insoluble reduction products away from the iron surface according to X-ray absorption fine structure (XAFS) study. Finally, Na-bent as the adsorbent to Fe(II) makes it more reactive with U(VI), which enhanced stoichiometrically the reduction capacity of NZVI besides accelerating the reaction rate.