Low Particle Number Research Articles

Influence of minor displacements in loops of the porcine parvovirus VP2 capsid on virus-like particles assembly and the induction of antibody responses

An antigen-delivery system based on hybrid virus-like particles (VLPs) formed by the self-assembly of the capsid VP2 protein of porcine parvovirus (PPV) and expressing foreign peptides offers an alternative method for vaccination. In this study, the three-dimensional structure of the PPV capsid protein and surface loops deletion mutants were analyzed to define essential domains in PPV VP2 for the assembly of VLPs. Electron microscopic analysis and SDS-PAGE analysis confirmed the presence of abundant VLPs in a loop2 deletion mutant of expected size and appropriate morphology. Loop4 and loop2-loop4 deletion mutants, however, resulted in a lower number of particles and the morphology of the particles was not well preserved. Furthermore, the green fluorescent protein (gfp) gene was used as a model. GFP was observed at the same level in displacements mutants. However, GFP displacement mutants in loop2 construct allowed better adaptation for the fusion GFP to be further displayed on the surface of the capsid-like structure. Immunogenicity study showed that there is no obvious difference in mice inoculated with rAd-VP2(Δloop2), rAd-VP2(Δloop4), rAd-VP2(Δloop2-Δloop4), and PPV inactivated vaccine. The results suggested the possibility of inserting simultaneously B and T cell epitopes in the surface loop2 and the N-terminus. The combination of different types of epitopes (B, CD4+, and CD8+) in different positions of the PPV particles opens the way to the development of highly efficient vaccines, able to stimulate at the same time the different branches of the immune system.

Water-based condensation particle counters comparison near a major freeway with significant heavy-duty diesel traffic

This study compares the instrumental performance of three TSI water-based condensation particle counter (WCPC) models measuring particle number concentrations in close proximity (15 m) to a major freeway that has a significant level of heavy-duty diesel traffic. The study focuses on examining instrument biases and performance differences by different WCPC models under realistic field operational conditions. Three TSI models (3781, 3783, and 3785) were operated for one month in triplicate (nine units in total) in parallel with two sets of Scanning Mobility Particle Sizer (SMPS) spectrometers for the concurrent measurement of particle size distributions. Inter-model bias under different wind directions were first evaluated using 1-min raw data. Although all three WCPC models agreed well in upwind conditions (lower particle number concentrations, in the range of 103–104 particles cm−3), the three models' responses were significantly different under downwind conditions (higher particle number concentrations, above 104 particles cm−3). In an effort to increase inter-model linear correlations, we evaluated the results of using longer averaging time intervals. An averaging time of at least 15 min was found to achieve R2 values of 0.96 or higher when comparing all three models. Similar results were observed for intra-model comparisons for each of the three models. This strong linear relationship helped identify instrument bias related to particle number concentrations and particle size distributions. The TSI 3783 produced the highest particle counts, followed by TSI 3785, which reported 11% lower during downwind conditions than TSI 3783. TSI 3781 recorded particle number concentrations that were 24% lower than those observed by TSI 3783 during downwind condition. We found that TSI 3781 underestimated particles with a count median diameter less than 45 nm. Although the particle size dependency of instrument performance was found the most significant in TSI 3781, both models 3783 and 3785 showed somewhat size dependency. In addition, within each tested WCPC model, one unit was found to count significantly different and be more sensitive to particle size than the other two. Finally, exponential regression analysis was used to numerically quantify instrumental inter-model bias. Correction equations are proposed to adjust the TSI 3781 and 3785 data to the most recent model TSI 3783.

IMPACTS OF AMBIENT TEMPERATURE AND PRESSURE ON PM2.5 EMISSION PROFILES OF LIGHT-DUTY DIESEL VEHICLES

The impact of the environmental factors on the emissions of particulate matter (PM) number, size distribution and mass size distribution from diesel passenger cars was evaluated. Particle measurements from five modern light-duty diesel vehicles (LDDV) were performed in June and November 2011. Commercial low sulfur diesel fuel (less than 50 ppm) was used during the testing of these vehicles which were not equipped with after-treatment devices. The dynamometer test was based on the Economic Commission of Europe (ECE) 15 cycles. The results indicate that PM2.5 emissions from LDDV are significantly affected by ambient temperature and pressure. A comparison of the emissions concentration of PM2.5 in these two different months showed that the number concentration in June was (3.8 ± 0.69) × 107 cm-3 and (2.5 ± 0.66) × 107 cm-3 in November. The PM concentration of about 30 nm diameter was 25 ± 6% of the total emissions in November while only 14 ± 3% of total emissions in June. In the 60 nm to 2.5 μm test range, November data shows less of a contribution for number than data from June testing. The concentration of mass emissions in June was (325 ± 44) mg/m3 and (92 ± 30) mg/m3 in November. The contribution of the number of PM particles in November testing is lower than testing in June by 34% and the mass concentration in November is 70% lower than that in June. With the decrease of ambient temperature and the increase of ambient pressure, both the oxygen concentration in cylinder and air–fuel ratio are increased, which caused lower particle number and mass emissions during November testing. The size distribution is also altered by these changes: the more efficient in-cylinder combustion resulted in a higher proportion of particles in the 30 nm and smaller range than for other particle sizes.

Stochastic simulations of pattern formation in excitable media.

We present a method for mesoscopic, dynamic Monte Carlo simulations of pattern formation in excitable reaction–diffusion systems. Using a two-level parallelization approach, our simulations cover the whole range of the parameter space, from the noise-dominated low-particle number regime to the quasi-deterministic high-particle number limit. Three qualitatively different case studies are performed that stand exemplary for the wide variety of excitable systems. We present mesoscopic stochastic simulations of the Gray-Scott model, of a simplified model for intracellular Ca oscillations and, for the first time, of the Oregonator model. We achieve simulations with up to particles. The software and the model files are freely available and researchers can use the models to reproduce our results or adapt and refine them for further exploration.

Levels of black carbon and their relationship with particle number levels—observation at an urban roadside in Taipei City

Information on the relationship between black carbon (BC) and particle number levels in urban areas is limited. Therefore, investigating the relationship between BC and particle number levels in different particle size ranges at an urban area is worthwhile. This study used an aethalometer and scanning mobility particle sizer to measure the levels of BC and particle number simultaneously at an urban roadside in Taipei City. Measurement results show that hourly BC levels are 0.62-8.80 μg m(-3) (mean=3.50 μg m(-3)) and hourly particle number levels are 4.21 × 10(3)-4.64 × 10(4) particles cm(-3) (mean=2.00 × 10(4) particles cm(-3)) in Taipei urban area. The BC and particle number levels peak during morning (7:00-9:00) and evening (16:00-18:00) rush hours on weekdays. Low BC and particle number levels exist in the early morning hours. Time variations in BC levels are the same as those of particle number levels in this study, clearly indicating that BC and particles are likely released from the same emission source. Additionally, BC levels in the urban area are more strongly associated with ultrafine particle levels than with total particle number levels, particularly in the size range of 56-180 nm. According to measurement results, most BC in aerosols in urban areas can be in the ultrafine size range.

Vegetable oils as core of cationic polymeric nanocapsules: influence on the physicochemical properties

Vegetable oils might be alternatives to mineral or synthetic oils used in nanostructured systems for cutaneous application, due to their advantages with regard to skin care and protection. In this study, we propose the use of vegetable oils (Brazil nut, sunflower seed, olive, rose hip, grape seed and carrot oils) as oily core of Eudragit RS100® nanocapsules and determine their influence on the physicochemical properties of those nanoparticles, in comparison with nanocapsules with capric/caprylic triglycerides as oily core. The formulations containing vegetable oils as core presented pH values suitable for topical application, average diameter close to 280 nm (SPAN around 2.5) and zeta potential close to +7 mV, due to the cationic properties of the polymer. Their viscosities were not affected by the type of oil used as core. By means of multiple light scattering, a reversible particle creaming phenomenon was observed for all the formulations. The nanocapsules prepared using Brazil nut, sunflower seed, olive, grape seed, rose-hip and carrot oils presented some distinct physicochemical properties when compared to nanocapsules obtained with capric/caprylic triglycerides: a higher size and SPAN value, a lower number of particles and a higher tendency to reversible creaming. Those findings are probably related to the lower density and higher viscosity of the vegetable oils.

Age-Dependent TLR3 Expression of the Intestinal Epithelium Contributes to Rotavirus Susceptibility

Rotavirus is a major cause of diarrhea worldwide and exhibits a pronounced small intestinal epithelial cell (IEC) tropism. Both human infants and neonatal mice are highly susceptible, whereas adult individuals remain asymptomatic and shed only low numbers of viral particles. Here we investigated age-dependent mechanisms of the intestinal epithelial innate immune response to rotavirus infection in an oral mouse infection model. Expression of the innate immune receptor for viral dsRNA, Toll-like receptor (Tlr) 3 was low in the epithelium of suckling mice but strongly increased during the postnatal period inversely correlating with rotavirus susceptibility, viral shedding and histological damage. Adult mice deficient in Tlr3 (Tlr3−/−) or the adaptor molecule Trif (TrifLps2/Lps2) exerted significantly higher viral shedding and decreased epithelial expression of proinflammatory and antiviral genes as compared to wild-type animals. In contrast, neonatal mice deficient in Tlr3 or Trif did not display impaired cell stimulation or enhanced rotavirus susceptibility. Using chimeric mice, a major contribution of the non-hematopoietic cell compartment in the Trif-mediated antiviral host response was detected in adult animals. Finally, a significant age-dependent increase of TLR3 expression was also detected in human small intestinal biopsies. Thus, upregulation of epithelial TLR3 expression during infancy might contribute to the age-dependent susceptibility to rotavirus infection.

Subhaloes going Notts: the subhalo-finder comparison project

We present a detailed comparison of the substructure properties of a single Milky Way sized dark matter halo from the Aquarius suite at five different resolutions, as identified by a variety of different (sub-)halo finders for simulations of cosmic structure formation. These finders span a wide range of techniques and methodologies to extract and quantify substructures within a larger non-homogeneous background density (e.g. a host halo). This includes real-space, phase-space, velocity-space and time- space based finders, as well as finders employing a Voronoi tessellation, friends-of-friends techniques, or refined meshes as the starting point for locating substructure.A common post-processing pipeline was used to uniformly analyse the particle lists provided by each finder. We extract quantitative and comparable measures for the subhaloes, primarily focusing on mass and the peak of the rotation curve for this particular study. We find that all of the finders agree extremely well on the presence and location of substructure and even for properties relating to the inner part part of the subhalo (e.g. the maximum value of the rotation curve). For properties that rely on particles near the outer edge of the subhalo the agreement is at around the 20 per cent level. We find that basic properties (mass, maximum circular velocity) of a subhalo can be reliably recovered if the subhalo contains more than 100 particles although its presence can be reliably inferred for a lower particle number limit of 20. We finally note that the logarithmic slope of the subhalo cumulative number count is remarkably consistent and <1 for all the finders that reached high resolution. If correct, this would indicate that the larger and more massive, respectively, substructures are the most dynamically interesting and that higher levels of the (sub-)subhalo hierarchy become progressively less important.

Competition between pairing and deformation in finite Fermi systems

The problem of the possible deformation of finite Fermi systems with particle numbers of ≤104 is studied within the Bardeen-Cooper-Schrieffer model. It is shown that for slight deformations of a spherical system consisting of one valence l shell (l ≫ 1), the results of the quasi-particle theory and the exact Richard- son theory virtually coincide. Analysis of the equilibrium deformation of a cluster shows that the system does not keep its spherical form even at a low number of valence particles.

Measurement of the Particle Velocity in a HVOF Spray with PIV Under Industrial Conditions

The particle velocity and the particle temperature are two of the most important parameters influencing the deposition process of high-velocity oxy-fuel (HVOF) spraying. This article addresses the application of particle image velocimetry to the measurement of the 2D particle velocity distribution in the flame of an industrial burner in a spraying booth under industrial conditions. Therefore, the measurement equipment has to be protected from overspray. Moreover, the low particle number density of HVOF sprays requires a technique called average correlation for image evaluation. The effect of the variation of process parameters, such as oxygen flow rate and mass loading of the flame, on the particle velocity is discussed.

In search of proof‐of‐concept: gene therapy for glycogen storage disease type Ia

The emergence of life threatening long-term complications in glycogen storage disease type Ia (GSD-Ia) has emphasized the need for new therapies, such as gene therapy, which could achieve biochemical correction of glucose-6-phosphatase deficiency and reverse clinical involvement. We have developed gene therapy with a novel adeno-associated virus (AAV) vector that: 1) prevented mortality and corrected glycogen storage in the liver, 2) corrected hypoglycemia during fasting, and 3) achieved efficacy with a low number of vector particles in G6Pase-deficient mice and dogs. However, the gradual loss of transgene expression from episomal AAV vector genomes eventually necessitated the administration of a different pseudotype of the AAV vector to sustain dogs with GSD-Ia. Further preclinical development of AAV vector-mediated gene therapy is therefore warranted in GSD-Ia.

Error Bound for Piecewise Deterministic Processes Modeling Stochastic Reaction Systems

Biological processes involving the random interaction of $d$ species with integer particle numbers are often modeled by a Markov jump process on $\mathbb{N}_{0}^{d}$. Realizations of this process can, in principle, be generated with Gillespie's classical stochastic simulation algorithm, but for very reactive systems this method is usually inefficient. Hybrid models based on piecewise deterministic processes offer an attractive alternative which can decrease the simulation time considerably in applications where species with rather low particle numbers interact with very abundant species. We investigate the convergence of the hybrid model to the original one for a class of reaction systems with two distinct scales. Our main result is an error bound which states that, under suitable assumptions, the hybrid model approximates the marginal distribution of the discrete species and the conditional moments of the continuous species up to an error of $\mathcal{O}\!\left(M^{-1}\right)$, where $M$ is the scaling pa...

An analysis of direct-injection spark-ignition (DISI) soot morphology

Abstract We have characterized particle emissions produced by a 4-cylinder, 2.0 L DISI engine using transmission electron microscopy (TEM) and image analysis. Analyses of soot morphology provide insight to particle formation mechanisms and strategies for prevention. Particle emissions generated by two fueling strategies were investigated, early injection and injection modified for low particle number concentration emissions. A blend of 20% ethanol and 80% emissions certification gasoline was used for the study given the likelihood of increased ethanol content in widely available fuel. In total, about 200 particles and 3000 primary soot spherules were individually measured. For the fuel injection strategy which produced low particle number concentration emissions, we found a prevalence of single solid sub-25 nm particles and fractal-like aggregates. The modal diameter of single solid particles and aggregate primary particles was between 10 and 15 nm. Solid particles as small as 6 nm were present. Although nanoparticle aggregates had fractal-like morphology similar to diesel soot, the average primary particle diameter per aggregate had a much wider range that spanned from 7 to 60 nm. For the early fuel injection strategy, liquid droplets were prevalent, and the modal average primary particle diameter was between 20 and 25 nm. The presence of liquid droplets may have been the result of unburned fuel and/or lubricating oil originating from fuel impingement on the piston or cylinder wall; the larger modal aggregate primary particle diameter suggests greater fuel-rich zones in-cylinder than for the low particle number concentration point. However, both conditions produced aggregates with a wide range of primary particle diameters, which indicates heterogeneous fuel and air mixing.

Effect of aerosol-radiation feedback on regional air quality – A case study with WRF/Chem

Numerical simulations were performed in order to investigate the impact of the direct effect of aerosol particles on radiation and the indirect aerosol effect on meteorological variables and subsequent distributions of near surface ozone and PM10 over Europe. The fully coupled meteorology-chemistry community model WRF/Chem has been applied for June and July 2006 for a baseline case without any aerosol feedback on meteorology, a simulation with the direct effect included, and a simulation including the direct as well as the indirect aerosol effect. The impact of the subsequent changes in temperature, boundary layer height, and clouds that were triggered by the direct effect of aerosol on radiation (“semi-direct effect”) was found to dominate the direct effect of aerosol particles on solar radiation. Over Central Europe the mean reduction of global radiation alone was mostly 3–7 W m −2 , but changes in cloud cover due to semi-direct effects resulted in monthly mean changes between ±50 W m −2 . The inclusion of the indirect aerosol effect resulted in a pronounced decrease of cloud water content by up to 70% and a significantly higher mean rain water content over the North Atlantic. Although generally plausible, the effect appears to be too strong due to too low simulated aerosol particle numbers in this area. Regional changes in precipitation between −100% and 100% were simulated over the European continent. For the simulation including only the direct aerosol effect these changes are almost entirely due to semi-direct effects. Mean ozone mixing ratios over Europe in July were modified by up to 4 ppb or 10% over continental Europe, mostly related to changes in cloud cover. For PM10 the inclusion of the direct effect resulted for the considered episode in a mean decrease by 20–50% due to an increased atmospheric boundary layer height except for the regions with high PM10 concentrations. When the indirect aerosol effect was additionally taken into account an increase of the monthly PM10 concentration by 1–3 μg m −3 was found for July 2006 over large parts of continental Europe.

A novel poisson distribution-based approach for testing boundaries of real-time PCR assays for food pathogen quantification.

The validation of quantitative real-time PCR systems and above all, proof of the detection limit of this method, is a frequently and intensively discussed topic in food pathogen detection. Among proper sample collection, assay design, careful experimental design, execution of real-time PCR, and data analysis, the validation of the method per se ensuring reliable quantification data is of prime importance. The purpose of this study was to evaluate a novel validation tool for real-time PCR assays, based on the theoretical possibility of the amplification of a single DNA target. The underlying mathematical basis for the work is Poisson distribution, which describes patterns of low particle numbers in a volume. In this context, we focused on the quantitative aspect of real-time PCR for the first time. This allowed for demonstration of the reliable amplification of a lone target DNA molecule and the demonstration of the distinct discrimination between integer molecular numbers when using low initial copy numbers. A real-time PCR assay amplifying a 274-bp fragment of the positive regulatory protein A locus of Listeria monocytogenes was used for this work. Evidence for a linear range of quantification from a single target copy to 10 ng of target DNA was experimentally demonstrated, and evidence for the significance of this novel validation approach is presented here.

Extension of the cumulant definition for low particle number systems

AbstractCumulants are usually interpreted as the connected components of density matrices, but this interpretation fails and practical problems arise when the rank n of cumulants is larger than the number of particles (N) in the system. In that case, cumulants defined in the traditional way become disconnected. To solve this problem, the definition of cumulants is extended by introducing a simple modulation factor. The modified cumulants reduce to the conventional definition, but they vanish when N &lt; n. Using the modified definition also eliminates the error in the approximation of density matrices by low‐rank cumulants, when N &lt; n. The problem assumes a slightly different form when we work with active space–based theories, and it can be solved by a similar approach. Another problem with cumulants, due to spin coupling [Herbert, Int J Quantum Chem 2007, 107, 703], can be solved via the introduction of a similar modulation factor. A related yet more serious issue, termed as the local particle number constraint problem, is also discussed. © 2010 Wiley Periodicals, Inc. Int J Quantum Chem, 2011

Targeted Vault Nanoparticles Engineered with an Endosomolytic Peptide Deliver Biomolecules to the Cytoplasm

Vault nanoparticles were engineered to enhance their escape from the endosomal compartment by fusing a membrane lytic peptide derived from adenovirus protein VI (pVI) to the N-terminus of the major vault protein to form pVI-vaults. We demonstrate that these pVI-vaults disrupt the endosomal membrane using three different experimental protocols including (1) enhancement of DNA transfection, (2) co-delivery of a cytosolic ribotoxin, and (3) direct visualization by fluorescence. Furthermore, direct targeting of vaults to specific cell surface epidermal growth factor receptors led to enhanced cellular uptake and efficient delivery of vaults to the cytoplasm. This process was monitored with fluorescent vaults, and morphological changes in the endosomal compartment were observed. By combining targeting and endosomal escape into a single recombinant vault, high levels of transfection efficiency were achieved using low numbers of vault particles. These results demonstrate that engineered vaults are effective, efficient, and nontoxic nanoparticles for targeted delivery of biomaterials to the cell cytoplasm.

Aerosol Behavior in the near Wake Region of the Vehicle

In the present study, the interaction effects of different vehicle speeds and vehicular exhaust tailpipe exit velocity conditions on the exhaust particle dynamic behaviors in the near-wake region behind the studied vehicle were numerically investigated. The results showed that the particles were drawn up into the recirculation region and then moved toward into the upper vortex by its lower vortex. A lower particle number concentration behind the studied vehicle is observed for a higher vehicle speed because of the rapidly increasing dilution ratio and the nucleation rate. The particle volume concentration is the highest in the recirculation region behind the studied vehicle and is diluted gradually beyond the recirculation region in the downstream.

TOF-SIMS analysis of exhaled particles from patients with asthma and healthy controls

Particles in exhaled air (PEx) may reflect the composition of respiratory tract lining fluid (RTLF); thus, there is a need to assess their potential as sources of biomarkers for respiratory diseases. In the present study, we compared PEx from patients with asthma and controls using time-of-flight-secondary ion mass spectrometry (TOF-SIMS) and multivariate analysis. Particles were collected using an instrument developed in-house. 15 nonsmoking subjects with physician-diagnosed asthma and 11 nonsmoking healthy controls performed 10 consecutive forced exhalations into the instrument. Particle concentrations were recorded and samples of particles collected on silicon plates were analysed by TOF-SIMS. Subjects with asthma exhaled significantly lower numbers of particles than controls (p=0.03) and the ratio of unsaturated to saturated phospholipids was significantly lower in samples from subjects with asthma (0.25 versus 0.35; p=0.036). Orthogonal partial least squares-discriminant analysis models showed good separation between both positive and negative spectra. Molecular ions from phosphatidylcholine and phosphatidylglycerol, and protein fragments were found to discriminate the groups. We conclude that analysis of PEx is a promising method to examine the composition of RTLF. In the present explorative study, we could discriminate between subjects with asthma and healthy controls based on TOF-SIMS spectra from PEx.

Comparisons of particulate matter, CO, and CO2 levels in underground and ground-level stations in the Taipei mass rapid transit system

As of 2010, the Taipei mass rapid transit (MRT) comprised 8 rapid transit lines stretching 90.5 km and linking 82 stations. Currently, over 1.4 million commuters a day travel on the Taipei MRT system. However, information related to the characteristics of indoor air quality at ground-level and in underground stations in the Taipei MRT system is limited. This study used portable monitors to measure the PM 10 , PM 2.5 , particle number (PN), CO, and CO 2 levels at an underground (UG) and two ground-level (AG1 and AG2) MRT stations on the same rapid transit line. All measurements were taken between August 20 and November 12, 2010. The mean PM 10 and PM 2.5 levels in underground environments (underground ticket hall and platform of the UG and underground platform of the AG1) are approximately 51–65 μg m −3 and 25–40 μg m −3 , respectively. Additionally, the mean PM 10 and PM 2.5 levels in ground-level indoor environments (ground-level ticket halls of the AG1 and AG2) are approximately 44–48 μg m −3 and 26–29 μg m −3 , respectively. Measurement results reveal that PM levels in underground environments are greater than those in ground-level indoor environments. PM levels in ground-level indoor environments are strongly associated with outdoor ambient PM levels, while PM levels in underground environments are less influenced by outdoor ambient conditions. The presence of coarse PM in the underground environments may have been generated internally through the mechanical friction processes or re-suspended by the movement of commuters, and fine PM is likely transferred from its source outside the station. The mean PM 2.5 -to-PM 10 ratios in underground environments are 0.45–0.79, and at ground-level indoor environments are approximately 0.58. Based on differences in the environment conditions among these selected locations, it is reasonable to assume that PM 2.5 -to-PM 10 ratios in underground and ground-level MRT stations will not be consistent, even those on the same metro line. Furthermore, low PN and CO levels are observed in underground environments. Mean PN and CO levels in underground environments are 9.5 × 10 3 –1.7 × 10 4 particles cm −3 and 0.30–0.48 ppm, respectively, and significantly lower than those in outdoor environments by approximately 0.3–0.6 times. However, the CO 2 levels in underground environments are 574–1051 ppm and significantly higher than those in outdoor environments, by approximately 1.3–2.4 times. The CO 2 in underground environments is likely elevated by the exhalation of commuters. ► Coarse PM in the underground environments may have been generated internally. ► Fine PM is likely transferred from its source outside the station. ► Low PN and CO levels are observed in underground environments.