Scale-independent model based on fractal theory for calculating the adhesion force between a particle and rough surfaces.

The elimination of the scale-dependent statistical parameters is a challenge in the estimation of the van der Waals force between a particle and rough surfaces. Herein, a scale-independent parameter, the fractal dimension, was introduced into the Rabinovich model to calculate the microadhesion force. First, a Weierstrass-Mandelbrot function is proposed to generate a random 2D contour, which has a simplified form and is more reasonable for spectrum analysis. And then, the roughness scale extraction method was employed to calculate the fractal-dimension value of the real aluminum (Al) surface. And last, the proposed scale-independent Rabinovich-Fractal model was used to calculate the adhesion forces, which are consistent with the results of the atomic force microscopy adhesion test. Aside from providing a more accurate estimation of the van der Waals forces, the proposed model is not influenced by the sampling scale of the surface roughness measurement, which eliminates the related error in van der Waals force estimation.

An Atomistic Level Investigation of the PFDTES-Graphene Interfacial Shear Strength and the Stick-Slip Mechanism.

1H,1H,2H,2H-Perfluorodecyltriethoxysilane (PFDTES) is the most widely used coating material with low surface energy and has the potential to be used as a dust-mitigating coating material during lunar landing missions. Graphene can be added to the PFDTES matrix to improve its mechanical properties. In this study, molecular dynamics simulations were performed to investigate the interfacial shear strength and friction mechanism between the PFDTES matrix and graphene. A systematic molecular dynamics (MD) simulation has been performed to calculate the interfacial shear strength of the PFDTES-graphene interface with considering the effect of graphene sliding velocity and vacancy defect density. For a pristine graphene layer with a size of 10 nm × 10 nm, the interfacial force between graphene and the PFDTES matrix is around 3 nN. Like other polymeric materials, the interfacial shear force exhibited stick-slip behavior under loading. The interfacial shear force will start to increase after the graphene starts sliding against the PFDTES matrix and reaches a stable plateau in a very short distance. It has been found that the influence of the interfacial shear strength from the sliding velocity of graphene is minimal. However, a significant increase in the interfacial shear strength has been observed after the graphene defect density increased; i.e., the magnitude of the shear force increased from 3 nN to around 14 nN after the defect density increased from 0% for pristine graphene to 40%. It has been found that vacancy defects will increase the fluctuation in the interfacial shear force, and it is due to not only the increased roughness near defects but also the stretched bonds in graphene under loading according to the distribution of the bond length. This study concluded that interfacial stick-slip behavior also exists in the PFDTES-graphene interface, and vacancy defects will have a significant improvement in the interfacial shear strength.

Rapid Fabrication of Antilunar Dust Aluminum Surface by Nanosecond Laser Etching.

Although a dust-repellent surface is desirable for lunar exploration missions, its fabrication process is complicated and time-consuming. Herein, we report a simple and fast method to fabricate a lunar dust-repellent surface by texturing on an Al substrate via nanosecond laser etching. The laser-induced photothermal effect can rapidly create hierarchical papillary structures on 25 × 25 mm Al substrates (within 30 s). Both atomic force microscopy (AFM) and in situ scanning electron microscopy (SEM) reveal that such structures enable a reduced contact area between the Al substrate and lunar dust and thus reduced adhesion. The reduced dust adhesion force of Al substrates facilitates improving their antidust performance. By optimizing processing parameters, the Al substrate etched with a laser scanning spacing of 80 µm exhibits a lower dust adhesion force (9.58 nN) due to the smallest contact area with dust. Accordingly, its static antilunar dust performance (dust coverage of 1.95%) is significantly improved compared to the pristine Al substrate (dust coverage of 12.98%). Besides, the accumulated dust on the laser-etched Al substrates with low surface adhesion force is easily cleaned up by flipping and gravity (the dust residual rates are less than 17%). The Al substrate with excellent antidust ability presents good potential for lunar exploration missions.

Characterization of Residual Stress in SOI Wafers by Using MEMS Cantilever Beams.

Silicon-on-insulator (SOI) wafers are crucial raw materials in the manufacturing process of microelectromechanical systems (MEMS). Residual stresses generated inside the wafers during the fabrication process can seriously affect the performance, reliability, and yield of MEMS devices. In this paper, a low-cost method based on mechanical modeling is proposed to characterize the residual stresses in SOI wafers in order to calculate the residual stress values based on the deformation of the beams. Based on this method, the residual strain of the MEMS beam, and thus the residual stress in the SOI wafer, were experimentally determined. The results were also compared with the residual stress results calculated from the deflection of the rotating beam to demonstrate the validity of the results obtained by this method. This method provides valuable theoretical reference and data support for the design and optimization of devices based on SOI-MEMS technology. It provides a lower-cost solution for the residual stress measurement technique, making it available for a wide range of applications.

BATF2 promotes HSC myeloid differentiation by amplifying IFN response mediators during chronic infection.

Basic leucine zipper ATF-like transcription factor 2 (BATF2), an interferon-activated immune response regulator, is a key factor responsible for myeloid differentiation and depletion of HSC during chronic infection. To delineate the mechanism of BATF2 function in HSCs, we assessed Batf2 KO mice during chronic infection and found that they produced less pro-inflammatory cytokines, less immune cell recruitment to the spleen, and impaired myeloid differentiation with better preservation of HSC capacity compared to WT. Co-IP analysis revealed that BATF2 forms a complex with JUN to amplify pro-inflammatory signaling pathways including CCL5 during infection. Blockade of CCL5 receptors phenocopied Batf2 KO differentiation defects, whereas treatment with recombinant CCL5 was sufficient to rescue IFNγ-induced myeloid differentiation and recruit more immune cells to the spleen in Batf2 KO mice. By revealing the mechanism of BATF2-induced myeloid differentiation of HSCs, these studies elucidate potential therapeutic strategies to boost immunity while preserving HSC function during chronic infection.

Lunar Dust-Mitigation Behavior of Aluminum Surfaces with Multiscale Roughness Prepared by a Composite Etching Method.

Reducing lunar dust adhesion to various material surfaces is important for protecting equipment from damage during lunar exploration missions. In this study, we investigate the lunar dust-mitigation ability and dust adhesion force of aluminum (Al) substrates prepared using different etching methods. Among them, composite etching methods (combining chemical and electrochemical steps) can result in multiscale structures with micro- and nanoroughness, reducing the contact area between the substrate and thus decreasing lunar dust adhesion. After composite etching, the dust adhesion force of the Al substrate was significantly reduced by 80% from 45.53 to 8.89 nN. The dust adhesion force of Al substrates dominates their dust-mitigation performance in floating dust environments. The lunar dust coverage (2.19%) of the Al substrate modified by composite etching (placed with a tilt angle of 90°) was 4-fold lower than that of the pristine Al substrate (9.11%), indicating excellent lunar-dust repellence. In addition, other factors such as tilt angle of the substrate and dust loading significantly affect dust-mitigation performance of the modified Al substrates. The Al substrate with an excellent dust-mitigation ability highlights good potential for lunar exploration missions.

Research Progress of MEMS Inertial Switches.

As a typical type of MEMS acceleration sensor, the inertial switch can alter its on-off state while the environmental accelerations satisfy threshold value. An exhaustive summary of the design concept, performance aspects, and fabrication methods of the micro electromechanical system (MEMS) inertial switch is provided. Different MEMS inertial switch studies were reviewed that emphasized acceleration directional and threshold sensitivity, contact characteristics, and their superiorities and disadvantages. Furthermore, the specific fabrication methods offer an applicability reference for the preparation process for the designed inertial switch, including non-silicon surface micromachining technology, standard silicon micromachining technology, and the special fabrication method for the liquid inertial switch. At the end, the main conclusions of the current challenges and prospects about MEMS inertial switches are drawn to assist with the development of research in the field of future engineering applications.

Conformational Changes of RORγ During Response Element Recognition and Coregulator Engagement.

The retinoic acid receptor-related orphan receptor γ (RORγ) is a ligand-dependent transcription factor of the nuclear receptor super family that underpins metabolic activity, immune function, and cancer progression. Despite being a valuable drug target in health and disease, our understanding of the ligand-dependent activities of RORγ is far from complete. Like most nuclear receptors, RORγ must recruit coregulatory protein to enact the RORγ target gene program. To date, a majority of structural studies have been focused exclusively on the RORγ ligand-binding domain and the ligand-dependent recruitment of small peptide segments of coregulators. Herein, we examine the ligand-dependent assembly of full length RORγ:coregulator complexes on cognate DNA response elements using structural proteomics and small angle x-ray scattering. The results from our studies suggest that RORγ becomes elongated upon DNA recognition, preventing long range interdomain crosstalk. We also determined that the DNA binding domain adopts a sequence-specific conformation, and that coregulatory protein may be able to 'sense' the ligand- and DNA-bound status of RORγ. We propose a model where ligand-dependent coregulator recruitment may be influenced by the sequence of the DNA to which RORγ is bound. Overall, the efforts described herein will illuminate important aspects of full length RORγ and monomeric orphan nuclear receptor target gene regulation through DNA-dependent conformational changes.

Acetylation of histone H3K27 signals the transcriptional elongation for estrogen receptor alpha.

As approximately 70% of human breast tumors are estrogen receptor α (ERα)-positive, estrogen and ERα play essential roles in breast cancer development. By interrupting the ERα signaling pathway, endocrine therapy has been proven to be an effective therapeutic strategy. In this study, we identified a mechanism by which Transcription Start Site (TSS)-associated histone H3K27 acetylation signals the Super Elongation Complex (SEC) to regulate transcriptional elongation of the ESR1 (ERα) gene. SEC interacts with H3K27ac on ESR1 TSS through its scaffold protein AFF4. Depletion of AFF4 by siRNA or CRISPR/Cas9 dramatically reduces expression of ESR1 and its target genes, consequently inhibiting breast cancer cell growth. More importantly, a AFF4 mutant which lacks H3K27ac interaction failed to rescue ESR1 gene expression, suggesting H3K27 acetylation at TSS region is a key mark bridging the transition from transcriptional initiation to elongation, and perturbing SEC function can be an alternative strategy for targeting ERα signaling pathway at chromatin level.

Refolding Proteins from Inclusion Bodies using Differential Scanning Fluorimetry Guided (DGR) Protein Refolding and MeltTraceur Web.

Differential Scanning Fluorimetry Guided Refolding (DGR) is a simple methodology that can be used to rapidly screen for and identify conditions capable of accurately refolding protein preparations, such as those obtained from Escherichia coli inclusion bodies. It allows for the production in E. coli of functional proteins that would otherwise require far more expensive production methods. This unit describes how to set up a DGR refolding assay, perform DGR refolding trials in microplate format, use MeltTraceur Web software to interactively analyze the resulting data, scale-up protein production via refolding, and lastly, validate that the protein is properly folded. © 2018 by John Wiley & Sons, Inc.

Pyrazolo[1,5-a]pyrimidine TRPC6 antagonists for the treatment of gastric cancer.

Transient receptor potential canonical 6 (TRPC6) proteins form receptor-operated Ca2+-permeable channels, which have been thought to bring benefit to the treatment of diseases, including cancer. However, selective antagonists for TRPC channels are rare and none of them has been tested against gastric cancer. Compound 14a and analogs were synthesized by chemical elaboration of previously reported TRPC3/6/7 agonist 4o. 14a had very weak agonist activity at TRPC6 expressed in HEK293 cells but exhibited strong inhibition on both 4o-mediated and receptor-operated activation of TRPC6 with an IC50 of about 1 µM. When applied to the culture media, 14a suppressed proliferation of AGS and MKN45 cells with IC50 values of 17.1 ±â€¯0.3 and 18.5 ±â€¯1.0 µM, respectively, and inhibited tube formation and migration of cultured human endothelial cells. This anti-tumor effect on gastric cancer was further verified in xenograft models using nude mice. This study has found a new tool compound which shows excellent therapeutic potential against human gastric cancer most likely through targeting TRPC6 channels.

Structure-activity relationships of 2-substituted phenyl-N-phenyl-2-oxoacetohydrazonoyl cyanides as novel antagonists of exchange proteins directly activated by cAMP (EPACs).

Exchange proteins directly activated by cAMP (EPACs) are critical cAMP-dependent signaling pathway mediators that play important roles in cancer, diabetes, heart failure, inflammations, infections, neurological disorders and other human diseases. EPAC specific modulators are urgently needed to explore EPAC's physiological function, mechanism of action and therapeutic applications. On the basis of a previously identified EPAC specific inhibitor hit ESI-09, herein we have designed and synthesized a novel series of 2-substituted phenyl-N-phenyl-2-oxoacetohydrazonoyl cyanides as potent EPAC inhibitors. Compound 31 (ZL0524) has been discovered as the most potent EPAC inhibitor with IC50 values of 3.6 µM and 1.2  µM against EPAC1 and EPAC2, respectively. Molecular docking of 31 onto an active EPAC2 structure predicts that 31 occupies the hydrophobic pocket in cAMP binding domain (CBD) and also opens up new space leading to the solvent region. These findings provide inspirations for discovering next generation of EPAC inhibitors.

A cell-based, quantitative and isoform-specific assay for exchange proteins directly activated by cAMP.

Extensive functional studies of the exchange protein directly activated by cAMP (EPAC) family of signaling molecules have demonstrated that EPAC proteins play a fundamental role in several physiological and pathophysiological responses, therefore are attractive drug targets. In this report, the development of a cell-based, medium to high throughput screening assay that is capable of monitoring EPAC-mediated activation of cellular Rap1 in an isoform-specific manner is described. This assay adapts a conventional ELISA format with immobilized RalGDS-RBD as a bait to selectively capture GTP-bound active Rap1. As a result, it fills an urgent need for a cell-based EPAC assay that can be conveniently performed using microtiter plates for the discovery and/or validation of isoform-specific EPAC agonists and antagonists.

Identification of novel 2-(benzo[d]isoxazol-3-yl)-2-oxo-N-phenylacetohydrazonoyl cyanide analoguesas potent EPAC antagonists.

Two series of novel EPAC antagonists are designed, synthesized and evaluated in an effort to develop diversified analogues based on the scaffold of the previously identified high-throughput (HTS) hit 1 (ESI-09). Further SAR studies reveal that the isoxazole ring A of 1 can tolerate chemical modifications with either introduction of flexible electron-donating substitutions or structurally restrictedly fusing with a phenyl ring, leading to identification of several more potent and diversified EPAC antagonists (e.g., 10 (NY0617), 14 (NY0460), 26 (NY0725), 32 (NY0561), and 33 (NY0562)) with low micromolar inhibitory activities. Molecular docking studies on compounds 10 and 33 indicate that these two series of compounds bind at a similar site with substantially different interactions with the EPAC proteins. The findings may serve as good starting points for the development of more potent EPAC antagonists as valuable pharmacological probes or potential drug candidates.

Pyrazolopyrimidines as Potent Stimulators for Transient Receptor Potential Canonical 3/6/7 Channels.

Transient receptor potential canonical 3/6/7 (TRPC3/6/7) are highly homologous receptor-operated nonselective cation channels. Despite their physiological significance, very few selective and potent agonists are available for functional examination of these channels. Using a cell-based high throughput screening approach, a lead compound with the pyrazolopyrimidine skeleton was identified as a TRPC6 agonist. Synthetic schemes for the lead and its analogues were established, and structural-activity relationship studies were carried out. A series of potent and direct agonists of TRPC3/6/7 channels were identified, and among them, 4m-4p have a potency order of TRPC3 > C7 > C6, with 4n being the most potent with an EC50 of <20 nM on TRPC3. Importantly, these compounds exhibited no stimulatory activity on related TRP channels. The potent and selective compounds described here should be suitable for evaluation of the roles of TRPC channels in the physiology and pathogenesis of diseases, including glomerulosclerosis and cancer.

Structure-Activity Relationship Studies with Tetrahydroquinoline Analogs as EPAC Inhibitors.

EPAC proteins are therapeutic targets for the potential treatment of cardiac hypertrophy and cancer metastasis. Several laboratories use a tetrahydroquinoline analog, CE3F4, to dissect the role of EPAC1 in various disease states. Here, we report SAR studies with tetrahydroquinoline analogs that explore various functional groups. The most potent EPAC inhibitor 12a exists as a mixture of inseparable E (major) and Z (minor) rotamers. The rotation about the N-formyl group indeed impacts the activity against EPAC.

Functionalized N,N-Diphenylamines as Potent and Selective EPAC2 Inhibitors.

N,N-Diphenylamines were discovered as potent and selective EPAC2 inhibitors. A study was conducted to determine the structure-activity relationships in a series of inhibitors of which several compounds displayed submicromolar potencies. Selectivity over the related EPAC1 protein was also demonstrated. Computational modeling reveals an allosteric site that is distinct from the cAMP binding domain shared by both EPAC isoforms, providing a theory with regards to subtype selectivity.

A Terrestrial Microbial Fuel Cell for Powering a Single-Hop Wireless Sensor Network.

Microbial fuel cells (MFCs) are envisioned as one of the most promising alternative renewable energy sources because they can generate electric current continuously while treating waste. Terrestrial Microbial Fuel Cells (TMFCs) can be inoculated and work on the use of soil, which further extends the application areas of MFCs. Energy supply, as a primary influential factor determining the lifetime of Wireless Sensor Network (WSN) nodes, remains an open challenge in sensor networks. In theory, sensor nodes powered by MFCs have an eternal life. However, low power density and high internal resistance of MFCs are two pronounced problems in their operation. A single-hop WSN powered by a TMFC experimental setup was designed and experimented with. Power generation performance of the proposed TMFC, the relationships between the performance of the power generation and the environment temperature, the water content of the soil by weight were measured by experiments. Results show that the TMFC can achieve good power generation performance under special environmental conditions. Furthermore, the experiments with sensor data acquisition and wireless transmission of the TMFC powering WSN were carried out. We demonstrate that the obtained experimental results validate the feasibility of TMFCs powering WSNs.

Critical role for Epac1 in inflammatory pain controlled by GRK2-mediated phosphorylation of Epac1.

cAMP signaling plays a key role in regulating pain sensitivity. Here, we uncover a previously unidentified molecular mechanism in which direct phosphorylation of the exchange protein directly activated by cAMP 1 (EPAC1) by G protein kinase 2 (GRK2) suppresses Epac1-to-Rap1 signaling, thereby inhibiting persistent inflammatory pain. Epac1(-/-) mice are protected against inflammatory hyperalgesia in the complete Freund's adjuvant (CFA) model. Moreover, the Epac-specific inhibitor ESI-09 inhibits established CFA-induced mechanical hyperalgesia without affecting normal mechanical sensitivity. At the mechanistic level, CFA increased activity of the Epac target Rap1 in dorsal root ganglia of WT, but not of Epac1(-/-), mice. Using sensory neuron-specific overexpression of GRK2 or its kinase-dead mutant in vivo, we demonstrate that GRK2 inhibits CFA-induced hyperalgesia in a kinase activity-dependent manner. In vitro, GRK2 inhibits Epac1-to-Rap1 signaling by phosphorylation of Epac1 at Ser-108 in the Disheveled/Egl-10/pleckstrin domain. This phosphorylation event inhibits agonist-induced translocation of Epac1 to the plasma membrane, thereby reducing Rap1 activation. Finally, we show that GRK2 inhibits Epac1-mediated sensitization of the mechanosensor Piezo2 and that Piezo2 contributes to inflammatory mechanical hyperalgesia. Collectively, these findings identify a key role of Epac1 in chronic inflammatory pain and a molecular mechanism for controlling Epac1 activity and chronic pain through phosphorylation of Epac1 at Ser-108. Importantly, using the Epac inhibitor ESI-09, we validate Epac1 as a potential therapeutic target for chronic pain.

Structure-Activity Relationship Studies of Substituted 2-(Isoxazol-3-yl)-2-oxo-N'-phenyl-acetohydrazonoyl Cyanide Analogues: Identification of Potent Exchange Proteins Directly Activated by cAMP (EPAC) Antagonists.

Exchange proteins directly activated by cAMP (EPAC) as guanine nucleotide exchange factors mediate the effects of the pivotal second messenger cAMP, thereby regulating a wide variety of intracellular physiological and pathophysiological processes. A series of novel 2-(isoxazol-3-yl)-2-oxo-N'-phenyl-acetohydrazonoyl cyanide EPAC antagonists was synthesized and evaluated in an effort to optimize properties of the previously identified high-throughput (HTS) hit 1 (ESI-09). Structure-activity relationship (SAR) analysis led to the discovery of several more active EPAC antagonists (e.g., 22 (HJC0726), 35 (NY0123), and 47 (NY0173)) with low micromolar inhibitory activity. These inhibitors may serve as valuable pharmacological probes to facilitate our efforts in elucidating the biological functions of EPAC and developing potential novel therapeutics against human diseases. Our SAR results have also revealed that further modification at the 3-, 4-, and 5-positions of the phenyl ring as well as the 5-position of the isoxazole moiety may allow for the development of more potent EPAC antagonists.