AP2M1 Supports TGF-β Signals to Promote Collagen Expression by Inhibiting Caveolin Expression

The extracellular matrix (ECM) is important for normal development and disease states, including inflammation and fibrosis. To understand the complex regulation of ECM, we performed a suppressor screening using Caenorhabditis elegans expressing the mutant ROL-6 collagen protein. One cuticle mutant has a mutation in dpy-23 that encodes the μ2 adaptin (AP2M1) of clathrin-associated protein complex II (AP-2). The subsequent suppressor screening for dpy-23 revealed the lon-2 mutation. LON-2 functions to regulate body size through negative regulation of the tumor growth factor-beta (TGF-β) signaling pathway responsible for ECM production. RNA-seq analysis showed a dominant change in the expression of collagen genes and cuticle components. We noted an increase in the cav-1 gene encoding caveolin-1, which functions in clathrin-independent endocytosis. By knockdown of cav-1, the reduced TGF-β signal was significantly restored in the dpy-23 mutant. In conclusion, the dpy-23 mutation upregulated cav-1 expression in the hypodermis, and increased CAV-1 resulted in a decrease of TβRI. Finally, the reduction of collagen expression including rol-6 by the reduced TGF-β signal influenced the cuticle formation of the dpy-23 mutant. These findings could help us to understand the complex process of ECM regulation in organism development and disease conditions.     

Multidirectional focus measure for accurate three-dimensional shape recovery of microscopic objects

Shape from focus (SFF) is a technique to recover the shape of an object from multiple images taken at various focus settings. Most of conventional SFF techniques compute focus value of a pixel by applying one of focus measure operators on neighboring pixels on the same image frame. However, in the optics with limited depth of field, neighboring pixels of an image have different degree of focus for curved objects, thus the computed focus value does not reflect the accurate focus level of the pixel. Ideally, an accurate focus value of a pixel needs to be measured from the neighboring pixels lying on tangential plane of the pixel in image space. In this article, a tangential plane on each pixel location (i, j) in image sensor is searched by selecting one of five candidate planes based on the assumption that the maximum variance of focus values along the optical axis is achieved from the neighborhood lying on tangential plane of the pixel (i, j). Then, a focus measure operator is applied on neighboring pixels lying on the searched plane. The experimental results on both the synthetic and real microscopic objects show the proposed method produces more accurate three-dimensional shape in comparison to conventional SFF method that applies focus measures on original image planes.     

A Polyallylamine Anchored Amine‐Rich Laser‐Ablated Graphene Platform for Facile and Highly Selective Electrochemical IgG Biomarker Detection

Current immunosensors have an insufficient number of binding sites for the recognition of biomolecules, which leads to false positive or negative results. In this research, a facile, cost‐effective, disposable, and highly selective electrochemical immunosensing platform is developed based on cationic polyelectrolyte polyallylamine (PAAMI) anchored laser‐ablated graphene (LAG). Here, for the first time, PAAMI is introduced to stabilize LAG flakes, while retaining the intrinsic thermal and electronic properties of the substrate by noncovalent π–π interaction and electrostatic physical absorption. The sensing platform offers a suitable number of anchoring sites for the immobilized antibodies by providing ? NH₂ functional groups. The proper grafting of PAAMI is confirmed through X‐ray photoelectron spectroscopy and Raman spectroscopy. The immunosensing platform is applied to detect immunoglobulin (IgG) biomarkers as a proof of concept. Under optimized conditions, the sensing platform exhibits a linear range of 0.012–15 and 15–352 ng mL^?1 with a limit of detection of 6 pg mL^?1 for IgG detection with high selectivity. Based on the analysis, the developed immunosensing platform can be used for point‐of‐care detection of IgG in clinical diagnostic centers. Furthermore, the developed strategy is well suited for the detection of other cancer biomarkers after immobilizing the relevant antibodies.     

Synthesis and luminescence properties of reddish-orange-emitting Ca2GdNbO6:Sm3+ phosphors with good thermal stability for high CRI white applications

Novel reddish-orange-emitting Ca₂GdNbO₆:Sm³⁺ phosphors based on the emission of ⁴G_5/2 → ⁶H_9/2 transition at 651 nm with the chromatic coordinate of (0.633, 0.366) were synthesized. The crystal structure and chemical purity were identified in detail. Under the 407 nm excitation, the optimum concentration of Sm³⁺ ion was found to be 5 mol% dominated by the dipole-dipole interaction in the Ca₂GdNbO₆ host material. The color purity of the sample with optimum doping was estimated to be about 78.38%. Besides, the thermal stability was also studied, and it was further found that the emission intensity remained 65.32% at 423 K. The packaged white LED device exhibited excellent CRI and CCT values of 92.43 and 4896 K. Finally, the polydimethylsiloxane film with a stable structure and flexible property was prepared. These above results reveal that novel reddish-orange-emitting Ca₂GdNbO₆:Sm³⁺ phosphors can be applied in high CRI white communication and flexible display applications.     

Phenotypic Discovery of SB1501, an Anti-obesity Agent,through Modulating Mitochondrial Activity

Obesity has become a pandemic that threatens the quality of life and discovering novel therapeutic agents that can reverse obesity and obesity-related metabolic disorders are necessary. Here, we aimed to identify new anti-obesity agents using a phenotype-based approach. We performed image-based high-content screening with a fluorogenic bioprobe (SF44), which visualizes cellular lipid droplets (LDs), to identify initial hit compounds. A structure-activity relationship study led us to yield a bioactive compound SB1501, which reduces cellular LDs in 3T3-L1 adipocytes without cytotoxicity. SB1501 induced the expression of gene products that regulate mitochondrial biogenesis and fatty acid oxidation in 3T3-L1 adipocytes. Daily treatment with SB1501 improved the metabolic states of db/db mice by reducing body fat mass, adipose tissue mass, food intake, and increasing glucose tolerance. The anti-obesity effect of SB1501 may result from perturbation of the PGC-1α–UCP1 regulatory axis in inguinal white adipose tissue and brown adipose tissue. These data suggest the therapeutic potential of SB1501 as an anti-obesity agent via modulating mitochondrial activities.     

Aqueous dispersion of 1D van der Waals Mo6S3I6 crystal using biocompatible tri-block copolymer

To achieve the stable dispersion of 1D van der Waals crystal Mo₆S₃I₆ in aqueous media, the tri-block copolymer (Poloxamer) is used as dispersant. The head group of Poloxamer, hydrophobic polypropylene oxide parts can be adsorbed to Mo₆S₃I₆ surface by hydrophobic interaction and the tail group with hydrophilic polyethylene oxide exposed to the outside of the Mo₆S₃I₆ is soluble in water and can form sufficient steric hindrance, resulting in stable aqueous dispersion in nm scale. The excellent biocompatibility of aqueous dispersed nm scale 1D Mo₆S₃I₆ was demonstrated by effective proliferation of C2C12 cells.     

Prelabeling Expansion Single-Molecule Localization Microscopy with Minimal Linkage Error

Expansion microscopy combined with single-molecule localization microscopy (ExSMLM) has a potential for approaching molecular resolution. However, ExSMLM faces multiple challenges such as loss of fluorophores and proteins during polymerization, digestion or denaturation, and an increase in linkage error arising from the distance between the fluorophore and the target molecule. Here, we introduce a trifunctional streptavidin to link the target, fluorophore and gel matrix via a biotinylizable peptide tag. The resultant ExSMLM images of vimentin filaments demonstrated high labeling efficiency and a minimal linkage error of ∼5 nm. Our ExSMLM provides a simple and practical means for fluorescence imaging with molecular resolution.     

Stimulation of densification during the reduction of U3O8 to UO2 by atmosphere control

A reduction process in the head-end for pyroprocessing has been adopted to avoid oxidation attack on the molybdenum crucible during sintering. The reduction process is employed to reduce U₃O₈ pellets to UO₂ prior to sintering. This allows elimination of the oxygen source, which causes oxidation attack during sintering, thereby permitting the use of a metallic crucible. However, little densification occurs due to the low reduction temperature limited by the INCONEL crucible. Consequently, the amount of material scraps from the pellets increases, thus creating an additional processing burden due to its high radioactivity. To reduce the amount of scraps, densification should be enhanced. This study suggests a simple atmospheric control strategy and clarifies its effects. With the atmospheric control, a higher bulk density and better attrition resistance were obtained in comparison to without this strategy. This can be explained in terms of O/U ratio dependent diffusion kinetics during the reduction of U₃O₈ to UO₂.