Rapid and Sensitive Detection of Antigen from SARS-CoV-2 Variants of Concern by a Multivalent Minibinder-Functionalized Nanomechanical Sensor.

New platforms for the rapid and sensitive detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants of concern are urgently needed. Here we report the development of a nanomechanical sensor based on the deflection of a microcantilever capable of detecting the SARS-CoV-2 spike (S) glycoprotein antigen using computationally designed multivalent minibinders immobilized on a microcantilever surface. The sensor exhibits rapid (<5 min) detection of the target antigens down to concentrations of 0.05 ng/mL (362 fM) and is more than an order of magnitude more sensitive than an antibody-based cantilever sensor. Validation of the sensor with clinical samples from 33 patients, including 9 patients infected with the Omicron (BA.1) variant observed detection of antigen from nasopharyngeal swabs with cycle threshold (Ct) values as high as 39, suggesting a limit of detection similar to that of the quantitative reverse transcription polymerase chain reaction (RT-qPCR). Our findings demonstrate the use of minibinders and nanomechanical sensors for the rapid and sensitive detection of SARS-CoV-2 and potentially other disease markers.

Single-Crystal Polycationic Polymers Obtained by Single-Crystal-to-Single-Crystal Photopolymerization.

The efficient preparation of single-crystalline ionic polymers and fundamental understanding of their structure-property relationships at the molecular level remains a challenge in chemistry and materials science. Here, we describe the single-crystal structure of a highly ordered polycationic polymer (polyelectrolyte) and its proton conductivity. The polyelectrolyte single crystals can be prepared on a gram-scale in quantitative yield, by taking advantage of an ultraviolet/sunlight-induced topochemical polymerization, from a tricationic monomer-a self-complementary building block possessing a preorganized conformation. A single-crystal-to-single-crystal photopolymerization was revealed unambiguously by in situ single-crystal X-ray diffraction analysis, which was also employed to follow the progression of molecular structure from the monomer, to a partially polymerized intermediate, and, finally, to the polymer itself. Collinear polymer chains are held together tightly by multiple Coulombic interactions involving counterions to form two-dimensional lamellar sheets (1 nm in height) with sub-nanometer pores (5 Å). The polymer is extremely stable under 254 nm light irradiation and high temperature (above 500 K). The extraordinary mechanical strength and environmental stability-in combination with its impressive proton conductivity (∼3 × 10-4 S cm-1)-endow the polymer with potential applications as a robust proton-conducting material. By marrying supramolecular chemistry with macromolecular science, the outcome represents a major step toward the controlled synthesis of single-crystalline polyelectrolyte materials with perfect tacticity.

Ethylenediammonium-Based "Hollow" Pb/Sn Perovskites with Ideal Band Gap Yield Solar Cells with Higher Efficiency and Stability.

The power conversion efficiency (PCE) of halide perovskite solar cells is now comparable to that of commercial solar cells. These solar cells are generally based on multication mixed-halide perovskite absorbers with nonideal band gaps of 1.5-1.6 eV. The PCE should be able to rise further if the solar cells could use narrower-band gap absorbers (1.2-1.4 eV). Reducing the Pb content of the semiconductors without sacrificing performance is also a significant driver in the perovskite solar cell research. Here, we demonstrate that mixed Pb/Sn-based perovskites containing the oversized ethylenediammonium ( en) dication, { en}FA0.5MA0.5Sn0.5Pb0.5I3 (FA = formamidinium, MA = methylammonium), can exhibit ideal band gaps of 1.27-1.38 eV, suitable for the assembly of single-junction solar cells with higher efficiencies. The use of en dication creates a three-dimensional (3D) hollow inorganic perovskite structure, which was verified through crystal density measurements and single-crystal X-ray diffraction structural analysis as well as nuclear magnetic resonance measurements. The { en}FA0.5MA0.5Sn0.5Pb0.5I3 structure has massive Pb/Sn vacancies and much higher chemical stability than the same structure without en and vacancies. This new property reduces the dark current and carrier trap density and increases the carrier lifetime of the Pb/Sn-based perovskite films. Therefore, solar cells using { en}FA0.5MA0.5Sn0.5Pb0.5I3 light absorbers have substantially enhanced air stability and around 20% improvement in efficiency. After overlaying a thin MABr top layer, we found that the {5% en}FA0.5MA0.5Sn0.5Pb0.5I3 material gives an optimized PCE of 17.04%. The results highlight the strong promise of 3D hollow mixed Pb/Sn perovskites in achieving ideal band gap materials with higher chemical stability and lower Pb content for high-performance single-junction solar cells or multijunction solar cells serving as bottom cells.

Energy landscapes and functions of supramolecular systems.

By means of two supramolecular systems--peptide amphiphiles engaged in hydrogen-bonded ß-sheets, and chromophore amphiphiles driven to assemble by π-orbital overlaps--we show that the minima in the energy landscapes of supramolecular systems are defined by electrostatic repulsion and the ability of the dominant attractive forces to trap molecules in thermodynamically unfavourable configurations. These competing interactions can be selectively switched on and off, with the order of doing so determining the position of the final product in the energy landscape. Within the same energy landscape, the peptide-amphiphile system forms a thermodynamically favoured product characterized by long bundled fibres that promote biological cell adhesion and survival, and a metastable product characterized by short monodisperse fibres that interfere with adhesion and can lead to cell death. Our findings suggest that, in supramolecular systems, functions and energy landscapes are linked, superseding the more traditional connection between molecular design and function.

Surface Heterogeneity at the Polymer-Food Interface Influences Ag Migration from Plastic Packaging Incorporating Ag-Exchanged Zeolites.

Silver-enabled polymers, with their antimicrobial properties, could prolong the shelf life and maintain quality in packaged foods. However, there is limited understanding about how the Ag form in the polymer, food chemistry, and other factors affect the transfer (migration) of Ag from the polymer to the food under the intended conditions of use. In this study, we investigated the release of Ag from polymer composites (PCs) incorporating two different Ag-exchanged zeolites (Ag-Y), which have been explored as potential scaffolds for loading high concentrations of Ag within common polymers. We manufactured two Ag-Y films based on low-density polyethylene (LDPE): one incorporating ionic Ag (Ag+) and one incorporating nanoparticulate Ag (AgNPs), each with similar initial Ag concentrations. Then, we assessed the migration of Ag out of these PCs into food simulants under accelerated room temperature storage conditions. In all simulants investigated, the Ag+-Y/LDPE film exhibited a higher migration of Ag compared to the AgNP-Y/LDPE film, suggesting a lower fraction of readily releasable Ag in the latter material. Total Ag migration from AgNP-Y/LDPE over 10 days at 40 °C was 11.10 ± 2.05 ng cm-2 of packaging surface area in water, 7.63 ± 1.59 ng cm-2 in a 9 wt % aqueous sucrose solution, and 21.29 ± 1.98 ng cm-2 in a commercial sweetened carbonated beverage (Squirt). In contrast, Ag migration from Ag+-Y/LDPE was measured at 49.61 ± 3.46, 57.48 ± 9.65, and 91.54 ± 5.58 ng cm-2 in water, sucrose solution, and Squirt drink, respectively. Surface characterization techniques, including atomic force microscopy (AFM), scanning electron microscopy (SEM), and conductivity measurements, revealed the presence of exposed zeolite particles at the surface of the films, suggesting that direct interactions between Ag-exchanged zeolites and food components at the simulant-polymer interface play an important role in determining Ag migration from Ag-Y/LDPE PCs.

Highly sensitive and ultra-rapid antigen-based detection of SARS-CoV-2 using nanomechanical sensor platform.

The rapid spread of COVID-19 including recent emergence of new variants with its extreme range of pathologies create an urgent need to develop a versatile sensor for a rapid, precise, and highly sensitive detection of SARS-CoV-2. Herein, we report a microcantilever-based optical detection of SARS-CoV-2 antigenic proteins in just few minutes with high specificity by employing fluidic-atomic force microscopy (f-AFM) mediated nanomechanical deflection method. The corresponding antibodies against the target antigens were first grafted on the gold-coated microcantilever surface pre-functionalized with EDC-NHS chemistry for a suitable antibody-antigen interaction. Rapid detection of SARS-CoV-2 nucleocapsid (N) and spike (S1) receptor binding domain (RBD) proteins was first demonstrated at a clinically relevant concentration down to 1 ng/mL (33 pM) by real-time monitoring of nanomechanical signal induced by antibody-antigen interaction. More importantly, we further show high specific detection of antigens with nasopharyngeal swab specimens from patients pre-determined with qRT-PCR. The results take less than 5 min (swab to signal ≤5 min) and exhibit high selectivity and analytical sensitivity (LoD: 100 copies/ ml; 0.71 ng/ml of N protein). These findings demonstrate potential for nanomechanical signal transduction towards rapid antigen detection for early screening of SARS-CoV-2 and its related mutants.

A cortactin CTTN coding SNP contributes to lung vascular permeability and inflammatory disease severity in African descent subjects.

The cortactin gene (CTTN), encoding an actin-binding protein critically involved in cytoskeletal dynamics and endothelial cell (EC) barrier integrity, contains single nucleotide polymorphisms (SNPs) associated with severe asthma in Black patients. As loss of lung EC integrity is a major driver of mortality in the Acute Respiratory Distress Syndrome (ARDS), sepsis, and the acute chest syndrome (ACS), we speculated CTTN SNPs that alter EC barrier function will associate with clinical outcomes from these types of conditions in Black patients. In case-control studies, evaluation of a nonsynonymous CTTN coding SNP Ser484Asn (rs56162978, G/A) in a severe sepsis cohort (725 Black subjects) revealed significant association with increased risk of sepsis mortality. In a separate cohort of sickle cell disease (SCD) subjects with and without ACS (177 SCD Black subjects), significantly increased risk of ACS and increased ACS severity (need for mechanical ventilation) was observed in carriers of the A allele. Human lung EC expressing the cortactin S484N transgene exhibited: (i) delayed EC barrier recovery following thrombin-induced permeability; (ii) reduced levels of critical Tyr486 cortactin phosphorylation; (iii) inhibited binding to the cytoskeletal regulator, nmMLCK; and (iv) attenuated EC barrier-promoting lamellipodia dynamics and biophysical responses. ARDS-challenged Cttn+/- heterozygous mice exhibited increased lung vascular permeability (compared to wild-type mice) which was significantly attenuated by IV delivery of liposomes encargoed with CTTN WT transgene but not by CTTN S484N transgene. In summary, these studies suggest that the CTTN S484N coding SNP contributes to severity of inflammatory injury in Black patients, potentially via delayed vascular barrier restoration.

Exploring the Factors Affecting the Mechanical Properties of 2D Hybrid Organic-Inorganic Perovskites.

Mechanical stability of hybrid organic-inorganic perovskites (HOIPs) is essential to achieve long-term durable HOIP-based devices. While HOIPs in two-dimensional (2D) form offer numerous options in the structure and composition to tune their mechanical properties, little is known about the structure-mechanical-property relationship in this family of materials. Here, we investigated a series of 2D lead halide HOIPs by nanoindentation to explore the impact of critical factors controlling the properties of both the organic and inorganic layers on the materials' out-of-plane mechanical performance. We find that the lead-halide bond in the inorganic framework can significantly influence the mechanical properties of 2D Ruddlesden-Popper (RP) HOIPs with n = 1. Like 3D HOIPs, stronger lead-halide bond strength leads to a higher Young's modulus in these 2D HOIPs, i.e., E⊥Cl ≳ E⊥Br > E⊥I. In contrast, the hardness of 2D RP HOIPs follows a trend of HBr2D > HCl2D > HI2D, which is different from that found in 3D HOIPs, probably due to the combined effects from the Pb-X bond strength and inorganic framework structural change (e.g., symmetry and distortion). We further show that the interface between the organic layers in 2D HOIPs can be an effective route to engineer the materials' mechanical properties. Replacing the weak CH3-CH3 van der Waals forces by covalent bonds or phenyl-phenyl interactions in the interface can lead to a much stiffer and harder 2D HOIPs. Finally, we discover that the mechanical performance of 2D HOIPs with linear aliphatic diammonium spacer molecules is affected by the two basic structural parameters, i.e., the thicknesses of the organic and inorganic layers, in a similar way compared to that of 2D RP HOIPs with linear aliphatic monoammonium spacer molecules. A thinner organic layer and a thicker inorganic layer can result in 2D HOIPs with larger elastic modulus and hardness values. Our results offer intriguing insights into the structure-property relationship of 2D HOIPs from a mechanical perspective, providing guidelines and inspirations to achieve material design with required mechanical properties for applications.

Ovarian stiffness increases with age in the mammalian ovary and depends on collagen and hyaluronan matrices.

Fibrosis is a hallmark of aging tissues which often leads to altered architecture and function. The ovary is the first organ to show overt signs of aging, including increased fibrosis in the ovarian stroma. How this fibrosis affects ovarian biomechanics and the underlying mechanisms are unknown. Using instrumental indentation, we demonstrated a quantitative increase in ovarian stiffness, as evidenced by an increase in Young's modulus, when comparing ovaries from reproductively young (6-12 weeks) and old (14-17 months) mice. This ovarian stiffness was dependent on collagen because ex vivo enzyme-mediated collagen depletion in ovaries from reproductively old mice restored their collagen content and biomechanical properties to those of young controls. In addition to collagen, we also investigated the role of hyaluronan (HA) in regulating ovarian stiffness. HA is an extracellular matrix glycosaminoglycan that maintains tissue homeostasis, and its loss can change the biomechanical properties of tissues. The total HA content in the ovarian stroma decreased with age, and this was associated with increased hyaluronidase (Hyal1) and decreased hyaluronan synthase (Has3) expression. These gene expression differences were not accompanied by changes in ovarian HA molecular mass distribution. Furthermore, ovaries from mice deficient in HAS3 were stiffer compared to age-matched WT mice. Our results demonstrate that the ovary becomes stiffer with age and that both collagen and HA matrices are contributing mechanisms regulating ovarian biomechanics. Importantly, the age-associated increase in collagen and decrease in HA are conserved in the human ovary and may impact follicle development and oocyte quality.

Structured silicon for revealing transient and integrated signal transductions in microbial systems.

Bacterial response to transient physical stress is critical to their homeostasis and survival in the dynamic natural environment. Because of the lack of biophysical tools capable of delivering precise and localized physical perturbations to a bacterial community, the underlying mechanism of microbial signal transduction has remained unexplored. Here, we developed multiscale and structured silicon (Si) materials as nongenetic optical transducers capable of modulating the activities of both single bacterial cells and biofilms at high spatiotemporal resolution. Upon optical stimulation, we capture a previously unidentified form of rapid, photothermal gradient-dependent, intercellular calcium signaling within the biofilm. We also found an unexpected coupling between calcium dynamics and biofilm mechanics, which could be of importance for biofilm resistance. Our results suggest that functional integration of Si materials and bacteria, and associated control of signal transduction, may lead to hybrid living matter toward future synthetic biology and adaptable materials.

Mapping Hot Spots at Heterogeneities of Few-Layer Ti3C2 MXene Sheets.

Structural defects and heterogeneities play an enormous role in the formation of localized hot spots in 2D materials used in a wide range of applications from electronics to energy systems. In this report, we employ scanning thermal microscopy (SThM) to spatially map the temperature rise across various defects and heterogeneities of titanium carbide (Ti3C2T x; T stands for surface terminations) MXene nanostructures under high electrical bias with sub-50 mK temperature resolution and sub-100 nm spatial resolution. We investigated several Ti3C2T x flakes having different thicknesses as well as heterogeneous MXene structures incorporating line defects or vertical heterojunctions. High-resolution temperature rise maps allow us to identify localized hot spots and to quantify the nonuniformity of the temperature fields across various morphological features. The results show that the local heating is most severe in vertical junctions of MXene flakes and is highly affected by nonuniform conduction due to the presence of line defects. These results provide a direct insight into the power dissipation of MXene-based devices and the roles of various heterogeneities that are inherent to the material synthesis process. This study provides a guideline for how a better understanding of the structure-property-processing correlations and further optimization of the synthesis routes could improve the lifetime, safety, and operation limits of the MXene-based devices.

Spatial Mapping of Hot-Spots at Lateral Heterogeneities in Monolayer Transition Metal Dichalcogenides.

Lateral heterogeneities in atomically thin 2D materials such as in-plane heterojunctions and grain boundaries (GBs) provide an extrinsic knob for manipulating the properties of nano- and optoelectronic devices and harvesting novel functionalities. However, these heterogeneities have the potential to adversely affect the performance and reliability of the 2D devices through the formation of nanoscopic hot-spots. In this report, scanning thermal microscopy (SThM) is utilized to map the spatial distribution of the temperature rise within monolayer transition metal dichalcogenide (TMD) devices upon dissipating a high electrical power through a lateral interface. The results directly demonstrate that lateral heterojunctions between MoS2 and WS2 do not largely impact the distribution of heat dissipation, while GBs of MoS2 appreciably localize heating in the device. High-resolution scanning transmission electron microscopy reveals that the atomic structure is nearly flawless around heterojunctions but can be quite defective near GBs. The results suggest that the interfacial atomic structure plays a crucial role in enabling uniform charge transport without inducing localized heating. Establishing such structure-property-processing correlation provides a better understanding of lateral heterogeneities in 2D TMD systems which is crucial in the design of future all-2D electronic circuitry with enhanced functionalities, lifetime, and performance.

Out-of-Plane Mechanical Properties of 2D Hybrid Organic-Inorganic Perovskites by Nanoindentation.

Two-dimensional (2D) layered hybrid organic-inorganic perovskites (HOIPs) have demonstrated improved stability and promising photovoltaic performance. The mechanical properties of such functional materials are both fundamentally and practically important to achieve both high performance and mechanically stable (flexible) devices. Here, we report the mechanical properties of a series of 2D layered lead iodide HOIPs and investigate the role of structural subunits (e.g., variation of the length of the organic spacer molecules, R and the number of inorganic layers, n) in the mechanical properties. Although 2D HOIPs have much lower nominal elastic modulus and hardness than 3D HOIPs, larger n number and shorter R lead to stiffer materials. Density functional theory simulations showed a trend similar to the experimental results. We compared these findings with other 2D layered crystals and shed light on routes to further tune the out-of-plane mechanical properties of 2D layered HOIPs.

Stretching and Breaking of Ultrathin 2D Hybrid Organic-Inorganic Perovskites.

Two-dimensional (2D) hybrid organic-inorganic perovskites (HOIPs) are recent members of the 2D materials family with wide tunability, highly dynamic structural features, and excellent physical properties. Ultrathin 2D HOIPs and their heterostructures with other 2D materials have been exploited for study of physical phenomena and device applications. The in-plane mechanical properties of 2D ultrathin HOIPs are critical for understanding the coupling between mechanical and other physical fields and for integrated devices applications. Here we report the in-plane mechanical properties of ultrathin freestanding 2D lead iodide perovskite membranes and their dependence on the membrane thickness. The in-plane Young's moduli of 2D HOIPs are smaller than that of conventional covalently bonded 2D materials. As the thickness increases from monolayer to three-layer, both the Young's modulus and breaking strength decrease, while three-layer and four-layer 2D HOIPs have almost identical in-plane mechanical properties. These thickness-dependent mechanical properties can be attributed to interlayer slippage during deformation. Our results show that ultrathin 2D HOIPs exhibit outstanding breaking strength/Young's modulus ratio compared to many other widely used engineering materials and polymeric flexible substrates, which renders them suitable for application into flexible electronic devices.

Mammary epithelium-specific inactivation of V-ATPase reduces stiffness of extracellular matrix and enhances metastasis of breast cancer.

Extracellular matrix (ECM) critically impacts tumor progression and is influenced by both cancer and host tissue cells. While our understanding of cancer cell ECM remodeling is widespread, the importance of host tissue ECM, which provides initial congenial environment for primary tumor formation, is partly understood. Here, we report a novel role of epithelial cell-associated vacuolar ATPase 'a2' isoform (a2V) in regulating breast tissue ECM stiffness to control metastasis. Using a mammary gland-specific a2V-knockout model, we show that in the absence of a2V, breast tumors exhibit atypically soft tumor phenotype, less tumor rigidity, and necrotic tumor microenvironment. These tumors contain a decreased number of cancer cells at primary tumor site, but showed extensive metastases compared to control. Nanomechanical evaluation of normal breast tissues revealed a decrease in stiffness and collagen content in ECM of a2V-deleted breast tissues. Mechanistically, inhibition of a2V expression caused dispersed Golgi morphology with relocation of glycosyltransferase enzymes to early endosomes in mammary epithelial cells. This resulted in defective glycosylation of ECM proteins and production of compromised ECM that further influenced tumor metastasis. Clinically, in patients with cancer, low a2V expression levels in normal breast tissue correlated with lymph node metastasis. Thus, using a new knockout mouse model, we have identified a2V expression in epithelial cells as a key requirement for proper ECM formation in breast tissue and its expression levels can significantly modulate breast tumor dissemination. Evaluation of a2V expression in normal breast tissues can help in identifying patients with high risk of developing metastases.

Micromachined Chip Scale Thermal Sensor for Thermal Imaging.

The lateral resolution of scanning thermal microscopy (SThM) has hitherto never approached that of mainstream atomic force microscopy, mainly due to poor performance of the thermal sensor. Herein, we report a nanomechanical system-based thermal sensor (thermocouple) that enables high lateral resolution that is often required in nanoscale thermal characterization in a wide range of applications. This thermocouple-based probe technology delivers excellent lateral resolution (∼20 nm), extended high-temperature measurements >700 °C without cantilever bending, and thermal sensitivity (∼0.04 °C). The origin of significantly improved figures-of-merit lies in the probe design that consists of a hollow silicon tip integrated with a vertically oriented thermocouple sensor at the apex (low thermal mass) which interacts with the sample through a metallic nanowire (50 nm diameter), thereby achieving high lateral resolution. The efficacy of this approach to SThM is demonstrated by imaging embedded metallic nanostructures in silica core-shell, metal nanostructures coated with polymer films, and metal-polymer interconnect structures. The nanoscale pitch and extremely small thermal mass of the probe promise significant improvements over existing methods and wide range of applications in several fields including semiconductor industry, biomedical imaging, and data storage.

Development of ultrasound bioprobe for biological imaging.

We report the development of an ultrasound bioprobe for in vitro molecular imaging. In this method, the phase of the scattered ultrasound wave is mapped to provide in vitro and intracellular imaging with nanometer-scale resolution under physiological conditions. We demonstrated the technique by successfully imaging a magnetic core in silica core shells and the stiffness image of intracellular fibers in endothelial cells that were stimulated with thrombin. The findings demonstrate a significant advancement in high-resolution ultrasound imaging of biological systems with acoustics under physiological conditions. These will open up various applications in biomedical and molecular imaging with subsurface resolution down to the nanometer scale.

Thickness Resonance Acoustic Microscopy for Nanomechanical Subsurface Imaging.

A nondestructive scanning near-field thickness resonance acoustic microscopy (SNTRAM) has been developed that provides high-resolution mechanical depth sensitivity and sharp phase contrast of subsurface features. In SNTRAM technology, we excited the sample at its thickness resonance, at which a sharp change in phase is observed and mapped with a scanning probe microscopy stage in near field to provide nanometer-scale nanomechanical contrast of subsurface features/defects. We reported here the remarkable subsubsurface phase contrast and sensitivity of SNTRAM by exciting the sample with a sinusoidal elastic wave at a frequency equal to the thickness resonance of the sample. This results in a large shift in phase component associated with the bulk longitudinal wave propagating through the sample thickness, thus suggesting the usefulness of this method for (a) generating better image contrast due to high S/N of the transmitted ultrasound wave to the other side of the sample and (b) sensitive detection of local variation in material properties at much better resolution due to the sharp change in phase. We demonstrated that the sample excited at the thickness resonance has a more substantial phase contrast and depth sensitivity than that excited at off-resonance and related acoustic techniques. Subsurface features down to 5-8 nm lateral resolution have been demonstrated using a standard sample.

Nano-Biomechanical Study of Spatio-Temporal Cytoskeleton Rearrangements that Determine Subcellular Mechanical Properties and Endothelial Permeability.

The endothelial cell (EC) lining of the pulmonary vascular system forms a semipermeable barrier between blood and the interstitium and regulates various critical biochemical functions. Collectively, it represents a prototypical biomechanical system, where the complex hierarchical architecture, from the molecular scale to the cellular and tissue level, has an intimate and intricate relationship with its biological functions. We investigated the mechanical properties of human pulmonary artery endothelial cells (ECs) using atomic force microscopy (AFM). Concurrently, the wider distribution and finer details of the cytoskeletal nano-structure were examined using fluorescence microscopy (FM) and scanning transmission electron microscopy (STEM), respectively. These correlative measurements were conducted in response to the EC barrier-disrupting agent, thrombin, and barrier-enhancing agent, sphingosine 1-phosphate (S1P). Our new findings and analysis directly link the spatio-temporal complexities of cell re-modeling and cytoskeletal mechanical properties alteration. This work provides novel insights into the biomechanical function of the endothelial barrier and suggests similar opportunities for understanding the form-function relationship in other biomechanical subsystems.

Elucidating molecular mass and shape of a neurotoxic Aß oligomer.

Alzheimer's disease (AD), the most prevalent type of dementia, has been associated with the accumulation of amyloid ß oligomers (AßOs) in the central nervous system. AßOs vary widely in size, ranging from dimers to larger than 100 kDa. Evidence indicates that not all oligomers are toxic, and there is yet no consensus on the size of the actual toxic oligomer. Here we used NU4, a conformation-dependent anti-AßO monoclonal antibody, to investigate size and shape of a toxic AßO assembly. By using size-exclusion chromatography and immuno-based detection, we isolated an AßO-NU4 complex amenable for biochemical and morphological studies. The apparent molecular mass of the NU4-targeted oligomer was 80 kDa. Atomic force microscopy imaging of the AßO-NU4 complex showed a size distribution centered at 5.37 nm, an increment of 1.5 nm compared to the size of AßOs (3.85 nm). This increment was compatible with the size of NU4 (1.3 nm), suggesting a 1:1 oligomer to NU4 ratio. NU4-reactive oligomers extracted from AD human brain concentrated in a molecular mass range similar to that found for in vitro prepared oligomers, supporting the relevance of the species herein studied. These results represent an important step toward understanding the connection between AßO size and toxicity.