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The biophysical properties of human metaphase chromosomes were directly measured by Kelvin probe force microscopy (KPFM) and PeakForce-quantitative nanomechanics (PF-QNM) with high resolution.

Nitrogen dioxide (NO2) is a representative toxicant in air pollution that mostly arises from the exhaust gas released by automobiles. It is related to various respiratory diseases such as pneumonia and sudden infant death syndrome. Additionally, because the toxicity of nitrogen dioxide is high in overpopulated areas (i.e., a capital or metropolis), the development of simple, practical, and facile sensors is highly needed. This work presents a flexible and disposable paper-based NO2 sensor based on a reduced graphene oxide/chitosan (rGO/CS) composite. The synthesized rGO/CS composite can be easily flexed and deformed into various shapes, which are attributed to chitosan molecules that function as a dispersion and reduction agent and support material. In addition, this composite can be attached to paper owing to its adhesive property; hence it can be utilized in versatile applications in a disposable manner. By analyzing the conductive change of the rGO/CS composite when it reacts with NO2, we can detect nitrogen dioxide in a concentration range of 0–100 ppm with a detection limit of 1 ppm. Moreover, we performed NO2 detection in the exhaust gas released by automobiles using the rGO/CS composite for practical application. The results indicated that the rGO/CS composite has the potential to be used in feasible gas sensing as a facile and disposable sensor under various conditions.

Colorimetric paper sensors are used in various fields due to their convenience and intuitive manner. However, these sensors present low accuracy in practical use because it is difficult to distinguish color changes for a minute amount of analyte with the naked eye. Herein, we demonstrate that a machine learning (ML)-based paper sensor platform accurately determines the color changes. We fabricated a colorimetric paper sensor by adsorbing polyaniline nanoparticles (PAni-NPs), whose color changes from blue to green when the ambient pH decreases. Adding glucose oxidase (GOx) to the paper sensor enables colorimetric glucose detection. Target analytes (10 μL) were aliquoted onto the paper sensors, and their images were taken with a smartphone under the same conditions in a darkroom. The red-green-blue (RGB) data from the images were extracted and used to train and test three regression models: support vector regression (SVR), decision tree regression (DTR), and random forest regression (RFR). Of the three regression models, RFR performed the best at estimating pH levels (

Cells secrete extracellular vesicles (EVs) carrying cell-of-origin markers to communicate with surrounding cells. EVs regulate physiological processes ranging from intercellular signaling to waste management. However, when senescent cells (SnCs) secrete EVs, the EVs, which are newly regarded as senescence-associated secretory phenotype (SASP) factors, can evoke inflammation, senescence induction, and metabolic disorders in neighboring cells. Unlike other soluble SASP factors, the biophysical properties of EVs, including small EVs (sEVs), derived from SnCs have not yet been investigated. In this study, sEVs were extracted from a human IMR90 lung fibroblast

Recurrent spillovers of α- and β-coronaviruses (CoV) such as severe acute respiratory syndrome (SARS)-CoV, Middle East respiratory syndrome-CoV, SARS-CoV-2, and possibly human CoV have caused serious morbidity and mortality worldwide. In this study, six receptor-binding domains (RBDs) derived from α- and β-CoV that are considered to have originated from animals and cross-infected humans were linked to a heterotrimeric scaffold, proliferating cell nuclear antigen (PCNA) subunits, PCNA1, PCNA2, and PCNA3. They assemble to create a stable mosaic multivalent nanoparticle, 6RBD-np, displaying a ring-shaped disk with six protruding antigens, like jewels in a crown. Prime-boost immunizations with 6RBD-np in mice induced significantly high Ab titers against RBD antigens derived from α- and β-CoV and increased interferon (IFN-γ) production, with full protection against the SARS-CoV-2 wild type and Delta challenges. The mosaic 6RBD-np has the potential to induce intergenus cross-reactivity and to be developed as a pan-CoV vaccine against future CoV spillovers.

The assembly of α-synuclein (αS) oligomers is recognized as the main pathological driver of synucleinopathies. While the elimination of toxic αS oligomers shows promise for the treatment of Parkinson's disease (PD), the discovery of αS oligomer degradation drugs has been hindered by the lack of proper drug screening tools. Here, we report a drug screening platform for monitoring the efficacy of αS-oligomer-degrading drugs using amyloid-shelled gold nanocomplexes (ASGNs). We fabricate ASGNs in the presence of dopamine, mimicking the in vivo generation process of pathological αS oligomers. To test our platform, the first of its kind for PD drugs, we use αS-degrading proteases and various small molecular substances that have shown efficacy in PD treatment. We demonstrate that the ASGN-based in vitro platform has strong potential to discover effective αS-oligomer-targeting drugs, and thus it may reduce the attrition problem in drug discovery for PD treatment.

We report that repeated thermal perturbation by thermal cycling (TC) accelerates the formation rate of amyloid filaments at microliter volumes (10-200 μL) and produces a new conformation of zigzag-shaped filaments. The amyloid filaments have been synthesized under different TC conditions, such as temperature variations (Δ

The conventional tissue biopsy method yields isolated snapshots of a narrow region. Therefore, it cannot facilitate comprehensive disease characterization and monitoring. Recently, the detection of tumor-derived components in body fluids─a practice known as liquid biopsy─has attracted increased attention from the biochemical research and clinical application viewpoints. In this vein, surface-enhanced Raman scattering (SERS) has been identified as one of the most powerful liquid-biopsy analysis techniques, owing to its high sensitivity and specificity. Moreover, it affords high-capacity spectral multiplexing for simultaneous target detection and a unique ability to obtain intrinsic biomolecule-fingerprint spectra. This paper presents the fabrication of silver nanosnowflakes (SNSFs) using the polyol method and their subsequent dropping onto a hydrophobic filter paper. The SERS substrate, which comprises the SNSFs and hydrophobic filter paper, facilitates the simultaneous detection of creatinine and cortisol in human sweat using a hand-held Raman spectrometer. The proposed SERS system affords Raman spectrometry to be performed on small sample volumes (2 μL) to identify the normal and at-risk creatinine and cortisol groups.

Highly sensitive rapid testing for COVID-19 is essential for minimizing virus transmission, especially before the onset of symptoms and in asymptomatic cases. Here, we report bioengineered enrichment tools for lateral flow assays (LFAs) with enhanced sensitivity and specificity (BEETLES2), achieving enrichment of SARS-CoV-2 viruses, nucleocapsid (N) proteins and immunoglobulin G (IgG) with 3-minute operation. The limit of detection is improved up to 20-fold. We apply this method to clinical samples, including 83% with either intermediate (35%) or low viral loads (48%), collected from 62 individuals (n = 42 for positive and n = 20 for healthy controls). We observe diagnostic sensitivity, specificity, and accuracy of 88.1%, 100%, and 91.9%, respectively, compared with commercial LFAs alone achieving 14.29%, 100%, and 41.94%, respectively. BEETLES2, with permselectivity and tunability, can enrich the SARS-CoV-2 virus, N proteins, and IgG in the nasopharyngeal/oropharyngeal swab, saliva, and blood serum, enabling reliable and sensitive point-of-care testing, facilitating fast early diagnosis.

Proteolysis of amyloids is related to prevention and treatment of amyloidosis. What if the conditions for proteolysis were the same to those for amyloid formation? For example, pepsin, a gastric protease is activated in an acidic environment, which, interestingly, is also a condition that induces the amyloid formation. Here, we investigate the competition reactions between proteolysis and synthesis of amyloid under pepsin-activated conditions. The changes in the quantities and nanomechanical properties of amyloids after pepsin treatment were examined by fluorescence assay, circular dichroism and atomic force microscopy. We found that, in the case of pepsin-resistant amyloid, a secondary reaction can be accelerated, thereby proliferating amyloids. Moreover, after this reaction, the amyloid became 32.4 % thicker and 24.2 % stiffer than the original one. Our results suggest a new insight into the proteolysis-driven proliferation and rigidification of pepsin-resistant amyloids.

The fast measurement of fibrinogen is essential in evaluating life-threatening sepsis and cardiovascular diseases. Here, we aim to utilize biomimetic plasmonic Au nanoparticles using red blood cell membranes (RBCM-AuNPs) and demonstrate nanoscale coagulation-inspired fibrinogen detection via cross-linking between RBCM-AuNPs. The proposed biomimetic RBCM-AuNPs are highly suitable for fibrinogen detection because hemagglutination, occurring in the presence of fibrinogen, induces a shift in the localized surface plasmon resonance of the NPs. Specifically, when the two ends of the fibrinogen protein are bound to receptors on separate RBCM-AuNPs, cross-linking of the RBCM-AuNPs occurs, yielding a corresponding plasmon shift within 10 min. This coagulation-inspired fibrinogen detection method, with a low sample volume, high selectivity, and high speed, could facilitate the diagnosis of sepsis and cardiovascular diseases.

Colorimetric glucose sensors using enzyme-coronated gold nanoparticles have been developed for high-throughput assays to monitor the blood glucose levels of diabetic patients. Although those sensors have shown sensitivity and wide linear detection ranges, they suffer from poor selectivity and stability in detecting blood glucose, which has limited their practical use. To address this limitation, herein, we functionalized glucose-oxidase-coronated gold nanoparticles with an erythrocyte membrane (EM-GOx-GNPs). Because the erythrocyte membrane (EM) selectively facilitates the permeation of glucose via glucose transporter-1 (GLUT1), the functionalization of GOx-GNPs with EM improved the stability, selectivity (3.3- to 15.8-fold higher), and limit of detection (LOD). Both membrane proteins, GLUT1 and aquaporin-1 (AQP1), on EM were shown to be key components for selective glucose detection by treatment with their inhibitors. Moreover, we demonstrated the stability of EM-GOx-GNPs in high-antioxidant-concentration conditions, under long-term storage (∼4 weeks) and a freeze-thaw cycle. Selectivity of the EM-GOx-GNPs against other saccharides was increased, which improved the LOD in phosphate-buffered saline and human serum. Our results indicated that the functionalization of colorimetric glucose sensors with EM is beneficial for improving selectivity and stability, which may make them candidates for use in a practical glucose sensor.

Graphene-based e-textile gas sensors have received significant attention as wearable electronic devices for human healthcare and environmental monitoring. Theoretically, more the attached graphene on the devices, better is the gas-sensing performance. However, it has been hampered by poor adhesion between graphene and textile platforms. Meanwhile, amyloid nanofibrils are reputed for their ability to improve adhesion between materials, including between graphene and microorganisms. Despite that fact, there has been no attempt to apply amyloid nanofibrils to fabricate graphene-based e-textiles. By biomimicking the adhesion ability of amyloid nanofibrils, herein, we developed a graphene-amyloid nanofibril hybrid e-textile yarn (RGO/amyloid nanofibril/CY) for the detection of NO2. Compared to traditional e-textile yarn, the RGO/amyloid nanofibril/CY showed better performance in response time, sensing efficiency, sensitivity, and selectivity for NO2. Last, we suggested a practical use of RGO/amyloid nanofibril/CY combined with a light-emitting diode as a wearable e-textile gas sensor.

Graphene-based electronic textile (e-textile) gas sensors have been developed for detecting hazardous NO2 gas. For the e-textile gas sensor, electrical conductivity is a critical factor because it directly affects its sensitivity. To obtain a highly conductive e-textile, biomolecules have been used for gluing the graphene to the textile surface, though there remain areas to improve, such as poor conductivity and flexibility. Herein, we have developed a dopamine-graphene hybrid electronic textile yarn (DGY) where the dopamine is used as a bio-inspired adhesive to attach graphene to the surface of yarns. The DGY shows improved electrical conductivity (∼40 times) compared to conventional graphene-based e-textile yarns with no glue. Moreover, it exhibited improved sensing performance in terms of short response time (∼2 min), high sensitivity (0.02 μA/ppm), and selectivity toward NO2. The mechanical flexibility and durability of the DGY were examined through a 1000-cycle bending test. For a practical application, the DGY was attempted to detect the NOx emitted from vehicles, including gasoline, diesel, and fuel cell electric vehicles. Our results demonstrated that the DGYs-as a graphene-based e-textile gas sensor for detecting NO2-are simple to fabricate, cheap, disposable, and mechanically stable.

Spiders synthesize their web using a liquid bridge-to-solidification mechanism at the end of their glands. Inspired by this process, in this work, we fabricated micro-glue threads (μGTs, polymer microwires) by a simple "pinch and spread" process using just two fingertips. The μGTs exhibited excellent tensile strength (∼50 GPa), comparable to those of spider silk and biological fibers. The chemical, physical, and mechanical properties of the μGTs were investigated, and it was confirmed that the thickness of the μGTs could be controlled by ethanol treatment in varying concentrations. Moreover, electrically conductive μGTs were easily fabricated by simply mixing them with various nanomaterials such as gold nanoparticles, zinc oxide nanowires, and reduced graphene oxide (rGO). Interestingly, the conductive μGTs, fabricated using rGO, exhibited remarkable electrical conductivity (0.45 μS) compared to those fabricated using other materials. The conductive μGTs are applicable not only to NO2 gas sensing but also as electrical fuselike materials that melt when the humidity increases. Collectively, the results present μGTs as cost-effective, simple, and versatile materials, which enables their application in a variety of sensors.

In the development of enzymatic glucose sensors, accurate glucose sensing has been a challenging task because of the existence of numerous interfering molecules in the blood. Meanwhile, red blood cells (RBCs) selectively uptake glucose via a membrane protein called glucose transporter-1. In this study, we developed the RBC membrane (RBCM)-coated enzymatic glucose sensors that mimic the glucose uptake. The RBCM-coated sensors were examined via scanning electron microscopy, atomic force microscopy, and ATR-FTIR. We optimized the glucose permeability of the RBCM filter by controlling the thickness of the filter. The sensing range of the optimized sensor was 1-15 mM, the detection limit was 0.66 mM, and the sensitivity was 2.978 μA mM-1. Intriguingly, the RBCM-coated sensor was highly accurate and precise, despite the coexistence of glucose and interfering molecules (e.g., mannose, galactose, ascorbic acid, uric acid, and cysteine). For each interfering molecule, the errors of our sensor were 0.8 to 2.3%, which was 4.8-14.2 times more accurate than the uncoated one. A similar result was verified for a human serum sample containing countless interfering molecules. Also, the sensing performance of the sensor was consistent after 4 weeks of storage. The results suggest that applying RBCM may improve the selectivity of various types of glucose sensors including the continuous monitoring system.

Recurrent spillovers of α- and β-coronaviruses (CoV) such as acute respiratory syndrome (SARS)-CoV, Middle East respiratory syndrome (MERS)-CoV, SARS-CoV-2, and possibly human CoV (NL63, 229E, OC43, and HKU1) have caused serious morbidity and mortality worldwide. Six receptor binding domains (RBDs) derived from α- and β-CoV that are considered to have originated from animals and cross-infected humans were linked to proliferating cell nuclear antigen (PCNA) heterotrimeric subunits, PCNA1, PCNA2, and PCNA3. These were used to form a scaffold-based mosaic multivalent antigen, 6RBD-np. Electron microscopic and atomic force microscopic images show a ring-shaped disk with six protruding RBDs, like jewels in a crown, with a size of 40 nm. Prime-boost immunizations with 6RBD-np in BALB/c mice elicited strong, dose-dependent antibody responses. In human angiotensin converting enzyme 2-transgenic mice, the same immunization induced full-protection against SARS-CoV-2 wild type and Delta challenges, resulting in a 100% survival rate. The mosaic 6RBD-np provides a potential platform for developing a pan-CoV vaccine against newly emerging SARS-CoV-2 variants and future CoV spillovers.SignificanceDespite the arsenal of COVID-19 vaccines, hospitalization and mortality associated with SARS-CoV-2 (acute respiratory syndrome coronavirus 2) variants remain high. There is an urgent need to develop next-generation COVID vaccines that provide broad protection against diseases by current and newly emerging SARS-CoV-2 variants. In this study, six receptor binding domains (RBDs) derived from α- and β-CoV were linked to proliferating cell nuclear antigen (PCNA) heterotrimeric scaffolds. They assemble to create a stable mosaic multivalent nanoparticle, 6RBD-np, displaying a ring-shaped disk with six protruding antigens. The prime-boost immunization in BALB/c and human angiotensin converting enzyme 2-transgenic mice with the 6RBD-np elicited strong, dose-dependent antibody responses and induced full-protection against both the SARS-CoV-2 wild type (WT) and Delta challenges. This study provides proof-of-concept that the mosaic 6RBD-np induces 100% protection against SARS-CoV-2 WT and Delta. It provides the potential of co-displaying heterologous antigens for novel vaccine designs, which can be deployed countering future pandemics.

Graphene electronic textiles (e-textiles) have attracted significant attention in various sensing applications owing to their strong advantages. During the fabrication of these textiles, there are factors to consider, such as electrical conductivity, mechanical flexibility, weight, and applicability in other practical applications. Bioinspired lotus fiber has appropriate advantages to be used as graphene e-textiles, including lightweight (2 gas molecules within a short exposure time (~3 min), including a low detection limit (~1 ppm), selectivity, and resistance to relative humidity. Moreover, we verified the mechanical flexibility of RGOLF through a 1,000-cycle bending test. These results suggest that the bioinspired RGOLF could be used as a gas sensor in environmental air with a strong potential for use in various wearable applications.

Melanoma is visible unlike other types of cancer, but it is still challenging to diagnose correctly because of the difficulty in distinguishing between benign nevus and melanoma. We conducted a robust investigation of melanoma, identifying considerable differences in local elastic properties between nevus and melanoma tissues by using atomic force microscopy (AFM) indentation of histological specimens. Specifically, the histograms of the elasticity of melanoma displayed multimodal Gaussian distributions, exhibiting the heterogeneous mechanical properties, in contrast with the unimodal distributions of elasticity in the benign nevus. We identified this notable signature was consistent regardless of blotch incidence by sex, age, anatomical site (e.g., thigh, calf, arm, eyelid, and cheek), or cancer stage (I, IV, and V). In addition, we found that the non-linearity of the force-distance curves for melanoma is increased compared to benign nevus. We believe that AFM indentation of histological specimens may technically complement conventional histopathological analysis for earlier and more precise melanoma detection.

Because fibrinogen plays an important role as a biomarker of many diseases, including blood coagulation-related and cardiovascular diseases, various methods for the detection of fibrinogen have been developed. Herein, we developed a simple and ultrasensitive electrochemical impedance biosensor (EIB) by draping an erythrocyte membrane (EM) for the detection of fibrinogen. Measurements with the EIB revealed that the specific (selective) adsorption of fibrinogen onto the EM caused a clear rise in the value of interfacial charge transfer resistance. The sensing ability of the EIB for fibrinogen detection showed a wide linear range from 0.0001 to 5 mg/mL, with a limit of detection of 49 ng/mL (144 pM). Moreover, the EIB was able to analyze the fibrinogen in the serum sample, which suggests that it is a facile, low-cost, and ultrasensitive method with great potential for use in the quantitative detection of fibrinogen.

A clear understanding of amyloid formation with diverse morphologies is critical to overcoming the fatal disease amyloidosis. Studies have revealed that monomer concentration is a crucial factor for determining amyloid morphologies, such as protofibrils, annular, or spherical oligomers. However, gaining a complete understanding of the mechanism of formation of the various amyloid morphologies has been limited by the lack of experimental devices and insufficient knowledge. In this study, we demonstrate that the monomer concentration is an essential factor in determining the morphology of beta-amyloid (Aβ) oligomers or protofibrils. By computational and experimental approaches, we investigated the strategies for structural stabilization of amyloid protein, the morphological changes, and amyloid aggregation. In particular, we found unprecedented conformations, e.g., single bent oligomers and segmented ring-shaped protofibrils, the formation of which was explained by the computational analysis. Our findings provide insight into the structural features of amyloid molecules formed at low concentrations of monomer, which will help determine the clinical targets (in therapy) to effectively inhibit amyloid formation in the early stages of the amyloid growth phase.

Enzymatic blood glucose detection with selectivity is one of the most important conundrums, because human blood contains many components that can hinder enzyme-substrate reactions. Meanwhile, cancer cells express much higher levels of glucose transporter-1 on their cell membrane to selectively and excessively uptake more α-D-glucose than do normal cells. Inspired by such cellular permselectivity for glucose, herein we significantly improved the selectivity of a glucose sensor by using a breast cancer cell membrane (BCCM). The BCCM was extracted from MDA-MB-231 cells and coated onto an enzyme-deposited electrode via a vesicle fusion method. We investigated BCCM-coated sensors using ATR-FTIR, SEM, AFM, and cyclic voltammetry. The exceptional permselectivity of BCCM-coated sensors was validated using glucose solutions containing various interfering molecules (e.g., D-(−)-fructose, D-(+)-xylose, D-(+)-maltose, L-cysteine, L-ascorbic acid, and uric acid) and human serum (4.35–7.35 mM of glucose), implying their high potential for practical use.

Fibrinogen, which is a glycoprotein that circulates in the blood, plays various important biological roles, e.g., in blood coagulation, fibroblast proliferation, angiogenesis, and wound healing. Abnormal levels of fibrinogen in plasma have been identified as a key biomarker of a variety of disorders from cardiovascular diseases to hemophilia. Therefore, the development of a quantitative assay for fibrinogen in the blood has emerged as an important issue for the prevention and diagnosis of these diseases. Meanwhile, it is well known that erythrocytes can selectively capture fibrinogen because of the fibrinogen receptor expressed on their plasma membrane. Inspired by these biological interactions, herein, we devised an erythrocyte membrane (EM)-blanketed biosensor based on localized surface plasmon resonance (LSPR) for highly sensitive detection of fibrinogen. By placing the EM onto a nanoparticle-on-substrate, we enhanced the LSPR signal, achieving highly sensitive and selective detection of fibrinogen. We demonstrated that fibrinogen detection is possible over a wide concentration range, 0.001–5.000 mg/mL, which can cover normal and pathological blood fibrinogen levels. In addition, it was verified that the biosensor selectively detects fibrinogen in comparison with other human-blood-plasma components. The nanoplasmonic sensor blanketed with the EM opens up new opportunities for the development of a robust fibrinogen-sensing technology.

MicroRNAs (miRNAs) are important diagnostic and prognostic biomarkers for various tumors. Currently, many diagnostic systems have been developed to detect miRNAs, but simple techniques for detecting miRNAs are still required. Recently, we reported that the expression of miRNA-135b is upregulated in gastric epithelial cells during gastric inflammation and carcinogenesis. Our aim was to develop an in vitro diagnostic platform to analyze the expression of gastric cancer-related biomarkers in the blood. The diagnostic platform comprised an isothermal amplification-based lateral flow biosensor (IA-LFB) that enables easy diagnosis of gastric cancer through visual observation. In this platform, trace amounts of biomarkers are isothermally amplified through rolling circle amplification (RCA), and the amplified product is grafted to the LFB. The performance of the IA-LFB was confirmed using RNAs extracted from in vitro and in vivo models. The platform could detect target miRNAs within 3 h with excellent sensitivity and selectivity. In particular, the IA-LFB could detect the overexpression of gastric cancer-related markers (miRNA-135b and miRNA-21) in RNAs extracted from the blood of patients with various stages (stages 1-4) of gastric cancer compared to that in healthy volunteers. Therefore, IA-LFB is a simple and sensitive in vitro diagnostic system for detecting gastric cancer-related biomarkers and can contribute to the early diagnosis and prognosis monitoring of gastric cancer. Furthermore, this technology can be applied to systems that can detect multiple biomarkers related to various diseases (such as infectious and genetic diseases).