Viral quantitative capillary electrophoresis for counting intact viruses

The quantification of a virus plays an important role in vaccine development, clinical diagnostics, and environmental contamination assays. In all these cases, it is essential to calculate the concentration or number of intact virus particles (ivp) and estimate the degree of degradation and contamination of virus samples. In this work, we propose a cost-efficient, robust method for the quantification and characterization of intact viruses based on capillary zone electrophoresis. This separation method is demonstrated on vaccinia virus (VV) with oncolytic properties. After virus sample preparation, the solution contains intact VV as well as broken viruses and residual DNA from the host cell used for preparation. Regulatory requirements limit the amount of the host cell DNA that can be present in vaccines or human therapeutics. We apply capillary electrophoresis to separate intact virus particles and the residual DNA and to measure the level of virus contamination with DNA impurities. Intercalating YOYO-1 dye is used to detect the encapsulated and free DNA by laser-induced fluorescence. After soft lysis of VV with proteinase K, all encapsulated DNA is dissolved to the free DNA. The change in peak areas and a DNA calibration curve help determine the initial concentration of intact viruses. This viral quantitative capillary electrophoresis (Viral qCE) is able to quantify the oncolytic vaccinia virus in the range of 10(6) to 10(12) ivp/mL.     

Temporal variations in road traffic fatalities in South Africa

The annual road traffic fatality (RTF) burden of 43 deaths per 100 000 inhabitants in South Africa (SA) is disproportionately high in comparison to the world average of 22 per 100 000 population. Recent research revealed strong geographical variations across district councils in the country, as well as a substantial peak in mortality occurring during December. In this study, the factors that explain temporal variations in RTFs in SA are examined. Using weekly data from the period 2002–2006 for the country's nine provinces, non-linear auto-regression exogenous (NARX) regression models were fitted to explain variations in RTFs and to assess the degree to which the variations between the provinces were associated with the temporal variations in risk factors. Results suggest that a proportion of the variations in weekly RTFs could be explained by factors other than the size of the province population, with both temporal and between-province residual variance remaining after accounting for the modelled risks. Policies directed at reducing the effects of the modifiable risks identified in our study will be important in reducing RTFs in SA.     

Rugged, portable tungsten coil atomic emission spectrometer

Tungsten coil atomic emission spectrometry is an ideal technique for field applications because of its simplicity, low cost, low power requirement, and independence from cooling systems. A new, portable, compact design is reported here. The tungsten coil is extracted from an inexpensive 24 V, 250 W commercial light bulb. The coil is housed in a small, aluminum cell. The emission signal exits from a small aperture in the cell, while the bulk of the blackbody emission from the tungsten coil is blocked. The resulting spectra exhibit extremely low background signals. The atomization cell, a single lens, and a hand-held charge coupled device (CCD) spectrometer are fixed on a 1 × 6 × 30 cm ceramic base. The resulting system is robust and easily transported. A programmable, miniature 400 W solid-state constant current power supply controls the temperature of the coil. Fifteen elements are determined with the system (Ba, Cs, Li, Rb, Cr, Sr, Eu, Yb, Mn, Fe, Cu, Mg, V, Al, and Ga). The precision ranges from 4.3% to 8.4% relative standard deviation for repetitive measurements of the same solution. Detection limits are in the 0.04 to 1500 μg/L range. Accuracy is tested using standard reference materials for polluted water, peach leaves, and tomato leaves. For those elements present above the detection limit, recoveries range from 72% to 147%.     

Silver bullet or trojan horse? The effects of inclusionary zoning on local housing markets in the United States

Many local governments are adopting inclusionary zoning (IZ) as a means of producing affordable housing without direct public subsidies. In this paper, panel data on IZ in the San Francisco metropolitan area and suburban Boston are used to analyse how much affordable housing the programmes produce and how IZ affects the prices and production of market-rate housing. The amount of affordable housing produced under IZ has been modest and depends primarily on how long IZ has been in place. Results from suburban Boston suggest that IZ has contributed to increased housing prices and lower rates of production during periods of regional house price appreciation. In the San Francisco area, IZ also appears to increase housing prices in times of regional price appreciation, but to decrease prices during cooler regional markets. There is no evidence of a statistically significant effect of IZ on new housing development in the Bay Area.     

Compositional heterogeneity may limit the usefulness of some commercial naphthenic acids for toxicity assays

Naphthenic acids are considered variously as monocarboxylic acids fitting the formula C_nH_2n + zO₂ (where z is a zero or negative even integer), as only alicyclic (i.e. non-aromatic) monocarboxylic acids fitting this formula (z ≤ 0), or simply as those carboxylic acids occurring in petroleum products or crude oils that have been formed through biodegradation of hydrocarbons. Such acids are known constituents of the process-affected water associated with some expanding oil sands industries, of some immature and biodegraded crude oils, of produced water discharges from oil production platforms and are used as biocides and as components in the manufacture of steel radial tyres.As a result of these potential vectors of the acids into the environment, various naphthenic acid mixtures which are available commercially have been used for a range of toxicity studies. However, as some manufacturers make clear, but which is not often emphasised in the toxicity studies, a range of different quality naphthenic acids is produced commercially. It has been suggested previously, and we showed recently and elucidate further herein, that such commercial mixtures therefore sometimes contain toxic components other than carboxylic acids. For example, we identify herein by two-dimensional comprehensive gas chromatography-mass spectrometry, a range of C_0-6 alkylphenols in a batch of commercial naphthenic acids. Since these compounds are known toxicants, the contribution of such non-carboxylic acids, if any, to the toxicity attributed previously to the acids, should also be considered. This will be reflected in the concentrations and effective toxicities of such components. In order to establish the toxicity of the acids per se, assays of pure synthetic carboxylic acids of the type now known to be present in naphthenic acids from petroleum or oil sands may be more appropriate than tests of the toxicity of largely unknown, heterogeneous, mixtures.     

Mechanism of Crystallization and Implications for Charge Transport in Poly(3‐ethylhexylthiophene) Thin Films

In this work, crystallization kinetics and aggregate growth of poly(3‐ethylhexylthiophene) (P3EHT) thin films are studied as a function of film thickness. X‐ray diffraction and optical absorption show that individual aggregates and crystallites grow anisotropically and mostly along only two packing directions: the alkyl stacking and the polymer chain backbone direction. Further, it is also determined that crystallization kinetics is limited by the reorganization of polymer chains and depends strongly on the film thickness and average molecular weight. Time‐dependent, field‐effect hole mobilities in thin films reveal a percolation threshold for both low and high molecular weight P3EHT. Structural analysis reveals that charge percolation requires bridged aggregates separated by a distance of ≈2–3 nm, which is on the order of the polymer persistence length. These results thus highlight the importance of tie molecules and inter‐aggregate distance in supporting charge percolation in semiconducting polymer thin films. The study as a whole also demonstrates that P3EHT is an ideal model system for polythiophenes and should prove to be useful for future investigations into crystallization kinetics.     

Understanding Local and Macroscopic Electron Mobilities in the Fullerene Network of Conjugated Polymer‐based Solar Cells: Time‐Resolved Microwave Conductivity and Theory

The efficiency of bulk heterojunction (BHJ) organic photovoltaics is sensitive to the morphology of the fullerene network that transports electrons through the device. This sensitivity makes it difficult to distinguish the contrasting roles of local electron mobility (how easily electrons can transfer between neighboring fullerene molecules) and macroscopic electron mobility (how well‐connected is the fullerene network on device length scales) in solar cell performance. In this work, a combination of density functional theory (DFT) calculations, flash‐photolysis time‐resolved microwave conductivity (TRMC) experiments, and space‐charge‐limit current (SCLC) mobility estimates are used to examine the roles of local and macroscopic electron mobility in conjugated polymer/fullerene BHJ photovoltaics. The local mobility of different pentaaryl fullerene derivatives (so‐called ‘shuttlecock’ molecules) is similar, so that differences in solar cell efficiency and SCLC mobilities result directly from the different propensities of these molecules to self‐assemble on macroscopic length scales. These experiments and calculations also demonstrate that the local mobility of phenyl‐C₆₀ butyl methyl ester (PCBM) is an order of magnitude higher than that of other fullerene derivatives, explaining why PCBM has been the acceptor of choice for conjugated polymer BHJ devices even though it does not form an optimal macroscopic network. The DFT calculations indicate that PCBM's superior local mobility comes from the near‐spherical nature of its molecular orbitals, which allow strong electronic coupling between adjacent molecules. In combination, DFT and TRMC techniques provide a tool for screening new fullerene derivatives for good local mobility when designing new molecules that can improve on the macroscopic electron mobility offered by PCBM.     

Optical and electrical properties and phonon drag effect in low temperature TEP measurements of AgSbSe2 thin films

Polycrystalline thin films of silver antimony selenide have been deposited using a reactive evaporation technique onto an ultrasonically cleaned glass substrate at a vacuum of 10-5 torr. The preparative parameters, like substrate temperature and incident fluxes, have been properly controlled in order to get stoichiometric, good quality and reproducible thin film samples. The samples are characterized by XRD, SEM, AFM and a UV-vis-NIR spectrophotometer. The prepared sample is found to be polycrystalline in nature. From the XRD pattern, the average particle size and lattice constant are calculated. The dislocation density, strain and number of crystallites per unit area are evaluated using the average particle size. The dependence of the electrical conductivity on the temperature has also been studied and the prepared AgSbSe2 samples are semiconducting in nature. The AgSbSe2 thin films exhibited an indirect allowed optical transition with a band gap of 0.64 eV. The compound exhibits promising thermoelectric properties, a large Seebeck coefficient of 30 mV/K at 48 K due to strong phonon electron interaction. It shows a strong temperature dependence on thermoelectric properties, including the inversion of a dominant carrier type from p to n over a low temperature range 9-300 K, which is explained on the basis of a phonon drag effect.     

Influence of Texture on Taste: Insights Gained During Studies of Hardness, Juiciness, and Sweetness of Apple Fruit

Apple flesh can be considered as a living structure within which juice and tastants are encapsulated in individual cells. The breakdown of this structure and release of juice varies between hard and soft apples. We hypothesized that the texture of the fruit and the way it releases juice during chewing would therefore influence the perception of sweetness. Time‐intensity (TI) techniques were used by a trained panel to study the temporal changes in the intensity of “juiciness” and “sweetness” during consumption of apples that varied in mechanical hardness (measured by puncture test). All apple samples had a similar water content, but the pattern of juice release was markedly different. Juice was released into the mouth in a pattern of increasing and then deceasing intensity. The maximum intensity (Imax), duration of the stimulus (DUR), and area under the TI curve (AUC) were significantly higher for hard compared with soft apples. The impact of juice release on perception of sweetness was examined using apples in which the Imax and AUC for juiciness of soft apples were half the value of those for firm apples. There was no consistent difference in Imax, DUR, or AUC for sweet taste in these 2 sets of apples. We concluded that sufficient juice was released from soft and less juicy apples to ensure that sweetness perception was maximized. These results are discussed in relation to earlier studies that demonstrated that texture (thickening agents) suppresses flavor perception in solutions and juices.     

Microbiological sampling of carcasses by excision or swabbing

Groups of 25 carcasses were obtained by random selection of carcasses at the end of each of eight commercial processes for the dressing or cooling of carcasses. Samples were collected from six groups of pig or beef carcasses by excision or swabbing with sponge, gauze, or cotton wool, with one sample obtained by each of the four methods from a separate, randomly selected site on each carcass. Total aerobic counts, coliforms, and Escherichia coli from each sample were enumerated. Values for the mean log10, log10 mean, and log10 total numbers recovered were calculated for each set of total aerobic counts. Those statistics indicated that the numbers of bacteria recovered by excision or swabbing with sponge or gauze were similar, while the numbers recovered by swabbing with cotton wool were at the lower end of or below the range of the numbers recovered by the other methods. The numbers of coliforms or E. coli recovered from carcasses by sampling areas up to 100 cm2 were too few for the estimation of log mean numbers. Sampling of two groups of carcasses by swabbing with gauze indicated that each 10-fold increase in the area sampled, from 10 to 1,000 cm2, approximately doubled the number of samples from which coliforms or E. coli were recovered. Sampling of six groups of carcasses from one process indicated that the sizes of swabs and volumes of diluent used for processing swabs did not have to be increased proportionally to the area of carcass surface sampled to recover numbers of E. coli proportional to the sampled area. It therefore appears that carcass sampling techniques can be varied widely without compromising the recovery of bacteria, and that the relative efficiencies with which bacteria are recovered by different techniques can be assessed by sampling each carcass in a group of 25 by each of the methods to be compared.