Effect of alcohol consumption on risk of hypertension based on alcohol-related facial flushing response: From the health examinees study.

BACKGROUND: Alcohol consumption is a significant public health concern in Korea, with many individuals engaging in risky drinking behaviors. This study aims to analyze the association between facial flushing responses and the progression of hypertension in Korean adults, stratified by gender, using a large-scale prospective cohort study. METHODS: This study included 39,868 participants (males 10,868 and females 29,000) from the health examinees cohort. Participants were divided into two groups according to their weekly drinking patterns (moderate and heavy) and facial flushing responses (non-flusher and flusher) for statistical analysis. A multivariate Cox proportional hazards regression model was used to calculate hazard ratios and 95% confidence intervals between flushing response after alcohol consumption and hypertension risk. RESULTS: In males, the flusher group with a moderate alcohol intake pattern had a decreased risk of incident hypertension compared with non-flushers. In females, the flusher group had relatively higher risk of pre-hypertension and hypertension compared to the non-flusher group. CONCLUSION: The association between facial flushing and hypertension varies depending on the level of alcohol consumption and should be considered in relation to gender differences. Further research is needed to understand the relationship between facial flushing response and the risk of hypertension based on alcohol consumption levels.

Design of an effective small expression tag to enhance GPCR production in E. coli-based cell-free and whole cell expression systems.

G protein-coupled receptors (GPCRs) play crucial roles in sensory, immune, and tumor metastasis processes, making them valuable targets for pharmacological and sensing applications in various industries. However, most GPCRs have low production yields in Escherichia coli (E. coli) expression systems. To overcome this limitation, we introduced AT10 tag, an effective fusion tag that could significantly enhance expression levels of various GPCRs in E. coli and its derived cell-free protein synthesis (CFPS) system. This AT10 tag consisted of an A/T-rich gene sequence designed via optimization of translation initiation rate. It is translated into a short peptide sequence of 10 amino acids at the N-terminus of GPCRs. Additionally, effector proteins could be utilized to suppress cytotoxicity caused by membrane protein expression, further boosting GPCR production in E. coli. Enhanced expression of various GPCRs using this AT10 tag is a promising approach for large-scale production of functional GPCRs in E. coli-based CFPS and whole cell systems, enabling their potential utilization across a wide range of industrial applications.

Advances in the Production of Olfactory Receptors for Industrial Use.

In biological olfactory systems, olfactory receptors (ORs) can recognize and discriminate between thousands of volatile organic compounds with very high sensitivity and specificity. The superior properties of ORs have led to the development of OR-based biosensors that have shown promising potential in many applications over the past two decades. In particular, newly designed technologies in gene synthesis, protein expression, solubilization, purification, and membrane mimetics for membrane proteins have greatly opened up the previously inaccessible industrial potential of ORs. In this review, gene design, expression and solubilization strategies, and purification and reconstitution methods available for modern industrial applications are examined, with a focus on ORs. The limitations of current OR production technology are also estimated, and future directions for further progress are suggested.

FET-based nanobiosensors for the detection of smell and taste.

Various nanobiosensors composed of biomaterials and nanomaterials have been developed, due to their demonstrated advantage of showing high performance. Among various biomaterials for biological recognition elements of the nanobiosensor, sensory receptors, such as olfactory and taste receptors, are promising biomaterials for developing nanobiosensors, because of their high selectivity to target molecules. Field-effect transistors (FET) with nanomaterials such as carbon nanotube (CNT), graphene, and conducting polymer nanotube (CPNT), can be combined with the biomaterials to enhance the sensitivity of nanobiosensors. Recently, many efforts have been made to develop nanobiosensors using biomaterials, such as olfactory receptors and taste receptors for detecting various smells and tastes. This review focuses on the biomaterials and nanomaterials used in nanobiosensor systems and studies of various types of nanobiosensor platforms that utilize olfactory receptors and taste receptors which could be applied to a wide range of industrial fields, including the food and beverage industry, environmental monitoring, the biomedical field, and anti-terrorism.

Visual detection of odorant geraniol enabled by integration of a human olfactory receptor into polydiacetylene/lipid nano-assembly.

A new polydiacetylene lipid/human olfactory receptor nano-assembly was fabricated for the visual detection of an odorant for the first time. The assembly consisted of phospholipid-mixed polydiacetylenes (PDAs) and human olfactory receptors (hORs) in detergent micelles. To overcome the limitations of bioelectronic noses, hOR-embedded chromatic complexes (PDA/hORs) were developed, introducing PDAs that showed color and fluorescence transitions against various stimuli. The chromatic nanocomplexes reacted with target molecules, showing a fluorescence intensity increase in a dose-dependent manner and target selectivity among various odorants. As a result, a color transition of the assembly from blue to purple occurred, allowing the visual detection of the odorant geraniol. Through circular dichroism (CD) spectroscopy and a tryptophan fluorescence quenching method, the structural and functional properties of the hORs embedded in the complexes were confirmed. Based on this first work, future array devices, integrating multiple nano-assemblies, can be substantiated and utilized in environmental assessment and analysis of food quality.

Co3O4/Au Hybrid Nanostructures as Efficient Peroxidase Mimics for Colorimetric Biosensing.

Nanomaterials with enzyme-like characteristics (nanozymes) have emerged as potential replacements for natural enzymes due to their potential to overcome several critical limitations of natural enzymes, including low stability as well as high costs in preparation and purification. Herein, we have developed hybrid nanostructures that incorporate cobalt oxide nanoparticles (Co3O4 NPs) and gold nanoclusters (AuNCs) through electrostatic attraction induced by simple incubation in an aqueous buffer for 2 hours. Owing to the synergistic effect of Co3O4 NPs and AuNCs, the constructed Co3O4/Au hybrid nanostructures yielded highly enhanced peroxidase-like activity and enabled rapid catalytic oxidation of a chromogenic substrate, 3,3',5,5'-tetramethylbenzidine (TMB), producing a blue colored solution depending on the amount of H2O2. Moreover, we observed catalytic activity of the Co3O4/Au hybrid over a broad pH range, especially at physiologically relevant pH in the range of 5.0-7.4, which is advantageous for applications in biological systems. Using the hybrid as peroxidase mimic, we successfully determined the level of target H2O2 or glucose by coupling with glucose oxidase with excellent specificity and sensitivity. Based on this study, we expect that Co3O4/Au hybrid nanostructures can serve as potent peroxidase mimics for the detection of clinically important target molecules.

N- and B-Codoped Graphene: A Strong Candidate To Replace Natural Peroxidase in Sensitive and Selective Bioassays.

The work describes a carbon-based peroxidase mimic, N- and B-codoped reduced graphene oxide (NB-rGO), which shows high peroxidase-like activity without oxidase-like activity and has a catalytic efficiency nearly 1000-fold higher than that of undoped rGO. The high catalytic activity of NB-rGO is explained by density functional theory by calculating Gibbs free energy change during the peroxide decomposition reaction. Acetylcholine and C-reactive protein are successfully quantified with high sensitivity and selectivity, which were comparable to or better than those obtained using natural peroxidase. Furthermore, NB-rGO, which does not have oxidase-like activity, is proven to have higher sensitivity toward acetylcholine than Pt nanoparticles having oxidase-like activity. This work will facilitate studies on development, theoretical analysis for rational design, and bioassay applications of enzyme mimics based on nanomaterials.

Highly Efficient Electrochemical Detection of Phenolic Compounds Utilizing Superior Catalytic Activity of Nanohybrids Consisting of Magnetic Nanoparticles and Gold Nanoclusters.

Although Fe3O4 magnetic nanoparticles (MNPs) have gathered particular interest as potent peroxidase mimetics, their practical utility has been critically limited by their low catalytic activity. Here, we have developed a nanohybrid material to significantly enhance the catalytic activity of MNPs by incorporating other enzyme mimetics, gold nanoclusters (AuNCs), through electrostatic attraction. Owing to the synergistic effect of MNPs and AuNCs, the constructed nanohybrid yielded highly enhanced peroxidase-like activity and higher resolution in electrochemical detection of H2O2 than bare MNPs. The nanohybrids were also successfully applied to detecting phenolic compounds including phenol and cresol, producing a concentration-dependent increase of cathodic current. Based on this result, we expect that the nanohybrids consisting of AuNCs and MNPs can serve as potent peroxidase mimetics for environmental monitoring.

Highly sensitive colorimetric detection of allergies based on an immunoassay using peroxidase-mimicking nanozymes.

Nanomaterials that exhibit enzyme-like characteristics, which are called nanozymes, have recently attracted significant attention due to their potential to overcome the intrinsic limitations of natural enzymes, such as low stability and relatively high cost for preparation and purification. In this study, we report a highly efficient colorimetric allergy detection system based on an immunoassay utilizing the peroxidase-mimicking activity of hierarchically structured platinum nanoparticles (H-Pt NPs). The H-Pt NPs had a diameter of 30 nm, and were synthesized by a seed-mediated growth method, which led to a significant amount of peroxidase-like activity. This activity mainly occurs because of the high catalytic power of the Pt element, and the fact that the H-Pt NPs have a large surface area available for catalytic events. The H-Pt NPs were conjugated to an antibody for the detection of immunoglobulin E (IgE) in the analytes; IgE is a representative marker for the diagnosis of allergies. They were then successfully integrated into a conventionally used allergy diagnostic test, the ImmunoCAP diagnostic test, as a replacement for natural signaling enzymes. Using this strategy, total and specific IgE levels were detected within 5 min at room temperature, with high specificity and sensitivity. The practical utility of the immunoassay was also successfully verified by correctly determining the levels of both total and specific IgE in real human serum samples with high precision and reproducibility. The present H-Pt NP-based immunoassay system would serve as a platform for rapid, robust, and convenient analysis of IgE, and can be extended to the construction of diagnostic systems for a variety of clinically important target molecules.

Pt-Decorated Magnetic Nanozymes for Facile and Sensitive Point-of-Care Bioassay.

Development of facile and sensitive bioassays is important for many point-of-care applications. In this study, we fulfilled such demand by synthesizing Pt-decorated magnetic nanozymes and developing a bioassay based on unique properties of the newly synthesized nanozymes. Fe3O4-Pt/core-shell nanoparticles (MPt/CS NPs) with various compositions were synthesized and characterized. Fe3O4 NP itself is a good nanozyme with catalytic activity superior to that of natural enzymes, but catalytic activity can be further improved by incorporating Pt to the outer layers of the Fe3O4 NPs and building heterogeneous nanostructures. Magnetic properties of MPt/CS NPs enable magnetic enrichment of liquid samples, whereas catalytic properties of MPt/CS NPs allow signal amplification by enzyme-like reactions. By integrating MPt/CS NPs in lateral flow immunoassay strips, one of the widely used point-of-care bioassay devices, and harnessing magnetic and enzyme-like properties of MPt/CS NPs, an increase in sensitivity of two orders of magnitude was achieved compared to the sensitivity of conventional lateral flow immunoassay based on Au nanoparticles.

Nanohybrids consisting of magnetic nanoparticles and gold nanoclusters as effective peroxidase mimics and their application for colorimetric detection of glucose.

Although protein-stabilized gold nanoclusters (AuNCs) have gathered recent attention as biocompatible peroxidase mimics, their practical utility has been critically limited by the low catalytic activity. Here, the authors have developed a nanohybrid material to significantly enhance the catalytic activity of AuNCs by combining them with other inorganic enzyme mimetics, Fe3O4 magnetic nanoparticles (MNPs), through electrostatic attraction. Owing to the synergistic effect by incorporating AuNCs and MNPs, the constructed nanohybrids yielded highly enhanced catalytic activity and enabled rapid catalytic oxidation of 3,3',5,5'-tetramethylbenzidine substrate to produce a blue-colored solution in proportional to the amount of H2O2. Moreover, a highly sensitive and selective glucose biosensing strategy was developed based on the coupled catalytic action between glucose oxidase and the nanohybrids. Using this method, target glucose was successfully detected in a linear concentration range from 150 to 750 µM with a detection limit as low as 100 µM. Along with excellent linearity, high precision and reproducibility were achieved by employing real human blood serums, which enables its use for the reliable quantification of glucose in practical use. Based on these results, the authors anticipate that the nanohybrids consisting of MNPs and AuNCs can serve as potent peroxidase mimics for the detection of clinically important target molecules.

Nitric oxide induces apoptosis in mouse C2C12 myoblast cells.

To investigate whether nitric oxide (NO) induces apoptosis in myoblast cells, the effect of the sodium nitroprusside (SNP), NO donor, on the apoptosis of mouse C2C12 myoblast cells was examined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, flow cytometry, 4,6-diamidino-2-phenylindole (DAPI) staining, terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) assay, DNA fragmentation assay, reverse transcription-polymerase chain reaction (RT-PCR), Western blot analysis, and caspase-3 enzyme assay. Mouse C2C12 myoblast cells treated with SNP exhibited several apoptotic features. SNP increased p53 expression and bax expression. SNP also enhanced caspase-3 enzyme activity. The data show that NO may induce apoptotic cell death in myoblast cells through the activation of p53-, bax-, and caspase-dependent intracellular death-related pathways.

Hypothermia inhibits cell proliferation and nitric oxide synthase expression in rats.

In the present study, the effects of cold-water immersion on cell proliferation and nitric oxide synthase expression in the dentate gyrus of rats were investigated. Sprague-Dawley rats were divided into four groups: the control-rest group; the control-heat group; the cold-rest group; and the cold-heat group. Cold-water immersion for 5 min at 4 degrees C suppressed the numbers of 5-bromo-2'-deoxyuridine-positive and nicotinamide adenine dinucleotide phosphate-diaphorase-positive cells in the dentate gyrus, and these numbers were increased by warming for 30 min at 30 degrees C. In the present study, it was demonstrated that warming protects against cold stress-induced suppression of new cell formation, and results suggest that nitric oxide, the synthesis of which is affected adversely by cold-water immersion, may play an important role in the regulation of cell proliferation.