Target gene overexpression and enhanced metabolism confer resistance to nicosulfuron in Eriochloa villosa (Thunb.).

Eriochloa villosa (Thunb.) Kunth is a troublesome weed widely distributed in maize (Zea mays L.) fields in Northeast China. Many populations of E. villosa have evolved resistance to nicosulfuron herbicides, which inhibit acetolactate synthase (ALS). The objectives of this research were to confirm that E. villosa is resistant to nicosulfuron and to investigate the basis of nicosulfuron resistance. Whole-plant dose-response studies revealed that the R population had not developed a high level of cross-resistance and exhibited greater resistant (25.62-fold) to nicosulfuron than that of the S population and had not yet developed a high level of cross-resistance. An in vitro ALS activity assay demonstrated that the I50 of nicosulfuron was 6.87-fold greater in the R population than the S population. However, based on ALS gene sequencing, the target ALS gene in the R population did not contain mutations. Quantitative real-time polymerase chain reaction (qRT-PCR) revealed that ALS gene expression between the R and S populations was significantly different after nicosulfuron application, but no differences were observed in the gene copy number. After the cytochrome P450 inhibitor malathion or the GST inhibitor NBD-Cl was applied, the resistant E. villosa population exhibited increased sensitivity to nicosulfuron. Based on the activities of GSTs and P450s, the activities of the R population were greater than those of the S population after nicosulfuron application. This is the first report that the resistance of E. villosa to ALS inhibitors results from increased target gene expression and increased metabolism. These findings provide a theoretical foundation for the effective control of herbicide-resistant E. villosa.

Comparative study of first-principles approaches for effective Coulomb interaction strength Ueff between localized f-electrons: Lanthanide metals as an example.

As correlation strength has a key influence on the simulation of strongly correlated materials, many approaches have been proposed to obtain the parameter using first-principles calculations. However, a comparison of the different Coulomb strengths obtained using these approaches and an investigation of the mechanisms behind them are still needed. Taking lanthanide metals as an example, we research the factors that affect the effective Coulomb interaction strength, Ueff, by local screened Coulomb correction (LSCC), linear response (LR), and constrained random-phase approximation (cRPA) in the Vienna Ab initio Simulation Package. The Ueff LSCC value increases from 4.75 to 7.78 eV, Ueff LR is almost stable at about 6.0 eV (except for Eu, Er, and Yb), and Ueff cRPA shows a two-stage decreasing trend in both light and heavy lanthanides. To investigate these differences, we establish a scheme to analyze the coexistence and competition between the orbital localization and the screening effect. We find that LSCC and cRPA are dominated by the orbital localization and the screening effect, respectively, whereas LR shows the balance of the competition between the two factors. Additionally, the performance of these approaches is influenced by different starting points from the Perdew-Burke-Ernzerhof (PBE) and PBE + U, especially for cRPA. Our results provide useful knowledge for understanding the Ueff of lanthanide materials, and similar analyses can also be used in the research of other correlation strength simulation approaches.

Concentration optimization of combinatorial drugs using Markov chain-based models.

BACKGROUND: Combinatorial drug therapy for complex diseases, such as HSV infection and cancers, has a more significant efficacy than single-drug treatment. However, one key challenge is how to effectively and efficiently determine the optimal concentrations of combinatorial drugs because the number of drug combinations increases exponentially with the types of drugs. RESULTS: In this study, a searching method based on Markov chain is presented to optimize the combinatorial drug concentrations. In this method, the searching process of the optimal drug concentrations is converted into a Markov chain process with state variables representing all possible combinations of discretized drug concentrations. The transition probability matrix is updated by comparing the drug responses of the adjacent states in the network of the Markov chain and the drug concentration optimization is turned to seek the state with maximum value in the stationary distribution vector. Its performance is compared with five stochastic optimization algorithms as benchmark methods by simulation and biological experiments. Both simulation results and experimental data demonstrate that the Markov chain-based approach is more reliable and efficient in seeking global optimum than the benchmark algorithms. Furthermore, the Markov chain-based approach allows parallel implementation of all drug testing experiments, and largely reduces the times in the biological experiments. CONCLUSION: This article provides a versatile method for combinatorial drug screening, which is of great significance for clinical drug combination therapy.

Atomic force microscopy for revealing micro/nanoscale mechanics in tumor metastasis: from single cells to microenvironmental cues.

Mechanics are intrinsic properties which appears throughout the formation, development, and aging processes of biological systems. Mechanics have been shown to play important roles in regulating the development and metastasis of tumors, and understanding tumor mechanics has emerged as a promising way to reveal the underlying mechanisms guiding tumor behaviors. In particular, tumors are highly complex diseases associated with multifaceted factors, including alterations in cancerous cells, tissues, and organs as well as microenvironmental cues, indicating that investigating tumor mechanics on multiple levels is significantly helpful for comprehensively understanding the effects of mechanics on tumor progression. Recently, diverse techniques have been developed for probing the mechanics of tumors, among which atomic force microscopy (AFM) has appeared as an excellent platform enabling simultaneously characterizing the structures and mechanical properties of living biological systems ranging from individual molecules and cells to tissue samples with unprecedented spatiotemporal resolution, offering novel possibilities for understanding tumor physics and contributing much to the studies of cancer. In this review, we survey the recent progress that has been achieved with the use of AFM for revealing micro/nanoscale mechanics in tumor development and metastasis. Challenges and future progress are also discussed.

Label-free characterization of different kinds of cells using optoelectrokinetic-based microfluidics.

We report a novel, to the best of our knowledge, method to rapidly characterize different kinds of cells and drug-treated cancer cells using a label-free biomarker of self-rotation in an optoelectrokinetics (OEK)-based microfluidic platform. OEK incorporates optics and electrokinetics into microfluidics, thereby offering a contact-free, label-free, and rapid approach to the cellular manipulation community. Self-rotational behaviors of four different kinds of cells were experimentally investigated by the frequency-sweeping of an AC bias potential in an optically induced nonuniform and irrotational electric field. The results revealed that these kinds of cells displayed a Gaussian distribution versus the AC frequency as well as different self-rotational speeds under the same conditions. Furthermore, the peak self-rotational speed varied from one kind of cell to another, with that of cancer cells higher than that of normal cells. In addition, MCF-7 cells treated by various concentrations of drug showed remarkably different self-rotational speeds. This finding suggests a high potential of developing a new label-free biomarker to rapidly distinguish different kinds of cancer cells and quantitatively monitor the response of cancer patients to various treatments.

Recent advances in microfluidic technologies for separation of biological cells.

Cell separation has always been a key topic in academic research, especially in the fields of medicine and biology, due to its significance in diagnosis and treatment. Accurate, high-throughput and non-invasive separation of individual cells is key to driving the development of biomedicine and cellular biology. In recent years, a series of researches on the use of microfluidic technologies for cell separation have been conducted to solve bio-related problems. Hence, we present here a comprehensive review on the recent developments of microfluidic technologies for cell separation. In this review, we discuss several cell separation methods, mainly including: physical and biochemical method, their working principles as well as their practical applications. We also analyze the advantages and disadvantages of each method in detail. In addition, the current challenges and future prospects of microfluidic-based cell separation were discussed.

Rotational Cluster Anion Enabling Superionic Conductivity in Sodium-Rich Antiperovskite Na3OBH4.

Sodium superionic conductors are keys to develop high safety and low cost all-solid-state sodium batteries. Among developed sodium ionic conductors, antiperovskite-type ionic conductors have attracted vast interest due to their high structural tolerance and good formability. Herein, we successfully synthesize Na3OBH4 with cubic antiperovskite structure by solid-state reaction from Na2O and NaBH4. Na3OBH4 exhibits ionic conductivity of 4.4 × 10-3 S cm-1 at room temperature (1.1 × 10-2 S cm-1 at 328 K) and activation energy of 0.25 eV. The ionic conductivity is 4 orders of magnitude higher than the existing antiperovskite Na3OX (X = Cl, Br, I). It is shown that such enhancement is not only due to the specific cubic antiperovskite structure of Na3OBH4 but also because of the rotation of BH4 cluster anion. This work deepens the understanding of the antiperovskite structure and the role of cluster anions for superionic conduction.

Local screened Coulomb correction approach to strongly correlated d-electron systems.

Materials with open-shell d or f-electrons are of great importance for their intriguing electronic, optical, and magnetic properties. Often termed as strongly correlated systems, they pose great challenges for first-principles studies based on density-functional theory (DFT) in the local density approximation or generalized gradient approximation (GGA). The DFT plus the Hubbard U correction (DFT + U) approach, which is widely used in first-principles studies of strongly correlated systems, depends on the local Coulomb interaction parameters (the Hubbard U and the Hund exchange J) that are often chosen empirically, which significantly limits its predictive capability. In this work, we propose a local screened Coulomb correction (LSCC) approach in which the on-site Coulomb interaction parameters are determined by the local electron density based on the Thomas-Fermi screening model in a system-dependent and self-consistent way. The LSCC approach is applied to several typical strongly correlated systems (MnO, FeO, CoO, NiO, ß-MnO2, K2CuF4, KCuF3, KNiF3, La2CuO4, NiF2, MnF2, KMnF3, K2NiF4, La2NiO4, and Sr2CuO2Cl2), and the results are compared to those obtained from the hybrid functional and GGA methods. We found that the LSCC method can provide an accurate description of electronic and magnetic properties of considered strongly correlated systems and its performance is less sensitive to the effective range of the local projection than the closely related DFT + U approach. Therefore, the LSCC approach provides a parameter-free first-principles approach to strongly correlated systems.

Dynamic Model for Characterizing Contractile Behaviors and Mechanical Properties of a Cardiomyocyte.

Studies on the contractile dynamics of heart cells have attracted broad attention for the development of both heart disease therapies and cardiomyocyte-actuated micro-robotics. In this study, a linear dynamic model of a single cardiomyocyte cell was proposed at the subcellular scale to characterize the contractile behaviors of heart cells, with system parameters representing the mechanical properties of the subcellular components of living cardiomyocytes. The system parameters of the dynamic model were identified with the cellular beating pattern measured by a scanning ion conductance microscope. The experiments were implemented with cardiomyocytes in one control group and two experimental groups with the drugs cytochalasin-D or nocodazole, to identify the system parameters of the model based on scanning ion conductance microscope measurements, measurement of the cellular Young's modulus with atomic force microscopy indentation, measurement of cellular contraction forces using the micro-pillar technique, and immunofluorescence staining and imaging of the cytoskeleton. The proposed mathematical model was both indirectly and qualitatively verified by the variation in cytoskeleton, beating amplitude, and contractility of cardiomyocytes among the control and the experimental groups, as well as directly and quantitatively validated by the simulation and the significant consistency of 90.5% in the comparison between the ratios of the Young's modulus and the equivalent comprehensive cellular elasticities of cells in the experimental groups to those in the control group. Apart from mechanical properties (mass, elasticity, and viscosity) of subcellular structures, other properties of cardiomyocytes have also been studied, such as the properties of the relative action potential pattern and cellular beating frequency. This work has potential implications for research on cytobiology, drug screening, mechanisms of the heart, and cardiomyocyte-based bio-syncretic robotics.

Determination of Cell Membrane Capacitance and Conductance via Optically Induced Electrokinetics.

Cell membrane capacitance and conductance are key pieces of intrinsic information correlated with the cellular dielectric parameters and morphology of the plasma membrane; these parameters have been used as electrophysiological biomarkers to characterize cellular phenotype and state, and they have many associated clinical applications. Here, we present our work on the non-invasive determination of cell membrane capacitance and conductance by an optically activated microfluidics chip. The model for determining the cell membrane capacitance and conductance was established by a single layer of the shell-core polarization model. Three-dimensional finite-element analyses of the positive and negative optically induced dielectrophoresis forces generated by the projected light arrays of spots were performed, thus providing a theoretical validation of the feasibility of this approach. Then, the crossover frequency spectra for four typical types of cells (Raji cells, MCF-7 cells, HEK293 cells, and K562 cells) were experimentally investigated by using a micro-vision based motion-tracking technique. The different responses of these cells to the positive and negative ODEP forces were studied under four different liquid conductivities by automatic observation and tracking of the cellular trajectory and texture during the cells' translation. The cell membrane capacitance and conductance were determined from the curve-fitted spectra, which were 11.1 ± 0.9 mF/m2 and 782 ± 32 S/m2, respectively, for Raji cells, 11.5 ± 0.8 mF/m2 and 114 ± 28 S/m2 for MCF-7 cells, 9.0 ± 0.9 mF/m2 and 187 ± 22 S/m2 for HEK293 cells, and 10.2 ± 0.7 mF/m2 and 879 ± 24 S/m2 for K562 cells. Furthermore, as an application of this technique, the membrane capacitances of MCF-7 cells treated with four different concentrations of drugs were acquired. This technique introduces a determination of cell membrane capacitance and conductance that yields statistically significant data while allowing information from individual cells to be obtained in a non-invasive manner.

High-Throughput Fabrication and Modular Assembly of 3D Heterogeneous Microscale Tissues.

3D hydrogel microstructures that encapsulate cells have been used in broad applications in microscale tissue engineering, personalized drug screening, and regenerative medicine. Recent technological advances in microstructure assembly, such as bioprinting, magnetic assembly, microfluidics, and acoustics, have enabled the construction of designed 3D tissue structures with spatially organized cells in vitro. However, a bottleneck exists that still hampers the application of microtissue structures, due to a lack of techniques that combined high-throughput fabrication and flexible assembly. Here, a versatile method for fabricating customized microstructures and reorganizing building blocks composed of functional components into a combined single geometric shape is demonstrated. The arbitrary microstructures are dynamically synthesized in a microfluidic device and then transferred to an optically induced electrokinetics chip for manipulation and assembly. Moreover, building blocks containing different cells can be arranged into a desired geometry with specific shape and size, which can be used for microscale tissue engineering.

Imaging and Force Recognition of Single Molecular Behaviors Using Atomic Force Microscopy.

The advent of atomic force microscopy (AFM) has provided a powerful tool for investigating the behaviors of single native biological molecules under physiological conditions. AFM can not only image the conformational changes of single biological molecules at work with sub-nanometer resolution, but also sense the specific interactions of individual molecular pair with piconewton force sensitivity. In the past decade, the performance of AFM has been greatly improved, which makes it widely used in biology to address diverse biomedical issues. Characterizing the behaviors of single molecules by AFM provides considerable novel insights into the underlying mechanisms guiding life activities, contributing much to cell and molecular biology. In this article, we review the recent developments of AFM studies in single-molecule assay. The related techniques involved in AFM single-molecule assay were firstly presented, and then the progress in several aspects (including molecular imaging, molecular mechanics, molecular recognition, and molecular activities on cell surface) was summarized. The challenges and future directions were also discussed.

Structurally Well-Defined Sigmoidal Gold Clusters: Probing the Correlation between Metal Atom Arrangement and Chiroptical Response.

Asymmetric arrangement of metal atoms is crucial for understanding the chirality origin of chiral metal nanoclusters and facilitating the design and development of new chiral catalysts and chiroptical devices. Here, we describe the construction of four asymmetric gold and gold-silver clusters by chirality transfer from diimido ligands. The acquired metal clusters show strong circular dichroism (CD) response with large anisotropy factors of up to 6 × 10(-3), larger than the values of most reported chiral gold nanoclusters. Regardless of the same absolute configuration of the applied three diimido ligands, sigmoidal and reverse-sigmoidal arrangements of gold atoms both can be achieved, which resultantly produce an opposite Cotton effect within a specific absorption range. On the basis of the detailed structural characterization via X-ray crystallography and contrast experiments, the chirality contribution of the imido ligand, the asymmetrically arranged metal cluster, and the chiral arrangement of aromatic rings of phosphine ligands have been qualitatively evaluated. Time-dependent DFT calculations reveal that the chiroptical property of the acquired metal clusters is mainly influenced by the asymmetrically arranged metal atoms. Correlation of asymmetric arrangements of metal atoms in clusters with their chiroptical response provides a viable means of fabricating a designable chiral surface of metal nanoclusters and opens a broader prospect for chiral cluster application.

Single-pixel camera with one graphene photodetector.

Consumer cameras in the megapixel range are ubiquitous, but the improvement of them is hindered by the poor performance and high cost of traditional photodetectors. Graphene, a two-dimensional micro-/nano-material, recently has exhibited exceptional properties as a sensing element in a photodetector over traditional materials. However, it is difficult to fabricate a large-scale array of graphene photodetectors to replace the traditional photodetector array. To take full advantage of the unique characteristics of the graphene photodetector, in this study we integrated a graphene photodetector in a single-pixel camera based on compressive sensing. To begin with, we introduced a method called laser scribing for fabrication the graphene. It produces the graphene components in arbitrary patterns more quickly without photoresist contamination as do traditional methods. Next, we proposed a system for calibrating the optoelectrical properties of micro/nano photodetectors based on a digital micromirror device (DMD), which changes the light intensity by controlling the number of individual micromirrors positioned at + 12°. The calibration sensitivity is driven by the sum of all micromirrors of the DMD and can be as high as 10(-5)A/W. Finally, the single-pixel camera integrated with one graphene photodetector was used to recover a static image to demonstrate the feasibility of the single-pixel imaging system with the graphene photodetector. A high-resolution image can be recovered with the camera at a sampling rate much less than Nyquist rate. The study was the first demonstration for ever record of a macroscopic camera with a graphene photodetector. The camera has the potential for high-speed and high-resolution imaging at much less cost than traditional megapixel cameras.

Characterization of the self-rotational motion of stored red blood cells by using optically-induced electrokinetics.

We report a label-free approach toward the object of characterizing the self-rotational motions of red blood cells (RBCs) during storage under the optically-induced electrokinetics-based microfluidics mechanism. A theoretical analysis of the transmembrane potential across RBCs was performed getting a threshold voltage for keeping cellular biological integrity. Then, by investigation of the self-rotational behaviors of the individual RBCs in larger population, the RBCs that were stored more than three weeks statistically showed the distinctive self-rotational speed. Results verified that the self-rotational biomarkers of the RBCs could be used to label-free reckon the qualities of the stored RBCs in this kind of microfluidics chip. This finding may be further developed as a new criterion to real-time and label-free monitoring of the banked blood qualities, thereby diminishing the blood transfusion venture.

Facile modulation of cell adhesion to a poly(ethylene glycol) diacrylate film with incorporation of polystyrene nano-spheres.

Poly(ethylene glycol) diacrylate (PEGDA) is a common hydrogel that has been actively investigated for various tissue engineering applications owing to its biocompatibility and excellent mechanical properties. However, the native PEGDA films are known for their bio-inertness which can hinder cell adhesion, thereby limiting their applications in tissue engineering and biomedicine. Recently, nano composite technology has become a particularly hot topic, and has led to the development of new methods for delivering desired properties to nanomaterials. In this study, we added polystyrene nano-spheres (PS) into a PEGDA solution to synthesize a nano-composite film and evaluated its characteristics. The experimental results showed that addition of the nanospheres to the PEGDA film not only resulted in modification of the mechanical properties and surface morphology but further improved the adhesion of cells on the film. The tensile modulus showed clear dependence on the addition of PS, which enhanced the mechanical properties of the PEGDA-PS film. We attribute the high stiffness of the hybrid hydrogel to the formation of additional cross-links between polymeric chains and the nano-sphere surface in the network. The effect of PS on cell adhesion and proliferation was evaluated in L929 mouse fibroblast cells that were seeded on the surface of various PEGDA-PS films. Cells density increased with a larger PS concentration, and the cells displayed a spreading morphology on the hybrid films, which promoted cell proliferation. Impressively, cellular stiffness could also be modulated simply by tuning the concentration of nano-spheres. Our results indicate that the addition of PS can effectively tailor the physical and biological properties of PEGDA as well as the mechanical properties of cells, with benefits for biomedical and biotechnological applications.

A rapid and automated relocation method of an AFM probe for high-resolution imaging.

The atomic force microscope (AFM) is one of the most powerful tools for high-resolution imaging and high-precision positioning for nanomanipulation. The selection of the scanning area of the AFM depends on the use of the optical microscope. However, the resolution of an optical microscope is generally no larger than 200 nm owing to wavelength limitations of visible light. Taking into consideration the two determinants of relocation-relative angular rotation and positional offset between the AFM probe and nano target-it is therefore extremely challenging to precisely relocate the AFM probe to the initial scan/manipulation area for the same nano target after the AFM probe has been replaced, or after the sample has been moved. In this paper, we investigate a rapid automated relocation method for the nano target of an AFM using a coordinate transformation. The relocation process is both simple and rapid; moreover, multiple nano targets can be relocated by only identifying a pair of reference points. It possesses a centimeter-scale location range and nano-scale precision. The main advantages of this method are that it overcomes the limitations associated with the resolution of optical microscopes, and that it is label-free on the target areas, which means that it does not require the use of special artificial markers on the target sample areas. Relocation experiments using nanospheres, DNA, SWCNTs, and nano patterns amply demonstrate the practicality and efficiency of the proposed method, which provides technical support for mass nanomanipulation and detection based on AFM for multiple nano targets that are widely distributed in a large area.

The local projection in the density functional theory plus U approach: A critical assessment.

Density-functional theory plus the Hubbard U correction (DFT + U) method is widely used in first-principles studies of strongly correlated systems, as it can give qualitatively (and sometimes, semi-quantitatively) correct description of energetic and structural properties of many strongly correlated systems with similar computational cost as local density approximation or generalized gradient approximation. On the other hand, the DFT + U approach is limited both theoretically and practically in several important aspects. In particular, the results of DFT + U often depend on the choice of local orbitals (the local projection) defining the subspace in which the Hubbard U correction is applied. In this work we have systematically investigated the issue of the local projection by considering typical transition metal oxides, ß-MnO2 and MnO, and comparing the results obtained from different implementations of DFT + U. We found that the choice of the local projection has significant effects on the DFT + U results, which are more significant for systems with stronger covalent bonding (e.g., MnO2) than those with more ionic bonding (e.g., MnO). These findings can help to clarify some confusion arising from the practical use of DFT + U and may also provide insights for the development of new first-principles approaches beyond DFT + U.

Experimental study and modeling of atomic-scale friction in zigzag and armchair lattice orientations of MoS2.

Physical properties of two-dimensional materials, such as graphene, black phosphorus, molybdenum disulfide (MoS2) and tungsten disulfide, exhibit significant dependence on their lattice orientations, especially for zigzag and armchair lattice orientations. Understanding of the atomic probe motion on surfaces with different orientations helps in the study of anisotropic materials. Unfortunately, there is no comprehensive model that can describe the probe motion mechanism. In this paper, we report a tribological study of MoS2 in zigzag and armchair orientations. We observed a characteristic power spectrum and friction force values. To explain our results, we developed a modified, two-dimensional, stick-slip Tomlinson model that allows simulation of the probe motion on MoS2 surfaces by combining the motion in the Mo layer and S layer. Our model fits well with the experimental data and provides a theoretical basis for tribological studies of two-dimensional materials.

Effects of methotrexate on the viscoelastic properties of single cells probed by atomic force microscopy.

Methotrexate is a commonly used anti-cancer chemotherapy drug. Cellular mechanical properties are fundamental parameters that reflect the physiological state of a cell. However, so far the role of cellular mechanical properties in the actions of methotrexate is still unclear. In recent years, probing the behaviors of single cells with the use of atomic force microscopy (AFM) has contributed much to the field of cell biomechanics. In this work, with the use of AFM, the effects of methotrexate on the viscoelastic properties of four types of cells were quantitatively investigated. The inhibitory and cytotoxic effects of methotrexate on the proliferation of cells were observed by optical and fluorescence microscopy. AFM indenting was used to measure the changes of cellular viscoelastic properties (Young's modulus and relaxation time) by using both conical tip and spherical tip, quantitatively showing that the stimulation of methotrexate resulted in a significant decrease of both cellular Young's modulus and relaxation times. The morphological changes of cells induced by methotrexate were visualized by AFM imaging. The study improves our understanding of methotrexate action and offers a novel way to quantify drug actions at the single-cell level by measuring cellular viscoelastic properties, which may have potential impacts on developing label-free methods for drug evaluation.