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Mucus secretions typically protect exposed surfaces of the eyes and respiratory, gastrointestinal and female reproductive tracts from foreign entities, including pathogens and environmental ultrafine particles. We hypothesized that excess exposure to some foreign particles, however, may cause disruption of the mucus barrier. Many synthetic nanoparticles are likely to be mucoadhesive due to hydrophobic, electrostatic or hydrogen bonding interactions. We therefore sought to determine whether mucoadhesive particles (MAP) could alter the mucus microstructure, thereby allowing other foreign particles to more easily penetrate mucus. We engineered muco-inert probe particles 1 µm in diameter, whose diffusion in mucus is limited only by steric obstruction from the mucus mesh, and used them to measure possible MAP-induced changes to the microstructure of fresh human cervicovaginal mucus. We found that a 0.24% w/v concentration of 200 nm MAP in mucus induced a ∼10-fold increase in the average effective diffusivity of the probe particles, and a 2- to 3-fold increase in the fraction capable of penetrating physiologically thick mucus layers. The same concentration of muco-inert particles, and a low concentration (0.0006% w/v) of MAP, had no detectable effect on probe particle penetration rates. Using an obstruction-scaling model, we determined that the higher MAP dose increased the average mesh spacing ("pore" size) of mucus from 380 nm to 470 nm. The bulk viscoelasticity of mucus was unaffected by MAP exposure, suggesting MAP may not directly impair mucus clearance or its function as a lubricant, both of which depend critically on the bulk rheological properties of mucus. Our findings suggest mucoadhesive nanoparticles can substantially alter the microstructure of mucus, highlighting the potential of mucoadhesive environmental or engineered nanoparticles to disrupt mucus barriers and cause greater exposure to foreign particles, including pathogens and other potentially toxic nanomaterials.
Citation: Wang Y-Y, Lai SK, So C, Schneider C, Cone R, Hanes J (2011) Mucoadhesive Nanoparticles May Disrupt the Protective Human Mucus Barrier by Altering Its Microstructure. PLoS ONE 6(6): e21547. https://doi.org/10.1371/journal.pone.0021547
Editor: Wei-Chun Chin, University of California, Merced, United States of America
Received: March 17, 2011; Accepted: June 1, 2011; Published: June 29, 2011
Copyright: © 2011 Wang et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Funding: This work was supported in part by the National Institutes of Health (http://www.nih.gov/) grants 5U01AI066726 (R.C.), P01HL51811 (J.H.), R21HL089816 and R01HD062844 (J.H. and R.C.) and by a National Science Foundation (http://www.nsf.gov/) graduate research fellowship (Y.-Y.W.). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Heart, Lung, and Blood Institute, the National Institute of Biomedical Imaging and Bioengineering, the National Cancer Institute or the National Institutes of Health. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Competing interests: The authors have declared that no competing interests exist.
Engineered or synthetic nanoparticles are increasingly used in diverse applications, ranging from ultra-light high strength materials to electronics, cosmetics and medicine. As the manufacturing and use of synthetic nanoparticles become more common, the potential for unintended human exposure also increases. Concerns over the safety of nanoparticles have centered primarily on their small size and, therefore, potential for deeper tissue penetration and higher reactivity (due to their high surface area to volume ratio) [1]–[3]. Recent studies have shown that some non-degradable environmental or engineered nanoparticles may trigger significant pulmonary inflammation and/or immunological responses in vivo [1]–[3]. Several investigators have also suggested certain synthetic nanoparticles can be directly toxic to cells through the generation of reactive oxygen species [2]–[3].
Exposure to synthetic nanoparticles may occur via percutaneous or trans-mucosal absorption. Nevertheless, the interactions between nanoparticles and mucus in the airways, as well as other mucosal organs, have yet to be fully characterized. Mucus is a viscoelastic and adhesive gel that coats and protects nearly all exposed surfaces of the human body not covered by skin, serving as the first line of defense against foreign particles that impinge on these surfaces. The dense mucin mesh network allows mucus to efficiently trap most foreign particles through steric obstruction and/or adhesion via hydrophobic, electrostatic or hydrogen bonding interactions [4]. Trapped particles are quickly eliminated in as little as seconds to minutes (surface of the eye) to minutes to hours (gastrointestinal, respiratory and female reproductive tracts) by normal mucus clearance mechanisms [4]–[6]. Synthetic nanoparticles typically feature hydrophobic, charged and/or hydrogen bonding surfaces [7]–[10] and are, therefore, likely to be strongly mucoadhesive due to interactions with periodic exposed hydrophobic domains or negatively charged glycosylated segments along mucin fibers [4]. Although these and other mucoadhesive particles are unlikely to perturb the microstructure of mucus at low exposure levels, at high concentrations, they may crosslink mucin fibers and cause them to bundle together, enlarging pores in the mucus gel (Figure 1). Larger pores may compromise the ability of mucus to trap foreign particles, and increase the likelihood that other foreign particles or pathogens will reach and potentially injure the underlying epithelia.
MAP may increase mucus pore sizes by bundling mucin fibers through polyvalent adhesive interactions. This would allow larger non-mucoadhesive particles to penetrate mucus more readily.
Here, we sought to investigate the ability of synthetic mucoadhesive particles (MAP) to alter the microstructure of human mucus, using 200 nm amine-modified polystyrene nanoparticles as model MAP. We performed our studies with freshly obtained and minimally altered ex vivo human cervicovaginal mucus (CVM), a physiological, highly viscoelastic gel that is difficult to reproduce in cell culture. To detect changes in the mucus microstructure, we use 1 µm muco-inert particles as probes whose diffusional speeds can be related to effective pore sizes in the mucus gel. This method involves minimal perturbation of the mucus sample, in contrast to electron microscopy, which is known to introduce artifacts due to dehydration and fixation steps during sample preparation [11]–[13]. We have recently employed this technique to characterize the pore size of native CVM [14] and cystic fibrosis sputum [15]. The speeds of the probe particles also allow us to estimate the fraction of foreign particles that may penetrate a physiologically thick mucus layer over time, providing another assessment of how MAP may compromise the mucus barrier.
Materials and Methods
Ethics statement
Research ethics approval for the collection of human CVM samples used in this study was received from the Institutional Review Board of the Johns Hopkins University. Written informed consent was obtained from each participant enrolled in the study.
Human cervicovaginal mucus collection
Mucus samples were obtained from women of reproductive age (ranging from 18 to 27 years old) with healthy vaginal micro-flora. Donors stated they had not used vaginal products nor participated in unprotected intercourse within 3 days prior to donating. Undiluted CVM, averaging 0.3 g per sample, was collected using a self-sampling menstrual collection device following protocols approved by the Institutional Review Board of the Johns Hopkins University [8], [16]. The device was inserted into the vagina for approximately 30 s, removed, and placed into a 50 mL centrifuge tube. Samples were centrifuged at 1000 rpm for 30 s to collect the secretions. Collected mucus was stored at 4 °C until used for microscopy the same day. The samples were collected at random times throughout the menstrual cycle, but none were obtained during ovulation based on the absence of spinnbarkeit by visual inspection. Samples that were non-uniform in color or consistency were discarded.
Nanoparticle preparation and characterization
For model MAP, we used amine-modified polystyrene nanoparticles sized 200 nm (Molecular Probes, Eugene, OR), which were concentrated to 8% w/v (∼1013 particles/mL) by centrifugation and resuspension in water. These particles feature a hydrophobic core and a positively charged surface at the native pH of CVM (pH∼4), and may adhere to mucus through hydrophobic interactions with oily domains along mucin fibers or through electrostatic and/or hydrogen bonding interactions with the negatively charged, glycosylated domains of mucins [4]. For muco-inert probe particles, we covalently modified fluorescent, carboxylated polystyrene particles sized 1 µm (Molecular Probes, Eugene, OR) with 2 kDa amine-modified poly(ethylene glycol) (PEG; Nektar Therapeutics, San Carlos, CA) via a carboxyl-amine reaction, as published previously [8]. PEG is a hydrophilic and uncharged polymer that, at high surface density and low MW, can effectively shield the hydrophobic polystyrene core from adhesive interactions with mucins, while also minimizing interpenetrating network effects (polymer chain entanglements) and hydrogen bonding between PEG chains and mucins [8], [17].
Particle size and ξ-potential (Table 1) were determined by dynamic light scattering and laser Doppler anemometry, respectively, using a Zetasizer Nano ZS90 (Malvern Instruments, Southborough, MA). Size measurements were performed at 25°C at a scattering angle of 90°. Samples were diluted in 10 mM NaCl solution (pH 4 or pH 7) and measurements performed according to instrument instructions. A near-neutral ξ-potential at pH 7 was used to confirm PEG conjugation onto the 1 µm probe particles [8]. Based on our previous findings, muco-inert particles 1 µm or larger in diameter are highly hindered in native human mucus by steric obstruction, whereas 500 nm muco-inert particles are nearly freely diffusive due to the availability of sufficiently large spaces or pores in the mucus mesh [14], [18]. As such, 1 µm muco-inert particles are sensitive probes for detecting increases in mucus pore size.
Multiple particle tracking
Each mucus sample was separated into aliquots and treated with 8% w/v MAP or, as controls, (i) saline, (ii) a diluted solution of MAP (0.02% w/v) or (iii) 8% w/v of 200 nm muco-inert particles (prepared and characterized as described above for 1 µm probe particles). Solutions were added to mucus at 3% v/v, with gentle stirring to achieve visually uniform particle distribution, yielding final particle concentrations of 0.24% w/v or 0.0006% w/v. The concentration of 0.24% w/v falls within dose limits (0.17–0.35% w/v) estimated based on the Occupational Safety and Health Administration's Permissible Exposure Limits for airborne particulates, such as metal oxide nanoparticles (∼10 mg/m3, 8 hr time-weighted average [19]); an average breathing rate of 3.2 m3/hr for heavily active adults [20]; a deposition fraction of ∼5–10% for 0.1–1 µm particles in the tracheobronchial (TB) region of the lungs [21]; a TB surface area of 2741 cm2 [22]; and an average TB mucus layer thickness of 30 µm [4]. Treated samples were incubated at least 15 min prior to addition of diluted probe particle solutions (∼1010 particles/mL) at 5% v/v. Samples were transferred to custom-made chambers holding 20–30 µL of mucus each and incubated 2 hr before microscopy. Samples incubated at room temperature or at 37°C yielded similar results. The trajectories of the fluorescent probe particles in CVM were recorded using a silicon-intensified target camera (VE-1000, Dage-MTI, Michigan, IN) mounted on an inverted epifluorescence microscope equipped with 100X oil-immersion objective (numerical aperture 1.3). Movies were captured with Metamorph software (Universal Imaging Corp., Downingtown, PA) at a temporal resolution of 66.7 ms for 20 s. The tracking resolution was 10 nm, as determined by tracking the displacements of particles immobilized with a strong adhesive [23]. Trajectories of n>100 probe particles were analyzed for each experiment, and six experiments were performed in independent CVM samples from different donors. The coordinates of particle centroids were transformed into time-averaged mean squared displacements (MSD), calculated as <Δr2(τ)> = [x(t + τ) – x(t)]2 + [y(t + τ) – y(t)]2 (where τ = time scale or time lag), from which individual particle and ensemble average MSD and effective diffusivities (Deff = MSD/(4τ) for 2D particle tracking) were calculated, as previously demonstrated [8], [24]. Immobile particles are defined as those with an average MSD below the 10 nm tracking resolution at a time scale of 1 s.
Mucus pore size analysis
The sizes of pores between mucin fibers of saline- and MAP-exposed CVM were estimated based on an obstruction-scaling model originally developed by Amsden and coworkers for covalently cross-linked hydrogels, but equally applicable to gels with physical entanglement cross-links, such as mucus [25]–[28]. The model is valid when there is no chemical interaction between probe particles and the gel mesh, and particles traveling through the pores of the gel experience the viscous drag of water. We have shown previously that particles well coated with low MW PEG exhibit minimal adhesive interactions with mucus constituents [8], [14], [17], as well as other synthetic particles (unpublished observations). The model describes the ratio of diffusion in a gel and diffusion in water as Dg/Do = exp((−π/4)((rs + rf)/(rg + rf))2), where Dg is the diffusion coefficient of the probe particle in the polymer gel, Do is its diffusion coefficient in water, rs is the particle radius, rf is the gel fiber radius, and rg is the effective radius of the pore. An rf of 3.5 nm was used as the current best estimate for the radius of individual mucin fibers from biochemical, electron microscopy and atomic force microscopy observations [27]–[28].
Simulation of particle penetration
To estimate particle penetration across mucus, we performed a Monte Carlo simulation of one-dimensional particle transport across a mucus slab of uniform thickness. The simulation consisted of ∼20,000 particles with effective diffusivities equal to those measured experimentally for the 1 µm PEG-coated probe particles in saline- or MAP-exposed CVM at a time scale of 1 s (30 particles simulated per measured diffusivity). Particles were initially located at one surface of the slab, and then allowed to undergo Brownian motion. Particle positions over time (in steps of 1 s) were recorded, and the fraction of penetrable particles was calculated as the fraction that reaches the opposite surface of the mucus slab.
Bulk rheological characterization of MAP-exposed CVM
Bulk rheological characterization of CVM was performed with a strain-controlled cone and plate rheometer (ARES-100, Rheometrics, Piscataway, NJ). CVM from 4–5 donor samples was pooled (total volume ∼1.3 mL) and stored at 4°C until use. The temperature of specimens was maintained at 37°C during measurements. Oscillatory deformations of small amplitude (1% strain) and controlled frequency were applied to extract the frequency-dependent viscoelastic properties with minimal shearing damage to the CVM samples. We report the frequency-dependent elastic and viscous moduli, G′(ω) and G″(ω), which are the in-phase and out-of-phase components, respectively, of the stress induced in the CVM samples divided by the maximum amplitude of the applied deformation. Phase angle is defined as arctan(G″/G′); a phase angle of 90° corresponds to a viscous liquid, while 0° corresponds to an elastic solid. The rheology of control and MAP-exposed CVM was evaluated in sequential order. As with particle tracking experiments, MAP (8% w/v) were added at 3% v/v, gently stirred to ensure exposure of the entire sample, and incubated at least 15 min prior to subsequent measurements.
Statistical analysis
All data are presented as a mean with standard error of the mean (mean ± SEM) indicated. Statistical significance between saline and MAP conditions was determined by a one-tailed, non-parametric Wilcoxon signed-rank test, since the response of mucus samples to MAP depends on a number of factors (see Discussion) and cannot be assumed to be normally distributed. p values less than 0.05 were considered statistically significant.
Results and Discussion
Effect of MAP on mucus microstructure
In agreement with our previous work [14], [18], the diffusion of 1 µm muco-inert (PEG-coated) probe particles was strongly hindered in control mucus samples treated with 3% v/v saline, as evident by their constrained and non-Brownian trajectories (Figure 2A and Video S1). In contrast, the probes exhibited more diffusive trajectories in aliquots of the same native mucus samples treated with 3% v/v MAP at a toxicologically relevant dose of 0.24% w/v final concentration (see Methods for details; Video S2). As we have previously observed [8], MAP were nearly completely immobilized in mucus (data not shown), whereas similarly sized and larger (up to at least 500 nm) muco-inert particles diffuse freely through mucus, suggesting MAP must be immobilized by adhesive interactions to the mucin mesh network. Using multiple particle tracking [8], [24], we quantified probe particle motions in terms of ensemble average mean square displacements (<MSD>; Figure 2B). Five out of six independent samples showed an increase in probe particle <MSD> upon exposure of mucus to MAP, while one showed no significant change. On average, MAP exposure increased probe particle effective diffusivity by ∼10-fold for each sample, at a time scale of 1 s. This improvement was also reflected by the slope α of the log-log <MSD> vs. time scale plot (α = 1 represents unconstrained Brownian transport, while decreasing α corresponds to increased obstruction to particle movement). The average α was 0.68 for probe particles in treated samples, compared to only 0.36 in control samples. The increase in particle transport rates was due in part to a significant drop in the fraction of immobile probes (from 33% to 20%; Figure 3).
(A) Sample trajectories of probe particles with effective diffusivities within one S.E.M. of the mean at a time scale of 1 s. (B) Ensemble-averaged geometric mean square displacements (<MSD>) as a function of time scale for probe particles. Data represent six independent experiments with samples from different donors (n≥100 particles per experiment). α represents the slope of the log-log <MSD> vs. time scale plot. * indicates statistical significance at p<0.05.
Since well-PEGylated particles have minimal affinity to mucin fibers [8], [17], the increase in probe particle speeds in MAP-exposed mucus suggests MAP enlarged the pores in the mucus mesh. To quantify this change in the mucus microstructure, we fitted the effective diffusivities of the probe particles to an obstruction-scaling model, which relates the obstruction experienced by non-interacting solute particles to the pore size of the surrounding mesh network [25]–[26]. The average pore size of mucus samples increased significantly from 380±30 nm in the saline control to 470±80 nm in the MAP-exposed condition (p<0.05). The fraction of pores with spacings ≥500 nm increased from 11% to 19%, and the fraction of pores ≥1 µm increased from 3% to 9%.
In contrast to our observations with a relatively high MAP dose (0.24% w/v), the same amount of 200 nm muco-inert nanoparticles (densely coated with PEG) or 400-fold less MAP did not significantly affect the mucus microstructure (data not shown), suggesting the effects of synthetic nanoparticles on mucus depend on both particle concentration and surface chemistry. Synthetic nanoparticles may adhere to mucins through hydrophobic or electrostatic interactions with hydrophobic and glycosylated domains along mucin fibers, respectively. At low concentrations, the number of MAP is likely insufficient to form the bonds with mucins (i.e., avidity) necessary to crosslink or bundle mucins, which are normally held apart by entanglements or terminal disulfide bridges. Likewise, particles that interact minimally with mucins would have little effect on the mucus microstructure, even at high concentrations. However, the collective binding interactions of large numbers of MAP with multiple mucin fibers may generate sufficient avidity to cause mucins to crosslink and bundle together, thus yielding larger pores in the mucus mesh. This notion is consistent with visual observations by Olmsted and coworkers of high concentrations of fluorescent polystyrene nanoparticles agglomerating mid-cycle cervical mucins into thick cables [27]. In that work, polystyrene nanoparticles caused a striking collapse of the mucus gel, to an extent greater than what we observed here. A key difference that may explain this discrepancy is the use of ovulatory cervical mucus (OCM) by Olmsted et al. compared to non-ovulatory cervicovaginal mucus here. OCM is more dilute [29], so there are fewer entanglements and crosslinks to resist the displacement of mucin fibers during bundling by MAP. In addition, electron microscopy studies have suggested ovulatory mucus contains long strands of mucin fibers arranged in parallel with minimal entanglements, which may promote bundling by MAP compared to non-ovulatory mucus that consists of a random mesh of highly entangled mucin fibers [30]. Other experimental differences include the volume of particles added (20% v/v by Olmsted vs. 3% v/v here), the total amount of particles added (final concentration 2% w/v vs. 0.24% w/v here) and the degree of particle mixing into mucus (relatively rigorous vs. gentle stirring here).
We observed substantial heterogeneity in MAP-induced changes to the microstructure, which may be attributed to compositional differences between CVM samples collected at random points throughout the menstrual cycle (excluding the ovulatory phase) and from different donors. Due to hormonal changes, the contents of mucins (both total mucin concentration and different MUC types) [29], [31], lipids [32] and other mucus constituents differ markedly throughout the menstrual cycle. As discussed above, mucin concentration may determine the extent of mucin bundling possible. Lipids, which naturally coat the hydrophobic domains of mucins [33], may also interfere with adhesive interactions between mucins and MAP. It is likely that the effect of a higher particle concentration in improving the mucus-altering ability of MAP may eventually saturate, since the number of adhesive interactions mucins can form with particles is finite. In addition, physical entanglements and disulfide crosslinks within the mucin mesh network may restrict the extent of mucin bundling possible.
Predicted particle penetration into MAP-exposed mucus layers
The increased mobility of the probe particles in MAP-exposed mucus suggests a greater fraction may penetrate physiologically thick mucus layers. To model the long range diffusion of the particles, we performed a Monte Carlo simulation of particles undergoing Brownian diffusion across a mucus slab, using the diffusivities of hundreds of probe particles measured above (Figure 4). The thickness of physiological mucus layers varies from tens to several hundred microns depending on anatomical location [4], [34]–[36] here, we assumed thicknesses ranging from 10 to 55 µm, which correspond to values reported for the large airways [36]. For a thickness of 10 µm, about 2-fold more of 1 µm probe particles were predicted to penetrate the mucus layer within 1 hr in MAP-exposed vs. saline-treated mucus (11% vs. 6%, respectively), and, for a thickness of 30 µm, 3-fold more probe particles were predicted to penetrate (3% vs. 1%, respectively). For comparison, if the particles were diffusing through pure water, 64% and 17% of probe particles would theoretically be able to penetrate 10 µm and 30 µm, respectively, over the same duration. The increase in probe particle penetration suggests exposure of mucus to MAP may similarly enhance penetration by pathogens or other foreign particles with muco-inert surfaces (e.g., Norwalk virus and human papilloma virus [27]), which may significantly increase risk of infection or toxicity. Nevertheless, the window of opportunity for harmful particles to penetrate the mucus barrier is likely limited, since MAP-exposed mucus is likely to be cleared by natural mucus clearance mechanisms as the mucus blanket is continuously renewed, on the order of minutes to hours depending on mucosal site [4].
The simulation consists of ∼20,000 particles undergoing random diffusion with diffusivities equal to the experimentally measured diffusivities of probe particles in either condition. The % of particles able to reach the opposite side of the mucus layer is reported.
Bulk rheology of MAP-exposed mucus
Rapid mucus clearance is critical to the effectiveness of the mucus barrier. Since proper mucus clearance depends strongly on the bulk rheological properties of the mucus gel [37], we also investigated whether MAP may alter the bulk viscoelasticity of mucus. We pooled mucus from several donors and performed strain-controlled cone and plate rheometric measurements on native and MAP-exposed samples sequentially. The bulk viscous and elastic moduli were quantified by applying a small, fixed-amplitude oscillatory stress at specified frequencies. Both the viscous and elastic moduli changed minimally upon MAP exposure (Figure 5), decreasing on average by ∼4% and 2%, respectively. The phase angle (δ) for both native and MAP-exposed samples was ∼14°, indicating that mucus remained a viscoelastic solid (δ = 0° indicates a Hookean solid, δ = 90° indicates a purely viscous fluid and 0°<δ<90° indicates a viscoelastic material). These values are comparable to those we have measured before for fresh mucus [18]. Thus, although MAP may alter mucus microstructure substantially, the bulk rheology of mucus appeared unperturbed.
Classical polymer physics models [18], [38]–[40] would predict a 35–65% decrease in the bulk elastic modulus as pore size increases from 380 nm to 470 nm, in contrast to the ∼2% decrease observed here. However, the essentially unchanged bulk rheology of mucus upon MAP exposure is in agreement with our previous finding that the microstructure of mucus can be selectively altered without perturbing bulk rheology [18]. In that work, we showed that treatment of mucus with a non-ionic detergent disrupted adhesive interactions between mucins, reducing natural mucin bundling and, hence, pore sizes. We postulated that the resulting increase in the number of entanglements between mucins (expected to increase bulk viscoelasticity) likely offset the decrease in adhesive interactions (expected to decrease bulk viscoelasticity), resulting in no detectable change in the bulk rheology. Similarly, an increase in mucin fiber bundling upon MAP exposure here may result in a decrease in the number of entanglements under shear, as well as a simultaneous increase in adhesive interactions between mucins. These two opposing effects may balance each other to the extent that no change in bulk rheology was detected even with a sensitive strain-controlled rheometer. Based on visual observation of the fluorescently-labeled particles, MAP appeared to mix uniformly into the entire mucus sample; thus, it is unlikely that non-uniformly distributed MAP caused only local changes to mucus microstructure that were not detectable by bulk rheological measurements. Our results suggest proper mucus clearance, which depends critically on the bulk rheological properties of mucus, may not be directly impaired by exposure to MAP, even at relatively high exposure levels. However, it should be noted that toxic nanoparticles that bypass the mucus barrier may cause toxicity to underlying epithelial cells, which may lead to mucus hypersecretion, altered mucus bulk rheology and poor mucus clearance, similar to the pathogenesis of inflammatory lung diseases [41]–[44].
Implications for drug delivery applications
Finally, our results may offer a unique approach for potentially enhancing drug or nucleic acid delivery to mucosal surfaces. Large drug carriers are preferred for improved drug loading and release kinetics, but even those engineered to have muco-inert surfaces are markedly slowed by steric obstruction from the mucus mesh, particularly in thicker secretions like cystic fibrosis sputum [15]. The use of biodegradable and biocompatible MAP that transiently enlarge mucus pores may significantly enhance the penetration of large drug carriers through mucus. These 'sacrificial' mucoadhesive nanoparticles may be employed prior to administration of the drug carriers. As mucus is continuously secreted, MAP will be displaced over time; however, mucus clearance in the vagina and other mucosal surfaces occurs on the order of several hours [4], providing sufficient time for drug carriers to penetrate MAP-altered mucus. In pilot studies, exposure of mucus to a high concentration of MAP did not cause existing MAP that were already immobilized to become mobile. This suggests that, in the absence of exposure to exogenous pathogens, the use of 'sacrificial' nanoparticles would not increase the risk of infection by already-present pathogens while facilitating sustained and/or targeted drug delivery to mucosal surfaces.
In practice, the amount of mixing of MAP into mucus that occurs during particle administration may be limited, depending on the mucosal surface of interest. For example, little to no mixing is expected in the respiratory tract, where aerosolized MAP would deposit and remain trapped on the surface of the mucus layer. In contrast, some mixing may occur in the cervicovaginal tract, where intra-abdominal pressure generates squeezing flows that can cause mixing of MAP into the mucus layer. Nevertheless, even if MAP are distributed only in the outermost, luminal mucus layer, they may still provide therapeutic benefits by sufficiently enlarging pores to enhance the penetration of drug carriers into deeper mucus layers. This is of particular significance at mucosal surfaces like the vagina, where secreted mucins form two distinct layers, a quickly cleared luminal mucus layer and a more slowly cleared adherent mucus layer adjacent to the epithelium. By reaching the slowly cleared adherent mucus layer, drug carriers may be retained at the mucosal surface in close proximity to target cells for prolonged periods of time, leading to enhanced efficacy.
In summary, we have characterized the effect of synthetic particles on the microstructure of native mucus gel, and demonstrated that both particle concentration and surface chemistry play a role in whether the particles disrupt the mucus barrier. High concentrations of MAP can increase pore sizes in mucus through the crosslinking or bundling of mucin fibers, allowing greater penetration of foreign particles across mucus. While we used CVM here, lung and other human mucus secretions have generally similar bulk rheological properties and compositions as CVM, with 2–5% w/w mucins (predominantly of the MUC5B mucin type [45]) and 90–98% w/w water [4], [33]. Thus, our findings may also extend to these other types of human mucus. These results highlight the importance of understanding nanoparticle-mucus interactions and their implications for nanotoxicology, as well as for biomedical applications.
Supporting Information
Video S1.
Transport of 1 µm muco-inert (PEG-coated) probe particles in human cervicovaginal mucus samples treated with 3% v/v saline, over the course of 20 s. The trajectories of the probe particles are strongly hindered.
Video S2.
Transport of 1 µm muco-inert (PEG-coated) probe particles in human cervicovaginal mucus samples treated with 3% v/v mucoadhesive particles (MAP; 0.24% w/v final concentration), over the course of 20 s. The trajectories of the probe particles are much more diffusive than in the same mucus samples treated with saline.
Author Contributions
Conceived and designed the experiments: YYW SL RC JH. Performed the experiments: YYW. Analyzed the data: YYW SL C. So C. Schneider. Wrote the paper: YYW SL RC JH.
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- 17. Wang YY, Lai SK, Suk JS, Pace A, Cone R, et al. (2008) Addressing the PEG mucoadhesivity paradox to engineer nanoparticles that "slip" through the human mucus barrier. Angew Chem Int Ed Engl 47: 9726–9729.YY WangSK LaiJS SukA. PaceR. Cone2008Addressing the PEG mucoadhesivity paradox to engineer nanoparticles that "slip" through the human mucus barrier.Angew Chem Int Ed Engl4797269729
- 18. Lai SK, Wang YY, Cone R, Wirtz D, Hanes J (2009) Altering mucus rheology to "solidify" human mucus at the nanoscale. PLoS ONE 4: e4294.SK LaiYY WangR. ConeD. WirtzJ. Hanes2009Altering mucus rheology to "solidify" human mucus at the nanoscale.PLoS ONE4e4294
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- 23. Apgar J, Tseng Y, Fedorov E, Herwig MB, Almo SC, et al. (2000) Multiple-particle tracking measurements of heterogeneities in solutions of actin filaments and actin bundles. Biophys J 79: 1095–1106.J. ApgarY. TsengE. FedorovMB HerwigSC Almo2000Multiple-particle tracking measurements of heterogeneities in solutions of actin filaments and actin bundles.Biophys J7910951106
- 24. Valentine MT, Perlman ZE, Gardel ML, Shin JH, Matsudaira P, et al. (2004) Colloid surface chemistry critically affects multiple particle tracking measurements of biomaterials. Biophys J 86: 4004–4014.MT ValentineZE PerlmanML GardelJH ShinP. Matsudaira2004Colloid surface chemistry critically affects multiple particle tracking measurements of biomaterials.Biophys J8640044014
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- 27. Olmsted SS, Padgett JL, Yudin AI, Whaley KJ, Moench TR, et al. (2001) Diffusion of macromolecules and virus-like particles in human cervical mucus. Biophys J 81: 1930–1937.SS OlmstedJL PadgettAI YudinKJ WhaleyTR Moench2001Diffusion of macromolecules and virus-like particles in human cervical mucus.Biophys J8119301937
- 28. Shen H, Hu Y, Saltzman WM (2006) DNA diffusion in mucus: effect of size, topology of DNAs, and transfection reagents. Biophys J 91: 639–644.H. ShenY. HuWM Saltzman2006DNA diffusion in mucus: effect of size, topology of DNAs, and transfection reagents.Biophys J91639644
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- 40. Palmer A, Xu J, Kuo SC, Wirtz D (1999) Diffusing wave spectroscopy microrheology of actin filament networks. Biophys J 76: 1063–1071.A. PalmerJ. XuSC KuoD. Wirtz1999Diffusing wave spectroscopy microrheology of actin filament networks.Biophys J7610631071
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Spiedis Recipe
This is our favorite cookout dish, and it's very good served with a salad. This recipe originated here in my hometown in the 1930s. Our meat preference for speidis is venison, but we use chicken and beef when it's not available. —Gertrude Skinner, Binghamton, New York
- 1 cup vegetable oil
- 2/3 cup cider vinegar
- 2 tablespoons Worcestershire sauce
- 1/2 medium onion, finely chopped
- 1/2 teaspoon salt
- 1/2 teaspoon sugar
- 1/2 teaspoon dried basil
- 1/2 teaspoon dried marjoram
- 1/2 teaspoon dried rosemary, crushed
- 2-1/2 pounds boneless lean pork, beef, lamb, venison, chicken or turkey, cut into 1-1/2- to 2-inch cubes
- Italian rolls or hot dog buns
- 1. In a glass or plastic bowl, combine the first nine ingredients. Add meat and toss to coat. Cover and let marinate for 24 hours, stirring occasionally.
- 2. When ready to cook, thread meat on metal skewers and grill over hot heat until meat reaches desired doneness, about 10-15 minutes. Remove meat from skewers and serve on long Italian rolls or hot dog buns. Yield: 8 servings.
Nutritional Facts
4 ounce-weight: 205 calories, 12g fat (0 saturated fat), 42mg cholesterol, 104mg sodium, 1g carbohydrate (0 sugars, 0 fiber), 22g protein. Diabetic Exchanges: 2 lean meat, 1 fat.
Reviews for Spiedis
Average Rating
Reviewed Mar. 5, 2014
© 2017 RDA Enthusiast Brands, LLC

subset: commoncrawl


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While the main star Maura Tierney is undergoing medical evaluation, NBC postpones the plan to air 'Parenthood' this Fall.
NBC is rooting for its star to heal first before coming back to work. "Parenthood", one of the new series that the Peacock offers this Fall, has been shelved for midseason premiere in order to give Maura Tierney time to recuperate from an undisclosed illness.
"Start of production on NBC Universal's new series 'Parenthood' has been postponed for eight weeks due to medical evaluation that valued cast member Maura Tierney is undergoing," NBC said in a statement released to THR. "The top priority for NBC, Universal Media Studios and Imagine Television is the well-being of Ms. Tierney. We are unable to release further details and ask that you respect her privacy at this time."
In "Parenthood", Tierney plays Sarah who is one of the four Braverman siblings. She is a financially-challenged single mother who returns home to her parents and siblings in Berkeley, Calif.
NBC will air Michelle Trachtenberg-starring medical drama "Mercy" as replacement. It portrays the lives of the staff at Mercy Hospital as seen through the eyes of those who know it best - its nurses.
Fall Promo of "Parenthood":


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Last week was pretty miserable, in terms of my picks, as I went an abysmal 1-2-1. Although I didn't predict the fire heard seen the world, I did pick the score of Somers' win over Arlington dead on, so there was that.
Lets hope for a better performance this week.
With the end of the season just around the corner, the theme this week is league title battles. All five games are between two of the top teams in their respective leagues.
Eastchester at Bronxville, 4:30 p.m.
Pick: Eastchester, 2-1. The Eagles won the first meeting between these rivals a few weeks ago. Eastchester has continued to roll while the Broncos stumbled again last week with a loss to Pelham. The Eagles get a huge road win as they all but secure a league title.
Clarkstown South at North Rockland, 4:30 p.m.
Pick: North Rockland, 2-1. North Rockland won the first game of the series in a defensive battle on the road. I think the Red Raiders' offense opens up a bit more, and they come away with a win in their friendly confines.
Haldane at Panas, 6:30 p.m.
Pick: Panas, 2-1. I picked Haldane to win at Panas last week. You all know what happened. While I still think the Blue Devils are the top squad in Class C, the Panthers have really impressed me this season. Call me a flip flopper, if you must, but I'm going with the Panthers at home.
Rye at Byram Hills, 4:30 p.m.
Pick: Byram Hills, 3-2. The Bobcats were able to come away with a 2-1 win at Rye on Saturday night. It seems like Byram is firing on all cylinders when you consider the win came a little over 24 hours after a 5-1 win against Cornwall, a Class A state semifinalist last season. I can't think of a reason to go against Byram at home.
Tappan Zee at Albertus Magnus, 8 p.m.
Pick: Tappan Zee, 2-1, 2 OT. These two played to a 2-2 tie in their first meeting. It seems like Tappan Zee keeps coming up on the short end of the stick in all these close games. Somethings gotta give eventually. I'll take the Dutchies on the road.


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Vermont tech entrepreneurs
show their stuff…
InnoVaTe is a tech-centric series that highlights some of Vermont's most innovative and creative businesses.
Vermont is a state that supports a culture of innovation, generating creative thinkers and doers around every corner – and the technology realm is no exception.
We're pleased to bring you InnoVaTe, a series that explores and celebrates Vermont's innovator culture. It's hosted by nationally recognized social media professional Victoria Taylor, whose engaging, personal style opens up thought-provoking conversations with some forward-thinking Vermont entrepreneurs – making invention and innovation approachable and exciting, and highlighting Vermont's creative culture.
Through InnoVaTe, you'll get to know some truly original Vermonters – like these folks who are pushing high-tech in new directions and putting Vermont business and creativity on the map.
Victoria Taylor has hosted more than 2,000 "Ask Me Anything" interviews on reddit.com during her time as the site's director of communications. Now at WeWork, Victoria has been recognized by LinkedIn as one of the 10 top professionals under age 35 who are helping shift the digital, cultural and business landscape.
InnoVaTe is supported in part by...
Additional funding for provided by...
New episodes
Changing how humans and robotics relate to one another, this progressive company based in Richmond, Vermont is honing technology that allows sophisticated interfacing between man and machine.
On the cutting edge of data analytics, this Vermont electric company has developed a sophisticated system of weather prediction.
John Churchman
The power of social media and crowdfunding have helped make artist and photographer John Churchman's adorable animals like Sweetpea the Sheep a shining example of how to engage an audience, get published and make one's dreams come true.
With the popular saying "sitting is the new smoking," it's widely accepted that a sedentary lifestyle contributes to many health problems. As a solution, UVM professor and inventor Dr. Turner Olser presents his innovative chair, the QOR360, which utilizes a unique shape to promote"active sitting."
The future of efficient homes, VERMOD's goal is to create modular housing units built in Vermont that combine beauty and functionality rolled into a cost-effective package that appreciates in value over time.
Changing the world through video games, this revolutionary company with Marguerite Dibble at the helm uses video games and game concepts to help support clients seeking to educate or enact social and behavioral change through the dynamics of gaming.
James Kochalka
James Kochalka is Vermont's first Cartoon Laureate and is shaping popular culture through art, music and video games.
To learn more about James follow him on Twitter @the_kochalka
Previous episodes
SeedSheets - produces a ready-to-plant prefab garden kit complete with organic seeds and soil.
Logic Supply
Logic Supply - whose cutting-edge technology is bringing hi-tech advancements to a diverse array of industries.
ThinkMD - whose revolutionary platform for sharing medical information is expanding global access to healthcare.
Yonder - a new social media app that allows recreation enthusiasts to connect, share and meet up.
Generator - Burlington's new maker-space where art and innovative technology meet in a vibrant community.
Vermont Tech Jam
Vermont Tech Jam - a synergistic Vermont job fair and technology conference that gathers together the state's leading technological innovators.
Follow us for more innovative fun!
More fun…
Guster's Ryan Miller visits high-functioning weirdos in Vermont.
Qulture explores Montreal hotspots!
Vermont PBS educates, informs, entertains and inspires Vermonters to be lifelong learners and engaged in their community.
Vermont PBS Brings Ken Burns' and Lynn Novick's Epic Series The Vietnam War to Broadcast and to Communities throughout Vermont. Learn more and register for free screenings today!
A state-wide event exploring what we have learned over the last decade about opiate addiction, what has changed, what treatments and preventions are having real impact, and what we can do better. Learn more and register!
Montréal's 375th celebration combines history, heritage, art, and culture for a year of events. We offer a taste of Montréal in our Montréal Minutes and MTL375 interviews and travels in the city.
Watch our Vermont PBS Kids' 24/7 channel. Enjoy the benefits of joining our Kids' Club! Explore our many free videos, games, and resources for educators and parents!
Upcoming and featured!
People talk about Vermont PBS!
Vermontpbs.org proudly supported in part by...


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The Lanvin designer on his limited-edition makeup collection, Lancôme Show.
We're already Lancôme devotees, especially when it comes to mascara, but leave it to fashion visionary and Lanvin designer Alber Elbaz to make us covet something we've been using for years. He's reimagined the packaging for Hypnôse Doll Lashes, Hypnôse Drama, Hypnôse Star, and Definicils with adorable (but chic!) graphics. And he didn't stop there: He also restyled the Color Design 5 Shadow & Liner Palette. Here, Elbaz talks about this foray into beauty.
What do you think makes a woman stand out from the crowd?
Alber Elbaz: It's her true self. When people think about makeup they often think about disguising or hiding the truth. But true style—and this holds true for Lancôme too—is about bringing a woman's individuality to the surface. Not hiding it or playing a role, but truly revealing it. That's how I see my profession: veiling the body to unveil the person.
Did you work on this collection in the same way you work on a show?
AE: First of all, you have to find the story you want to tell. That's what this business is all about, so the process has to start there. I wanted to think about the stories you can tell with mascaras: a story of eyes, and their shape. A musician tells a story through the notes he plays, a painter transforms his canvas, and a fashion designer works with shape and color. So I took the four mascaras and built a story around them, with polka dots for Hypnôse Drama, stars for their namesake Hypnôse Star, hearts for Hypnôse Doll Lashes, and for the fourth mascara, Definicils itself, I took the legendary Lancôme rose and mixed it up with eyes. I wanted to maintain a whimsical, feel-good style. Mascaras tend to come in just gold and black. I wanted to create more of a personal touch. When you apply mascara you're almost touching your eyes, so it's a very intimate moment. It needed something truly personal that you can make your own.
It's quite rare to find joy and humor in the world of luxury…
AE: Yes, humor isn't generally in phase with luxury. But your key aim has to be to inspire emotion. A luxury boutique where everything is perfect and the staff look like models is rather like a pharmacy. You can make people feel with smell, taste, and above all with humor. I need to have my dreams and imagination inspired if I'm to actually buy a product. True luxury is about making people laugh and smile, saying, "I want that!" That's how luxury becomes democratic. It's not about creating an army of clones that eat the same things in the same places and share the same aspirations: It's about giving people a reason to feel that bit happier.


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Gloria Estefan "Miss Little Havana" Track-by-Track
www.billboard.com Gloria Estefan discusses each track of her new album "Miss Little Havana"
Video Rating: 4 / 5
www.billboard.com Gloria Estefan discusses each track of her new album "Miss Little Havana"
Video Rating: 4 / 5
her tits brought me here
go gloria !
Brillaint ! love it , love the Albumx
She looks good for mid-50s.
Im sorry but she kept it in lower key because gloria cant hit most of the notes she used to and she never really had a high range still like her
Hi there everybody! Come and enjoy the Gloria Estefan Official Fan TV on Youtube!!! (G's peeps)
is the best….
Amazing album ¡¡¡¡ fresh & sensual ¡¡¡ i love MLV ¡¡
she is back,u lioke or not,eat it
She supports a scumbag like Perez Hilton. Enough said.
This old lady's career ended in the 80s.
The rhythm has always been there and the rhythm is you!
Amen to that! I couldn't agree more!
ella es una leyenda… ya nadie la saca de ser una diva.
Te amo Gloria, eres mi Diva!
No doubt Gloria is the Latin Queen. She is brilliant. There would be no Shakira, Ricky Martin, Enrique without her.
am about to turn 20
You must be very young… LOL
@PaperP1ate She is really good
i've never heard of her before


text:
ED Magedson – Founder
Ripoff Report
LowesLOWE'S HOME IMPROVEMENT WAREHOUSE OF SANFORD Sanford, North Carolina U.S.A.
Does your business have a bad reputation?
Fix it the right way.
Corporate Advocacy Program™
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Arbitration Program
This report was posted on Ripoff Report on 09/30/2008 06:01 PM and is a permanent record located here: http://www.ripoffreport.com/reports/lowes/sanford-north-carolina/lowes-home-improvement-warehouse-of-sanford-after-spending-over-5000-cash-dollars-at-one-t-377445. The posting time indicated is Arizona local time. Arizona does not observe daylight savings so the post time may be Mountain or Pacific depending on the time of year.
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text:
Narrated by one of the funniest, wisest literary observers of all time this is a brilliant, charming and heartwarming novel about illicit love, sewing, blackouts and Belfast.
In a minute she'd beg me to do anything I could to save the garment. That's what they always did. Begged and pleaded. There was usually a lover involved, and a cheated-upon spouse. I, as the mender, would be saving their life. People had actually said I was worth my weight in gold . . .
But it wasn't my skill the clients were grateful for. No. It was my collusion. What lies are worth: their weight in gold.
You'd think that mending clothes would be an uneventful, uncomplicated occupation. No drama, no unnecessary explanations, no personal involvement. But people love to talk, and as they make their excuses to GoGo Sligo, of Megan Sligo Mending and Alterations, they reveal the holes in their stories as well. It doesn't take long for GoGo to get to the truth behind the rips and tears they've brought her to fix.
As GoGo listens and sews, she realises she is also helping her clients cheat and lie to their husbands and wives. She's covering their tracks so they won't be found out.
A five-week blackout brings the city to its knees, and a drama to her doorstep. A lover, a wife, and finally the cheating husband all come to claim a vintage Irish costume that GoGo's been mending. She doesn't want to like the guilty husband, but can't resist being drawn into the enticing web of his deceit, and then into his story of heartbreak and death on the streets of Belfast.
To keep him coming back to the blacked-out house and to prolong the telling of his family's story, GoGo pretends the costume isn't finished. As she makes him return to her, day after day, it becomes clear that another kind of spell is being woven, and GoGo must face the truth about herself and her own life and marriage.
An intoxicating, entrancing, gripping novel of illicit love, passion and embroidery, told in an inimitable voice by a brilliant writer.
Read Caroline Baum's Review:
As Australians, we know too little about contemporary writing from New Zealand. It's our loss. So it's a pleasure to discover this refreshingly original new voice from across the Tasman. Megan Sligo is an eccentric mender of garments. She can repair any tear. When a power outage darkens her home town of Auckland, her work is thrown into disarray. And when a woman brings a torn gorgeous beaded dress to her, things get complicated. It seems that several people want the dress… What makes this so captivating is the narrator's voice: warm yet sly, playful, (she makes jokes about post modern poets) and eccentric. There's a lot going on here, stitched into the narrative like hidden seams and concealed pockets, but above all this idiosyncratic debut is a contemporary reminder that storytelling is the most timeless seduction of all.
About the Author
Anne Kennedy's first novel was an experimental work titled 100 Traditional Smiles (1988). She then went on to write Musica Ficta (1993) and A Boy and His Uncle (1998). Her most recent work was to write the screenplay for The Monkey's Mask, a highly successful film based on Dorothy Porter's verse novel. Anne is also a well known and award winning poet. She lives in Auckland and has also lived in Ireland and Hawaii.
REVIEW SNAPSHOT®by PowerReviews
Best Uses
Reviewed by 3 customers
Best Uses
Comments about The Last Days of the National Costume:
Somewhat weird tale set during Auckland's major power blackout, about a mender of clothes who turns an heirloom Irish dancing costume into a prolonged personal saga. Some knowledge of inner Auckland helpful in appreciating the setting of the drama.
Best Uses
Comments about The Last Days of the National Costume:
I loved this story. Author has "stitched" the characters and their stories together with her own as she works.
Don't see the need for colloquial idiom to include four-letter words, but overall I was rapt in THAT DRESS.
Best Uses
Comments about The Last Days of the National Costume:
A novel for those interested in variety of story lines. This one is quirky and amazingly imaginative
Audience: General
Format: Paperback
Language: English
Number Of Pages: 400
Published: 29th June 2013
Publisher: Allen & Unwin
Country of Publication: AU
Dimensions (cm): 23.4 x 15.3 x 3.1
Weight (kg): 0.5
Edition Number: 1


text:
We confirm close to 50% of member vacations before inventory is released on this website, because we match our member vacation requests prior to stock being uploaded onto our website.Our team's Number 1 focus is getting you on vacation - if the vacation you are looking for is not available when you search, simply place a request in our system and our team will work hard to source a suitable option for you.
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Fantastic skiing is close by as are rejuvenating natural hot springs. Lake Louise, just north of Banff, is known as 'the jewel of the Rockies' and is a must visit. Inhale crisp fresh air as you absorb the beauty of this natural wonderland.
Banff Rocky Mountain Resort is nestled at the base of picturesque Rundle and Cascade Mountains in Banff National Park just over two miles from downtown Banff.
There's recreation for all seasons in this natural paradise including skiing, dog sledding, golf, hiking, biking, canoeing and excellent wildlife viewing. Banff is renowned for its dining excellence.
The resort offers easy access to many winter activities, such as downhill skiing at Sunshine Village, Mount Norquay and Lake Louise, plus cross country skiing, hele-skiing and sleigh rides.
In the summer, enjoy heli-skiing, hot springs, fishing, golfing, horseback riding and boating. The Sulphur Mountain Gondola Lift is one of the areas many attractions.
The resort is well appointed with Owners Lounge/Club House, Restaurant/Bistro, indoor pool, spa pool, fitness club, squash and tennis courts.
There are full concierge services for planning and booking excursions, and a complimentary shuttle to Banff town center.
The resort has a fully developed activities schedule with guided hikes, nature walks, showshoeing or tobogganing, broomball, walleyball, beach volleyball, horsehoes, basketball and Frisbee golf.
The resort features a restaurant, Pepper's Bar and Deli offering lively cocktails, coffees and deli delights. Eat in and enjoy a game of darts or billiards, or grab a delicious snack to go.
Banff Rocky Mountain Resort offers 171 guest rooms from standard suites to condominium style suites with kitchens. All suites feature comfortable living room areas with gas or wood-burning fireplaces, cable television, hairdryers and data port phones. Unfortunately the Resort is not handicap accessible- there are stairs.
If the spectacular scenery and many activities offered by Banff and Lake Louise appeal then Banff Rocky Mountain Resort is your ticket to fun.
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text:
The Hall of Honor pays tribute to those who have been the most generous supporters of the School-as evidenced by their outstanding leadership and dedicated service in building Johnson's strength.
These alumni and friends are honored for their devotion and commitment to the Samuel Curtis Johnson Graduate School of Management.
For professionals seeking leadership development and management expertise
Professional certifications or advanced degrees may qualify for this accelerated MBA program.
A one-year program for students who want to advance their careers by mastering the field of accounting
Six options to concurrently earn two top-ranked Cornell degrees
For professionals who desire leadership opportunities in the digital economy
Our newest program: a dual degree MBA program offered in China
A 17-month program offered in more than 20 cities
Offered on weekends in the Metro NY area
Advancing leadership and innovation in healthcare delivery
Our top-tier journal on organizational studies.


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FOUR men involved in a plot to blackmail a businessman – which culminated in the man's car being torched – have been sentenced.
The gang was involved in a two-month conspiracy to blackmail the victim over a debt his business partner owed to Tommy Roye, 39, of Flixton Road, Urmston.
Following a trial, Roye was convicted of blackmail, conspiracy to commit arson and threatening to destroy property.
Jamie Keegan, 31, of Epsom Avenue, Sale, and Marcel Lauder, 44, of Morningside Close, Derby pleaded guilty to conspiracy to commit arson and blackmail at an earlier hearing.
Ashley Guishard, 39, of Totnes Road, Ashton-on-Mersey, was convicted of blackmail following a trial.
Today, February 13, Roye was sentenced to six years, Guishard to three years, plus a further four years for an unrelated possession of Class A drugs with intent to supply charge, Lauder to 28 months (which includes four months for production of Class B drugs) and Keegan to 24 months at Manchester Crown Court, Crown Square.
The victim was involved in the running of a Cheshire nightclub with his business partner, who owed up to £30,000 to various people, one of whom was Roye.
These two men had a tense altercation in a Stretford pub on August 13 2012, which resulted in the man in debt calling the victim and asking for help.
Concerned, the victim drove to the pub and spoke to his partner, who said he feared for his life.
In desperation, the victim told Roye he would pay £5,000 towards settling the debt in order to buy his business partner time to raise the money himself.
The victim was assured by his partner that he himself would not need to pay Roye any money.
However, just before 11pm on October 1 2012, as the victim and a friend left the Lord Nelson pub in Urmston, they were ambushed by Roye and Guishard.
Roye threatened the victim's life, so he gave Roye the keys to his Audi in the hope it would placate him.
Roye told the victim that if he went to the police his car would be torched.
The car was eventually returned.
But on October 10, Roye again bombarded the victim with texts demanding payment.
When the victim said he would call the police, Rowe replied: "POLICE, CALL THE FIRE SERVICE ASWEL (sic)."
Roye recruited Keegan and Lauder, who went to the victim's home at around 8pm on October 12 and torched the victim's people carrier.
Roye and Guishard were arrested the next day, and during a search of Roye's home, officers seized two ornamental Samurai swords and a machete.
Det Sgt Patrick Goodrich said: "This case involved a totally innocent and legitimate businessman who unwittingly became embroiled in a blackmail plot driven by a gang of violent criminals.
"All he wanted to do was help out a colleague who had got himself into debt, and for his troubles endured two months of hell at the hands of an extremely intimidating and aggressive band of criminals.
"That they went so far as to torch this man's car as it was parked on his driveway, bearing in mind this man has a wife and daughter, shows how dangerous these men were and you can only imagine the anguish this man suffered for those two months, being bombarded with violent threats simply for trying to help a colleague.
"I have no doubt whatsoever that the streets are a lot safer now that these four men are behind bars. I would also like to praise the tenacity of the victim himself who hung in there despite the hell he was subjected to and helped us to convict these men.
"Organised crime groups like these cause real misery and hardship for law-abiding and innocent members of our communities.
"However, in Trafford we have been targeting organised crime groups at all levels for the last two years and have achieved some fantastic results by working in collaboration with a host of partner agencies.
"We are tackling these groups with every tactic open to us, taking away their liberty, their homes, their cars and their cash.
"We will continue to target these organised crime groups and people in Trafford can help us by reporting any criminal activity they see in their neighbourhood."


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Best Reasons to Own Cooking Books
Best Reasons to Own Cooking Books from Overstock.com. Even with the internet, traditional cookbooks are still useful for casual and serious cooks alike.
ITEM#: 10870846Advanced Bread & Pastry has a unique approach to providing advanced level concepts, techniques and formulas to those aspiring to be professional bakers and professional pastry chefs. It is the only available resource covering complex bread making, viennoiserie and pastry into one book. Exquisite photographs are throughout to further inspire learners and professionals of the unlimited potential of the craft. Advanced Bread and Pastry provides in depth information and troubleshooting strategies for addressing the complex techniques of the advanced level of bread and pastry arts.
|COOKING / Methods / Baking
|Publisher Date
|2008-04-04 00:00:00.0
|Delmar Pub
|Copyright Year
|11.0 in
|8.75 in
|1.75 in
|Unit weight
|7.05 lb
|A Professional Approach
|Edition Number
|Suas, Michel
|Country of Origin
|United States
Best Reasons to Own Cooking Books from Overstock.com. Even with the internet, traditional cookbooks are still useful for casual and serious cooks alike.
Learn about good books that will empower young girls, tweens and teens. Shop books for girls on Overstock.com.
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Twilight: Beyond the Novels from Overstock.com. Our guides provide customers with information about Twilight, the novel.
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10 Fun Facts about Disney from Overstock.com. Disney has a long, exciting history, and here are some fun and lesser known facts.
Tips on Selecting the Perfect Entertainment Gift from Overstock.com. Our guides provide customers with tips on selecting the perfect entertainment gift.
This item will be delivered to you via USPS Trackable Media Mail or UPS Mail Innovations and will take from 2 days to 3 weeks from the time the item leaves our warehouse. *
This date is subject to change. In order to assure you receive Overstock.com's low price on this item, your credit card will be charged upon order placement. The item will ship immediately upon release.
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Advanced Bread and Pastry: A Professional Approach (Hardcover)
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Atlanta Ballet Adds SWAN LAKE to Season
For its 12-13 season finale, Atlanta Ballet explores the theme of love through both classic and innovative choreography. Highlighting the athleticism and passion of Atlanta Ballet's dancers, these performances find new meaning in the world's greatest love stories. Love Stories will feature the Atlanta Ballet premiere of "Requiem for a Rose" by choreographer Annabelle Lopez Ochoa and the return of "Prayer of Touch" by Atlanta Ballet's new resident choreographer Helen Pickett. The program will also feature the White Swan pas de deux from choreographer Matthew Bourne's revolutionary "Swan Lake" - regarded as a modern day classic since its world premiere in 1995.
Atlanta Ballet Premiere of Annabelle Lopez Ochoa's "Requiem for a Rose" - Music by Franz Schubert
In the tradition of introducing its audiences to the pioneers and trailblazers in dance, Atlanta Ballet brings another strong choreographic voice to its stage with the premiere of Annabelle Lopez Ochoa's "Requiem for a Rose."
Set to an achingly romantic and longing adagio by Schubert, "Requiem for a Rose" is a zealous work that explores the transient nature of love. The dynamic piece envisions 12 dancers as a bouquet of roses, with bold and fluid movement hinging on a single heartbeat. Dance View Times described the ballet as a piece filled with "tender" moments that is "deftly and honestly crafted."
The Belgian-Colombian choreographer began her dance making career in 2003 after 12 years as a professional dancer for Holland's Scapino Ballet. In that same year she was hailed as a "rising star of the Dutch dance scene" (NRC newspaper) and, only 7 years later, the Temecula Performing Arts Examiner wrote: "Ochoa is truly a masterful choreographer with an edge for what dance can and should be in this constantly changing industry."


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Software Developer - C++
Software Developer - C++
Nov 7, 2012 at 11:51pm UTC
Location: Hayward, CA
Job Description:
Interactive Data has an immediate opening for world class developer to join a technology team that builds software interacting with major exchanges around the globe! In this role, you will write and tune software for class-leading performance, design and implement improvements to live market data systems, and monitor and analyze the performance of the system.
Perform real-time and multi-threaded programming in support of company's online products, with emphasis on researching, creating and maintaining high-performance, object-oriented, light-weight and highly efficient commercial enterprise applications (Linux, C++)
Be responsible for all phases of software development, including analyses and generation of system and component requirements, schedules, design, implementation, integration, unit and acceptance testing of new and enhanced applications and features
Work with a team of highly skilled software developers in a fast-paced, customer-driven environment
Required Experience:
Bachelor's Degree in Computer Science, Electrical Engineering, or related degree, or equivalent work experience
7+ years of professional C++ programming experience, including server side, multi-threaded programming for high-performance real-time applications
Exceptional programming skills, a penchant for collaboration, and an interest in working on high-performance, mission-critical systems.
Solid experience with C/C++ in linux environment
experience working with large-scale multi-threaded, realtime systems
Keen eye for writing latency sensitive code, and solid understanding of hardware architecture, algorithms, data structures, high performance software, and cutting edge IPC mechanisms.
Effectively work under strict deadlines, and be a team player with good people skills.
Experience with Linux g++, multi-threading, TCP / IP
Experience with real-time application engineering, debugging, performance analysis and tuning
Experience developing 24 x 7-available, mission-critical server applications that run in an operations center
Strong oral and written communication skills
Strong problem-solving skills
Ability to be independent and highly self-motivated
Ability to set and achieve goals, requiring minimal supervision
Experience writing or maintaining applications that handle securities exchange feed data (SIAC, NYSE Euronext, NASDAQ OMX, CME, ARCA, Eurex, LME etc.)
Familiarity with applications that handle various securities types (equities, options, futures, fixed income)
Experience with Java programming (server-side / high-performance).
Last edited on
Nov 7, 2012 at 11:53pm UTC
Nov 8, 2012 at 9:22am UTC
Hi, I have done B.E Computer Engineering. I have 2.9 year of Exp in C++ Windows/Linux. Also i have worked in multi-threadeding, Financial Trading Algorithm development and Socket Programming also. I am interested for Job. Let me know if anything for me..
Skype: mota.pratik
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Northern Tier Energy (NTI)
Speaking of those end-users, refiner Northern Tier Energy LP (NTI) might be an interesting high yielding pick.
The key for NTI is that it is a variable-distribution master limited partnership. Essentially, NTI plans on distributing all of its cash flows back to investors as dividends. That has resulted in some huge swings in its underlying yield. Just a few quarters ago, NTI was yielding a monster 19%.
However, as crack spreads began to dwindle, so did NTI's yield. Today, Northern Tier pays out 5.1% in distributions.
Yet, NTI's refinery is located in one of the best areas to take advantage of cheap WTI and Canadian crude. That means as crack spreads begin to widen — which they currently are — NTI will be able to increase its margins on the products it refines. All in all, that could send NTI's dividend higher in the future.
While it most likely won't yield 19% again anytime soon, NTI's current 5.1% dividend yield is still pretty juicy and makes the MLP an interesting choice.


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Apple's contract manufacturer in China, Foxconn, has been attacked by organizations for hiring underage workers and for generally poor working conditions. Cook said in the interview that companies like Apple have their assembly work done in China not just because of the lower salaries, but also because the manufacturing skills required to build, say, an Apple iPhone, aren't present in the U.S. anymore.
This doesn't mean that Apple will do it ourselves, but we'll be working with people and we'll be investing our money." The production being moved to the U.S. is a small part of the Cupertino based firms' overall business. Sales of the Apple iPhone and Apple iPad swamp sales of the company's Mac line of computers.
Under Cook's watch, Apple's shares have risen 43% even after Wednesday's huge decline, and the company has has released two new Apple iPhone models, two new versions of the Apple iPad and the Apple iPad mini. The CEO said it was time to bring some jobs back to the United States. While things have been going well for Apple, the executive did admit that the company made a mistake with Apple Maps. "We screwed up and we are putting the weight of the company behind correcting it," the executive said.
The interview with Tim Cook will be shown on Thursday night on Rock Center broadcast at 10pm/9pm central time on NBC.
source: NBCNews via NYPost


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LESLIE HERMELIN: firstname.lastname@example.com NOAH BETHKE: firstname.lastname@example.com
Findlay Brown is a Yorkshire born, Brooklyn based singer-songwriter and producer. He grew up in the countryside and sold his Grandfathers Beatles autographs to buy his first guitar. Findlay is a modern day troubadour. His music is melodic, gentle and intimate drawing on influences from the rural settings where he grew up. His songs are metaphoric often using nature's imagery to illustrate the inner conflict in the search for a more truthful way of living.
His first solo album Separated By the Sea (2007) Peacefrog, was a critically acclaimed release, receiving high praise such as Five Stars in the Guardian newspaper, whose music columnist hailed it as: "the most unlikely classic of the year", while the international music magazine Mojo said the album "Sets the benchmark for gorgeous troubadour folk." The single Loosing the Will to Survive (2007) earned Brown playtime on the prestigious Radio 1 in the UK with support from radio hosts Zane Lowe and Jo Whiley.
Findlay released Promised Land (2010) a 10" gold vinyl cover of Joe Smooth's House hit on the French label, Record Makers, with remixes from Pilooski and Acid Washed among others.
Love Will Find You (2010) Verve, was Findlay's highly anticipated second solo album, produced by Bernard Butler with an epic, Phil Spector wall of sound production and likened to the 50s and 60s pop crooners Roy Orbinson and Scott Walker. Soon after the US release on Verve Forecast Findlay moved to New York and appeared on The Late Show with David Letterman. He did a nationwide tour of America."Full of gorgeous melodies and lush, organic instrumentation" - LA Times, "Findlay proves he is a craftsman and a chameleon". Critic's Choice - New York Times.
Slow Light (2015) DPC Records, is Findlay Brown's 3rd solo album release.
Melancholic and heartfelt Slow Light goes beyond folk. To his fans it marks a return to the more intimate stripped down nature ofSeparated by The Sea, but with cinematic and epic elements. Co-produced with Danish producer Tobias Wilner with influences drawing from Minimalist music, Sound Tracks, African Music and classic song-writing, the sound is glacial and filmic, yet manages to remain personal.
Findlay Brown has toured and collaborated with: Bat for Lashes, Beyond the Wizard Sleeve, Lovefingers, Fionn Regan, Bert Janch, John Renbourne, Alberta Cross, Au Revoir Simone, Bernard Butler, Sleepy Jackson, Simon Lord, Richard Swift, Taylor Swift, Passenger, Duffy, Barbarossa, Paul Weller, Angus and Julia Stone, at the Warchild Benefit Concert at Brixton Academy with PetShop Boys, Lilly Allen, Brendon Benson and Keane. Blue Foundation, Shelby Lynne, Luca C and Brigante, Brendon Lynch, Drivin n Cryin, Carl Barret, Johnny Flynn and many more.
In 2013 Findlay Brown teamed up with Xander Ferreira to form the New York based dj-duo The Happy Show hosting parties bringing African groove to NYC nightlife. At their Output residency in Brooklyn and parties around the world they have created a joyous, ritualistic, dance gathering.


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We offer free, tracked UK delivery on orders over £10, with delivery starting from £1.99 for orders below this value. We also have a selection of premium shipping options available including timed slots, Saturday delivery and Collect+
All of our bicycles are fully built by our experienced and certified mechanics prior to delivery; all you will need to do is turn the handlebars and fit some pedals.
Bicycles are shipped from a different warehouse to other items and so may arrive in advance of any other items you have ordered.
View all Delivery Options
We want you to be 100% satisfied with your shopping experience at Rutland Cycling, so we offer a 90-day exchange or refund on any unused item (within the UK).
We've chosen to offer products at the very lowest prices, rather than provide a free returns service. If you do need to return an item, we provide printable postage labels and collection services at a small cost for all items - including bicycles.
See our full Returns policy.
All the convenience of buying online, without the risk
When you buy a new bike at full RRP from rutlandcycling.com, you can now benefit from the added reassurance of our Rutland 30-day test ride. Once your new bike arrives, you can ride it as your own for 30 days, and we're confident you'll love it! However, if it's not right for you, we'll exchange it for another model. (There's a nominal charge of £10 for us to collect your bike - just make sure you keep the box your bike arrived in.)
On which bikes is the 30-day test ride available?
The Rutland 30-day test ride is available on all full price, non-discounted bikes available for home delivery on rutlandcycling.com. It is not available on discounted or special offer bikes, click and collect bikes, bikes bought on finance, bikes bought through Cyclescheme, or bikes bought in store. This offer is only available on bikes delivered to mainland UK addresses.
How many times can I use the 30-day test ride?
We will allow up to two exchanges within this scheme.
Full T's & C's
Get a confidential decision online in seconds.
At Rutland Cycling, we offer Finance on orders over £389, with 0% finance up to 24 months available on selected bikes, including some sale bikes. Visit the product page to see which finance products are available.
You can apply for Finance online or in store. To apply online, add your selected items to your basket, then checkout as normal. Select Finance as your payment method, fill in the application form, then complete your order.
See our Finance page for more information.
During June, we're offering 10% Rutland Rewards on 2017 bikes. Earn your cashback instantly and spend it on anything, online & in store, over the next 12 months.
For a limited time, save up to £50 using codes:
use code: 'SAVE5' on £50
use code: 'SAVE15' on £150
use code: 'SAVE25' on £250
use code: 'SAVE50' on £500
We have found 378 Items in 'Road Bikes'
For the purest cycling experience, a fast and furious road bike takes some beating. In the cockpit of a quality road bike, you'll feel every twist and turn and experience the high-speed thrill of the open road. So whether you're a Tour de France junkie, or like to commute with a little more pace (and a lot more style), then a road bike is just what you're looking for.
Road bikes are easy to spot thanks to their skinny tyres, drop handlebars and aerodynamic appearance. Their frames are typically made from two materials: aluminium on road bikes under £1000 or on a sturdy commuter, and carbon for higher-end lightweight speed machines.
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My husband & I are 60 yr. old & in average physical condition or better. We need early departure off the Spirit in New Orleans. We understand we will be carrying our luggage off the ship. It is rolling luggage & not overly packed. Will we encounter steps? Can anyone tell us what to expect @ the port there when we get off the ship? email@example.com
Hi sommar, welcome to Cruise-Chat! Congratulations on your up-coming cruise! The only stairs you will encounter will be if the elevators are jammed, then you will end up walking down the stairs. If you're doing the self-debarkation, get an early start to avoid the elevators being crammed. You have all these people, with all their luggage, trying to get off at once. Many times, we've just taken the stairs. The terminal will have the escalators and elevators, they too will be crowded, but doable.
Best wishes for a fantastic cruise!
Just remember, the ships' crew can't help you with your luggage off the ship...and yes, there will be many people trying to get off early too....
Bob E
|All times are GMT -7. The time now is 05:20 PM.


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Press release
from Urban Dharma NC
Dr. Hun Y. Lye, Founder and President of Urban Dharma NC, has had the title and position of "Dorjé Lopön" (Vajra Master) bestowed upon him by His Holiness Drikung Kyabgön Chetsang Rinpoché, the leader of the Drikung Kagyü lineage of Tibetan Buddhism at Drikung Kagyü Institute in Dehradun, India, on March 10, 2013.
This is the first instance the title of Dorjé Lopön has been given to a non-monastic teacher in the Drikung Kagyü in the West. While Dorjé Lopön is a title traditionally used by high-level monks who preside over vajrayāna ceremonies, His Holiness the Drikung Kyabgön has bestowed this title on Dr. Lye in recognition of his extensive studies and years of practice of Buddhism and in particular, Drikung Kagyü. Furthermore, this title formalizes his responsibility for passing on Drikung Kagyü teachings and transmissions that he has properly received and practiced.
On March 17, His Holiness the Drikung Kyabgön's designated regent –His Eminence Thritsab Gyabra Rinpoché – enthroned Dr. Lye at a Drikung Kagyü center in Penang, Malaysia (Dr. Lye's birthplace). This event was witnessed by a gathering of dozens of monastics of the Thai Theravada, Chinese Mahāyāna and Tibetan Vajrayāna traditions and hundreds of lay followers.
Dr. Lye's Buddhist education took place both in Asia and the West, academic as well as religious. Having grown up in Malaysia and studied under Buddhist teachers of various traditions in and from Asia, Dr. Hun Lye also has a doctoral degree in Religious Studies from the University of Virginia. As such, Dr. Lye's teaching style incorporates the best of both worlds – often offering a combination of a fresh, engaging and critical look at the material with an honest, heartfelt and personal approach.
Dr. Lye leads Urban Dharma NC, an innovative, growing Buddhist community in Asheville, NC. Urban Dharma is the first public Buddhist temple in western NC. All its programs are offered on a donation basis and it also runs an online Tibetan Buddhist supplies store (www.tibetanspirit.com). Urban Dharma offers weekly activities (secularly-oriented "Introduction to Meditation" class, classic vajrayāna deity-yoga practices and weekend service) and regular teachings and workshops by visiting teachers.
Please visit the Urban Dharma website, http://www.udharmanc.com, or the Urban Dharma Facebook page,http://urbandharmancpodcast.wordpress.com to learn more.


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Welcome to Bird Holdings Ltd.
: Our company direction is to seek out opportunities for the development of new real estate projects or redevelopment of existing buildings, while at the same time pursuing investments in other fields of business to build diversification. We have successfully applied the concept of partnership to a number of new ventures over recent years, and have found it to be an ideal combination for a healthy and productive business.
Click here to learn more about
A residential development by
Bird Lands Limited
Did you know we are closely associated with
Click here for more information
Please have a look – leasing and
development opportunities worth pursuing!
47,000 sqft commercial/industrial building
Vanier Industrial Park, Fredericton
32' ceiling height for maximum cube utilization
Climate controlled, both industrial and office space
Air system in place for pneumatic equipment
Digital security controls with infrared motion sensors
Almost 6 acres of land, and more available
See more about 40 Blizzard Street
6 acres commercial development land
McAllister Industrial Park, Saint John
Prime location adjacent Irving Oil Refinery
Excellent neighbours, Hertz and Sancton
Paved streets with concrete curbs and gutters
Fully serviced, natural gas available
Land cleared and level for immediate construction
See more about 125 McIlveen Drive
4 acres commercial development land
Doak/Alison corner, Fredericton
Prime location with direct access from Trans Canada Highway
Rapidly growing area situated between two industrial parks
Land cleared and level for immediate construction
See more about 311 Doak Road
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