Development of a synthetic oxytetracycline-inducible expression system for streptomycetes using de novo characterized genetic parts

Precise control of gene expression using exogenous factors is of great significance. To develop ideal inducible expression systems for streptomycetes, new genetic parts, oxytetracycline responsive repressor OtrR, operator otrO, and promoter otrBp from Streptomyces rimosus, were selected de novo and characterized in vivo and in vitro. OtrR showed strong affinity to otrO (KD = 1.7 × 10–10 M) and oxytetracycline induced dissociation of the OtrR/DNA complex in a concentration-dependent manner. On the basis of these genetic parts, a synthetic inducible expression system Potr* was optimized. Induction of Potr* with 0.01–4 μM of oxytetracycline triggered a wide-range expression level of gfp reporter gene in different Streptomyces species. Benchmarking Potr* against the widely used constitutive promoters ermE* and kasOp* revealed greatly enhanced levels of expression when Potr* was fully induced. Finally, Potr* was used as a tool to activate and optimize the expression of the silent jadomycin biosynthetic gene cluster in Streptomyces venezuelae. Altogether, the synthetic Potr* presents a new versatile tool for fine-tuning gene expression in streptomycetes

Micropore evolution in additively manufactured aluminum alloys under heat treatment and inter-layer rolling

The application of wire + arc additively manufactured (WAAM) aluminum alloys has been restricted by the porosity defect, which is generally detrimental to the mechanical properties. Suppressing of micropores in the WAAM components has attracted considerable attention in recent years. Inter-layer rolling was introduced to eliminate micropores during the WAAM deposition of the Al–Cu6.3 and Al–Mg4.5 alloys. The distribution characteristics and individual morphology of micropores were revealed by the X-ray diffraction tomography. Key findings demonstrated that the number, volume, size, and roundness of micropores in rolled alloys decreased similarly with increasing loads, eventually achieving a density of over 99.9%. After the heat treatment, the homogeneous distribution of fine (around 5.3 μm) and spherical (0.70–0.74) micropores was realized in the 45 kN rolled alloys. All the evaluated indicators of micropores in the 45 kN rolled + heat treated alloys were superior to the post-deposition heat treated state. The evolution mechanisms include the reprecipitation of hydrogen pores, formation of vacant voids, and re-opening of unclosed pores. The hybrid technique of WAAM + rolling + heat treatment has great potential in promoting mechanical properties of WAAM alloys. The results will provide a theoretical guidance for the design of high-performance WAAM aluminum alloy components

Microstructure, defects, and mechanical properties of wire + arc additively manufactured AlCu4.3-Mg1.5 alloy

The wire with a composition of AlCu4.3%Mg1.5% was customized and used to deposit the WAAM alloy with the power source of cold metal transfer. The microstructure, defect, and mechanical properties of the as-deposited and heat-treated WAAM alloys were studied. Key findings demonstrated that the microstructure of the as-deposited alloy was characterized by a hierarchical distribution of dendrites, equiaxed grains, and a slight number of columnar grains. The volume fraction of the network-like scattered coarse particles of second phases θ + S reduced by 95% after the T6 heat treatment. With an average microhardness of 161.4 HV, the mean yield strength and ultimate tensile strength of the WAAM alloy increased by 116% and 66% achieving 399 MPa and 485 MPa in the horizontal direction after heat treatment. The precipitation of a high density of needle-shaped metastable S′ precipitates was responsible for the significantly enhanced mechanical properties. However, this WAAM alloy has exhibited an anisotropic tensile property. A considerable number of sharp-angled defects like linear and chain-like micropores, which generally depress the mechanical properties, were formed in the WAAM alloys

Coating of manganese functional polyetheretherketone implants for osseous interface integration

Polyetheretherketone (PEEK) has been used extensively in biomedical engineering and it is highly desirable for PEEK implant to possess the ability to promote cell growth and significant osteogenic properties and consequently stimulate bone regeneration. In this study, a manganese modified PEEK implant (PEEK-PDA-Mn) was fabricated via polydopamine chemical treatment. The results showed that manganese was successfully immobilized on PEEK surface, and the surface roughness and hydrophilicity significantly improved after surface modification. Cell experiments in vitro demonstrated that the PEEK-PDA-Mn possesses superior cytocompatibility in cell adhesion and spread. Moreover, the osteogenic properties of PEEK-PDA-Mn were proved by the increased expression of osteogenic genes, alkaline phosphatase (ALP), and mineralization in vitro. Further rat femoral condyle defect model was utilized to assess bone formation ability of different PEEK implants in vivo. The results revealed that the PEEK-PDA-Mn group promoted bone tissue regeneration in defect area. Taken together, the simple immersing method can modify the surface of PEEK, giving outstanding biocompatibility and enhanced bone tissue regeneration ability to the modified PEEK, which could be applied as an orthopedic implant in clinical

RNA-Sequencing Analyses Demonstrate the Involvement of Canonical Transient Receptor Potential Channels in Rat Tooth Germ Development

Tooth development depends on multiple molecular interactions between the dental epithelium and mesenchyme, which are derived from ectodermal and ectomesenchymal cells, respectively. We report on a systematic RNA sequencing analysis of transcriptional expression levels from the bud to hard tissue formation stages of rat tooth germ development. We found that GNAO1, ENO1, EFNB1, CALM1, SIAH2, ATP6V0A1, KDELR2, GTPBP1, POLR2C, SORT1, and members of the canonical transient receptor potential (TRPC) channel family are involved in tooth germ development. Furthermore, Cell Counting Kit 8 (CCK8) and Transwell migration assays were performed to explore the effects of these differentially expressed genes (DEGs) on the proliferation and migration of dental pulp stem cells. Immunostaining revealed that TRPC channels are expressed at varying levels during odontogenesis. The identified genes represent novel candidates that are likely to be vital for rat tooth germ development. Together, the results provide a valuable resource to elucidate the gene regulatory mechanisms underlying mammalian tooth germ development

A Hyper-Chaotically Encrypted Robust Digital Image Watermarking Method with Large Capacity Using Compress Sensing on a Hybrid Domain

The digital watermarking technique is a quite promising technique for both image copyright protection and secure transmission. However, many existing techniques are not as one might have expected for robustness and capacity simultaneously. In this paper, we propose a robust semi-blind image watermarking scheme with a high capacity. Firstly, we perform a discrete wavelet transformation (DWT) transformation on the carrier image. Then, the watermark images are compressed via a compressive sampling technique for saving storage space. Thirdly, a Combination of One and Two-Dimensional Chaotic Map based on the Tent and Logistic map (TL-COTDCM) is used to scramble the compressed watermark image with high security and dramatically reduce the false positive problem (FPP). Finally, a singular value decomposition (SVD) component is used to embed into the decomposed carrier image to finish the embedding process. With this scheme, eight 256×256 grayscale watermark images are perfectly embedded into a 512×512 carrier image, the capacity of which is eight times over that of the existing watermark techniques on average. The scheme has been tested through several common attacks on high strength, and the experiment results show the superiority of our method via the two most used evaluation indicators, normalized correlation coefficient (NCC) values and the peak signal-to-noise ratio (PSNR). Our method outperforms the state-of-the-art in the aspects of robustness, security, and capacity of digital watermarking, which exhibits great potential in multimedia application in the immediate future