Transcriptional Characteristics Showed That miR-144-y/FOXO3 Participates in Embryonic Skin and Feather Follicle Development in Zhedong White Goose.

Skin and feather follicle development are essential processes for goose embryonic growth. Transcriptome and next-generation sequencing (NGS) network analyses were performed to improve the genome of Zhedong White goose and discover the critical genes, miRNAs, and pathways involved in goose skin and feather follicle morphogenesis. Sequencing output generated 6,002,591,668 to 8,675,720,319 clean reads from fifteen libraries. There were 1234, 3024, 4416, and 5326 different genes showing differential expression in four stages, E10 vs. E13, E10 vs. E18, E10 vs. E23, and E10 vs. E28, respectively. The differentially expressed genes (DEGs) were found to be implicated in multiple biological processes and pathways associated with feather growth and development, such as the Wnt signaling pathway, cell adhesion molecules, ECM-receptor interaction signaling pathways, and cell cycle and DNA replication pathways, according to functional analysis. In total, 8276 DEGs were assembled into twenty gene profiles with diverse expression patterns. The reliability of transcriptome results was verified by real-time quantitative PCR by selecting seven DEGs and five miRNAs. The localization of forkhead box O3 (FOXO3), connective tissue growth factor (CTGF), protein parched homolog1 (PTCH1), and miR-144-y by in situ hybridization showed spatial-temporal expression patterns and that FOXO3 and miR-144-y have an antagonistic targeting relationship. The correlation coefficient of FOXO3 and miR-144-y was -0.948, showing a strong negative correlation. Dual-luciferase reporter assay results demonstrated that miR-144-y could bind to the expected location to suppress the expression of FOXO3, which supports that there is a targeting relationship between them. The detections in this report will provide critical insight into the complex molecular mechanisms and breeding practices underlying the developmental characteristics of skin and feather follicles in Zhedong white geese.

Ginger (Zingiber officinale) extract mediated green synthesis of silver nanoparticles and evaluation of their antioxidant activity and potential catalytic reduction activities with Direct Blue 15 or Direct Orange 26.

The green synthesis of silver nanoparticles (AgNPs) using a water extract of Ginger (Zingiber officinale) root by microwave irradiation and its antibacterial activities have been reported. However, AgNPs prepared from different parts of ginger root water or ethanol extract by ultrasound synthesis and their antioxidant activity and whether the biogenic could be used to catalyze the reduction of hazardous dye are unknown. This study concentrated on the facile green synthesis of AgNPs prepared from different parts (unpeeled ginger, peeled ginger, and ginger peel) of ginger root water or ethanol extract by the ultrasound-assisted method. We studied their antioxidant activity and catalytic degradation of hazardous dye Direct Orange 26 (DO26) and Direct Blue 15 (DB15). The surface plasmon resonance (SPR) peak of AgNPs was at 428-443 nm. The biogenic AgNPs were approximately 2 nm in size with a regular spherical shape identified from TEM analysis. The ethanol extracts of dried unpeeled ginger and peeled ginger, fresh peeled ginger and ginger peel. The Z. officinale AgNPs synthesized by dried unpeeled ginger ethanol extract showed the best antioxidant activity. Their scavenging activities were significantly better than BHT (p <0.05). The different parts of ginger extracts showed no catalytic degradation activities of DB15 and DO26. Still, the synthesized Z. officinale AgNPs exhibited good catalytic degradation activities, while their ability to catalytic degradation to DB15 was better than DO26. In the additive ratio of 3 mL DB15, 0.1 mL NaBH4 and 0.1 mL AgNPs, the degradation rates of DB15 (or DO26) at 15 min, 30 min and 60 min were only 1.8% (0.9%), 2.8% (1.4%) and 3.5% (1.6%) in the absence of AgNPs. When adding Z. officinale AgNPs prepared from dried ginger peel ethanol extract or fresh ginger peel water extract, the degradation rates of DB15 sharply increased to 97% and 93% after 30 min, respectively. In conclusion, ginger extract has good antioxidant properties. Z. officinale AgNPs biosynthesis from ginger extract exhibit excellent catalytic degradation activities, especially for the ginger peel extract. They have application value in the treatment of textile effluents and provide a new idea and method for the comprehensive development and utilization of ginger resources.

In Ovo Injection of CHIR-99021 Promotes Feather Follicle Development via Modulating the Wnt Signaling Pathway and Transcriptome in Goose Embryos (Anser cygnoides).

Feather performs important physiological functions in birds, and it is also one of the economic productions in goose farming. Understanding and modulating feather follicle development during embryogenesis are essential for bird biology and the poultry industry. CHIR-99021 is a potent Wnt/ß-catenin signaling pathway activator associated with feather follicle development. In this study, goose embryos (Anser cygnoides) received an in ovo injection of CHIR-9902, which was conducted at the beginning of feather follicle development (E9). The results showed that feather growth and feather follicle development were promoted. The Wnt signaling pathway was activated by the inhibition of GSK-3ß. Transcriptomic analyses showed that the transcription changes were related to translation, metabolism, energy transport, and stress in dorsal tissue of embryos that received CHIR-99021, which might be to adapt and coordinate the promoting effects of CHIR-99021 on feather follicle development. This study suggests that in ovo injection of CHIR-99021 is a potential strategy to improve feather follicle development and feather-related traits for goose farming and provides profiling of the Wnt signaling pathway and transcriptome in dorsal tissue of goose embryos for further understanding of feather follicle development.

In ovo injection of CHIR-99021 promotes feather follicles development via activating Wnt/ß-catenin signaling pathway during chick embryonic period.

The Wingless-types/beta-catenin (Wnt/ß-catenin) signaling pathway plays an important role in embryonic development and affects the physiological development processes of feather follicles. To investigate the role of Wnt/ß-catenin pathway in regulating feather follicles morphogenesis, in ovo injection of CHIR-99021, an activator of the Wnt/ß-catenin signaling pathway, was conducted in chick embryo model. Initially, a total of 40 embryos were used to assess feather follicles morphogenesis and the expression of ß-catenin (E9-E17). The histological results showed that feather follicle morphogenesis was mainly completed from E9 to E17. ß-catenin was involved in the processing of the appearance of dermal cell condensation (E9) and the completion of the feather follicles morphogenesis (E17). Next, a total of 160 fertilized eggs were randomly divided into 8 groups for in ovo injection at E9, including a Normal Saline injected group (CON) and the 500, 1,000, 2,000, 5,000, 10,000, 50,000, and 100,000 ng CHIR-99021 groups. Dorsal skin tissue samples were collected at E17 for investigating feather follicles morphology and expressions of ß-catenin and lymphoid enhancerbinding factor-1 (LEF1) at gene and protein levels. The results showed that feather follicle diameter in the injected groups were significantly (P < 0.05) increased with limit dose-independence compared to the CON group. CHIR-99021 significantly (P < 0.05) influenced the mRNA expressions of catenin beta-1 (CTNNB1) and downstream target LEF1. In ovo injection of CHIR-99021 caused that ß-catenin and LEF1 were significantly (P < 0.05) increased followed the increased doses as determined by western blotting. The immunochemical results showed that ß-catenin was detected in the dermal papilla of feather follicles. Given these results, this study suggests to developmental biology that in ovo injection of CHIR-99021 promoted feather follicles morphogenesis and development via activating Wnt/ß-catenin signaling pathway and upregulating downstream target LEF1 during embryonic period in chick embryo model. Moreover, CHIR-99021 may be a strong candidate to promote the animal feather/hair industry, especially as a reference for bird feather production.

Asymmetric free-standing film with multifunctional anti-bacterial and self-cleaning properties.

A superhydrophobic/hydrophilic asymmetric free-standing film has been created using layer-by-layer assembly technique. Poly(ethylene-imine)-Ag(+) complex (PEI-Ag(+)) at pH 9.0 was assembled with poly(acrylic acid) (PAA) at pH 3.2 on a Teflon substrate to yield a micronanostructured surface that can be turned to be superhydrophobic after being coated with a low surface energy compound. Silver nanoparticle loaded free-standing film with one surface being superhydrophobic while the other surface is hydrophilic was then obtained after detachment from the substrate. The superhydrophobicity enabled the upper surface with anti-adhesion and self-cleaning properties and the hydrophilic bottom surface can release silver ions as antibiotic agent. The broad-spectrum antimicrobial capability of silver ions released from the bottom surface coupled with superhydrophobic barrier protection of the upper surface may make the free-standing film a new therapy for open wound.

Humidity responsive asymmetric free-standing multilayered film.

A humido-responsive free-standing film has been created using layer-by-layer assembly technique. Polyethylenimine (PEI) at high pH was assembled with poly(acrylic acid) (PAA) at low pH on a Teflon substrate to yield a micro-nanostructured surface that can be made superhydrophobic after being coated with a low surface energy compound. The resulting asymmetric free-standing film with one surface being superhydrophobic while the other is hydrophilic after detachment from the substrate can undergo reversible bending/unbending shape transitions when the environmental humidity is changed. The bending/unbending movement of the free-standing film can be ascribed to the different responses of these two surfaces to humidity.

Self-assembled chitosan/heparin multilayer film as a novel template for in situ synthesis of silver nanoparticles.

Chitosan and heparin multilayer films were successfully constructed via layer-by-layer self assembly. These films were used as a polymeric template to synthesize silver nanoparticles. The silver concentration and nanoparticle size can be simply controlled by the assembly pH and loading pH, as demonstrated by UV-visible spectroscopy, transmission electron microscopy and atomic absorbance spectroscopy. The pH tunable uncompensated charge density within the multilayer films is believed to have great effect on the loading of silver ions, and then control the size and amount of silver nanoparticles within multilayer films. The antibacterial experiment shows that the silver nanoparticle-loaded chitosan/heparin multilayer films exhibit greatly enhanced antibacterial performance compared to the chitosan/heparin multilayer films without silver nanoparticles. In addition, the strong antibacterial property of silver nanoparticle-loaded films can last more than 1 month. Our method of in situ synthesis of metal nanoparticles in biocompatible multilayer films might provide great potential to design biofunctional nanocomposite films.

pH-amplified exponential growth multilayers: a facile method to develop hierarchical micro- and nanostructured surfaces.

We report a direct method to amplify the exponential growth of multilayers significantly by the alternating deposition of polyethylenimine (PEI) at high pH and poly(acrylic acid) (PAA) at low pH. The alternating pH switches the degree of ionization of the polyelectrolytes in the multilayers, which enhances the diffusion of PEI into and out of the film and hence increases the deposited mass per cycle. The synergetic action of the pH-tunable charge density and diffusivity of the weak polyelectrolytes provides a new method for the enhanced growth of multilayers with hierarchal micro- and nanostructured surfaces.

A facile method to construct hybrid multilayered films as a strong and multifunctional antibacterial coating.

A multifunctional multilayered film containing TiO(2) nanoparticles as contact-active antibacterial agent and nanosilver as a release-active antibacterial agent was fabricated via layer-by-layer assembly. TiO(2) nanoparticles with the anatase crystalline dominant structure were synthesized via a sol-gel method. The QCM, AFM, and contact angle measurement results indicated that the TiO(2) nanoparticle-chitosan was successfully assembled with heparin via layer-by-layer assembly. The UV visible spectroscopy demonstrated that the silver ions could be loaded into the multilayers and in situ synthesize silver nanoparticles in the multilayers template. The short-term antibacterial assay showed the TiO(2)-chitosan/heparin multilayers loaded with nanosilver was bactericidal both in the low intensity UV light and in the dark. The long-term antibacterial assay indicated although the antibacterial in dark decreased with the PBS immersion time, the hybrid multilayered films sustained the long-term antibacterial in the low intensity UV light.

Construction of antibacterial multilayer films containing nanosilver via layer-by-layer assembly of heparin and chitosan-silver ions complex.

Antibacterial multilayer films containing nanosilver were prepared via layer-by-layer fashion. PET film was aminolyzed with 1,6-hexanediamine to introduce amino groups on PET film surface; chitosan-silver nitrate complex and heparin were alternately deposited onto an aminolyzed PET film surface, and subsequently, the silver ions within the multilayer films were reduced with ascorbic acid to form silver nanoparticles. UV-visible spectroscopy and transmission electron microscopy confirmed the formation of well-dispersed nanosilver particles with sizes (10-40 nm) that depended on the initial concentration of silver ions in chitosan solution and the pH of ascorbic acid solution. The chitosan/heparin multilayer films were possessed of bactericidal effect on Escherichia coli (E. coli), and this antibacterial effect could be significantly enhanced by the incorporation of silver nanoparticles into the multilayer films. The multilayer films containing nanosilver were not only effective as antibacterial but also as anticoagulant coating. And cell toxicity evaluation suggested that the multilayer films containing nanosilver did not show any cytotoxicity. The multilayer films containing nanosilver may have good potentials for surface modification of medical devices, especially for cardiovascular implants.

Construction of anti-adhesive and antibacterial multilayer films via layer-by-layer assembly of heparin and chitosan.

Chitosan as an antibacterial agent and heparin as an anti-adhesive agent were alternatively deposited onto aminolyzed poly(ethylene terephthalate) (PET) films to construct anti-adhesive and antibacterial multilayer films. The contact-angle and UV data verified the progressive buildup of the multilayer film by alternate deposition of the polyelectrolytes. The properties of multilayer films were investigated by contact angle, atomic force microscopy (AFM), lateral force microscopy (LFM) and UV spectra. The results of initial adhesion of Escherichia coli (E. coli) on PET substrates showed that the number of E. coli adhered onto the control PET was in a much greater extent than onto the chitosan/heparin multilayer films, and the number of adhesive bacteria decreased with a decrease in assembly pH. The in vitro antibacterial test indicated that a multilayer of chitosan/heparin could kill the bacteria effectively. The number of viable bacteria decreased by 7% after 7 h in contact with the control PET films, but by 46-68% for the multilayer-modified PET films. Only 3-8% of the cells were viable for the multilayer-modified PET films after 24h. It is interesting to find the assembly pH has a remarkable effect on the antibacterial property of the multilayer. The number of viable bacteria on the multilayer assembled at pH = 3.8, 2.9 and 6.0 decreased by 68%, 58% and 46%, respectively. Such an easy processing and shape-independent method to prepare an anti-adhesive and antibacterial surface may have good potential for surface modification of cardiovascular devices.