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Toolkit for integrating millimeter-sized microfluidic biomedical devices with multiple membranes and electrodes
In recent years, microfluidic systems have evolved to incorporate increasingly complex multi-layer and multi-material structures. While conventional 2-dimensional microfluidic systems are typically fabricated with lithographic techniques, the increase in system complexity necessitates a more versatile set of fabrication techniques. Similarly, although 3D printing can easily produce intricate microfluidic geometries, integrating multiple membranes and electrode components remains challenging. This study proposes a toolkit for fabricating free-standing 3-dimensional microfluidic systems for biomedical devices, incorporating flow channels, electrodes, and membranes. The fabrication techniques include molding separation using 3D printed molds, laser-based processing, and component assembly, each achieving micron resolution. Here, we introduce a novel approach to integrate membranes into microfluidics by directly curing elastomer-based microfluidics with the membrane through replica molding, while preserving membrane functionality by effectively removing elastomer residues through reactive ion etching. The resulting membrane-elastomer microfluidic component significantly simplifies the assembly of intricate microfluidic systems, reducing the device size to millimeter dimensions, suitable for implantable applications. The toolkit’s versatility is demonstrated by a redox flow iontophoretic drug delivery prototype at the millimeter scale, featuring two electrodes, four membranes, and four microfluidic channels.
ACOT12, a novel factor in the pathogenesis of kidney fibrosis, modulates ACBD5
Lipid metabolism, particularly fatty acid oxidation dysfunction, is a major driver of renal fibrosis. However, the detailed regulatory mechanisms underlying this process remain unclear. Here we demonstrated that acyl-CoA thioesterase 12 (Acot12), an enzyme involved in the hydrolysis of acyl-CoA thioesters into free fatty acids and CoA, is a key regulator of lipid metabolism in fibrotic kidneys. A significantly decreased level of ACOT12 was observed in kidney samples from human patients with chronic kidney disease as well as in samples from mice with kidney injuries. Acot12 deficiency induces lipid accumulation and fibrosis in mice subjected to unilateral ureteral obstruction (UUO). Fenofibrate administration does not reduce renal fibrosis in Acot12−/− mice with UUO. Moreover, the restoration of peroxisome proliferator-activated receptor α (PPARα) in Acot12−/−Pparα−/− kidneys with UUO exacerbated lipid accumulation and renal fibrosis, whereas the restoration of Acot12 in Acot12−/− Pparα−/− kidneys with UUO significantly reduced lipid accumulation and renal fibrosis, suggesting that, mechanistically, Acot12 deficiency exacerbates renal fibrosis independently of PPARα. In Acot12−/− kidneys with UUO, a reduction in the selective autophagic degradation of peroxisomes and pexophagy with a decreased level of ACBD5 was observed. In conclusion, our study demonstrates the functional role and mechanistic details of Acot12 in the progression of renal fibrosis, provides a preclinical rationale for regulating Acot12 expression and presents a novel means of preventing renal fibrosis.
Anti-icing properties of nonionic/hydrophilic concentrated polymer brushes and mechanistic insights via their swollen-state analysis
Anti-icing surfaces are important to prevent snow and ice accumulation, which can pose significant risks. Here, we analyze the anti-icing performance of concentrated polymer brushes (CPBs) consisting of a versatile nonionic/hydrophilic monomer and discuss the low-temperature properties of the CPB-retaining water. The anti-icing functionality is evaluated by measuring the ice adhesion strength as a function of the temperature and the structural parameters (e.g., density and length) of the polymer brushes. We demonstrate that only the CPB region (σ* ≥ 0.15) exhibits both high anti-icing functionality and excellent durability. Furthermore, the thickening of the CPBs is key to achieving a detailed characterization of the water present in the CPBs at low temperatures using in situ microscopic Fourier-transform infrared spectroscopy and differential scanning calorimetry. These results suggest that the water effectively remaining via quasi-equilibrium partial deswelling formed a lubricating layer, contributing to high anti-icing functionality and durability.
Gut microbiomes of cycad-feeding insects tolerant to β-methylamino-L-alanine (BMAA) are rich in siderophore biosynthesis
Ingestion of the cycad toxins β-methylamino-L-alanine (BMAA) and azoxyglycosides is harmful to diverse organisms. However, some insects are specialized to feed on toxin-rich cycads with apparent immunity. Some cycad-feeding insects possess a common set of gut bacteria, which might play a role in detoxifying cycad toxins. Here, we investigated the composition of gut microbiota from a worldwide sample of cycadivorous insects and characterized the biosynthetic potential of selected bacteria. Cycadivorous insects shared a core gut microbiome consisting of six bacterial taxa, mainly belonging to the Proteobacteria, which we were able to isolate. To further investigate selected taxa from diverging lineages, we performed shotgun metagenomic sequencing of co-cultured bacterial sub-communities. We characterized the biosynthetic potential of four bacteria from Serratia, Pantoea, and two different Stenotrophomonas lineages, and discovered a suite of biosynthetic gene clusters notably rich in siderophores. Siderophore semi-untargeted metabolomics revealed a broad range of chemically related yet diverse iron-chelating metabolites, including desferrioxamine B, suggesting the occurrence of an unprecedented desferrioxamine-like biosynthetic pathway that remains to be identified. These results provide a foundation for future investigations into how cycadivorous insects tolerate diets rich in azoxyglycosides, BMAA, and other cycad toxins, including a possible role for bacterial siderophores.
Phylogenetically and metabolically diverse autotrophs in the world’s deepest blue hole
The world’s deepest yongle blue hole (YBH) is characterized by sharp dissolved oxygen (DO) gradients, and considerably low-organic-carbon and high-inorganic-carbon concentrations that may support active autotrophic communities. To understand metabolic strategies of autotrophic communities for obtaining carbon and energy spanning redox gradients, we presented finer characterizations of microbial community, metagenome and metagenome-assembled genomes (MAGs) in the YBH possessing oxic, hypoxic, essentially anoxic and completely anoxic zones vertically. Firstly, the YBH microbial composition and function shifted across the four zones, linking to different biogeochemical processes. The recovery of high-quality MAGs belonging to various uncultivated lineages reflected high novelty of the YBH microbiome. Secondly, carbon fixation processes and associated energy metabolisms varied with the vertical zones. The Calvin–Benson–Bassham (CBB) cycle was ubiquitous but differed in affiliated taxa at different zones. Various carbon fixation pathways were found in the hypoxic and essentially anoxic zones, including the 3-hyroxypropionate/4-hydroxybutyrate (3HP/4HB) cycle affiliated to Nitrososphaeria, and Wood-Ljungdahl (WL) pathway affiliated to Planctomycetes, with sulfur oxidation and dissimilatory nitrate reduction as primary energy-conserving pathways. The completely anoxic zone harbored diverse taxa (Dehalococcoidales, Desulfobacterales and Desulfatiglandales) utilizing the WL pathway coupled with versatile energy-conserving pathways via sulfate reduction, fermentation, CO oxidation and hydrogen metabolism. Finally, most of the WL-pathway containing taxa displayed a mixotrophic lifestyle corresponding to flexible carbon acquisition strategies. Our result showed a vertical transition of microbial lifestyle from photo-autotrophy, chemoautotrophy to mixotrophy in the YBH, enabling a better understanding of carbon fixation processes and associated biogeochemical impacts with different oxygen availability.
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