Different replication characteristics of historical pseudorabies virus strains in porcine respiratory nasal mucosa explants.

Different alphaherpesviruses, including pseudorabies virus (PRV), are able to cross the basement membrane barrier in nasal respiratory epithelium. As a first step in investigating this invasion process, a detailed quantitative analysis system was set up to determine the kinetics of horizontal and vertical virus spread in nasal explants, using the virulent PRV strain 89V87. Plaque latitudes, total depths, depths measured from the basement membrane and volumes were determined at 0, 12, 24 and 36h post inoculation (pi). PRV 89V87 was found to spread in a plaquewise manner and to cross the basement membrane between 12 and 24hpi. During the 1960s-1970s, an increase in PRV virulence has been reported. To analyse potential differences in efficiency of infection and spread for different historical PRV strains, single infected cells and plaques of infected cells were quantified at 12 and 36hpi in nasal mucosa explants for seven European PRV strains, isolated in the 1960s (Becker, NIA1), the 1970s (NS374, NIA3, 75V19) and later (89V87, 00V72). All viruses were used at second passage in cell culture, except for the Becker strain, which had an unknown passage history. Older strains, Becker, NIA1 and/or NS374, showed lower numbers of primary infectious centers, lower capacity to form plaques and/or lower capacity to cross the basement membrane. The observed differences in virus-mucosa interactions may aid in understanding the virulence increase of PRV. The quantitative assay established here will be of use in unravelling the mechanism of alphaherpesvirus-mediated invasion through the basement membrane.

The compact disk as radon detector--a laboratory study of the method.

The radon absorption ability and the track etch properties of the polycarbonate material of commercial compact disks make them very useful as sensitive retrospective 222Rn detectors. The basic idea is to remove, after exposure, a surface layer that is thicker than the range of the alpha particles of the 222Rn and 220Rn progenies and to count the electrochemically etched tracks at the corresponding depths (>80 microm). The effects on the response due to differences in pressure, temperature, and humidity have been studied experimentally. The effect of the growing of 210Po after long-term exposures was also estimated. The effect of all listed factors except the temperature is either absent or restricted to maximum--about 10% for the very extreme cases. The variation of the response at 83 microm depth over the temperature interval 15-25 degrees C is +/-12% around the 20 degrees C value. The dependence of the calibration factor on the etched depth beneath the surface was studied at 4 different temperatures within the range expected indoors. The results show that the depth dependence is exponential with the parameters of the exponent also being dependent on the temperature. In practice, using the track density obtained in two or more depths beneath the compact disk's front surface, an a posteriori temperature correction could be made. By this correction it is possible to substantially reduce the bias in the results due to the unknown temperature during exposure. The results imply that by using home stored compact disks long-term retrospective 222Rn measurements could be made with an uncertainty that could be potentially better than 10%. The useful range of the method starts at about 3 Bq m(-3) (for 10 y exposure time) and appears to cover practically the whole range of indoors 222Rn concentrations.

Short-lived alpha sources of energies 6.0 MeV and 7.69 MeV for calibration purposes.

An approach for preparation of short-lived alpha sources of energy 6.0 MeV and 7.69 MeV is proposed. The sources are prepared by taking a sample of 222Rn progeny on an alpha spectrometric filter. The activities (or related parameters) of 218Po, 214Pb and 214Bi at the end of sampling are precisely determined by a reference measurement with an alpha spectrometer. Further they are used as input values to calculate with a sufficient precision the number of emitted alpha particles of any energy and at any time interval of interest. Theoretical modelling and experimental results demonstrated that such sources could be prepared with a sufficient purity. There is a potential for the number of alpha particles emitted in a given time interval to be certified with an accuracy of 1-2%.

3-D confocal microscopy of etched nuclear tracks in CR-39.

Solid State Nuclear Track Detectors (SSNTD's) are used in a wide range of applications such as Geological Dating, Environmental Sciences (radon), life Sciences (Radiobiology, Dosimetry...), as well as Nuclear and Astro-Physics. In order to be observable under a microscope, the nanometric latent damaged trails due to the slowing down of charged particles into the SSNTD have to be specifically etched. In our laboratory, we are studying this chemical action and propose models that enable simulations to be performed. In the literature, the basic model uses two distinct etch-rates that are considered constant, VB; the Bulk and VT the Track etch-rate. A little bit more sophisticated and realistic, a model with a variable track etch-rate was established, taking into account the variation of energy deposition along the particle's trajectory. Up to now, the known methods used for determining the response function of CR-39 are very time consuming and strenuous. The method we present here is based on the use of the confocal microscope, which provides three-dimensional track images. The obtained set of 3-D co-ordinates can be treated mathematically, giving, in the framework of the two etch-velocity model, the response function. With this new approach, tracks are analysed one by one; response functions are obtained for each track and can be compared to fundamental characteristics of the charged Particle-Matter interactions. Moreover, the method we propose is applied semi-automatically and could easily be automated in the near future.

Theoretical study of the relation between radon and its long-lived progeny in a room.

We present a theoretical study of the complex relation between radon and its long-lived progeny implanted in glass surfaces. The well known (extended) Jacobi room model, which is normally used to describe radon and its progeny in a room, was transformed into a two-parameter model revealing a linear correlation between long term radon exposure and surface activity due to implanted radon decay products. Furthermore, this new approach made integration into a Monte Carlo simulation possible so that the large variation of different room model parameters could be taken into account. This allowed the calculation of a probability distribution for radon exposure from the measurement of the implanted 210Po activity. The availability of a 95% confidence interval for the radon exposure is valuable in the application of retrospective radon assessment in epidemiological studies.

Single cell and tissue specific methods for evaluation of radiation and microgravity effects.

A discussion of different methods to evaluate dose/response and biological effects of ionizing radiation is given. Confocal scanning laser microscopy (CSLM) is presented as a high performing observation method for evaluating different cytological effects. Standard cytochemical techniques can be used to analyse the cell in situ with minimal disturbance of morphology and structure. If a relatively small number of cells are affected by the treatment, the use of confocal microscope observations is fast and has a better resolution than conventional fluorescence microscopy. The optical sectioning capability of the CSLM makes it possible to analyse stacks of cells on detectors up to a depth of 200 micrometer with a resolution of 0.7 micrometer. This is used to analyse single cell electrophoresis results and nuclear track analysis in poly allyl diglycol carbonate (PADC). Consecutive analysis of cells cultivated on PADC, and analysis of nuclear tracks after chemical etched tracks in the PADC, will make it possible to correlate physical dose with direct cellular effects. This is a promising method for single cell analysis and the study of the effects of ionizing radiation at low particle flux density.