Effect of calcium ions on in vitro pellicle formation from parotid and whole saliva.

The salivary pellicle is a protein-rich, bacteria-free, self-assembling film that adsorbs to all surfaces within the oral cavity. The pellicle has numerous functions that are vital for maintaining oral health. Currently however, there are no commercially available artificial salivas that accurately mimic the complex film forming properties (i.e. film thickness and viscoelasticity) of human saliva. To understand these properties further we have examined the in vitro formation of the salivary pellicle, by adsorbing stimulated parotid saliva (PS) and whole mouth saliva (WMS) from 14 healthy volunteers, onto oxidised silicon surfaces, using a quartz crystal microbalance with dissipation monitoring (QCMD) and a dual polarisation interferometer (DPI). A dramatic impact on the hydrated mass, polymer mass, thickness and polymer concentration of the pellicle for both WMS and PS was observed when the natural calcium concentration of the respective salivas was increased from 0 mM to 10mM. In addition, QCMD data showed that on addition of 10mM calcium the salivary pellicle formed by both PS and WMS became more predominantly elastic. The results presented here also suggest that calcium can easily diffuse in and out of the pellicle, permitting free calcium exchange between the saliva and the adsorbed pellicle under physiological conditions, which may potentially facilitate the mineralisation of enamel.

Kinetic models of guanidine hydrochloride-induced curing of the yeast [PSI+] prion.

A population of [PSI(+)] Saccharomyces cerevisiae cells can be cured of the [PSI(+)] prion by the addition of guanidine hydrochloride (GdnHCl). In this paper we extend existing nucleated polymerisation simulation models to investigate the mechanisms that might underlie curing. Our results are consistent with the belief that prions are dispersed through the cells at division following GdnHCl addition. A key feature of the simulation model is that the probability that a polymer is transmitted from mother to daughter during cell division is dependent upon the length of the polymer. The model is able to reproduce the essential features of data from several different experimental protocols involving addition and removal of GdnHCl.

The effect of physiological conditions on the surface structure of proteins: setting the scene for human digestion of emulsions.

Understanding and manipulating the interfacial mechanisms that control human digestion of food emulsions is a crucial step towards improved control of dietary intake. This article reports initial studies on the effects of the physiological conditions within the stomach on the properties of the film formed by the milk protein (ß-lactoglobulin) at the air-water interface. Atomic force microscopy (AFM), surface tension and surface rheology techniques were used to visualize and examine the effect of gastric conditions on the network structure. The effects of changes in temperature, pH and ionic strength on a preformed interfacial structure were characterized in order to simulate the actual digestion process. Changes in ionic strength had little effect on the surface properties. In isolation, acidification reduced both the dilatational and the surface shear modulus, mainly due to strong repulsive electrostatic interactions within the surface layer and raising the temperature to body temperature accelerated the rearrangements within the surface layer, resulting in a decrease of the dilatational response and an increase of surface pressure. Together pH and temperature display an unexpected synergism, independent of the ionic strength. Thus, exposure of a pre-formed interfacial ß-lactoglobulin film to simulated gastric conditions reduced the surface dilatational modulus and surface shear moduli. This is attributed to a weakening of the surface network in which the surface rearrangements of the protein prior to exposure to gastric conditions might play a crucial role.

Immunisation status of dental practice staff in Kent.

AIM: To determine the hepatitis B, tuberculosis (TB), varicella and rubella immunisation status of dental practice workers in Kent. METHOD: A cross-sectional survey using a) a dental practice questionnaire sent to all 275 registered dental practices in Kent in February 2005, to determine the numbers of staff employed and their job titles, and b) a confidential personal health questionnaire for every staff member employed by each practice, to determine past history of infections and immunisation history. RESULTS: Two hundred out of 257 (78%) dental practices took part in the survey, and 1,415 staff (76% of known participants) returned completed personal health questionnaires. Three hundred and eighty-four out of 395 dentists (97%) indicated previous immunisation against hepatitis B. The corresponding percentages for other occupational groups were dental hygienists (94%), nurses (89%), dental therapists (75%), and other non-clinical staff (65%). 1,197 (85%) of participants reported previous chicken pox and/or shingles; 1,208 (85%) gave a history of previous immunisation against TB; and 823 (58%) had either had rubella or were immunised against rubella. Male participants were less likely to have had rubella immunisation. CONCLUSIONS: The study has demonstrated the variations in knowledge about personal immunity status amongst dental practice staff for some infectious diseases. Improvement in establishing personal immunity status of individual dental care workers and provision of a vaccination programme could be facilitated. This preventive measure could be arranged through occupational health providers.

Approximations for expected generation number.

A deterministic formula is commonly used to approximate the expected generation number of a population of growing cells. However, this can give misleading results because it does not allow for natural variation in the times that individual cells take to reproduce. Here we present more accurate approximations for both symmetric and asymmetric cell division. Based on the first two moments of the generation time distribution, these approximations are also robust. We illustrate the improved approximations using data that arise from monitoring individual yeast cells under a microscope and also demonstrate how the approximations can be used when such detailed data are not available.

New approximations to the Malthusian parameter.

Approximations to the Malthusian parameter of an age-dependent branching process are obtained in terms of the moments of the lifetime distribution, by exploiting a link with renewal theory. In several examples, the new approximations are more accurate than those currently in use, even when based on only the first two moments. The new approximations are extended to include a form of asymmetric cell division that occurs in some species of yeast. When used for inference, the new approximations are shown to have high efficiency.

Single-locus gametophytic incompatibility in autotetraploids.

It is known that a single-locus gametophytic self-incompatibility (GSI) system can persist with just two distinct alleles in an autotetraploid population, in contrast to diploid GSI systems, assuming "competitive interaction" in which heteroallelic pollen is universally compatible. The steady-state population structure of a GSI system in autotetraploids was investigated in an undivided population assuming "competitive interaction." A deterministic model was developed to predict the frequencies of genotypes with two, three, or four distinct S alleles, assuming no mutation or population subdivision. The model showed that unlike in diploid GSI systems, the limiting values of the frequencies of genotype classes do not minimize pollen wastage.

Disruption of viscoelastic beta-lactoglobulin surface layers at the air-water interface by nonionic polymeric surfactants.

Nonequilibrium interfacial layers formed by competitive adsorption of beta-lactoglobulin and the nonionic triblock copolymer PEO99-PPO65-PEO99 (F127) to the air-water interface were investigated in order to explain the influence of polymeric surfactants on protein film surface rheology and foam stability. Surface dilatational and shear rheological methods, surface tension measurements, dynamic thin-film measurements, diffusion measurements (from fluorescence recovery after photo bleaching), and determinations of foam stability were used as methods. The high surface viscoelasticity, both the shear and dilatational, of the protein films was significantly reduced by coadsorption of polymeric surfactant. The drainage rate of single thin films, in the presence of beta-lactoglobulin, increased with the amount of added F127, but equilibrium F127 films were found to be thicker than beta-lactoglobulin films, even at low concentration of the polymeric surfactant. It is concluded that the effect of the nonionic triblock copolymer on the interfacial rheology of beta-lactoglobulin layers is similar to that of low molecular weight surfactants. They differ however in that F127 increases the thickness of thin liquid films. In addition, the significant destabilizing effect of low molecular weight surfactants on protein foams is not found in the investigated system. This is explained as due to long-range steric forces starting to stabilize the foam films at low concentrations of F127.

Estimating the number of prions in yeast cells.

Certain yeast cells contain proteins that behave like the mammalian prion PrP and are called yeast prions. The yeast prion protein Sup35p can exist in one of two stable forms, giving rise to phenotypes [PSI(+)] and [psi(-)]. If the chemical guanidine hydrochloride (GdnHCl) is added to a culture of growing [PSI(+)] cells, the proportion of [PSI(+)] cells decreases over time. This process is called curing and is due to a failure to propagate the prion form of Sup35p. We describe how curing can be modelled, and improve upon previous models for the underlying processes of cell division and prion segregation; the new model allows for asymmetric cell division and unequal prion segregation. We conclude by outlining plans for future experimentation and modelling.

Modelling the establishment and spread of autotetraploid plants in a spatially heterogeneous environment.

The establishment and spread of autotetraploids from an original diploid population in a heterogeneous environment were studied using a stochastic simulation model. Specifically, we investigated the effects of heterogeneous habitats and nonrandom pollen/seed dispersal on the critical value (micro) of unreduced 2n gamete production necessary for the establishment of autotetraploids as predicted by deterministic models. Introduction of a heterogeneous environment with random pollen/seed dispersal had little effect on the micro value. In contrast, incorporating nonrandom pollen/seed dispersal into a homogeneous environment considerably reduced the micro value. Incorporating both heterogeneous habitats and nonrandom pollen/seed dispersal may lead either to an increase or to a decrease in the micro value compared to that with random dispersal, indicating that the two factors interact in a complex way.

Models for yeast prions.

The cytoplasmic heritable determinant [PSI+] of the yeast Saccharomyces cerevisiae exhibits prion-like properties. The properties of yeast prions are studied in the hope that this will enhance the understanding of mammalian prions, which cause mad-cow, Creutzfeldt-Jakob, and related neurodegenerative diseases. When host cells divide, the yeast prions distribute themselves without loss over the daughter cells. Experimental data provide information on how the proportion of cells with prions decreases over time when priori replication is inhibited. One feature of scientific interest is the unknown mean number, n0, of prions assumed to be present in the cells at the start of the experiment. We develop several stochastic models and by fitting them to the data, we obtain substantially larger estimates of n0 compared with a previous analysis. An interesting feature of a model with constant cell generation times is that the predicted proportion of cells with prions varies over time as a sequence of linked hyperbolic curves. Avenues for future research are outlined, which relax simplifying assumptions made in the models. We make several recommendations for the design of future experiments.

In situ measurement of the displacement of protein films from the air/water interface by surfactant.

The displacement of spread protein films from the air/water interface by surfactant was followed using Brewster angle microscopy (BAM) and interfacial rheology. The displacement of beta-lactoglobulin and beta-casein by a nonionic surfactant was monitored as a function of both surface pressure and time. In both cases, protein displacement occurred over the same surface pressure range that had been observed previously by atomic force microscopy (AFM). In the case of the beta-lactoglobulin, surfactant domains grew large enough in the protein film to be visible in the BAM images. The shapes of the domains were very similar to those seen previously by AFM in the late stages of displacement. The results from both proteins confirm the results published previously while highlighting some implications for the application of the "orogenic" model of displacement for large protein films. The surface rheological data showed that the beta-lactoglobulin/surfactant mixed film retained much of its elasticity until the latter stages of displacement. This indicates that at least in the early stages of displacement, the mixed film was dominated by the behavior of the protein in the film.

The effect of peptide-pectin interactions on the gelation behaviour of a plant cell wall pectin.

The effect of basic peptides on the gelation of a pectin from the cell wall of tomato was examined through the determination of gel stiffness, and swelling behaviour of the gel in water. Poly-L-lysine, poly-L-arginine, and a synthetic peptide, designed to mimic a sequence of basic amino acids found in a plant cell wall extensin, act as crosslinking agents. Circular dichroism studies on the interaction of synthetic extensin peptides with sodium polygalacturonate demonstrated that a conformational change was induced as a result of their complexation. In addition to their effect as crosslinking agents, the polycationic peptides reduced the swelling of the pectin network in water.

Polylink: to support two-point linkage analysis in autotetraploids.

SUMMARY: Polylink runs under Microsoft Windows (95 or later). It performs various calculations that are useful for investigating two-point linkage analysis for autopolyploids, based on the random chromosome pairing model. These include calculation of offspring phenotypic probabilities as functions of the recombination fraction, calculation of theoretical standard errors for the maximum likelihood estimator of and numerical computation of maximum likelihood estimates. It also includes simulation facilities. AVAILABILITY: Polylink is free and available from Xiangming Xu via email

A score test for testing a zero-inflated Poisson regression model against zero-inflated negative binomial alternatives.

Count data often show a higher incidence of zero counts than would be expected if the data were Poisson distributed. Zero-inflated Poisson regression models are a useful class of models for such data, but parameter estimates may be seriously biased if the nonzero counts are overdispersed in relation to the Poisson distribution. We therefore provide a score test for testing zero-inflated Poisson regression models against zero-inflated negative binomial alternatives.

Analysis of segregation data from selfed progeny of allopolyploids.

This paper discusses the inference of parental genotype based on segregation data from selfed progeny of allopolyploids when there is incomplete information about genotypes and when alleles are codominant or null. The distinct alleles that are present in a genotype are assumed to be known, but not the frequency with which they occur. These assumptions may be appropriate when genotypes are deduced from DNA or protein banding patterns on electrophoretic gels. A computer program, SELF, is described that can generate all possible parental genotypes and rank them on the basis of their agreement with the progeny data. The program caters for tetraploids, hexaploids and octoploids. The methods are illustrated using data from a study of the inheritance of isoenzymes in selfed progeny of octoploid strawberry cultivars.

Cation permeability and selectivity of a root plasma membrane calcium channel.

Calcium channels in the plasma membrane of root cells fulfill both nutritional and signaling roles. The permeability of these channels to different cations determines the magnitude of their cation conductances, their effects on cell membrane potential and their contribution to cation toxicities. The selectivity of the rca channel, a Ca2+-permeable channel from the plasma membrane of wheat (Triticum aestivum L.) roots, was studied following its incorporation into planar lipid bilayers. The permeation of K+, Na+, Ca2+ and Mg2+ through the pore of the rca channel was modeled. It was assumed that cations permeated in single file through a pore with three energy barriers and two ion-binding sites. Differences in permeation between divalent and monovalent cations were attributed largely to the affinity of the ion binding sites. The model suggested that significant negative surface charge was present in the vestibules to the pore and that the pore could accommodate two cations simultaneously, which repelled each other strongly. The pore structure of the rca channel appeared to differ from that of L-type calcium channels from animal cell membranes since its ion binding sites had a lower affinity for divalent cations. The model adequately accounted for the diverse permeation phenomena observed for the rca channel. It described the apparent submillimolar Km for the relationship between unitary conductance and Ca2+ activity, the differences in selectivity sequences obtained from measurements of conductance and permeability ratios, the changes in relative cation permeabilities with solution ionic composition, and the complex effects of Ca2+ on K+ and Na+ currents through the channel. Having established the adequacy of the model, it was used to predict the unitary currents that would be observed under the ionic conditions employed in patch-clamp experiments and to demonstrate the high selectivity of the rca channel for Ca2+ influx under physiological conditions.

Improved estimation of the proportion of triploids in populations with diploid and triploid individuals.

We consider the estimation of the proportion of triploids in populations of plants or animals in which diploid and triploid individuals coexist, using data from electrophoretic analysis of isozyme or microsatellite markers. Individuals that have three distinct alleles at a locus are unambiguously triploid. However, other individuals cannot be classified with certainty as diploid or triploid, unless allelic dosage can be determined reliably. This is impossible for microsatellite markers, and for many isozyme markers. We therefore present a maximum likelihood method of estimating the proportion of triploids based only on the presence or absence of different alleles.

Effects of quadrat size and shape, initial epidemic conditions, and spore dispersal gradient on spatial statistics of plant disease epidemics.

ABSTRACT The spatiotemporal spread of plant diseases was simulated using a stochastic model to study the effects of initial conditions (number of plants initially infected and their spatial pattern), spore dispersal gradient, and size and shape of sampling quadrats on statistics describing the spatiotemporal dynamics of epidemics. The spatial spread of disease was simulated using a half-Cauchy distribution with median dispersal distance mu (units of distance). A total of 54 different quadrat types, including 23 distinct sizes ranging from 4 to 144 plants, were used to sample the simulated epidemics. A symmetric form of the binary power law with two parameters (alpha, beta) was fitted to the sampled epidemic data using each of the 54 quadrats for each replicate simulation run. The alpha and beta estimates were highly correlated positively with each other, and their estimates were comparable to those estimated from observed epidemics. Intraclass correlation (kappa) was calculated for each quadrat type; kappa decreased exponentially with increasing quadrat size. An asymmetric form of the binary power law with three parameters (alpha (1), beta(1), beta(2)) was used to relate kappa to the disease incidence (p); beta1 was highly correlated to beta: beta1 approximately beta - 1. In general, initial conditions and quadrat size affected alpha, beta, alpha(1), beta(1), and beta(2) greatly. The parameter estimates increased as quadrat size increased, and the relationships were described well by a linear regression model on the logarithm of quadrat size with the slope or intercept parameters dependent on initial conditions and mu. Compared with initial conditions and quadrat size, the overall effects of mu and quadrat shape were generally small, although within each quadrat size and initial condition they could be substantial. Quadrat shape had the greatest effect when the quadrat was long and thin. The relationship of the index of dispersion (D) to p and quadrat size was determined from the alpha and beta estimates. D was greatest when p was 0.5 and decreased when p approached 0 or 1. It increased with quadrat size and the rate of the increase was maximum when p was 0.5 and decreased when p approached 0 or 1.

Relationships between several quadrat-based statistical measures used to characterize spatial aspects of disease incidence data.

ABSTRACT This article investigates the relationships between various statistical measures that are used to summarize spatial aspects of disease incidence data. The focus is on quadrat data in which each plant in a quadrat is classified as diseased or healthy. We show that spatial autocorrelation plays a central role via the mean intraclass correlation, rho, which is defined as the average correlation of the disease status of all pairs of plants within the quadrat. The value of rho determines the variance of the number of infected plants in the quadrat and, if this variable follows a beta-binomial distribution, the heterogeneity parameter of the beta-binomial distribution is directly related to the mean intraclass correlation. We consider in detail a model in which the spatial autocorrelation depends only on the distance between the plants. For illustration, we consider a specific autocorrelation model that was derived from simulated data. We show that this model leads, approximately, to the binary form of the power law relating the variance of the number of infected plants per quadrat to the mean. Using an approximation technique, we then show how the index of dispersion is related to quadrat size and shape. The index of dispersion increases with quadrat size. The rate of increase is dependent on quadrat shape, but the effect of quadrat shape is small in comparison to the effect of quadrat size. Finally, we note that if the spatial autocorrelation depends on the relative orientation of the plants, as well as the distance between them, there are connections with distance class methods.