Image Sensing of Gaseous Acetone Using Secondary Alcohol Dehydrogenase-Immobilized Mesh for Exhaled Air.

Human-borne acetone is a potent marker of lipid metabolism. Here, an enzyme immobilization method for secondary alcohol dehydrogenase (S-ADH), which is suitable for highly sensitive and selective biosensing of acetone, was developed, and then its applicability was demonstrated for spatiotemporal imaging of concentration distribution. After various investigations, S-ADH-immobilized meshes could be prepared with less than 5% variation by cross-linking S-ADH with glutaraldehyde on a cotton mesh at 40 °C for 15 min. Furthermore, high activity was obtained by adjusting the concentration of the coenzyme nicotinamide adenine dinucleotide (NADH) solution added to the S-ADH-immobilized mesh to 500 µM and the solvent to a potassium phosphate buffer solution at pH 6.5. The gas imaging system using the S-ADH-immobilized mesh was able to image the decrease in NADH fluorescence (ex 340 nm, fl 490 nm) caused by the catalytic reaction of S-ADH and the acetone distribution in the concentration range of 0.1-10 ppm-v, including the breath concentration of healthy people at rest. The exhaled breath of two healthy subjects at 6 h of fasting was quantified as 377 and 673 ppb-v, which were consistent with the values quantified by gas chromatography-mass spectrometry.

Mouthguard-Type Wearable Sensor for Monitoring Salivary Turbidity to Assess Oral Hygiene.

Salivary turbidity is a promising indicator for evaluating oral hygiene. This study proposed a wearable mouthguard-type sensor for continuous and unconstrained measurement of salivary turbidity. The sensor evaluated turbidity by measuring the light transmittance of saliva with an LED and a phototransistor sealed inside a double-layered mouthguard. The sensor was also embedded with a Bluetooth wireless module, enabling the wireless measurement of turbidity. The mouthguard materials (polyethylene terephthalate-glycol and ethylene-vinyl acetate) and the wavelength of the LED (405 nm) were experimentally determined to achieve high sensitivity in salivary turbidity measurement. The turbidity quantification characteristic of the proposed sensor was evaluated using a turbidity standard solution, and the sensor was capable of turbidity quantification over a wide dynamic range of 1-4000 FTU (formazine turbidity unit), including reported salivary turbidity (400-800 FTU). In vitro turbidity measurement using a saliva sample showed 553 FTU, which is equivalent to the same sample measured with a spectrophotometer (576 FTU). Moreover, in vivo experiments also showed results equivalent to that measured with a spectrophotometer, and wireless measurement of salivary turbidity was realized using the mouthguard-type sensor. Based on these results, the proposed mouthguard-type sensor has promising potential for the unconstrained continuous evaluation of oral hygiene.

In Vitro Performance of a Long-Range Surface Plasmon Hydrogel Aptasensor for Continuous and Real-Time Vancomycin Measurement in Human Serum.

This study investigated the suitability of surface modification for a long-range surface plasmon (LRSP) aptasensor using two different hydrogels, aiming at real-time monitoring of vancomycin (VCM) in undiluted serum and blood. Three different layer structures were formed on a gold surface of LRSP sensor chip using poly[2-methacryloyloxyethyl phosphorylcholine (MPC)-co-N-methacryloyl-(L)-tyrosinemethylester (MAT)] (PMM) and poly[MPC-co-2-ethylhexyl methacrylate (EHMA)-co-MAT] (PMEM). The peptide aptamer for VCM was immobilized in PMM and PMEM via MAT. Among four differently prepared sensor chips, the LRSP hydrogel aptasensor with PMM, referred to as the PMM hydrogel, exhibited the highest sensor output and superior antifouling properties. Following the optimization of the PMM hydrogel preparation conditions, the shelf life of the PMM hydrogel was determined to exceed 2 weeks, and the same sensor chip could be used for 102 days without significant performance deterioration. The PMM hydrogel was then applied for VCM measurement in undiluted serum in vitro, where it demonstrated a limit of detection of 0.098 µM and a dynamic range of 0.18-100 µM, covering the therapeutic range. Additionally, the PMM hydrogel enabled the continuous measurement of various VCM concentrations in serum without rinsing and showed a concentration-dependent output in undiluted blood. These findings underscore the potential of the PMM hydrogel for real-time and direct monitoring of VCM in body fluids.

Biofluorometric Gas-Imaging System for Evaluating the Ripening Stages of "La France" Pear Based on Ethanol Vapor Emitted via the Epicarp.

Fruits can emit ethanol, which is generated through fermentation during hypoxic storage. We imaged spatiotemporal changes in the gaseous ethanol emitted by "La France" pear via its epicarp. The gas-imaging system utilized enzymes to transduce the ethanol concentration into fluorescence intensity. Initially, the uniformity of the enzyme and coenzyme distribution was evaluated to validate the imaging capability. Subsequently, two surface-fitting methods were compared to accurately image ethanol emitted from three-dimensional (3D) objects with a double-curved surface. The imaging results of ethanol emitted from the pear indicated that the distribution of ethanol was related to lenticels, which have been reported to possess high ethanol diffusivity, on the epicarp. As quantified by the system (uniformity of coenzyme and enzymes was 93.2 and 98.8%, respectively; dynamic range was 0.01-100 ppm), ethanol concentration increased with the storage period under hypoxic conditions (0.4-5.3 ppm, from day 1 to 10). The system enables the observation of the location, quantity, and temporal pattern of ethanol release from fruit, which could be a useful technology for agricultural applications.

Fabrication of 3D engineered intestinal tissue producing abundant mucus by air-liquid interface culture using paper-based dual-layer scaffold.

Fabrication of engineered intestinal tissues with the structures and functions as humans is crucial and promising as the tools for developing drugs and functional foods. The aim of this study is to fabricate an engineered intestinal tissue from Caco-2 cells by air-liquid interface culture using a paper-based dual-layer scaffold and analyze its structure and functions. Just by simply placing on a folded paper soaked in the medium, the electrospun gelatin microfiber mesh as the upper cell adhesion layer of the dual-layer scaffold was exposed to the air, while the lower paper layer worked to preserve and supply the cell culture medium to achieve stable culture over several weeks. Unlike the flat tissue produced using the conventional commercial cultureware, Transwell, the engineered intestinal tissue fabricated in this study formed three-dimensional villous architectures. Microvilli and tight junction structures characteristic of epithelial tissue were also formed at the apical side. Furthermore, compared to the tissue prepared by Transwell, mucus production was significantly larger, and the enzymatic activities of drug metabolism and digestion were almost equivalent. In conclusion, the air-liquid interface culture using the paper-based dual-layer scaffold developed in this study was simple but effective in fabricating the engineered intestinal tissue with superior structures and functions.