Engineered geranyl diphosphate methyltransferase produces 2-methyl-dimethylallyl diphosphate as a noncanonical C6 unit for terpenoid biosynthesis.

Terpenoids constitute the largest class of natural products with complex structures, essential functions, and versatile applications. Creation of new building blocks beyond the conventional five-carbon (C5) units, dimethylallyl diphosphate (DMAPP) and isopentenyl diphosphate, expands significantly the chemical space of terpenoids. Structure-guided engineering of an S-adenosylmethionine-dependent geranyl diphosphate (GPP) C2-methyltransferase from Streptomyces coelicolor yielded variants converting DMAPP to a new C6 unit, 2-methyl-DMAPP. Mutation of the Gly residue at the position 202 resulted in a smaller substrate-binding pocket to fit DMAPP instead of its native substrate GPP. Replacement of Phe residue at the position 222 with a Tyr residue contributed to DMAPP binding via hydrogen bond. Furthermore, using Escherichia coli as the chassis, we demonstrated that 2-methyl-DMAPP was accepted as a start unit to generate noncanonical trans- and cis-prenyl diphosphates (C5n+1) and terpenoids. This work provides insights into substrate recognition of prenyl diphosphate methyltransferases, and strategies to diversify terpenoids by expanding the building block portfolio.

DNA origami-based artificial antigen-presenting cells for adoptive T cell therapy.

Nanosized artificial antigen-presenting cells (aAPCs) with efficient signal presentation hold great promise for in vivo adoptive cell therapy. Here, we used DNA origami nanostructures as two-dimensional scaffolds to regulate the spatial presentation of activating ligands at nanoscale to construct high-effective aAPCs. The DNA origami-based aAPC comprises costimulatory ligands anti-CD28 antibody anchored at three vertices and T cell receptor (TCR) ligands peptide-major histocompatibility complex (pMHC) anchored at three edges with varying density. The DNA origami scaffold enables quantitative analysis of ligand-receptor interactions in T cell activation at the single-particle, single-molecule resolution. The pMHC-TCR-binding dwell time is increased from 9.9 to 12.1 s with increasing pMHC density, driving functional T cell responses. In addition, both in vitro and in vivo assays demonstrate that the optimized DNA origami-based aAPCs show effective tumor growth inhibiting capability in adoptive immunotherapy. These results provide important insights into the rational design of molecular vaccines for cancer immunotherapy.

Experimental and Numerical Study of a Rebar-Prestressed Cylinder Concrete Pipe (RPCCP) under Internal Load.

In order to study the load-bearing failure characteristics of a RPCCP under internal load, a field prototype test was designed, and a finite element model was established. An internal load was applied up to 2.0 MPa step by step and the force variation law of each part was obtained. During the production of the RPCCP, by wrapping prestressed steel bars around the concrete core with a cylinder, the core was subjected to an initial precompression stress. In the loading process, the protective cover cracked first, from where the concrete core gradually changed from the initial compression state to a tension state, finally cracking from the inner and outer diameter. The stresses of the cylinder and steel bars increased steadily with the internal load and did not yield. The finite element calculation results were in good agreement with the test results, and the influence characteristics of the tension control stress of the steel bar and the concrete strength on the failure of the RPCCP under internal load were discussed. The results showed that the internal load of the protective cover was independent of the tension control stress, but decreases with a decrease in concrete strength, while the load corresponding to the concrete core entering plasticity is related to the tension control stress and the concrete strength, and the relationships were basically linear.

The impact of job-related stress on township teachers' professional well-being: A moderated mediation analysis.

This study aimed to explore the relationship between job-related stress and township teachers' professional well-being. Based on Job Demand-Resource Model, this study examined the mediating role of teachers' professional identity and the moderating role of perceived organizational support in this relationship. A total of 24,276 township teachers in China responded to the Teacher Stress Scale, the Teachers' Professional Identity Scale, the Teachers' Professional Well-Being Structure Questionnaire, and the Perceived Organizational Support Scale. Results showed that: (1) the professional well-being of township teachers differed significantly in terms of school type and demographic characteristics like age and gender; (2) job-related stress negatively predicted township teachers' professional well-being, with teachers' professional identity playing a mediating role; (3) the relation between job-related stress and teachers' professional identity was moderated by perceived organizational support; and (4) in the moderated mediation analysis, job-related stress positively predicted township teachers' professional well-being. These findings indicated that township teachers' professional well-being was influenced by both organizational and individual factors, which provided a theoretical basis and intervention pathways for improving township teachers' professional well-being.

Protective effects of apple polyphenols on bone loss in mice with high fat diet-induced obesity.

Obesity-induced inflammation can lead to an imbalance in bone formation and resorption. Our previous studies have demonstrated that apple polyphenols (APs) can reduce body weight and inflammation. But their effect on bone is still unclear. In this study, we investigated the protective effects of APs on bone loss in mice with high-fat-diet (HFD)-induced obesity. Forty male C57BL/6J mice were divided into control group (10% fat diet), HFD group (60% fat diet), resveratrol group (60% fat diet), and AP group (60% fat diet). Micro-computed tomography revealed a significant increase in bone volume fraction and bone mineral density, and more trabecular bone and less trabecular bone separation in the AP group compared with the HFD group. In addition, serum tumor necrosis factor (TNF)-α, interleukin-6 (IL-6), and tartrate-resistant acid phosphatase (TRACP) levels were decreased; runt-related transcription factor 2 (Runx2) levels were increased; the collagen area was enlarged; and femur biomechanical property was enhanced in the AP group compared with the HFD group. APs significantly increased the ratio of osteoprotegerin to the receptor activator for the nuclear factor-κB ligand (OPG/RANKL) compared with the HFD group. Resveratrol could also improve the glucolipid regulation, but poorer osteogenic promotion was found compared to APs. The present study demonstrated that APs prevent loss of bone mass induced by obesity, which has potential implications for the prevention and treatment of obesity-related osteoporosis.

Nanoscale organization of two-dimensional multimeric pMHC reagents with DNA origami for CD8+ T cell detection.

Peptide-MHC (pMHC) multimers have excelled in the detection of antigen-specific T cells and have allowed phenotypic analysis using other reagents, but their use for detection of low-affinity T cells remains a challenge. Here we develop a multimeric T cell identifying reagent platform using two-dimensional DNA origami scaffolds to spatially organize pMHCs (termed as dorimers) with nanoscale control. We show that these dorimers enhance the binding avidity for low-affinity antigen-specific T cell receptors (TCRs). The dorimers are able to detect more antigen-specific T cells in mouse CD8+ T cells and early-stage CD4+CD8+ double-positive thymocytes that express less dense TCRs, compared with the equivalent tetramers and dextramers. Moreover, we demonstrate dorimer function in the analysis of autoimmune CD8+ T cells that express low-affinity TCRs, which are difficult to detect using tetramers. We anticipate that dorimers could contribute to the investigation of antigen-specific T cells in immune T cell function or immunotherapy applications.

The Genome-Wide Identification of Long Non-Coding RNAs Involved in Floral Thermogenesis in Nelumbo nucifera Gaertn.

The sacred lotus (Nelumbo nucifera Gaertn.) can maintain a stable floral chamber temperature when blooming, despite ambient temperature fluctuations; however, the long non-coding RNAs (lncRNAs) involved in floral thermogenesis remain unclear. In the present study, we obtain comprehensive lncRNAs expression profiles from receptacles at five developmental stages by strand-specific RNA sequencing to reveal the lncRNAs regulatory mechanism of the floral thermogenesis of N. nucifera. A total of 22,693 transcripts were identified as lncRNAs, of which approximately 44.78% had stage-specific expression patterns. Subsequently, we identified 2579 differential expressed lncRNAs (DELs) regulating 2367 protein-coding genes mainly involved in receptacle development and reproductive process. Then, lncRNAs with floral thermogenesis identified by weighted gene co-expression network analysis (WGCNA) were mainly related to sulfur metabolism and mitochondrial electron transport chains. Meanwhile, 70 lncRNAs were predicted to act as endogenous target mimics (eTMs) for 29 miRNAs and participate in the regulation of 16 floral thermogenesis-related genes. Our dual luciferase reporter assays indicated that lncRNA LTCONS_00068702 acted as eTMs for miR164a_4 to regulate the expression of TrxL2 gene. These results deepen our understanding of the regulation mechanism of floral thermogenesis by lncRNAs and accumulate data for further research.

Nucleic Acid-Based Cell Surface Engineering Strategies and Their Applications.

The cell membrane is a biological interface regulating the communications between cells and their environment. The ability to functionalize the cell membrane with molecules or nanomaterials allows us to manipulate cellular behaviors and to expand cellular functions. Due to their unique merits of synthetic accessibility, flexible design, and precise programmability, nucleic acids provide an emerging and promising molecular toolkit for cell surface engineering. In this review, the recent progress in nucleic acid-based cell surface engineering are summarized. We first introduce approaches to nucleic acid-based cell surface engineering, including monovalent and polyvalent surface engineering strategies. Then, the biological applications of nucleic acid-based cell surface engineering in biosensing of extracellular microenvironment, programming cell-cell interactions, and mimicking cellular behaviors are reviewed. Finally, we analyze the current challenges existing in this area and discuss the prospects for the future development.

DIA-Based Quantitative Proteomics Reveals the Protein Regulatory Networks of Floral Thermogenesis in Nelumbo nucifera.

The sacred lotus (Nelumbo nucifera) can maintain a stable floral chamber temperature between 30 and 35 °C when blooming despite fluctuations in ambient temperatures between about 8 and 45 °C, but the regulatory mechanism of floral thermogenesis remains unclear. Here, we obtained comprehensive protein profiles from receptacle tissue at five developmental stages using data-independent acquisition (DIA)-based quantitative proteomics technology to reveal the molecular basis of floral thermogenesis of N. nucifera. A total of 6913 proteins were identified and quantified, of which 3513 differentially abundant proteins (DAPs) were screened. Among them, 640 highly abundant proteins during the thermogenic stages were mainly involved in carbon metabolism processes such as the tricarboxylic acid (TCA) cycle. Citrate synthase was identified as the most connected protein in the protein-protein interaction (PPI) network. Next, the content of alternative oxidase (AOX) and plant uncoupling protein (pUCP) in different tissues indicated that AOX was specifically abundant in the receptacles. Subsequently, a protein module highly related to the thermogenic phenotype was identified by the weighted gene co-expression network analysis (WGCNA). In summary, the regulation mechanism of floral thermogenesis in N. nucifera involves complex regulatory networks, including TCA cycle metabolism, starch and sucrose metabolism, fatty acid degradation, and ubiquinone synthesis, etc.

DNA-Scaffolded Disulfide Redox Network for Programming Drug-Delivery Kinetics.

In response to specific stimuli, dynamic covalent materials enable the generation of new structures by reversibly forming/breaking chemical bonds, thus showing great potential for application in controlled drug release. However, using dynamic covalent chemistry to program drug-delivery kinetics remains challenging. Herein, an in situ polymerization-generated DNA-scaffolded disulfide redox network (DdiSRN) is reported in which nucleic acids are used as a scaffold for dynamic disulfide bonds. The constructed DdiSRN allows selective release of loading cargos inside cancer cells in response to redox stimuli. Moreover, the density of disulfide bonds in network can be tuned by precise control over their position and number on DNA scaffolds. As a result, drug-delivery kinetics can be programmed with a half-life, t1/2 , decreasing from 8.3 to 4.4 h, thus facilitating keeping an adequate drug concentration within the therapeutic window. Both in vitro and in vivo studies confirm that co-delivery of DOX and siRNA in combination with fast drug release inside cells using this DdiSRN enhances the therapeutic effect on multidrug-resistant cancer. This nontrivial therapeutic platform enabling kinetic control provides a good paradigm for precision cancer medicine.

The transcription factor GATA10 regulates fertility conversion of a two-line hybrid tms5 mutant rice via the modulation of UbL40 expression.

The thermosensitive genic male sterile 5 (tms5) mutation causes thermosensitive genic male sterility in rice (Oryza sativa) through loss of RNase ZS1 function, which influences ubiquitin fusion ribosomal protein L40 (UbL40 ) messenger RNA levels during male development. Here, we used ATAC-seq, combined with analysis of H3K9ac and H3K4me2, to identify changes in accessible chromatin during fertility conversion of the two-line hybrid rice Wuxiang S (WXS) derived from a mutant tms5 allele. Furthermore, RNA-seq and bioinformatic analyses identified specific transcription factors (TFs) in differentially accessible chromatin regions. Among these TFs, only GATA10 targeted UbL40 . Osgata10 knockout mutations, which resulted in low expression of UbL40 and a tendency toward male fertility, confirmed that GATA10 regulated fertility conversion via the modulation of UbL40 . Meanwhile, GATA10 acted as a mediator for interactions with ERF65, which revealed that transcriptional regulation is a complex process involving multiple complexes of TFs, namely TF modules. It appears that the ERF141/MADS7/MADS50/MYB modules affect metabolic processes that control anther and pollen development, especially cell wall formation. Our analysis revealed that these modules directly or indirectly affect metabolic pathway-related genes to coordinate plant growth with proper anther development, and furthermore, that GATA10 regulates fertility conversion via the modulation of UbL40 expression.

Fe, N-doped carbon spheres prepared by electrospinning method as high efficiency oxygen reduction catalyst.

Electrocatalysts for the oxygen reduction reaction (ORR) are crucial in metal-air batteries, fuel cells and other electrochemical devices. In this study, iron and nitrogen co-doped carbon sphere electrocatalysts were synthesized by electrospinning and thermal treatment. According to the results, the catalyst marked as Fe-N/MCS-181 (Fe, N-doped mesoporous carbon spheres, iron nitrate nonahydrate as the iron source) has not only the highest iron content, which reaches up to 0.13%, but also a spherical shape. And its pore sizes are 11 and 35 nm. For the electrochemical performance, the onset potential (E onset) of Fe-N/MCS-181 is -0.018 V, while the half-wave potential (E 1/2) of Fe-N/MCS-181 is -0.145 V, which is better than the commercial Pt/C catalyst (E 1/2 is -0.18 V). The durability of the Fe-N/MCS-181 catalyst is better than commercial Pt/C. After 10 000 s, the retention ratio of current density is 86.4%, while that of the commercial Pt/C catalyst is 84.2%. At the same time, the methanol tolerance of the Fe-N/MCS-181 catalyst is also excellent. After adding methanol, the current density of the Fe-N/MCS-181 catalyst has no obvious change. This study provides an easy method to fabricate a highly efficient and durable Fe, N-doped carbon catalyst for the oxygen reduction reaction.

The Effect of Inner Friction on Concrete Fracture Behavior under Biaxial Compression: A 3D Mesostructure Study.

The mechanical behavior of concrete under biaxial loading condition (especially biaxial compression) is one of the most important indexes to evaluate the quality of concrete. To study the mechanical behavior of concrete under biaxial compression at mesoscale, we adopted our recently developed 3D numerical model based on Voronoi tessellation and cohesive elements. A constitutive model considering the friction effect is used in the model to characterize the fracture behavior of all potential fracture surfaces inside the concrete. A series of numerical experiments with different biaxial compression stress ratios were carried out. It was found that with the increase of the biaxial compression ratio, the proportion of energy increment caused by friction stress increases. The effect of inner friction coefficient on the biaxial relative strength was also investigated, and this kind of study is hard to be carried out through laboratory experiments. The results show that the inner friction coefficient has a great influence on the biaxial relative strength of concrete, and there is a positive correlation between these two parameters. Based on the above rules, a conservative biaxial relative compression strength envelope is obtained by setting the inner friction coefficient as zero.

Circular RNA profiling of the rice photo-thermosensitive genic male sterile line Wuxiang S reveals circRNA involved in the fertility transition.

BACKGROUND: Circular RNAs (circRNAs) are known to play an important role in the regulation of gene expression in eukaryotes. Photo-thermosensitive genic male sterile (PTGMS) is a very important germplasm resource in two-line hybrid rice breeding. Although many circRNAs have been identified in rice (Oryza sativa L.), little is known about the biological roles of circRNAs in the fertility transition of the PTGMS rice line. RESULTS: In the present study, RNA-sequencing libraries were constructed from the young panicles of the Wuxiang S sterile line rice (WXS (S)) and its fertile line rice (WXS (F)) at three development stages with three biological replicates. A total of 9994 circRNAs were obtained in WXS rice based on high-throughput strand-specific RNA sequencing and bioinformatic approaches, of which 5305 were known circRNAs and 4689 were novel in rice. And 14 of 16 randomly selected circRNAs were experimentally validated with divergent primers. Our results showed that 186 circRNAs were significantly differentially expressed in WXS (F) compared with WXS (S), of which 97, 87 and 60 circRNAs were differentially expressed at the pollen mother cell (PMC) formation stage (P2), the meiosis stage (P3) and the microspore formation stage (P4), respectively. Fertility specific expression patterns of eight circRNAs were analysis by qRT-PCR. Gene ontology (GO) and KEGG pathway analysis of the parental genes of differentially expressed circRNAs (DECs) revealed that they mainly participated in various biological processes such as development, response to stimulation, hormonal regulation, and reproduction. Furthermore, 15 DECs were found to act as putative miRNA sponges to involved in fertility transition in PTGMS rice line. CONCLUSION: In the present study, the abundance and characteristics of circRNAs were investigated in the PTGMS rice line using bioinformatic approaches. Moreover, the expression patterns of circRNAs were different between WXS (F) and WXS (S). Our findings primarily revealed that circRNAs might be endogenous noncoding regulators of flower and pollen development, and were involved in the fertility transition in the PTGMS rice line, and guide the production and application of two-line hybrid rice.

[Study on interaction between ginsenosides Rg_1,Rb_1 and Ro and bovine serum albumin].

Small molecules with physiological or pharmacological activities need to interact with biological macromolecules in order to function in the body. As the protein with the highest proportion of plasma protein,serum albumin is the main protein binding to various endogenous or exogenous small molecules. Serum albumin interacts with small molecules in a reversible non-covalent manner and transports small molecules to target sites. Bovine serum albumin( BSA) is an ideal target protein for drug research because of its low cost and high homology with human serum albumin. The research on the interaction between drugs and BSA has become a hotspot in the fields of pharmacy,medicine,biology and chemistry. In this research,molecular docking method was used to study the interaction between three small ginsenosides with high pharmacological value( Rg_1,Rb_1,Ro) and bovine serum albumin( BSA),and the binding mode information of three ginsenosides interacting with BSA was obtained. The results of molecular docking showed that ginsenosides and amino acid residues in the active pocket of proteins could be combined by hydrophobic action,hydrogen bonding and electrostatic action. The interaction between small ginsenosides and bovine serum albumin is not the only form,and their interaction has many forms of force. The interaction between these molecules and various weak forces is the key factor for the stability of the complex. The results of this study can provide the structural information of computer simulation for the determination of the interaction patterns between active components and proteins of ginseng.

Fracture Behavior and Energy Analysis of 3D Concrete Mesostructure under Uniaxial Compression.

In order to investigate the fracture behavior of concrete mesostructure and reveal the inner failure mechanisms which are hard to obtain from experiments, we develop a 3D numerical model based on the Voronoi tessellation and cohesive elements. Specifically, the Voronoi tessellation is used to generate the aggregates, and the cohesive elements are applied to the interface transition zone (ITZ) and the potential fracture surfaces in the cement matrix. Meanwhile, the mechanical behavior of the fracture surfaces is described by a modified constitutive which considers the slips and friction between fracture surfaces. Through comparing with the experiments, the simulated results show that our model can accurately characterize the fracture pattern, fracture propagation path, and mechanical behaviors of concrete. In addition, we found that the friction on the loading surfaces has a significant effect on the fracture pattern and the strength of concrete. The specimens with low-friction loading surfaces are crushed into separate fragments whereas those with high-friction loading surfaces still remain relatively complete. Also, the strength of concrete decreases with the increase of the specimen height in the high friction-loading surfaces condition. Further, the energy analysis was applied to estimate the restraint impact of loading surfaces restraint on the compressive strength of concrete. It shows that the proportion of the friction work increases with the increase of the restraint degree of loading surfaces, which finally causes a higher compressive strength. Generally, based on the proposed model, we can characterize the complicated fracture behavior of concrete mesostructure, and estimate the inner fracture mode through extracting and analyzing the energies inside the cohesive elements.

Extracting Cross-Sectional Clinical Images Based on Their Principal Axes of Inertia.

Cross-sectional imaging is considered the gold standard in diagnosing a range of diseases. However, despite its widespread use in clinical practice and research, no widely accepted method is available to reliably match cross-sectional planes in several consecutive scans. This deficiency can impede comparison between cross-sectional images and ultimately lead to misdiagnosis. Here, we propose and demonstrate a method for finding the same imaging plane in images obtained during separate scanning sessions. Our method is based on the reconstruction of a "virtual organ" from which arbitrary cross-sectional images can be extracted, independent of the axis orientation in the original scan or cut; the key is to establish unique body coordinates of the organ from its principal axes of inertia. To verify our method a series of tests were performed, and the same cross-sectional plane was successfully extracted. This new approach offers clinicians access, after just a single scanning session, to the morphology and structure of a lesion through cross-sectional images reconstructed along arbitrary axes. It also aids comparable detection of morphological and structural changes in the same imaging plane from scans of the same patient taken at different times-thus potentially reducing the misdiagnosis rate when cross-sectional images are interpreted.