Exploring the multi-level regulation of lignocellulases in the filamentous fungus Trichoderma guizhouense NJAU4742 from an omics perspective.

BACKGROUND: Filamentous fungi are highly efficient at deconstructing plant biomass by secreting a variety of enzymes, but the complex enzymatic regulation underlying this process is not conserved and remains unclear. RESULTS: In this study, cellulases and xylanases could specifically respond to Avicel- and xylan-induction, respectively, in lignocellulose-degrading strain Trichoderma guizhouense NJAU4742, however, the differentially regulated cellulases and xylanases were both under the absolute control of the same TgXyr1-mediated pathway. Further analysis showed that Avicel could specifically induce cellulase expression, which supported the existence of an unknown specific regulator of cellulases in strain NJAU4742. The xylanase secretion is very complex, GH10 endoxylanases could only be induced by Avicel, while, other major xylanases were significantly induced by both Avicel and xylan. For GH10 xylanases, an unknown specific regulator was also deduced to exist. Meanwhile, the post-transcriptional inhibition was subsequently suggested to stop the Avicel-induced xylanases secretion, which explained the specifically high xylanase activities when induced by xylan in strain NJAU4742. Additionally, an economical strategy used by strain NJAU4742 was proposed to sense the environmental lignocellulose under the carbon starvation condition, that only slightly activating 4 lignocellulose-degrading genes before largely secreting all 33 TgXyr1-controlled lignocellulases if confirming the existence of lignocellulose components. CONCLUSIONS: This study, aiming to explore the unknown mechanisms of plant biomass-degrading enzymes regulation through the combined omics analysis, will open directions for in-depth understanding the complex carbon utilization in filamentous fungi.

Plasma Treatment Conversion of Phenolic Compounds into Fluorescent Organic Nanoparticles for Cell Imaging.

Fluorescent organic nanoparticles (FONs) are promising alternatives for biological imaging applications owing to the increasing concerns over the potential toxicity and poor degradability of inorganic particles-based probes. However, synthesis of stable, small-sized FONs in aqueous media remains challenging. Inspired by the self-polymerization chemistry of phenolic compounds, we demonstrate ultrafast synthesis of FONs (phenolic compound-derived FONs, PhFONs) from a variety of molecular building blocks including dopamine, norepinephrine, pyrogallol, and gallic acid, simply by nontherml plasma treatment at the aqueous interface. Specifically, using dopamine as the precursor, poly(dopamine) (PD)-FONs featuring a small size of 3 nm are obtained within 1 min. Compositional and structural characterizations confirm the polymeric architectures in PD-FONs. The PhFONs, with multicolor emissions, excellent biocompatibility, high stability, and size-dependent access into cell nucleus, are suitable for live cell imaging and developing nucleus-targeting imaging platforms.

A Self-Contained Chemiluminescent Lateral Flow Assay for Point-of-Care Testing.

Immunoassays whose readouts rely on chemiluminescence are increasingly useful for a broad range of analytical applications, but they are rarely made into point-of-care (POC) format because of the complex reagents required (some reagents have to be stored in low temperatures, and some reagents have to be freshly made right before the assay). This study reports a self-contained chemiluminescent lateral flow assay (CLFA), which prestores all necessary reagents. This CLFA contains three parts: the normal lateral flow assay (LFA) strip, the chemiluminescence substrate pad, and the polycarbonate (PC) holder. On the LFA strip, we simultaneously labeled horseradish peroxidase (HRP) and antibody on the gold nanoparticles (AuNPs) for the conjugate pad. For the substrate pad, we used sodium perborate as the oxidant and lyophilized the chemiluminescence substrate on the glass fiber, which allows long-term storage. After the transfer of substrate from the substrate pad to the nitrocellulose (NC) membrane, we captured the chemiluminescence signal for the quantification of the targets. The HRP on the AuNPs can amplify the chemiluminescence signal efficiently. We used this CLFA system to detect both macromolecules and small molecules successfully. This self-contained and easily processable device is exceedingly appropriate for rapid detection and is a convenient platform for POC testing.

Reverse Reconstruction and Bioprinting of Bacterial Cellulose-Based Functional Total Intervertebral Disc for Therapeutic Implantation.

The degradation of intervertebral discs (IVD), a typical hierarchical structured tissue, causes serious neck and back pain. The current methods cannot fully reconstitute the unique structure and function of native IVD. In this study, by reverse reconstruction of the structure of native IVD and bioprinting bacterial cellulose (BC) nanofibers with a high-throughput optimized micropattern screening microchip, a total IVD is created that contained type II collagen-based nucleus pulposus (NP) and hierarchically organized and micropatterned BC-based annulus fibrosus (AF), mimicking native IVD tissue. The artificial NP contains rat NP cells, whereas the AF contains concentrically arranged BC layers with aligned micropatterns and attached AF cells in +/-30° alternate directions between adjacent layers. Long-term (3 months) implantation experiments on rats demonstrate the excellent structural (shape maintenance, hydration, tissue integration) and functional (mechanical support and flexibility) performance of the artificial IVD. This study provides a novel strategy for creating highly sophisticated artificial tissues.

Peroxidase-like activity of amino-functionalized magnetic nanoparticles and their applications in immunoassay.

Amino-functionalized Fe3O4 magnetic nanoparticles with high peroxidase-like activity (MNPs-NH2) are reported in this work. The peroxidase-like activity is approximately the same as naked Fe3O4 magnetic nanoparticles (MNPs). MNPs-NH2 displayed much better thermal stability and pH tolerance than horseradish peroxidase (HRP). The optimal pH and temperature are approximately pH 4 and 40 °C, respectively. We showed that the MNPs-NH2 were successfully applied in a double-antibody sandwich "enzyme"-linked immunosorbent assay to replace of HRP, in which MNPs-NH2 were not only separation carriers, but also detection indicator.