Magnetically Responsive Superhydrophobic Surface with Switchable Adhesivity Based on Electrostatic Air Spray Deposition.

A new method was reported for preparing a magnetically responsive superhydrophobic surface by electrostatic air spray deposition (EASD) and magnetic induction. The mixture was fully atomized under the combined action of the electrostatic field and the high-speed airflow field, and a dense array of micropillars was formed. The atomization mechanism of EASD was explored. The distribution and physical parameters of the micropillars were evaluated and counted. Switchable adhesion characteristics of the surface and the reversibility in 10 cycles were examined. The influences of different electrostatic voltages, component concentration, spray distance, air pressure, and magnetic field intensity on the surface morphology and hydrophobicity were analyzed. The prepared surface can be reversibly transformed between the high-adhesion state (with a contact angle of 108°) and the low-adhesion state (with a contact angle of 154°) by on/off switching of an external magnetic field. After a 2.2 kPa pressure load was applied, the surface contact angle was 144° with an applied magnetic field of 0.4 T. After heated at 90 °C for more than 90 min, the surface can almost obtain superhydrophobicity (with a contact angle of 148°) in the absence of a magnetic field. By utilizing the switchable surface adhesion characteristics, various kinds of droplet transmissions were realized. When the cured surface was spray-coated with carbon nanoparticles (CNPs), active droplet manipulation can be achieved by simply moving the magnet. The advantages of this method include a simple preparation process without chemical surface modification.

Precision glass molding of diffractive optical elements with high surface quality.

Diffractive optical surfaces have attractive properties for use in optical systems, like reducing weight and correcting for chromatic aberrations, but fabrication of high-quality glass diffractive optics is challenging, preventing it from being widely adopted in commercial applications. In this Letter, we report on a fabrication method to address molding challenges for high-surface-quality diffractive glass optics at molding temperatures up to 550°C, including selection of mold material, mold fabrication, precision glass molding, durability, and stability of the mold. To enable optimal mold machining and easy mold release, nickel phosphorous (NiP) is chosen as the plating material for its cutting performance and anti-adhesion properties, and copper-nickel C71500 (CuNi) is selected as the mold substrate because its coefficient of thermal expansion (CTE) is close to NiP. By the proposed method, diffractive glass optics with 2 nm Sa surface roughness is demonstrated.

Discovery of a New Neisseria gonorrhoeae Type IV Pilus Assembly Factor, TfpC.

Neisseria gonorrhoeae relies on type IV pili (T4p) to promote colonization of their human host and to cause the sexually transmitted infection gonorrhea. This organelle cycles through a process of extension and retraction back into the bacterial cell. Through a genetic screen, we identified the NGO0783 locus of N. gonorrhoeae strain FA1090 as containing a gene encoding a protein required to stabilize the type IV pilus in its extended, nonretracted conformation. We have named the gene tfpC and the protein TfpC. Deletion of tfpC produces a nonpiliated colony morphology, and immuno-transmission electron microscopy confirms that the pili are lost in the ΔtfpC mutant, although there is some pilin detected near the bacterial cell surface. A copy of the tfpC gene expressed from a lac promoter restores pilus expression and related phenotypes. A ΔtfpC mutant shows reduced levels of pilin protein, but complementation with a tfpC gene restored pilin to normal levels. Bioinformatic searches show that there are orthologues in numerous bacterial species, but not all type IV pilin-expressing bacteria contain orthologous genes. Coevolution and nuclear magnetic resonance (NMR) analysis indicates that TfpC contains an N-terminal transmembrane helix, a substantial extended/unstructured region, and a highly charged C-terminal coiled-coil domain.IMPORTANCE Most bacterial species express one or more extracellular organelles called pili/fimbriae that are required for many properties of each bacterial cell. The Neisseria gonorrhoeae type IV pilus is a major virulence and colonization factor for the sexually transmitted infection gonorrhea. We have discovered a new protein of Neisseria gonorrhoeae called TfpC that is required to maintain type IV pili on the bacterial cell surface. There are similar proteins found in other members of the Neisseria genus and many other bacterial species important for human health.

Bamboo-joint-like platforms for fast, long-distance, directional, and spontaneous transport of fluids.

Spontaneous transport of fluids without external force offers an enabling tool for a wide spectrum of fields. However, the development of a universal spontaneous transport platform for liquids remains a challenge. In this work, a novel bamboo-joint-like platform with tapered micro-tubes as transport units is presented, which not only enables the spontaneous transport and extrusion of liquids but also enables customized and optional assembly of transport devices. Spontaneous transport characterized with long-distance, anti-gravity transport, directional transport, and liquid extrusion characteristics was found to show excellent transport capacity. The results indicated that both transport distance and speed varied periodically with time, which was mainly due to the difference in curvature caused by asymmetric structure and capillary force. The desired spontaneous transportation was successfully obtained even when the supply rate speed was up to 632.5 µl/min and length of platform reached a scale of hundreds of millimeters. Transport units were easily fabricated via a commercially available 3D printing technique, so that the customized and directional spontaneous directional transport can be realized for liquid distribution, serpentine loop transportation, and speed control. With the comprehensive use of transport units and connectors, it is very easy to implement self-service construction of a universal complex multi-functional transportation platform.

New testing and calculation method for determination viscoelasticity of optical glass.

Viscoelastic properties of glass within molding temperatures, such as shear relaxation modulus and bulk relaxation modulus, are key factors to build successful numerical model, predict forming process, and determine optimal process parameters for precision glass molding. However, traditional uniaxial compression creep tests with large strains are very limited in obtaining high-accuracy viscoelastic data of glass, due to the declining compressive stress caused by the increasing cross-sectional area of specimen in testing process. Besides, existing calculation method has limitation in transforming creep data to viscoelasticity data, especially when Poisson's ratio is unknown at molding temperature, which further induces a block to characterize viscoelastic parameter. This study proposes a systematic acquisition method for high-precision viscoelastic data, including creep testing, viscoelasticity calculation, and finite element verification. A minimal uniaxial creep testing (MUCT) method based on thermo-mechanical analysis (TMA) instrument is first built to obtain ideal and accurate creep data, by keeping compressive stress as a constant. A new calculation method on viscoelasticity determination is then proposed to derive shear relaxation modulus without the need of knowing bulk modulus or Poisson's ratio, which, compared with traditional method, extends the application range of viscoelasticity calculation. After determination, the obtained viscoelastic data are further incorporated into a numerical simulation model of MUCT to verify the accuracy of the determined viscoelasticity. Base on the great consistence between simulated and measured results (uniaxial creep displacement), the proposed systematic acquisition method can be used as a high accuracy viscoelasticity determination method.

Altering the Neisseria gonorrhoeae pilE Guanine Quadruplex Loop Bases Affects Pilin Antigenic Variation.

Neisseria gonorrhoeae possesses a programmed recombination system that allows the bacteria to alter the major subunit of the type IV pilus, pilin or PilE. An alternate DNA structure known as a guanine quadruplex (G4) is required for pilin antigenic variation (pilin Av). The G-C base pairs within the G4 motif are required for pilin Av, but simple mutation of the loop bases does not affect pilin Av. We show that more substantial changes to the loops, in both size and nucleotide composition, with the core guanines unchanged, decrease or abrogate pilin Av. We investigated why these loop changes might influence the efficiency of pilin Av. RecA is a recombinase required for pilin Av that can bind the pilE G4 in vitro. RecA binds different G4 structures with altered loops with varied affinities. However, changes in RecA binding affinities to the loop mutants do not absolutely correlate with the pilin Av phenotypes. Interestingly, the yeast RecA ortholog, Rad51, also binds the pilE G4 structure with a higher affinity than it binds single-stranded DNA, suggesting that RecA G4 binding is conserved in eukaryotic orthologs. The thermal stability the pilE G4 structure and its loop mutants showed that the parental G4 structure had the highest melting temperature, and the melting temperature of the loop mutants correlated with pilin Av phenotype. These results suggest that the folding kinetics and stability of G4 structures are important for the efficiency of pilin Av.

PacBio Amplicon Sequencing Method To Measure Pilin Antigenic Variation Frequencies of Neisseria gonorrhoeae.

Gene diversification is a common mechanism pathogens use to alter surface structures to aid in immune avoidance. Neisseria gonorrhoeae uses a gene conversion-based diversification system to alter the primary sequence of the gene encoding the major subunit of the pilus, pilE Antigenic variation occurs when one of the nonexpressed 19 silent copies donates part of its DNA sequence to pilE We have developed a method using Pacific Biosciences (PacBio) amplicon sequencing and custom software to determine pilin antigenic variation frequencies. The program analyzes 37 variable regions across the strain FA1090 1-81-S2 pilE gene and can be modified to determine sequence variation from other starting pilE sequences or other diversity generation systems. Using this method, we measured pilin antigenic variation frequencies for various derivatives of strain FA1090 and showed we can also analyze pilin antigenic variation frequencies during macrophage infection.IMPORTANCE Diversity generation systems are used by many unicellular organism to provide subpopulations of cell with different properties that are available when needed. We have developed a method using the PacBio DNA sequencing technology and a custom computer program to analyze the pilin antigenic variation system of the organism that is the sole cause of the sexually transmitted infection, gonorrhea.

Identification of Small Molecules Exhibiting Oxacillin Synergy through a Novel Assay for Inhibition of vraTSR Expression in Methicillin-Resistant Staphylococcus aureus.

Methicillin-resistant Staphylococcus aureus (MRSA) strains that are resistant to all forms of penicillin have become an increasingly common and urgent problem threatening human health. They are responsible for a wide variety of infectious diseases ranging from minor skin abscesses to life-threatening severe infections. The vra operon that is conserved among S. aureus strains encodes a three-component signal transduction system (vraTSR) that is responsible for sensing and responding to cell wall stress. We developed a novel and multifaceted assay to identify compounds that potentiate the activity of oxacillin, essentially restoring efficacy of oxacillin against MRSA, and performed high-throughput screening (HTS) to identify oxacillin potentiators. HTS of 13,840 small-molecule compounds from an antimicrobial-focused Life Chemicals library, using the MRSA cell-based assay, identified three different inhibitor scaffolds. Checkerboard assays for synergy with oxacillin, reverse transcriptase PCR (RT-PCR) assays against vraR expression, and direct confirmation of interaction with VraS by surface plasmon resonance (SPR) further verified them to be viable hit compounds. A subsequent structure-activity relationship (SAR) study of the best scaffold with diverse analogs was utilized to improve potency and provides a strong foundation for further development.

Novel cavitation fluid jet polishing process based on negative pressure effects.

Traditional abrasive fluid jet polishing (FJP) is limited by its high-pressure equipment, unstable material removal rate, and applicability to ultra-smooth surfaces because of the evident air turbulence, fluid expansion, and a large polishing spot in high-pressure FJP. This paper presents a novel cavitation fluid jet polishing (CFJP) method and process based on FJP technology. It can implement high-efficiency polishing on small-scale surfaces in a low-pressure environment. CFJP uses the purposely designed polishing equipment with a sealed chamber, which can generate a cavitation effect in negative pressure environment. Moreover, the collapse of cavitation bubbles can spray out a high-energy microjet and shock wave to enhance the material removal. Its feasibility is verified through researching the flow behavior and the cavitation results of the negative pressure cavitation machining of pure water in reversing suction flow. The mechanism is analyzed through a computational fluid dynamics simulation. Thus, its cavitation and surface removal mechanisms in the vertical CFJP and inclined CFJP are studied. A series of polishing experiments on different materials and polishing parameters are conducted to validate its polishing performance compared with FJP. The maximum removal depth increases, and surface roughness gradually decreases with increasing negative outlet pressures. The surface becomes smooth with the increase of polishing time. The experimental results confirm that the CFJP process can realize a high material removal rate and smooth surface with low energy consumption in the low-pressure environment, together with compatible surface roughness to FJP.

Optimized pre-thinning procedures of ion-beam thinning for TEM sample preparation by magnetorheological polishing.

Ion-beam-thinning is a well-established sample preparation technique for transmission electron microscopy (TEM), but tedious procedures and labor consuming pre-thinning could seriously reduce its efficiency. In this work, we present a simple pre-thinning technique by using magnetorheological (MR) polishing to replace manual lapping and dimpling, and demonstrate the successful preparation of electron-transparent single crystal silicon samples after MR polishing and single-sided ion milling. Dimples pre-thinned to less than 30 microns and with little mechanical surface damage were repeatedly produced under optimized MR polishing conditions. Samples pre-thinned by both MR polishing and traditional technique were ion-beam thinned from the rear side until perforation, and then observed by optical microscopy and TEM. The results show that the specimen pre-thinned by MR technique was free from dimpling related defects, which were still residual in sample pre-thinned by conventional technique. Nice high-resolution TEM images could be acquired after MR polishing and one side ion-thinning. MR polishing promises to be an adaptable and efficient method for pre-thinning in preparation of TEM specimens, especially for brittle ceramics.

The role of the Staphylococcal VraTSR regulatory system on vancomycin resistance and vanA operon expression in vancomycin-resistant Staphylococcus aureus.

Vancomycin is often the preferred treatment for invasive methicillin-resistant Staphylococcus aureus (MRSA) infection. With the increase in incidence of MRSA infections, the use of vancomycin has increased and, as feared, isolates of vancomycin-resistant Staphylococcus aureus (VRSA) have emerged. VRSA isolates have acquired the entercoccal vanA operon contained on transposon (Tn) 1546 residing on a conjugal plasmid. VraTSR is a vancomycin and ß-lactam-inducible three-component regulatory system encoded on the S. aureus chromosome that modulates the cell-wall stress response to cell-wall acting antibiotics. Mutation in vraTSR has shown to increase susceptibility to ß-lactams and vancomycin in clinical VISA strains and in recombinant strain COLVA-200 which expresses a plasmid borne vanA operon. To date, the role of VraTSR in vanA operon expression in VRSA has not been demonstrated. In this study, the vraTSR operon was deleted from the first clinical VRSA strain (VRS1) by transduction with phage harvested from a USA300 vraTSR operon deletion strain. The absence of the vraTSR operon and presence of the vanA operon were confirmed in the transductant (VRS1Δvra) by PCR. Broth MIC determinations, demonstrated that the vancomycin MIC of VRS1Δvra (64 µg/ml) decreased by 16-fold compared with VRS1 (1024 µg/ml). The effect of the vraTSR operon deletion on expression of the van gene cluster (vanA, vanX and vanR) was examined by quantitative RT-PCR using relative quantification. A 2-5-fold decreased expression of the vanA operon genes occured in strain VRS1Δvra at stationary growth phase compared with the parent strain, VRS1. Both vancomycin resistance and vancomycin-induced expression of vanA and vanR were restored by complementation with a plasmid harboring the vraTSR operon. These findings demonstrate that expression in S. aureus of the horizontally acquired enterococcal vanA gene cluster is enhanced by the staphylococcal three-component cell wall stress regulatory system VraTSR, that is present in all S. aureus strains.

Improved oxacillin treatment outcomes in experimental skin and lung infection by a methicillin-resistant Staphylococcus aureus isolate with a vraSR operon deletion.

Methicillin-resistant Staphylococcus aureus (MRSA) strains are major pathogens causing infections of the skin and soft tissues and more serious, life-threatening diseases, including sepsis and necrotizing pneumonia. The vraSR operon encodes the key regulatory system that modulates the stress response of S. aureus elicited upon exposure to cell wall antibiotics. Mutation of vraS and vraR results in decreased oxacillin resistance in vitro. We investigated the effect of oxacillin treatment in experimental models employing a clinical USA300 MRSA strain (strain 923) and an isogenic vraSR deletion mutant (strain 923-M23). In a murine model of S. aureus necrotizing pneumonia, animals were treated with oxacillin, beginning 15 min after inoculation. Among mice infected with mutant strain 923-M23, oxacillin treatment significantly improved survival compared with saline treatment, whereas oxacillin treatment had no effect in mice infected with strain 923. Similarly, treatment with oxacillin decreased the bacterial burden among animals infected with strain 923-M23 but not among animals infected with strain 923. In a murine skin infection model, oxacillin eliminated the development of dermonecrosis among 923-M23-infected mice and decreased the bacterial burden in the lesions, but not among strain 923-infected mice. We conclude that deletion of the vraSR operon allowed an oxacillin regimen to be effective in murine models of MRSA pneumonia and skin infection. These findings provide proof-of-principle for development of a new antibiotic that could restore the usefulness of oxacillin against MRSA by inhibiting VraS or VraR.

ELID grinding characteristics of large stamping die for Wolter mirror.

An ultra-precision ELID grinding of large stamping dies of Wolter mirror for X-ray telescope was presented in this paper. The large stamping dies (S55C) with confocal paraboloid and hyperboloid was ground by ELID arc-enveloped grinding. In this ELID grinding system, cast iron fiber bonded (CIFB) diamond wheels were controlled by 3-dimentional ways to scan the work-piece and generate required surfaces. Its grinding characteristics such as attainable form accuracy, surface roughness were investigated. Furthermore, some measures to improve form accuracy were discussed and verified such as truing, compensating, and on-machine measuring.

The VraS/VraR two-component regulatory system required for oxacillin resistance in community-acquired methicillin-resistant Staphylococcus aureus.

Methicillin/oxacillin (Oxa) resistance in Staphylococcus aureus is primarily mediated by the acquired penicillin-binding protein (PBP2a) encoded by mecA. PBP2a acts together with native PBP2 to mediate oxacillin resistance by contributing complementary transpeptidase and transglycosylase activities, respectively. The VraS/VraR two-component regulatory system is inducible by cell-wall antimicrobials (beta-lactams, glycopeptides) and controls transcriptional induction of many cell-wall genes including pbp2 and itself. We investigated the role of VraS/VraR in the phenotypic expression of oxacillin resistance by inactivating vraS in community-acquired MRSA clinical isolates that lack functional genes encoding the mecA regulatory sequences mecI and mecR1. Inactivation of vraS abrogated oxacillin resistance, and complementation with the vraS operon restored the resistance phenotype. mecA transcription increased in the vraS mutants; however, PBP2a abundance was similar to that of the wild type. Although pbp2 transcription decreased in the vraS mutants, overexpression of the pbp2 operon did not restore resistance. These data demonstrate that although expressions of mecA and pbp2 are required for oxacillin resistance, they are not sufficient. Therefore, the vraS/vraR regulatory system plays a crucial role in allowing MRSA to respond to beta-lactams by regulation of a gene target other than the known effectors of methicillin resistance.

VraSR two-component regulatory system and its role in induction of pbp2 and vraSR expression by cell wall antimicrobials in Staphylococcus aureus.

The VraS/VraR two-component system (VraSR) regulates transcriptional induction of penicillin-binding protein 2 (encoded by pbp2) by vancomycin in Staphylococcus aureus. We have now defined the vraSR operon and determined that its induction by beta-lactams as well as by vancomycin is autoregulated. Induction of the pbp2 and vraSR operons by beta-lactams and related compounds within 1 hour after exposure to the antimicrobials was dependent on vraS. However, when a disk diffusion assay that can detect induction of genes over an extended time period was used, induction of the pbp2 operon was mediated by some beta-lactams, including oxacillin; this induction was independent of vraS.

Transcriptional induction of the penicillin-binding protein 2 gene in Staphylococcus aureus by cell wall-active antibiotics oxacillin and vancomycin.

We found an increased abundance of pbpB-specific transcripts in vancomycin intermediate-resistant Staphylococcus aureus (VISA) isolates compared with that found in paired, genetically identical, susceptible isolates. This difference in expression cannot be explained by differences in the pbpB promoter sequence. Since the factors controlling pbpB gene expression have remained largely unexplored, various conditions that might affect pbpB transcript abundance were examined. In both vancomycin-susceptible and VISA strains, pbpB expression varied with the growth phase, with the highest abundance of pbpB-specific transcripts detected during mid-log phase. Interestingly, both vancomycin and oxacillin were able to induce pbpB transcription above a constitutive level. When vancomycin was absent, one of the three pbpB-specific transcripts that were usually faintly detected in non-VISA strains was more readily detected in VISA strains during mid-log but not stationary phase. This transcript was enhanced in non-VISA strains by vancomycin induction. Gel shift assays indicated that an increased amount of the putative transcription factor that binds to both P1 and P1' promoter regions is present in the cytosol of vancomycin-induced cells. Neither the SigB sigma factor nor the quorum-sensing agr locus was required for growth phase-variable pbpB expression or transcriptional induction of pbpB by vancomycin or oxacillin. Also, MecI, MecR1, BlaI, and BlaR1, regulatory proteins that mediate beta-lactam-inducible expression of mecA and beta-lactamase, were not required for antibiotic induction of pbpB transcription. These data support the idea that pbpB expression is modulated by a trans-acting factor in response to the presence of the cell wall-active antibiotics vancomycin and oxacillin.