Direct Experimental Observation of Facet-Dependent SERS of Cu2 O Polyhedra.

Semiconductor-based surface enhanced Raman scattering (SERS) has attracted great attention due to its excellent spectral reproducibility, high uniformity, and good anti-interference ability. However, its relatively low SERS sensitivity still hinders its further developments in both performance and applications. Since the SERS is a peculiar surface effect, investigating the facet-dependent SERS activity of semiconductor nanostructures is crucial to boost their SERS signals. Although the semiconductor facet-dependent SERS effect is predicted via numerical calculations, convincing experimental evidence is scarce due to complicated and undefined surface conditions. In this work, three facet-defined ({100}, {110}, and {111} facets) Cu2 O microcrystals (MCs) with clear surface atomic configuration are utilized to investigate the facet-dependent SERS effect. The results from the Kelvin probe force microscopy measurements on single Cu2 O polyhedron, demonstrate that the facet-dependent work function plays a crucial role in the interfacial charge transfer process. Comparing with the {110} and {111} facets, the {100} facet possesses the lowest electronic work function, which enables more efficient interfacial charge transfer. The simulation results further confirm that the {100}-facets can transfer the most electrons from Cu2 O MCs to molecules due to its lowest facet work function, resulting in the largest increment of the molecular polarization.

Biomimetic Bone-like Hydroxyapatite by Mineralization on Supramolecular Porous Fiber Networks.

Hydroxyapatite (HA), the main inorganic component of bone tissue, is mineralized with collagen fibril scaffolds during bone formation. Inspired by the process, a self-assembled porous network architecture was designed and synthesized by using the 2-ureido-4[1H]-pyrimidone (UPy) modified glycerol molecule UPy-Gly, which was further utilized as a template for biomimetic mineralization. When incubated in simulated body fluid (SBF), the HA nucleus first formed in the holes of the template by the induction of hydroxyls on the surface, grew along the nanofibers, and fused with the template to fabricate hydroxyapatite composites (UPy-Gly/HA). Transmission electron microscopic observation demonstrates that the mineral clusters are accumulated by lamella-like nano hydroxyapatite and the elasticity modulus measured by atomic force microscopy is about 5.5 GPa, which is quite close to the natural cancellous bone tissue of human both in structure and in mechanical properties. The Cell Counting Kit 8 (CCK-8) assay of UPy-Gly and UPy-Gly/HA shows noncytotoxicity to mouse fibroblast L-929 cells. This bioinspired composite will be a promising material for potential use in bone tissue implantation and regeneration engineering.

Free-Radical-Assisted Rapid Synthesis of Graphene Quantum Dots and Their Oxidizability Studies.

This work reports a modified electrochemical method for rapid and large-scale preparing graphene quantum dots (GQDs) by introduction of active free radicals, which were produced by hydrogen peroxide or ultraviolet radiation. These free radicals can deepen the oxidized or reduced level of working electrode in electrochemical process and thus lead to GQDs with high concentration and small size, but different surface oxidized degree. The improved oxidation and reduction mechanism were analyzed in this work. Meanwhile, the optical properties and oxidizability of GQDs with different surface oxidized degree were investigated. It is found that these GQDs can be used as an oxidizing agent and their oxidizability is related to the degree being oxidized.

Highly Sensitive CO2-Responsive Polymeric Microgels That Respond Within Seconds.

In this work, polymeric microgels with swift response to CO2 are synthesized by polymerization of tertiary-amine containing methacrylate monomers (N,N-diethylaminoethyl methacrylate, DEAEMA) and polyethylene glycol monomethyl ether acrylate (PEGMA) as stabilizers. The obtained microgels are stable but very sensitive to CO2, which can rapidly swell and further collapse within 5 s upon bubbling of CO2, or within minutes in an atmosphere of gaseous CO2. The protonation of the tertiary amine groups in the presence of CO2 induces sensitive swelling and further irreversible collapse of the microgels due to the internal charge repulsion and relatively low cross-linking density in the core area of microgels. This rapid response to CO2 may find further applications in the fields of sensitive detection or responsive loading and release upon CO2 stimulus.

Why a lotus-like superhydrophobic surface is self-cleaning? An explanation from surface force measurements and analysis.

The unique self-cleaning feature of the lotus-like superhydrophobic (SH) surface attracted worldwide interest in recent years. However, the mechanism of the self-cleaning phenomena remains unclear. Here, we attempt to provide a comprehensive understanding of why self-cleaning of the particles with a broad range of size can be realized on the lotus-like SH surfaces. After measurements and analysis of the force involved at the interface, we conclude that there are four main preconditions for self-cleaning: (1) contact angle (CA) > 90°, (2) low enough sliding angle, (3) low enough adhesion force, and (4) proper particle size. However, as far as the lotus-like SH surface and typical dust are concerned, all the preconditions will be satisfied automatically. We also observe that the particles with a broad range of size (from submicron level to the millimeter level) and density (virtually no limit) can be driven by a water droplet on the lotus-like SH surface. This interesting finding may be helpful for the design of novel engineering system at the micron-millimeter scale in the future.

DNA origami as a carrier for circumvention of drug resistance.

Although a multitude of promising anti-cancer drugs have been developed over the past 50 years, effective delivery of the drugs to diseased cells remains a challenge. Recently, nanoparticles have been used as drug delivery vehicles due to their high delivery efficiencies and the possibility to circumvent cellular drug resistance. However, the lack of biocompatibility and inability to engineer spatially addressable surfaces for multi-functional activity remains an obstacle to their widespread use. Here we present a novel drug carrier system based on self-assembled, spatially addressable DNA origami nanostructures that confronts these limitations. Doxorubicin, a well-known anti-cancer drug, was non-covalently attached to DNA origami nanostructures through intercalation. A high level of drug loading efficiency was achieved, and the complex exhibited prominent cytotoxicity not only to regular human breast adenocarcinoma cancer cells (MCF 7), but more importantly to doxorubicin-resistant cancer cells, inducing a remarkable reversal of phenotype resistance. With the DNA origami drug delivery vehicles, the cellular internalization of doxorubicin was increased, which contributed to the significant enhancement of cell-killing activity to doxorubicin-resistant MCF 7 cells. Presumably, the activity of doxorubicin-loaded DNA origami inhibits lysosomal acidification, resulting in cellular redistribution of the drug to action sites. Our results suggest that DNA origami has immense potential as an efficient, biocompatible drug carrier and delivery vehicle in the treatment of cancer.

Traceless cross-linker for photocleavable bioconjugation.

Photoresponsive bioconjugation empowers the development of novel methods for drug discovery, disease diagnosis, and high-throughput screening, among others. In this paper, we report on the characteristics of a traceless photocleavable cross-linker, di-6-(3-succinimidyl carbonyloxymethyl-4-nitro-phenoxy)-hexanoic acid disulfide diethanol ester (SCNE). The traceless feature and the biocompatibility of this photocleavable cross-linking reagent were corroborated. Consequently, we demonstrated its application in reversible phage particle immobilization that could provide a platform for direct single-phage screening. We also applied it in protein-photoprinting, where SCNE acts as a "photo-eraser" to remove the cross-linked protein molecules at a desired region in a simple, clean, and light-controllable fashion. We further demonstrated the two-tier atomic force microscopic (AFM) method that uses SCNE to carry out two subsequent AFM tasks in situ. The approach allows guided protein delivery and subsequent high-resolution imaging at the same local area, thus opening up the possibility of monitoring protein functions in live cells. The results imply that SCNE is a versatile cross-linker that can be used for a wide range of applications where photocleavage ensures clean and remote-controllable release of biological molecules from a substrate.

Analysis of affinity maps of membrane proteins on individual human embryonic stem cells.

The heterogeneity found in many cell types has greatly inspired research in single-cell gene and protein profiling for discovering the origin of heterogeneity and its role in cell fate decisions. Among the existing techniques to probe heterogeneity, atomic force microscopy (AFM) utilizes an antibody/ligand-modified tip to explore the distribution of a target membrane protein on individual cells in their native environment. In this paper, we establish a practical model to analyze the data systematically, and attempt the quantification of membrane protein abundance on single cells by taking account issues, such as the level of nonspecific interaction, the probe resolution, and the reproducibility of detecting protein distribution. We demonstrated the application in examining the heterogeneous distribution and the local protein abundance of TRA-1-81 antigen on human embryonic stem (hES) cells at the subcellular level. Heterogeneity in TRA-1-81 expression was also detected at the single cell level, suggesting the presence of subpopulation cells within an undifferentiated hES cell colony. The method provides a platform to unveiling the correlation between heterogeneity of membrane proteins and cell development in a complex cell community.

Microphase separation of a quadruple hydrogen bonding supramolecular polymer: effect of the steric hindrance of the ureido-pyrimidone on their viscoelasticity.

Supramolecular polymers based on 2-ureido-4[1H]-pyrimidone (UPy) units with extremely high dimerization constants and adjustable properties have received significant attention. In this work, we attempt to discuss the relationship between the micro-phase separation and the viscoelastic properties of the supramolecular polymers. For this reason, polymers with different UPy moieties structures and different UPy moieties contents were prepared and studied. It was found that the UPy moiety with little hindrance at the six-position of the pyrimidone could self-assemble into a nano-fiber structure and the degree of the micro-phase separation increased with the content of the UPy moiety. With the enlargement of the steric hindrance of the six-position of the pyrimidone, the nano-fiber structure gradually disappeared, meaning the degree of the micro-phase separation decreased astonishingly. More importantly, with the degree of the micro-phase separation increased, the storage modulus or the elasticity modulus increased exponentially and the T m and the loss modulus area increased linearly. These results would lead a new way to study and develop novel polymeric materials.

Profiling TRA-1-81 antigen distribution on a human embryonic stem cell.

Human embryonic stem (hES) cells hold great promise in regenerative medicine. Although hES cells have unlimited self-renewal potential, they tend to differentiate spontaneously in culture. TRA-1-81 is a biomarker of undifferentiated hES cells. Quantitative characterization of TRA-1-81 expression level in a single cell helps capture the "turn-on" signal and understand the mechanism of early differentiation. Here, we report on our examination of TRA-1-81 distribution and association on a hES cell membrane using an atomic force microscope (AFM). Our results suggest that aggregated distribution of TRA-1-81 antigen is characteristic for undifferentiated hES cells. We also evaluated the TRA-1-81 expression level at approximately 17,800 epitopes and approximately 700 epitopes per cell on an undifferentiated cell and a spontaneously differentiated cell, respectively. The method in this study can be adapted in examining other surface proteins on various cell types, thus providing a general tool for investigating protein distribution and association at the single cell level.

Characterization of a Bacillus anthracis spore coat-surface protein that influences coat-surface morphology.

Bacterial spores are encased in a multilayered proteinaceous shell, called the coat. In many Bacillus spp., the coat protects against environmental assault and facilitates germination. In Bacillus anthracis, the spore is the etiological agent of anthrax, and the functions of the coat likely contribute to virulence. Here, we characterize a B. anthracis spore protein, called Cotbeta, which is encoded only in the genomes of the Bacillus cereus group. We found that Cotbeta is synthesized specifically during sporulation and is assembled onto the spore coat surface. Our analysis of a cotbeta null mutant in the Sterne strain reveals that Cotbeta has a role in determining coat-surface morphology but does not detectably affect germination. In the fully virulent Ames strain, a cotbeta null mutation has no effect on virulence in a murine model of B. anthracis infection.

Using a biocompatible diazidecrosslinker to fabricate a robust polyelectrolyte multilayer film with enhanced effects on cell proliferation.

Due to the noncovalent interactions between the layers of polyelectrolyte films, the layer-by-layer assembled multilayered films always face the challenge of low film stiffness and chemical stability under extreme conditions. To handle this issue, we incorporated 4,4'-diazostilbene-2,2'-disulfonic acid disodium salt (DAS) as a crosslinker and subsequently photocrosslinked the layers of a poly(allylamine hydrochloride)/catalase multilayered film. The results showed that DAS could stabilize the prepared film in a manner similar to the traditional cross-linker glutaraldehyde. The multilayered film showed good biocompatibility with a positive effect on cell proliferation. Therefore, by using the commercially available DAS crosslinker, we provide a synthesis-free and biocompatible method to stabilize polyelectrolyte multilayers for broad and significant applications in biological fields, e.g., varying crosslinking density to adjust the mechanical strength of biomaterials, stabilizing susceptible biofilms, or introducing functional groups onto cell membranes.

Mechanical regulation of organ asymmetry in leaves.

How appendages, such as plant leaves or animal limbs, develop asymmetric shapes remains a fundamental question in biology. Although ongoing research has revealed the genetic regulation of organ pattern formation, how gene activity ultimately directs organ shape remains unclear. Here, we show that leaf dorsoventral (adaxial-abaxial) polarity signals lead to mechanical heterogeneity of the cell wall, related to the methyl-esterification of cell-wall pectins in tomato and Arabidopsis. Numerical simulations predicate that mechanical heterogeneity is sufficient to produce the asymmetry seen in planar leaves. Experimental tests that alter pectin methyl-esterification, and therefore cell wall mechanical properties, support this model and lead to polar changes in gene expression, suggesting the existence of a feedback mechanism for mechanical signals in morphogenesis. Thus, mechanical heterogeneity within tissue may underlie organ shape asymmetry.

Hole-enhanced Raman scattering.

Stokes and anti-Stokes non-resonant hole-enhanced Raman scattering (HERS) spectra with high signal-to-noise ratio (S/N) are reported for the first time for aqueous phase R6G molecules adsorbed onto random nanoholes in thin gold films. Compared to conventional surface-enhanced Raman scattering from nanometric gold colloid particles, HERS exhibits higher strength gain, exceptional reproducibility, simple and reliable substrate preparation, and excellent mechanical stability. By correlating the hole density with Raman scattering gain, we determined optimum HERS gain for 50 nm diameter holes at approximately 100 holes/microm(2). Providing a Raman substrate with uniform "hot spots", we expect that HERS will make the quantitative Raman analysis of biological molecules possible.

Confined supramolecular nanostructures of mesogen-bearing amphiphiles.

Stable surface nanostructures with different morphology have been successfully constructed by modifying the chemical structure of synthetic amphiphiles; by introducing mesogenic groups into bolaform amphiphiles, stable spaghetti-like or stripe-like nanostructures can be obtained; it is believed that such a kind of surface structure could be used for templating synthesis and assembly.

Spatially resolved quantification of E-cadherin on target hES cells.

The local expression and distribution pattern of protein on a cell play essential roles in signal transduction within a cell or between cells. Here we report on the development of a spatially resolved quantification method, which was applied in the study of E-cadherin local expression in identified undifferentiated and differentiated human embryonic stem (hES) cells in their native cellular environment. This was achieved by a novel immunofluorescence assisted affinity mapping (IF-AM) method, in which immunofluorescence provides the guidance to locate a desired type of cell in a cell community for performing affinity mapping to quantify the local protein density. The results unveiled the crucial role of E-cadherin in mediating hES cell proliferation and differentiation: the expression of E-cadherin is markedly higher on undifferentiated cells, and the growth of hES cells in unique colonies is contingent on the homogeneous distribution of E-cadherin. Due to the ability of directly assessing individual proteins of a cell, the IF-AM method is shown to be a sensitive tool for resolving subtle differences in the local expression of membrane proteins even at low abundance.

Localization and assembly of proteins comprising the outer structures of the Bacillus anthracis spore.

Bacterial spores possess a series of concentrically arranged protective structures that contribute to dormancy, survival and, ultimately, germination. One of these structures, the coat, is present in all spores. In Bacillus anthracis, however, the spore is surrounded by an additional, poorly understood, morphologically complex structure called the exosporium. Here, we characterize three previously discovered exosporium proteins called ExsFA (also known as BxpB), ExsFB (a highly related paralogue of exsFA/bxpB) and IunH (similar to an inosine-uridine-preferring nucleoside hydrolase). We show that in the absence of ExsFA/BxpB, the exosporium protein BclA accumulates asymmetrically to the forespore pole closest to the midpoint of the sporangium (i.e. the mother-cell-proximal pole of the forespore), instead of uniformly encircling the exosporium. ExsFA/BxpB may also have a role in coat assembly, as mutant spore surfaces lack ridges seen in wild-type spores and have a bumpy appearance. ExsFA/BxpB also has a modest but readily detected effect on germination. Nonetheless, an exsFA/bxpB mutant strain is fully virulent in both intramuscular and aerosol challenge models in Guinea pigs. We show that the pattern of localization of ExsFA/BxpB-GFP is a ring, consistent with a location for this protein in the basal layer of the exosporium. In contrast, ExsFB-GFP fluorescence is a solid oval, suggesting a distinct subcellular location for ExsFB-GFP. We also used these fusion proteins to monitor changes in the subcellular locations of these proteins during sporulation. Early in sporulation, both fusions were present throughout the mother cell cytoplasm. As sporulation progressed, GFP fluorescence moved from the mother cell cytoplasm to the forespore surface and formed either a ring of fluorescence, in the case of ExsFA/BxpB, or a solid oval of fluorescence, in the case of ExsFB. IunH-GFP also resulted in a solid oval of fluorescence. We suggest the interpretation that at least some ExsFB-GFP and IunH-GFP resides in the region between the coat and the exosporium, called the interspace.

Stabilizing bolaform amphiphile interfacial assemblies by introducing mesogenic groups.

We describe the synthesis and characterization of the mesogen-bearing bolaform amphiphile 4,4'-dihydroxybiphenylbis(11-pyridinium-N-yl-undecanoic ester) dibromide (BP-10) and its solid/liquid interfacial self-assembly. Cylindrical micelles are directly observed by atomic force microscopy (AFM) at the interface between mica and the aqueous solution above the critical micelle concentration (cmc). In situ and ex situ AFM studies indicate that the cylindrical micelles are stable both at the mica/solution interface and in the dry state. The enhanced stability of the micellar structures enables a detailed investigation of their self-assembly behavior and supramolecular structures at the interface. The adsorption model proposed here is supported by the variation of the interfacial self-assemblies on changing the solution concentration and substrate temperature.