Sulfated hyaluronic acid/collagen-based biomimetic hybrid nanofiber skin for diabetic wound healing: Development and preliminary evaluation.

Diabetic foot ulcers (DFUs) are one of the most serious and devastating complication of diabetes, manifesting as foot ulcers and impaired wound healing in patients with diabetes mellitus. To solve this problem, sulfated hyaluronic acid (SHA)/collagen-based nanofibrous biomimetic skins was developed and used to promote the diabetic wound healing and skin remodeling. First, SHA was successfully synthetized using chemical sulfation and incorporated into collagen (COL) matrix for preparing the SHA/COL hybrid nanofiber skins. The polyurethane (PU) was added into those hybrid scaffolds to make up the insufficient mechanical properties of SHA/COL nanofibers, the morphology, surface properties and degradation rate of hybrid nanofibers, as well as cell responses upon the nanofibrous scaffolds were studied to evaluate their potential for skin reconstruction. The results demonstrated that the SHA/COL, SHA/HA/COL hybrid nanofiber skins were stimulatory of cell behaviors, including a high proliferation rate and maintaining normal phenotypes of specific cells. Notably, SHA/COL and SHA/HA/COL hybrid nanofibers exhibited a significantly accelerated wound healing and a high skin remodeling effect in diabetic mice compared with the control group. Overall, SHA/COL-based hybrid scaffolds are promising candidates as biomimetic hybrid nanofiber skin for accelerating diabetic wound healing.

Cable bacteria accelerate the anaerobic removal of pyrene in black odorous river sediments.

Cable bacteria play an essential role in biogeochemical processes in sediments by long-distance electron transport (LDET). A potential relationship has been found between cable bacteria and organic contaminant removal; however, the mechanisms remain unclear. In this study, the response of cable bacteria to pyrene was investigated in sediments with and without pyrene, and the effect of cable bacteria on pyrene removal was explored by connecting and blocking the paths of cable bacteria to the suboxic zones. The results showed that pyrene significantly influenced the microbial community structure and the composition of cable bacteria. The pyrene removal efficiencies significantly increased with the enrichment of cable bacteria, while sulfur-reducing microorganisms and aromatic compound degraders were also significantly enriched and correlated with cable bacteria abundance. Metagenomic analysis showed that cable bacteria have a potential LDET-bound acetate/formate respiratory pathway to gain energy. The presence of pyrene probably selects and enriches cable bacteria with a high tolerance to organic contaminants and changes the related functional microbial community, leading to the acceleration of pyrene removal. This study provides new insights into the interaction mechanisms between contaminants and cable bacteria, shedding light on the applications of cable bacteria in the bioremediation of contaminants in sediments.

Construction of enzyme-laden vascular scaffolds based on hyaluronic acid oligosaccharides-modified collagen nanofibers for antithrombosis and in-situ endothelialization of tissue-engineered blood vessels.

The current use of synthetic grafts often yields low patency in the reconstruction of small-diameter blood vessels owing to the deposition of thrombi and imperfect coverage of the endothelium on the graft lumen. Therefore, the design of vascular scaffolds with antithrombotic performance and endothelialization is greatly required. Herein, we developed an enzyme-laden scaffold based on hyaluronic acid oligosaccharides-modified collagen nanofibers (labeled HA-COL) to improve the anti-platelet capacity and endothelialization of vascular grafts. In this study, HA-COL nanofibers not only encouraged the endothelialization of vascular scaffolds, but acted as an antiplatelet enzyme-laden platform. Apyrase (Apy) and 5'-nucleotidase (5'-NT) were covalently grafted onto the nanofibers, which in turn converted the platelet-sensitive substance: adenosine diphosphate (ADP) into adenosine monophosphate (AMP) and adenosine, thereby, improving the antithrombotic performance of the scaffolds. Notably, the catalytic end-product: adenosine would work in coordination with HA-COL to synergistically enhance the endothelialization of the vascular scaffolds. The results demonstrated that the enzyme-laden scaffolds maintained catalytic performance, reduced platelet adhesion and aggregation, and guaranteed higher patency after 1-month in situ transplantation. Moreover, these scaffolds showed optimal cytocompatibility, tissue compatibility, scaffold biodegradability and tissue regenerative capability during in vivo implantation. Overall, these engineered vascular scaffolds demonstrated their capacity for endothelialization and antithrombotic performance, suggesting their potential for small-diameter vascular tissue engineering applications. STATEMENT OF SIGNIFICANCE: Considering the critical problems in small-diameter vascular reconstruction, the enzyme-laden vascular scaffolds were prepared for improving in-situ endothelialization and antithrombotic performances of artificial blood vessels. The electrospun HA-COL nanofibers were used as the main matrix materials, which provided favorable structural templates for the regeneration of vasculature and functioned as a platform for the loading of enzymes. The enzyme-laden scaffolds with the biomimetic cascading reaction would convert ADP into adenosine, thereby, decreasing the sensitivity of platelets and improving the antithrombotic performance of tissue-engineered blood vessels (TEBVs). The nanofibrous scaffolds exhibited optimal cytocompatibility, tissue compatibility and regenerative capability, working together with catalytic products of dual-enzyme reaction that would synergistically contribute to TEBVs endothelialization. This study provides a new method for the improvement of in-situ endothelialization of small-diameter TEBVs while qualified with antithrombotic performance.

Multi-scale Fusion of Stretched Infrared and Visible Images.

Infrared (IR) band sensors can capture digital images under challenging conditions, such as haze, smoke, and fog, while visible (VIS) band sensors seize abundant texture information. It is desired to fuse IR and VIS images to generate a more informative image. In this paper, a novel multi-scale IR and VIS images fusion algorithm is proposed to integrate information from both the images into the fused image and preserve the color of the VIS image. A content-adaptive gamma correction is first introduced to stretch the IR images by using one of the simplest edge-preserving filters, which alleviates excessive luminance shifts and color distortions in the fused images. New contrast and exposedness measures are then introduced for the stretched IR and VIS images to achieve weight matrices that are more in line with their characteristics. The IR and luminance components of the VIS image in grayscale or RGB space are fused by using the Gaussian and Laplacian pyramids. The RGB components of the VIS image are finally expanded to generate the fused image if necessary. Comparisons experimentally demonstrate the effectiveness of the proposed algorithm to 10 different state-of-the-art fusion algorithms in terms of computational cost and quality of the fused images.

Precise Regulation of Differential Transcriptions of Various Catabolic Genes by OdcR via a Single Nucleotide Mutation in the Promoter Ensures the Safety of Metabolic Flux.

Synergistic regulation of the expression of various genes in a catabolic pathway is crucial for the degradation, survival, and adaptation of microorganisms in polluted environments. However, how a single regulator accurately regulates and controls differential transcriptions of various catabolic genes to ensure metabolic safety remains largely unknown. Here, a LysR-type transcriptional regulator (LTTR), OdcR, encoded by the regulator gene odcR, was confirmed to be essential for 3,5-dibromo-4-hydroxybenozate (DBHB) catabolism and simultaneously activated the transcriptions of a gene with unknown function, orf419, and three genes, odcA, odcB, and odcC, involved in the DBHB catabolism in Pigmentiphaga sp. strain H8. OdcB further metabolized the highly toxic intermediate 2,6-dibromohydroquinone, which was produced from DBHB by OdcA. The upregulated transcriptional level of odcB was 7- to 9-fold higher than that of orf419, odcA, or odcC in response to DBHB. Through an electrophoretic mobility shift assay and DNase I footprinting assay, DBHB was found to be the effector and essential for OdcR binding to all four promoters of orf419, odcA, odcB, and odcC. A single nucleotide mutation in the regulatory binding site (RBS) of the promoter of odcB (TAT-N11-ATG), compared to those of odcA/orf419 (CAT-N11-ATG) and odcC (CAT-N11-ATT), was identified and shown to enable the significantly higher transcription of odcB. The precise regulation of these genes by OdcR via a single nucleotide mutation in the promoter avoided the accumulation of 2,6-dibromohydroquinone, ensuring the metabolic safety of DBHB. IMPORTANCE Prokaryotes use various mechanisms, including improvement of the activity of detoxification enzymes, to cope with toxic intermediates produced during catabolism. However, studies on how bacteria accurately regulate differential transcriptions of various catabolic genes via a single regulator to ensure metabolic safety are scarce. This study revealed a LysR-type transcriptional activator, OdcR, which strongly activated odcB transcription for the detoxification of the toxic intermediate 2,6-dibromohydroquinone and slightly activated the transcriptions of other genes (orf419, odcA, and odcC) for 3,5-dibromo-4-hydroxybenozate (DBHB) catabolism in Pigmentiphaga sp. strain H8. Interestingly, the differential transcription/expression of the four genes, which ensured the metabolic safety of DBHB in cells, was determined by a single nucleotide mutation in the regulatory binding sites of the four promoters. This study describes a new and ingenious regulatory mode of ensuring metabolic safety in bacteria, expanding our understanding of synergistic transcriptional regulation in prokaryotes.

New injectable chitosan-hyaluronic acid based hydrogels for hemostasis and wound healing.

It is a challenge to develop hemostatic and wound dressings that are used for irregular shape and deep wound. Herein, a series of novel N-succinyl chitosan-oxidized hyaluronic acid based (NSC-OHA-based) hydrogels were fabricated, while calcium ions (Ca2+) and/or four-armed amine-terminated poly(ethylene glycol) (4-arm-PEG-NH2, labeled as PEG1) were introduced to regulate the mechanical behavior and bioactivities. We found all NSC-OHA-based hydrogels displayed self-healing and injectable performances. Besides, the addition of Ca2+ or PEG1 exhibited a positive effect on the adjustable mechanical behavior of hydrogels, providing the possibility to meet different mechanical requirements. Furthermore, Ca2+ or PEG1 significantly improved the biocompatibility, hemostasis and wound healing abilities of NSC-OHA hydrogel. Notably, compared with the commercial hemostatic agent (Arista™), hydrogels containing Ca2+ showed comparable hemostatic effects and significantly accelerated wound healing. Overall, the calcium-containing NSC-OHA hydrogels are promising for hemostasis and accelerating wound healing.

Hyaluronic acid oligosaccharide-collagen mineralized product and aligned nanofibers with enhanced vascularization properties in bone tissue engineering.

Considering the structural complexity of natural bone and the limitations of current treatment options, designing a biomimetic and functional tissue-engineered bone graft has been an urgent need for the replacement and regeneration of defected bone tissue. In light of the cell recruitment to the defect region, scaffold-guided bone tissue engineering has proven to be a viable strategy that is poised to deliver effective osseointegration and vascularization during bone remodeling. Herein, we provide an engineered bone scaffold based on aligned poly(lactic-co-glycolide) (PLGA) nanofibers incorporated with hyaluronic acid oligosaccharide-collagen mineralized microparticles (labeled oHA-Col/HAP) to guide the cell-specific orientation and osseointegration in bone healing. The aligned nanofibers were successfully prepared by a custom-made rotating mandrel with separating railings and HAs-Col/HAP mineralized microparticles were uniformly distributed in the composite scaffolds that acted as temporary templates for bone remodeling. The morphology, physicochemical properties and tensile strength of the scaffolds were characterized, the cell responses and in vivo biocompatibility and biodegradability of the scaffolds were also studied to evaluate the potential for bone tissue engineering. The experimental results illustrated that such anisotropic scaffolds loaded with oHA-Col/HAP microparticles mediated cell orderly arrangement conducive to the migration and recruitment of osseointegration-related cells and were stimulatory of cell proliferation. Those oHA-Col/HAP@PLGA scaffolds exhibited ideal biocompatibility and tissue regenerative capacity in vivo through a higher expression of vascularization-related genes. Overall, the novel engineered bone scaffold promises to serve as alternative candidates for bone tissue engineering applications.

The fate of erythromycin in soils and its effect on soil microbial community structure.

Erythromycin is one of the most commonly used macrolide antibiotics. However, little is known currently about the environmental behavior and fate of erythromycin in soils. Here erythromycin was 14C-labeled to investigate its degradation, mineralization and bound residues (BRs) in three typical agricultural soils. Results indicated the fate of 14C-erythromycin in soils varied greatly with soils types. Erythromycin was rapidly mineralized in black soil (BS) and fluvo-aquic soil (FS), whereas it rapidly formed large amounts of BRs in red soil (RS) with slow mineralization. At 120 d, about 90% of the introduced 14C-erythromycin was mineralized as 14CO2 in BS and FS, but only 30% in RS. There was still a certain proportion of BRs in all soils, especially in RS, up to 50%. Erythromycin residues (ERs) may be underestimated if its residues are only assessed by extractable residues. We recommend to include a practical silylation procedure to quantify Type I BRs in regular erythromycin residue monitoring, which can be used as signal of the need to initiate further laboratory BRs experiments. The degradation of erythromycin was mainly attributed to soil microorganisms, which promote erythromycin mineralization and lead to the re-release of BRs. Microbial analysis showed that erythromycin persisted longer in soils with lower microbial diversity and richness. Erythromycin at 2.5 mg kg-1 showed no significant impact on soil microbial diversity in all treatments, but caused changes in soil community composition. This study provides a reference for scientific evaluation and pollution remediation of erythromycin in soils.

Bioaugmentation of Atrazine-Contaminated Soil With Paenarthrobacter sp. Strain AT-5 and Its Effect on the Soil Microbiome.

Atrazine, a triazine herbicide, is widely used around the world. The residue of atrazine due to its application in the fore-rotating crop maize has caused phytotoxicity to the following crop sweet potato in China. Bioaugmentation of atrazine-contaminated soil with atrazine-degrading strains is considered as the most potential method to remove atrazine from soil. Nevertheless, the feasibility of bioaugmentation and its effect on soil microbiome still need investigation. In this study, Paenarthrobacter sp. AT-5, an atrazine-degrading strain, was inoculated into agricultural soils contaminated with atrazine to investigate the bioaugmentation process and the reassembly of the soil microbiome. It was found that 95.9% of 5 mg kg-1 atrazine was removed from the soils when inoculated with strain AT-5 with 7 days, and the phytotoxicity of sweet potato caused by atrazine was significantly alleviated. qRT-PCR analysis revealed that the inoculated strain AT-5 survived well in the soils and maintained a relatively high abundance. The inoculation of strain AT-5 significantly affected the community structure of the soil microbiome, and the abundances of bacteria associated with atrazine degradation were improved.

Reducing the Environmental Risk of Chlorpyrifos Application through Appropriate Agricultural Management: Evidence from Carbon-14 Tracking.

Chlorpyrifos (CPF) is one of the most critical insecticides in the world. However, many countries are gradually banning its use due to its reported hazardous impacts on humans. This study explored the possibility of reducing the environmental risk of CPF through appropriate agricultural management practices. Results showed that the environmental risk of CPF is lower under drainage conditions because there is more mineralization and less bound residues (BRs) than under submerged conditions. Bioaugmentation significantly enhanced the CPF mineralization and inhibited the formation of CPF-BRs. Biochar adsorbed CPF and thus reduced its bioavailability, but it could not completely eliminate the toxicity of CPF. In addition, bioaugmentation did not significantly affect the native microbial community of CPF-contaminated soil, suggesting its safety in reducing the environmental risk of CPF. The study indicated that the environmental risk of CPF could be reduced by appropriate agricultural management such as water management, bioaugmentation, soil biochar amendment, and selecting suitable soil types.

A two-component monooxygenase for continuous denitration and dechlorination of chlorinated 4-nitrophenol in Ensifer sp. strain 22-1.

The environmental fates of chlorinated 4-nitrophenols, 2,6-dichloro-4-nitrophenol (2,6-DCNP) and 2-chloro-4-nitrophenol (2C4NP), mediated via microbial catabolism have attracted great attention due to their high toxicity and persistence in the environment. In this study, a strain of Ensifer sp. 22-1 that was capable of degrading both 2,6-DCNP and 2C4NP was isolated from a halogenated aromatic-contaminated soil sample. A gene cluster cnpBADCERM was predicted to be involved in the catabolism of 2,6-DCNP and 2C4NP based on genome sequence analysis. A two-component monooxygenase CnpAB, composed of an oxygenase component (CnpA) and a reductase component (CnpB), was confirmed to catalyze the continuous denitration and dechlorination of 2,6-DCNP and 2C4NP to 6-chlorohydroxyquinol (6-CHQ) and hydroxyquinol (HQ), respectively. Knockout of cnpA resulted in the complete loss of the capacity for strain 22-1 to degrade 2,6-DCNP and 2C4NP. Homologous modeling and docking showed that Val155~Ala159, Phe206~Pro209 and Phe446~Arg461 of CnpA participated in the formation of the FAD-binding pocket, and Arg101, Val155 and Asn447 formed hydrogen bonds with 2,6-DCNP/2C4NP in the substrate-binding pocket. This work characterized a new two-component monooxygenase for 2,6-DCNP and 2C4NP, and enriched our understanding of the degradation mechanism of chlorinated nitrophenols (CNPs) by microorganisms.

Hyaluronic acid oligosaccharides modified mineralized collagen and chitosan with enhanced osteoinductive properties for bone tissue engineering.

In this study, we prepared a biomimetic hyaluronic acid oligosaccharides (oHAs)-based composite scaffold to develop a bone tissue-engineered scaffold for stimulating osteogenesis and endothelialization. The functional oHAs products were firstly synthesized, namely collagen/hyaluronic acid oligosaccharides/hydroxyapatite (Col/oHAs/HAP), chitosan/hyaluronic acid oligosaccharides (CTS/oHAs), and then uniformly distributed in poly (lactic-co-glycolic acid) (PLGA) solution followed by freeze-drying to obtain three-dimensional interconnected scaffolds as temporary templates for bone regeneration. The morphology, physicochemical properties, compressive strength, and degradation behavior of the fabricated scaffolds, as well as in vitro cell responses seeded on these scaffolds and in vivo biocompatibility, were investigated to evaluate the potential for bone tissue engineering. The results indicated that the oHAs-based scaffolds can promote the attachment of endothelial cells, facilitate the osteogenic differentiation of MC3T3-E1 and BMSCs, and have ideal biocompatibility and tissue regenerative capacity, suggesting their potential to serve as alternative candidates for bone tissue engineering applications.

Enhanced mineralization of chlorpyrifos bound residues in soil through inoculation of two synergistic degrading strains.

Bioaugmentation methods are frequently employed for pesticide pollution remediation; however, it is not clear whether the introduced bacteria affect the pesticide bound residue (BRs) composition and whether the BRs can be catabolized by the introduced strains. This study aimed at answering these questions by using 14C-chlorpyrifos (14C-CPF) and two CPF-degrading strains (Pseudomonas sp. DSP-1 and Cupriavidus sp. P2). The results showed that the BRs can be up to 83.0%, and that the CPF-BRs formed can be further transformed into 14CO2 by the strains. Indeed, the microbial inoculation can increase the CPF mineralization by 1.0-22.1 times and can decrease the BRs by up to ~50% of the control (on day 20). Compared with the control without bioaugmentation, microbial inoculation enhanced the release of BRs by 2.2-18.0 times. Adding biochar to the soil can greatly inhibit CPF mineralization and maintain the BR content at a relatively stable level. The CPF residue can affect the composition of the indigenous soil microbial community, but the introduction of bacteria for remediation did not have a significant effect. The results indicate that Pseudomonas sp. DSP-1 and Cupriavidus sp. P2 are useful for remediating both CPF extractable and bound residues.

Fabrication and assessment of chondroitin sulfate-modified collagen nanofibers for small-diameter vascular tissue engineering applications.

Chondroitin sulfate (ChS) has shown promising results in promoting cell proliferation and antithrombogenic activity. To engineered develop a dual-function vascular scaffold with antithrombosis and endothelialization, ChS was tethered to collagen to accelerate the growth of endothelial cells and prevent platelet activation. First, ChS was used to conjugate with collagen to generate glycosylated products (ChS-COL) via reductive amination. Then, the fabricated ChS-COL conjugates were electrospun into nanofibers and their morphologies and physicochemical characteristics, cell-scaffold responses and platelet behaviors upon ChS-COL nanofibers were comprehensively characterized to evaluate their potential use for small-diameter vascular tissue-engineered scaffolds. The experimental results demonstrated that the ChS modified collagen electrospun nanofibers were stimulatory of endothelial cell behavior, alleviated thrombocyte activation and maintained an antithrombotic effect in vivo in 10-day post-transplantation. The ChS-COL scaffolds encouraged rapid endothelialization, thus probably ensuring the antithrombotic function in long-term implantation, suggesting their promise for small-diameter vascular tissue engineering applications.

Molecular Sizes and Antibacterial Performance Relationships of Flexible Ionic Liquid Derivatives.

Cationic agents, such as ionic liquids (ILs)-based species, have broad-spectrum antibacterial activities. However, the antibacterial mechanisms lack systematic and molecular-level research, especially for Gram-negative bacteria, which have highly organized membrane structures. Here, we designed a series of flexible fluorescent diketopyrrolopyrrole-based ionic liquid derivatives (ILDs) with various molecular sizes (1.95-4.2 nm). The structure-antibacterial activity relationships of the ILDs against Escherichia coli (E. coli) were systematically studied thorough antibacterial tests, fluorescent tracing, morphology analysis, molecular biology, and molecular dynamics (MD) simulations. ILD-6, with a relatively small molecular size, could penetrate through the bacterial membrane, leading to membrane thinning and intracellular activities. ILD-6 showed fast and efficient antimicrobial activity. With the increase of molecular sizes, the corresponding ILDs were proven to intercalate into the bacterial membrane, leading to the destabilization of the lipid bilayer and further contributing to the antimicrobial activities. Moreover, the antibacterial activity of ILD-8 was limited, where the size was not large enough to introduce significant membrane disorder. Relative antibacterial experiments using another common Gram-negative bacteria, Pseudomonas aeruginosa (PAO1), further confirmed the proposed structure-antibacterial activity relationships of ILDs. More impressively, both ILD-6 and ILD-12 displayed significant in vivo therapeutic effects on the PAO1-infected rat model, while ILD-8 performed poorly, which confirmed the antibacterial mechanism of ILDs and proved their potentials for future application. This work clarifies the interactions between molecular sizes of ionic liquid-based species and Gram-negative bacteria and will provide useful guidance for the rational design of high-performance antibacterial agents.

Microbial catabolism of lindane in distinct layers of acidic paddy soils combinedly affected by different water managements and bioremediation strategies.

The environmental fate of the recalcitrant organic chlorine insecticide lindane and its removal from contaminated soils are still of great concern. However, the key factors influencing microbial removal of lindane from paddy soils with intermittent flooding and draining remain largely unknown. Here, we conducted laboratory experiments to investigated lindane biodegradation in different layers of typical acidic paddy soils under different water managements and bioremediation strategies, together with the changes of functional bacterial consortium, key genes and metabolic pathways. It was found that under flooded conditions, lindane spiking significantly stimulated the growth of some bacterial genera with potential anaerobic catabolic functions in both top- (0-20 cm depth) and subsoil (20-40 cm depth), leading to the shortest half-life of lindane with 7.6-9.0 d in the topsoil. In contrary, lindane spiking dramatically stimulated the growth of bacterial members with aerobic catabolic functions under drained conditions, exhibiting half-lives of lindane with 85-131 d and 18-23 d in the top- and subsoil, respectively. Overall, biostimulation coupled with flooding management would be the better combination for increased lindane bioremediation. Functional genes involved in lindane degradation and retrieved from metagenomic data further supported the anaerobic and aerobic biodegradation of lindane under flooded and drained conditions, respectively. Moreover, the integrated network analysis suggested water management and organic matter were the primary factors shaped the assembly of functional bacteria in lindane degradation, among which Clostridium and Rhodanobacter were the key anaerobic and aerobic functional genera, respectively. Taken together, our study provides a comprehensive understanding of lindane biodegradation in distinct layers of acidic paddy soils that were combinedly affected by different water managements and bioremediation strategies.

Design and comprehensive assessment of a biomimetic tri-layer tubular scaffold via biodegradable polymers for vascular tissue engineering applications.

Considering the structural complexity of the native artery wall and the limitations of current treatment strategies, developing a biomimetic tri-layer tissue-engineered vascular graft is a major developmental direction of vascular tissue regeneration. Biodegradable polymers exhibit adequate mechanical characteristics and feasible operability, showing potential prospects in the construction of tissue engineering scaffold. Herein, we present a bio-inspired tri-layer tubular graft using biodegradable polymers to simulate natural vascular architecture. The inner layer made of polycaprolactone (PCL) nanofiber possesses high tensile strength and contributed to endothelial cell adhesion and proliferation. The middle layer consisted of poly(lactic-co-glycolide) (PLGA) with a three-dimensional porous structure is appropriate for vascular smooth muscle cells (SMCs) penetration. The polyurethane (PU) was selected to be the outer layer, aiming to hold the entire tubular structure, suggesting superior mechanical properties and ideal biocompatibility. Adhesion between independent layers is achieved by thermal crosslinking. The compliance, burst pressure and suture retention force of the tubular scaffold were 2.50 ± 1.60%, 2737.73 ± 583.41 mmHg and 13.06 ± 1.89 N, respectively. The in vivo study of subcutaneous implantation for 8 weeks demonstrated the biomimetic tri-layer vascular graft could maintain intimal integrity, cell infiltration, collagen deposition and scaffold biodegradation. Overall, the biomimetic tri-layer vascular graft promises to be a potential candidate for vascular replacement and regeneration.

Pigmentiphaga sp. Strain D-2 Uses a Novel Amidase To Initiate the Catabolism of the Neonicotinoid Insecticide Acetamiprid.

Acetamiprid, a chloronicotinyl neonicotinoid insecticide, is among the most commonly used insecticides worldwide, and its environmental fate has caused considerable concern. The compound 1-(6-chloropyridin-3-yl)-N-methylmethanamine (IM 1-4) has been reported to be the main intermediate during acetamiprid catabolism in microorganisms, honeybees, and spinach. However, the molecular mechanism underlying the hydrolysis of acetamiprid to IM 1-4 has not yet been elucidated. In this study, a novel amidase (AceAB) that initially hydrolyzes the C-N bond of acetamiprid to generate IM 1-4 was purified and characterized from the acetamiprid-degrading strain Pigmentiphaga sp. strain D-2. Based on peptide profiling of the purified AceAB and the draft genome sequence of strain D-2, aceA (372 bp) and aceB (2,295 bp), encoding the α and ß subunits of AceAB, respectively, were cloned and found to be necessary for acetamiprid hydrolysis in strain D-2. The characteristics of AceAB were also systematically investigated. Though AceA and AceB showed 35% to 56% identity to the α and ß subunits of the N,N-dimethylformamidase from Paracoccus aminophilus, AceAB was specific for the hydrolysis of acetamiprid and showed no activities to N,N-dimethylformamide or its structural analogs.IMPORTANCE Acetamiprid, among the top neonicotinoid insecticides used worldwide, is one of the most important commercial insecticides. Due to its extensive use, the environmental fate of acetamiprid, especially its microbial degradation, has caused considerable concern. Although the catabolic pathways of acetamiprid in microorganisms have been extensively studied, the molecular mechanisms underlying acetamiprid biodegradation (except for a nitrile hydratase) remain largely unknown, and the enzyme responsible for the biotransformation of acetamiprid into its main intermediate, IM 1-4, have not yet been elucidated. The amidase AceAB and its encoding genes, aceA and aceB, characterized in this study, were found to be necessary and specific for the initial hydrolysis of the C-N bond of acetamiprid to generate IM 1-4 in Pigmentiphaga sp. strain D-2. The finding of the novel amidase AceAB will greatly enhance our understanding of the microbial catabolism of the widely used insecticide acetamiprid at the molecular level.