Cholesterol Modulates the Formation of the Aß Ion Channel in Lipid Bilayers.

The misfolding of amyloid beta (Aß) is one of the predominant hallmarks in the pathology of Alzheimer's disease (AD). In this study, we showed that the formation of the Aß ion channel on the membrane depended on the cholesterol concentration. From a mechanical aspect, we found that cholesterol levels affected the stability and assembly of lipid bilayers. Measurements on planar lipid bilayers indicated that a small amount of cholesterol interacted with Aß proteins and promoted the insertion process. Conversely, high cholesterol integrated the lipid bilayer and exerted an opposite effect on Aß insertion. The Aß ion channel was then detected by graphene-based field-effect transistors. Results demonstrated that the Aß ion channel promoted a Ca2+ flux in the presence of 15% cholesterol but prevented a Ca2+ flux in high cholesterol. Thus, cholesterol had a complex impact on the Aß ion channel that can be described as two different effects. First, a small amount of cholesterol interacted with Aß and facilitated the Aß ion channel formation in the membrane. Second, a large amount of cholesterol did not induce the ion flux in the membrane, which can be explained by the cholesterol damage to the regular distribution of the lipid bilayer. Overall, this study suggested a possible approach to consider cholesterol levels for the treatment of AD patients.

Interaction of Sp1 and APP promoter elucidates a mechanism for Pb2+ caused neurodegeneration.

A ubiquitously expressed transcription factor, specificity protein 1 (Sp1), interacts with the amyloid precursor protein (APP) promoter and likely mediates APP expression. Promoter-interaction strengths variably regulate the level of APP expression. Here, we examined the interactions of finger 3 of Sp1 (Sp1-f3) with a DNA fragment containing the APP promoter in different ionic solutions using atomic force microscope (AFM) spectroscopy. Sp1-f3 molecules immobilized on an Si substrate were bound to the APP promoter, which was linked to the AFM tips via covalent bonds. The interactions were strongly influenced by Pb2+, considering that substituting Zn2+ with Pb2+ increased the binding affinity of Sp1 for the APP promoter. The results revealed that the enhanced interaction force facilitated APP expression and that APP overexpression could confer a high-risk for disease incidence. An increased interaction force between Sp1-f3 and the APP promoter in Pb2+ solutions was consistent with a lower binding free energy, as determined by computer-assisted analysis. The impact of Pb2+ on cell morphology and related mechanical properties were also detected by AFM. The overexpression of APP caused by the enhanced interaction force triggered actin reorganization and further resulted in an increased Young's modulus and viscosity. The correlation with single-force measurements revealed that altered cellular activities could result from alternation of Sp1-APP promoter interaction. Our AFM findings offer a new approach in understanding Pb2+ associated neurodegeneration.

Simulation of Graphene Field-Effect Transistor Biosensors for Bacterial Detection.

Foodborne illness is correlated with the existence of infectious pathogens such as bacteria in food and drinking water. Probe-modified graphene field effect transistors (G-FETs) have been shown to be suitable for Escherichia coli (E. coli) detection. Here, the G-FETs for bacterial detection are modeled and simulated with COMSOL Multiphysics to understand the operation of the biosensors. The motion of E. coli cells in electrolyte and the surface charge of graphene induced by E. coli are systematically investigated. The comparison between the simulation and experimental data proves the sensing probe size to be a key parameter affecting the surface charge of graphene induced by bacteria. Finally, the relationship among the change in source-drain current (∆Ids), graphene-bacteria distance and bacterial concentration is established. The shorter graphene-bacteria distance and higher bacterial concentration give rise to better sensing performance (larger ∆Ids) of the G-FETs biosensors. The simulation here could serve as a guideline for the design and optimization of G-FET biosensors for various applications.

Compressive Video Recovery Using Block Match Multi-Frame Motion Estimation Based on Single Pixel Cameras.

Compressive sensing (CS) theory has opened up new paths for the development of signal processing applications. Based on this theory, a novel single pixel camera architecture has been introduced to overcome the current limitations and challenges of traditional focal plane arrays. However, video quality based on this method is limited by existing acquisition and recovery methods, and the method also suffers from being time-consuming. In this paper, a multi-frame motion estimation algorithm is proposed in CS video to enhance the video quality. The proposed algorithm uses multiple frames to implement motion estimation. Experimental results show that using multi-frame motion estimation can improve the quality of recovered videos. To further reduce the motion estimation time, a block match algorithm is used to process motion estimation. Experiments demonstrate that using the block match algorithm can reduce motion estimation time by 30%.

Cellular level robotic surgery: Nanodissection of intermediate filaments in live keratinocytes.

We present the nanosurgery on the cytoskeleton of live cells using AFM based nanorobotics to achieve adhesiolysis and mimic the effect of pathophysiological modulation of intercellular adhesion. Nanosurgery successfully severs the intermediate filament bundles and disrupts cell-cell adhesion similar to the desmosomal protein disassembly in autoimmune disease, or the cationic modulation of desmosome formation. Our nanomechanical analysis revealed that adhesion loss results in a decrease in cellular stiffness in both cases of biochemical modulation of the desmosome junctions and mechanical disruption of intercellular adhesion, supporting the notion that intercellular adhesion through intermediate filaments anchors the cell structure as focal adhesion does and that intermediate filaments are integral components in cell mechanical integrity. The surgical process could potentially help reveal the mechanism of autoimmune pathology-induced cell-cell adhesion loss as well as its related pathways that lead to cell apoptosis.

Probing for chemotherapy-induced peripheral neuropathy in live dorsal root ganglion neurons with atomic force microscopy.

Chemotherapy-induced peripheral neuropathy (CIPN) remains a major reason for cancer patients to withdraw from their lifesaving therapy. CIPN results in irreversible sensory and motor impairments; however, the epidemiology is largely unknown. Here, we report for the first time that chemotherapy drug vincristine not only reduced axonal regeneration in primary dorsal root ganglion neuron but also induced substantial changes in cell mechanical properties detected by atomic force microscopy (AFM). Confocal imaging analysis revealed vincristine-induced microtubule depolymerization. By using AFM for high-resolution live cell imaging and quantitative analysis, we observed significant changes in cell surface roughness and stiffness of vincristine-treated neurons. Elastic modulus was decreased (21-45%) with increasing dosage of vincristine. Further study with paclitaxel, another well-known CIPN drug, confirmed the link between cell mechanics and cytoskeleton organization. These data support that our system can be used for probing potential CIPN drugs that are of enormous benefit to new chemotherapy drug development. FROM THE CLINICAL EDITOR: This study concludes that reduced cell elasticity in dorsal root ganglion neurons accompanies the development of chemotherapy-induced peripheral neuropathy, providing a model system that enables testing of upcoming chemotherapy agents for this particularly inconvenient and often treatment-limiting complication.

Characterization of mechanical behavior of an epithelial monolayer in response to epidermal growth factor stimulation.

Cell signaling often causes changes in cellular mechanical properties. Knowledge of such changes can ultimately lead to insight into the complex network of cell signaling. In the current study, we employed a combination of atomic force microscopy (AFM) and quartz crystal microbalance with dissipation monitoring (QCM-D) to characterize the mechanical behavior of A431 cells in response to epidermal growth factor receptor (EGFR) signaling. From AFM, which probes the upper portion of an individual cell in a monolayer of cells, we observed increases in energy dissipation, Young's modulus, and hysteresivity. Increases in hysteresivity imply a shift toward a more fluid-like mechanical ordering state in the bodies of the cells. From QCM-D, which probes the basal area of the monolayer of cells collectively, we observed decreases in energy dissipation factor. This result suggests a shift toward a more solid-like state in the basal areas of the cells. The comparative analysis of these results indicates a regionally specific mechanical behavior of the cell in response to EGFR signaling and suggests a correlation between the time-dependent mechanical responses and the dynamic process of EGFR signaling. This study also demonstrates that a combination of AFM and QCM-D is able to provide a more complete and refined mechanical profile of the cells during cell signaling.

Cellular biophysical dynamics and ion channel activities detected by AFM-based nanorobotic manipulator in insulinoma ß-cells.

Distinct biochemical, electrochemical and electromechanical coupling processes of pancreatic ß-cells may well underlie different response patterns of insulin release from glucose and capsaicin stimulation. Intracellular Ca(2+) levels increased rapidly and dose-dependently upon glucose stimulation, accompanied with about threefold rapid increases in cellular stiffness. Subsequently, cellular stiffness diminished rapidly and settled at a value about twofold of the baseline. Capsaicin caused a similar transient increase in intracellular Ca(2+) changes. However, cellular stiffness increased gradually to about twofold until leveling off. The current study characterizes for the first time the biophysical properties underlying glucose-induced biphasic responses of insulin secretion, distinctive from the slow and single-phased stiffness response to capsaicin despite similar changes in intracellular Ca(2+) levels. The integrated AFM nanorobotics and optical investigation enables the fine dissection of mechano-property from ion channel activities in response to specific and non-specific agonist stimulation, providing novel biomechanical markers for the insulin secretion process. FROM THE CLINICAL EDITOR: This study characterizes the biophysical properties underlying glucose-induced biphasic responses of insulin secretion. Integrated AFM nanorobotics and optical investigations provided novel biomechanical markers for the insulin secretion process.

High-Fidelity sEMG Signals Recorded by an on-Skin Electrode Based on AgNWs for Hand Gesture Classification Using Machine Learning.

The human forearm is one of the most densely distributed parts of the human body, with the most irregular spatial distribution of muscles. A number of specific forearm muscles control hand motions. Acquiring high-fidelity sEMG signals from human forearm muscles is vital for human-machine interface (HMI) applications based on gesture recognition. Currently, the most commonly used commercial electrodes for detecting sEMG or other electrophysiological signals have a rigid nature without stretchability and cannot maintain conformal contact with the human skin during deformation, and the adhesive hydrogel used in them to reduce skin-electrode impedance may shrink and cause skin inflammation after long-term use. Therefore, developing elastic electrodes with stretchability and biocompatibility for sEMG signal recording is essential for developing HMI. Here, we fabricated a nanocomposite hybrid on-skin electrode by infiltrating silver nanowires (AgNWs), a one-dimensional (1D) nano metal material with conductivity, into polydimethylsiloxane (PDMS), a silicone elastomer with a similar Young's modulus to that of the human skin. The AgNW on-skin electrode has a thickness of 300 µm and low sheet resistance of 0.481 ± 0.014 Ω/sq and can withstand the mechanical strain of up to 54% and maintain a sheet resistance lower than 1 Ω/sq after 1000 dynamic strain cycles. The AgNW on-skin electrode can record high signal-to-noise ratio (SNR) sEMG signals from forearm muscles and can reflect various force levels of muscles by sEMG signals. Besides, four typical hand gestures were recognized by the multichannel AgNW on-skin electrodes with a recognition accuracy of 92.3% using machine learning method. The AgNW on-skin electrode proposed in this study has great potential and promise in various HMI applications that employ sEMG signals as control signals.

Nanomechanical analysis of insulinoma cells after glucose and capsaicin stimulation using atomic force microscopy.

AIM: Glucose stimulates insulin secretion from pancreatic islet ß cells by altering ion channel activity and membrane potential in the ß cells. TRPV1 channel is expressed in the ß cells and capsaicin induces insulin secretion similarly to glucose. This study aims to investigate the biophysical properties of the ß cells upon stimulation of membrane channels using an atomic force microscopic (AFM) nanoindentation system. METHODS: ATCC insulinoma cell line was used. Cell stiffness, a marker of reorganization of cell membrane and cytoskeleton due to ion channel activation, was measured in real time using an integrated AFM nanoindentation system. Cell height that represented structural changes was simultaneously recorded along with cell stiffness. RESULTS: After administration of glucose (16, 20 and 40 mmol/L), the cell stiffness was markedly increased in a dose-dependent manner, whereas cell height was changed in an opposite way. Lower concentrations of capsaicin (1.67 × 10(-9) and 1.67 × 10(-8) mol/L) increased the cell stiffness without altering cell height. In contrast, higher concentrations of capsaicin (1.67 × 10(-6) and 1.67 × 10(-7) mol/L) had no effect on the cell physical properties. CONCLUSION: A unique bio-nanomechanical signature was identified for characterizing biophysical properties of insulinoma cells upon general or specific activation of membrane channels. This study may deepen our understanding of stimulus-secretion coupling of pancreatic islet cells that leads to insulin secretion.

Detection of Bacterial Metabolic Volatile Indole Using a Graphene-Based Field-Effect Transistor Biosensor.

The existence of bacteria is a great threat to food safety. Volatile compounds secreted by bacteria during their metabolic process can be dissected to evaluate bacterial contamination. Indole, as a major volatile molecule released by Escherichia coli (E. coli), was chosen to examine the presence of E. coli in this research. In this work, a graphene field-effect transistor (G-FET) was employed to detect the volatile molecule-indole based on a π-π stacking interaction between the indole and the graphene. The exposure of G-FET devices to the indole provokes a change in electrical signal, which is ascribed to the adsorption of the indole molecule onto the graphene surface via π-π stacking. The adsorption of the indole causes a charge rearrangement of the graphene-indole complex, which leads to changes in the electrical signal of G-FET biosensors with a different indole concentration. Currently, the indole biosensor can detect indole from 10 ppb to 250 ppb and reach a limit of detection of 10 ppb for indole solution detection. We believe that our detection strategy for detecting bacterial metabolic gas molecules will pave a way to developing an effective platform for bacteria detection in food safety monitoring.

Investigation of human keratinocyte cell adhesion using atomic force microscopy.

Desmosomal junctions are specialized structures critical to cellular adhesion within epithelial tissues. Disassembly of these junctions is seen consequent to the development of autoantibodies directed at specific desmosomal proteins in blistering skin diseases such as pemphigus. However, many details regarding cell junction activity under normal physiological and disease conditions remain to be elucidated. Because of their complex structure, desmosomal junctions are not well suited to existing techniques for high-resolution three-dimensional structure-function analyses. Here, atomic force microscopy (AFM) is used for detailed characterization and visualization of the cell junctions of human epithelial cells. We demonstrate the ability to image the detailed three-dimensional structure of the cell junction at high magnification. In addition, the effect of specific antibody binding to desmosomal components of the cell junction is studied in longitudinal analyses before and after antibody treatment. We show that antibodies directed against desmoglein 3 (a major component of the desmosomal structural unit, and the major target of autoantibodies in patients with pemphigus vulgaris) are associated with changes at the cell surface of the human keratinocytes and alterations within keratinocyte intercellular adhesion structures, supporting the assertion that cell structures and junctions are modified by antibody binding. The present study indicates that the molecular structure of gap junctions can be more completely analyzed and characterized by AFM, offering a new technological approach to facilitate a better understanding of disease mechanisms and potentially monitor therapeutic strategies in blistering skin diseases. FROM THE CLINICAL EDITOR: Disassembly of desmosomal junctions is seen in blistering skin diseases such as Pemphigus. This present study demonstrates that the molecular structure of gap junctions can be more completely analyzed and characterized by atomic force microscopy.

An ultraflexible polyurethane yarn-based wearable strain sensor with a polydimethylsiloxane infiltrated multilayer sheath for smart textiles.

Waterproof fiber-based strain sensors with a high gauge factor and outstanding stability are essential for smart textiles, wearable devices and biomedical electronics. In this work, we demonstrate a highly flexible, stretchable, sensitive, and waterproof core-sheath structure strain sensor with a relatively wide strain-sensing range fabricated by a simple approach. Such a core-sheath structure is composed of a superelastic core material polyurethane (PU) yarn; a highly conductive multilayer sheath material, namely, graphene nanosheets/thin gold film/graphene nanosheets (GNSs/Au/GNSs); and a thin polydimethylsiloxane (PDMS) wrapping layer. The combination of the PU yarn, multilayer GNSs/Au/GNSs, and PDMS wrapping layer enables the strain sensor to achieve high flexibility and stretchability, high sensitivity, broad strain-sensing range, and good waterproof property simultaneously due to the infiltration of PDMS into the multilayer during stretching. Particularly, the yarn strain sensor exhibits a high gauge factor (GF: 661.59), outstanding stability with an applied strain of 50% for approximately 10 000 stretch/release cycles, and superior water resistance. Moreover, it can be readily integrated into textiles, including medical textile bandages and textile gloves, for monitoring various human motions (e.g., phonation, pulse, finger bending, and walking) and effectively control a hand robot. Therefore, strain sensors show considerable potential in textile, wearable, and biomedical electronics for healthcare-related applications, such as disease diagnosis, preventive healthcare, and rehabilitation care, and robot controlling-related applications (e.g., controlling a hand robot to catch some objects).

Topography induced stiffness alteration of stem cells influences osteogenic differentiation.

Topography-driven alterations in cell morphology tremendously influence cell biological processes, particularly stem cell differentiation. Aligned topography is known to alter the cell shape, which we anticipated to also induce altered physical properties of the cell. Here, we show that topography has a significant influence on single cell stiffness of human bone marrow derived-Mesenchymal Stem Cells (hBM-MSCs) and the osteogenic differentiation of these. Aligned topographies were used to control the cell elongation, depicted as the cell aspect ratio (CAR). Intriguingly, an equal CAR elicited from different topographies, resulted in highly altered differentiation behavior and the underlying single cell mechanics was found to be critical. The cell behavior was found to be focal adhesion-mediated and induced stiffness alterations rather than just influencing the cell elongation. The effect was further corroborated by investigations of the transcriptional regulators YAP. Our study provides insight into how mechanical properties of the cell, which are stimulated by topography, modulate the osteogenesis of hBM-MSCs, which is beneficial for improving the understanding of interactions between stem cells and topography for developing applications of tissue engineering and regenerative medicine.

Reversible reconfiguration of high-order DNA nanostructures by employing G-quartet toeholds as adhesive units.

G-quadruplex structures are becoming useful alternative interaction modules for the assembly of DNA nanomaterials because of their unique inducibility by cations. In this study, we demonstrated a new strategy for the assembly of polymeric DNA nanoarchitectures in the presence of cations, such as K+ and Na+, by employing G-quartet toeholds at the edges of discrete mini-square DNA building blocks as adhesive units. In comparison with the Watson-Crick base-paired duplex linkers, G-quadruplex arrays embedded in the self-assembled DNA system exhibit higher thermal stability. The morphology of these doughnut-shaped or spherical-shaped DNA nanostructures is highly regulated by the orientation of the folded G-quadruplexes either in parallel or antiparallel orientation in response to different cations. Furthermore, this G-quadruplex-mediated assembly strategy is able to manipulate the cycling of DNA assemblies between discrete and polymeric states by means of introducing cations and chelating agents sequentially. This property enables the reversible manipulation of the DNA-based nanosystems for at least 4 cycles. The G-quadruplex array embedded in this self-assembled DNA system can become a scaffold for functional molecules, as a number of organic molecules and proteins exhibit specific binding to these G-quadruplex structures. Besides, embedded G-quadruplexes are also considered as functional components of nanoscale electronic materials due to their electron transport through the stacked orientation of the G-quartet. Therefore, this work is an important step towards obtaining reversible, responsive G-quadruplex-induced DNA-based nanomaterials with versatile functionalities which will be highly useful in further electronic, biomedical and drug-delivery applications.

Nanoresonant signal boosters for carbon nanotube based infrared detectors.

We report the development of a sensitive carbon nanotube (CNT) infrared detector whose signals are boosted by nanoantenna-like features. This assembly is fabricated using nanoassembly of CNTs and a standard photolithographic process, together with nanoantenna-like features that are designed to create a resonance structure necessary to boost the electric field intensity at the CNT sensor. A novel approach is employed to find the near-field effect of the antenna. As a result, these effects are verified and demonstrated experimentally in this paper. The first experimental demonstration of a practical infrared device with nanoantenna-like structures is reported; it shows that the photocurrent is increased by an order of magnitude. The proposed fabrication and design process enables a ready integration of resonance structures into the manufacture of infrared devices, and opens the possibility of developing high fidelity infrared sensors with wide sensing range.

Extracellular Nanomatrix-Induced Self-Organization of Neural Stem Cells into Miniature Substantia Nigra-Like Structures with Therapeutic Effects on Parkinsonian Rats.

Substantia nigra (SN) is a complex and critical region of the brain wherein Parkinson's disease (PD) arises from the degeneration of dopaminergic neurons. Miniature SN-like structures (mini-SNLSs) constructed from novel combination of nanomaterials and cell technologies exhibit promise as potentially curative cell therapies for PD. In this work, a rapid self-organization of mini-SNLS, with an organizational structure and neuronal identities similar to those of the SN in vivo, is achieved by differentiating neural stem cells in vitro on biocompatible silica nanozigzags (NZs) sculptured by glancing angle deposition, without traditional chemical growth factors. The differentiated neurons exhibit electrophysiological activity in vitro. Diverse physical cues and signaling pathways that are determined by the nanomatrices and lead to the self-organization of the mini-SNLSs are clarified and elucidated. In vivo, transplantation of the neurons from a mini-SNLS results in an early and progressive amelioration of PD in rats. The sculptured medical device reported here enables the rapid and specific self-organization of region-specific and functional brain-like structures without an undesirable prognosis. This development provides promising and significant insights into the screening of potentially curative drugs and cell therapies for PD.

Facile construction of a DNA tetrahedron in unconventional ladder-like arrangements at room temperature.

A DNA tetrahedron as the most classical and simplest three-dimensional DNA nanostructure has been widely utilized in biomedicine and biosensing. However, the existing assembly approaches usually require harsh thermal annealing conditions, involve the formation of unwanted by-products, and have poor size control. Herein, a facile strategy to fabricate a discrete DNA tetrahedron as a single, thermodynamically stable product in a quantitative yield at room temperature is reported. This system does not require a DNA trigger or thermal annealing treatment to initiate self-assembly. This DNA tetrahedron was made of three chemically ligated triangular-shaped DNAs in unconventional ladder-like arrangements, with measured heights of ∼4.16 ± 0.04 nm, showing extra protections for enzymatic degradation in biological environment. They show substantial cellular uptake in different cell lines via temperature, energy-dependent and clathrin-mediated endocytosis pathways. These characteristics allow our DNA tetrahedron to be used as vehicles for the delivery of very small and temperature-sensitive cargos. This novel assembly strategy developed for DNA tetrahedra could potentially be extended to other highly complex polyhedra; this indicated its generalizability.

Biophysical Characteristics of Human Neuroblastoma Cell in Oligomeric $\beta $ -Amyloid (1-40) Cytotoxicity.

Beta amyloid ( ) peptide, which is a common neuropathological hallmark deposit in the brain of patients with Alzheimer's disease, typically comprises 39-43 amino acid residues. peptides exist as isoforms of and with various lengths. In this research, atomic force microscopy (AFM) was applied to investigate aggregations in Hank's Balanced Salt Solution. Toxic effect of oligomer was investigated in live SH-SY5Y neuroblastoma cells by characterizing cell morphology and cell mechanics using high-resolution AFM scanning. oligomer-induced cytoskeleton reorganization was also observed under confocal microscopy, and it can account for reduction in Young's modulus of cells. Meanwhile, phosphorylation of tau increased after oligomer treatment, possibly resulting in microtubule disassembly. This paper demonstrates the linkage between cellular mechanical changes and neurodegeneration mediated by . The method used implies promising applications of real-time monitoring of cellular mechanical properties given the toxic effects of on living neuronal cells.

Development of the Electric Equivalent Model for the Cytoplasmic Microinjection of Small Adherent Cells.

A novel approach utilizing current feedback for the cytoplasmic microinjection of biological cells is proposed. In order to realize the cytoplasmic microinjection on small adherent cells (diameter < 30 µm and thickness < 10 µm), an electrical model is built and analyzed according to the electrochemical properties of target cells. In this study, we have verified the effectiveness of the current measurement for monitoring the injection process and the study of ion channel activities for verifying the cell viability of the cells after the microinjection.