Cyclic dimeric GMP (c-di-GMP) is a bacterial second messenger that modulates many biological processes. Although its role in bacterial pathogenesis during mammalian infection has been documented, the role of c-di-GMP in a pathogen's life cycle within a vector host is less understood. The enzootic cycle of the Lyme disease pathogen Borrelia burgdorferi involves both a mammalian host and an Ixodes tick vector. The B. burgdorferi genome encodes a single copy of the diguanylate cyclase gene (rrp1), which is responsible for c-di-GMP synthesis. To determine the role of c-di-GMP in the life cycle of B. burgdorferi, an Rrp1-deficient B. burgdorferi strain was generated. The rrp1 mutant remains infectious in the mammalian host but cannot survive in the tick vector. Microarray analyses revealed that expression of a four-gene operon involved in glycerol transport and metabolism, bb0240-bb0243, was significantly downregulated by abrogation of Rrp1. In vitro, the rrp1 mutant is impaired in growth in the media containing glycerol as the carbon source (BSK-glycerol). To determine the contribution of the glycerol metabolic pathway to the rrp1 mutant phenotype, a glp mutant, in which the entire bb0240-bb0243 operon is not expressed, was generated. Similar to the rrp1 mutant, the glp mutant has a growth defect in BSK-glycerol medium. In vivo, the glp mutant is also infectious in mice but has reduced survival in ticks. Constitutive expression of the bb0240-bb0243 operon in the rrp1 mutant fully rescues the growth defect in BSK-glycerol medium and partially restores survival of the rrp1 mutant in ticks. Thus, c-di-GMP appears to govern a catabolic switch in B. burgdorferi and plays a vital role in the tick part of the spirochetal enzootic cycle. This work provides the first evidence that c-di-GMP is essential for a pathogen's survival in its vector host.

A computational model is proposed for the prediction of friction-related mechanical efficiency losses of parallel-axis gear pairs. The model incorporates a gear load distribution model, a friction model, and a mechanical efficiency formulation to predict the instantaneous mechanical efficiency of a gear pair under typical operating, surface, and lubrication conditions. The friction model uses a new friction coefficient formula obtained by using a validated non-Newtonian thermal elastohydrodynamic lubrication (EHL) model in conjunction with a multiple linear regression analysis. The load and friction coefficient distribution predictions are used to compute instantaneous torque/ power losses and the mechanical efficiency of a gear pair at any given rotational position. Efficiency measurements from gear pairs having various gear designs and surface treatments are compared to model predictions. Mechanical efficiency predictions are shown to be within 0.1% of the measured values, indicating that the proposed efficiency model is accurate. Results of a parametric study are presented at the end to highlight the influence of key basic gear geometric parameters, tooth modifications, operating conditions, surface finish, and lubricant properties on mechanical efficiency losses.

Alteration of surface lipoprotein profiles is a key strategy that the Lyme disease pathogen, Borrelia burgdorferi, has evolved to be maintained within its enzootic cycle between arthropods and mammals. Accumulated evidence indicates that the central regulatory pathway controlling differential gene expression by B. burgdorferi is the RpoN-RpoS pathway (the 54 -S sigma factor cascade). It was previously shown that activation of the RpoN-RpoS pathway is controlled by Rrp2, a two-component response regulator and 54 -dependent transcriptional activator. The role of Rrp2 in the infectious cycle of B. burgdorferi has not been determined heretofore. In this report, we demonstrate that an rrp2 mutant defective in activating 54 -dependent transcription was unable to establish infection in mice, but the rrp2 mutant was capable of surviving within ticks during and after tick feeding. Because the rrp2 mutant was defective in the production of OspC, an outer surface lipoprotein essential for mammalian host infection, we further examined whether the loss of infectivity of the rrp2 mutant was solely due to the inability to produce OspC. While transformation with a shuttle vector carrying ospC under the control of a constitutive flaB promoter restored infection to an ospC mutant in immunodeficient SCID mice, it could not rescue the avirulent phenotype of the rrp2 mutant. These data indicate that, in addition to controlling OspC, Rrp2 controls another factor(s) essential for B. burgdorferi to establish infection in mammals. Furthermore, microarray analyses revealed that 125 and 19 genes were positively and negatively regulated, respectively, by Rrp2, which provides a foundation for future identification of additional Rrp2-dependent virulence determinants in B. burgdorferi.

Borrelia burgdorferi, the Lyme disease spirochete, dramatically alters its transcriptome and proteome as it cycles between the arthropod vector and mammalian host. During this enzootic cycle, a novel regulatory network, the Rrp2-RpoN-RpoS pathway (also known as the σ54–σS sigma factor cascade), plays a central role in modulating the differential expression of more than 10% of all B. burgdorferi genes, including the major virulence genes ospA and ospC. However, the mechanism(s) by which the upstream activator and response regulator Rrp2 is activated remains unclear. Here, we show that none of the histidine kinases present in the B. burgdorferi genome are required for the activation of Rrp2. Instead, we present biochemical and genetic evidence that supports the hypothesis that activation of the Rrp2-RpoN-RpoS pathway occurs via the small, high-energy, phosphoryl-donor acetyl phosphate (acetyl∼P), the intermediate of the Ack-Pta (acetate kinase-phosphate acetyltransferase) pathway that converts acetate to acetyl-CoA. Supplementation of the growth medium with acetate induced activation of the Rrp2-RpoN-RpoS pathway in a dose-dependent manner. Conversely, the overexpression of Pta virtually abolished acetate-induced activation of this pathway, suggesting that acetate works through acetyl∼P. Overexpression of Pta also greatly inhibited temperature and cell density-induced activation of RpoS and OspC, suggesting that these environmental cues affect the Rrp2-RpoN-RpoS pathway by influencing acetyl∼P. Finally, overexpression of Pta partially reduced infectivity of B. burgdorferi in mice. Taken together, these findings suggest that acetyl∼P is one of the key activating molecule for the activation of the Rrp2-RpoN-RpoS pathway and support the emerging concept that acetyl∼P can serve as a global signal in bacterial pathogenesis.

A silicon hierarchical structure, silicon nanoporous pillar array (Si-NPA), was prepared by a hydrothermal etching method. The architecture of Si-NPA was characterized to be a regular array of micron-sized, quasi-identical and nanoporous silicon pillars with an additional porous layer beneath the array. The pore walls were proved to be consisted of a SiO(x) matrix and dispersive silicon nanocrystallites. An integral reflectivity below 4% was achieved in the wavelength range of 240-2400 nm. Three photoluminescence bands, one blue and two red, were observed at room temperature and attributed to the recombination processes through band-to-band transition and luminescent centers, respectively. The structural and physical properties indicate that Si-NPA might be as both a functional silicon nanostructure and a template for assembling silicon-based nanocomposites in fabricating optoelectronic nanodevices.

Rrp2 is the sole 54 -dependent transcriptional activator present in the Borrelia burgdorferi genome. We showed that recombinant Rrp2 binds to DNA in a sequence-nonspecific manner. During infection, Rrp2 activates 54 -dependent rpoS expression without an apparent upstream enhancer element commonly associated with other 54 -dependent transcriptional activators.

Graphene-based membranes have great potential to revolutionize nanofiltration technology, but achieving high solute rejections at high water flux remains extremely challenging. Herein, a family of ultrafine metal oxide/reduced graphene oxide (rGO) nanocomposites are synthesized through a heterogenous nucleation and diffusion-controlled growth process for dye nanofiltration. The synthesis is based on the utilization of oxygen functional groups on GO surface as preferential active sites for heterogeneous nucleation, leading to the formation of sub-3 nm size, monodispersing as well as high-density loading of metal oxide nanoparticles. The anchored ultrafine nanoparticles could inhibit the wrinkling of the rGO nanosheet, forming highly stable colloidal solutions for the solution processing fabrication of nanofiltration membranes. By functioning as pillars, the nanoparticles remarkably increase both vertical interlayer spacing and lateral tortuous paths of the rGO membranes, offering a water permeability of 225 L m−2 h−1 bar−1 and selectivity up to 98% in the size-exclusion separation of methyl blue.

A model to predict friction-related mechanical efficiency losses of hypoid gear pairs is proposed in this study. The model includes a gear contact model, a friction prediction model, and a mechanical efficiency formulation. The friction model uses a friction coefficient formula obtained by applying multiple linear regression analysis to a large number of elastohydrodynamic lubrication analyses covering typical ranges of key parameters associated with surface roughness, geometry, load, kinematics, and the lubricant. Formulations regarding the kinematic and geometric properties of the hypoid gear contact are presented. The load and friction coefficient distribution predictions are used to compute instantaneous torque/power losses and the mechanical efficiency of a hypoid gear pair at any given position. Results of a parametric study are presented at the end to highlight the influence of key operating conditions, surface finish, and lubricant properties on mechanical efficiency losses of hypoid gears.