Leaf water stable isotopes and water transport outside the xylem.

How water moves through leaves, and where the phase change from liquid to vapour occurs within leaves, remain largely mysterious. Some time ago, we suggested that the stable isotope composition of leaf water may contain information on transport pathways beyond the xylem, through differences in the development of gradients in enrichment within the various pathways. Subsequent testing of this suggestion provided ambiguous results and even questioned the existence of gradients in enrichment within the mesophyll. In this review, we bring together recent theoretical developments in understanding leaf water transport pathways and stable isotope theory to map a path for future work into understanding pathways of water transport and leaf water stable isotope composition. We emphasize the need for a spatially, anatomically and isotopically explicit model of leaf water transport.

In vitro validation of endovascular Doppler-derived flow rates in models of the cerebral circulation.

This study presents validation of endovascular Doppler velocimetry-based volumetric flow rate measurements conducted in a pulsatile flow loop simulating conditions in both the internal carotid and basilar artery. In vitro models of cerebral vessels, each containing an aneurysm, were fabricated from patient anatomies extracted from 3D rotational angiography. Flow velocity measurements were collected with three different experimental techniques: an endovascular Doppler wire, Particle Image Velocimetry, and a time-resolved ultrasonic flow meter. Womersley's theory of pulsatile flow in a cylindrical vessel was used to compute time-resolved volumetric flow rates from the endovascular Doppler velocity. The volumetric flow rates computed from the Doppler measurements were compared to those from the Particle Image Velocimetry profile measurements, and the direct measurements from the ultrasonic flow meter. The study establishes confidence intervals for any systematic or random errors associated with the wire-derived flow rates as benchmarked to the other two modalities. There is an approximately 10% random error in the Doppler-derived peak and time-averaged flow rates. There is a measurable uniform bias, about 15% too low, in the time-averaged Doppler-derived flow rates. There is also a small proportional bias in the peak systolic Doppler-derived flow rates. Potential sources of error are also discussed.

Nanoparticles for controlled delivery and sustained release of chlorhexidine in the oral environment.

OBJECTIVES: Chlorhexidine (CHX) is in widespread use as a topical antimicrobial agent. Within the field of oral medicine, it is used in the prevention of ventilator-associated pneumonia as well as in the treatment of oral candidosis and microbial-associated lichenoid reactions. The objective of this study was to develop a strategy for controlled, sustained topical delivery of CHX using nanoparticle technology. METHODS: Chlorhexidine was applied to hydroxyapatite, selected as a tooth analogue, as conventional CHX digluconate solutions and as aqueous suspensions of CHX hexametaphosphate nanoparticles with total CHX concentrations of 1, 2.2 and 5 mM. Soluble CHX release from the treated hydroxyapatite was monitored over a period of 7 days. RESULTS: A repeated-measures ANOVA with post hoc LSD test indicated that CHX release was 2-3× greater, and sustained for longer, when CHX was delivered as CHX hexametaphosphate nanoparticles than in aqueous solution with 2.2 and 5 mM CHX (P = 0.020 and 0.013, respectively), but there was no statistically significant difference at 1 mM CHX (P = 0.172). CONCLUSIONS: Chlorhexidine hexametaphosphate nanoparticles increased both the local dose and duration of soluble CHX delivery when applied to hydroxyapatite surfaces. This may provide a means to deliver a sustained dose of CHX with less frequent interventions.

Effects of buffering properties and undissociated acid concentration on dissolution of dental enamel in relation to pH and acid type.

To quantify the relationships between buffering properties and acid erosion and hence improve models of erosive potential of acidic drinks, a pH-stat was used to measure the rate of enamel dissolution in solutions of citric, malic and lactic acids, with pH 2.4-3.6 and with acid concentrations adjusted to give buffer capacities (ß) of 2-40 (mmol·l(-1))·pH(-1) for each pH. The corresponding undissociated acid concentrations, [HA], and titratable acidity to pH 5.5 (TA5.5) were calculated. In relation to ß, the dissolution rate and the strength of response to ß varied with acid type (lactic > malic ≥ citric) and decreased as pH increased. The patterns of variation of the dissolution rate with TA5.5 were qualitatively similar to those for ß, except that increasing pH above 2.8 had less effect on dissolution in citric and malic acids and none on dissolution in lactic acid. Variations of the dissolution rate with [HA] showed no systematic dependence on acid type but some dependence on pH. The results suggest that [HA], rather than buffering per se, is a major rate-controlling factor, probably owing to the importance of undissociated acid as a readily diffusible source of H(+) ions in maintaining near-surface dissolution within the softened layer of enamel. TA5.5 was more closely correlated with [HA] than was ß, and seems to be the preferred practical measure of buffering. The relationship between [HA] and TA5.5 differs between mono- and polybasic acids, so a separate analysis of products according to predominant acid type could improve multivariate models of erosive potential.

The effect of monoalkyl phosphates and fluoride on dissolution of hydroxyapatite, and interactions with saliva.

The aims were to investigate the effect of monoalkyl phosphates (MAPs) and fluoride on dissolution rate of native and saliva-coated hydroxyapatite (HA). Fluoride at 300 mg/l (as NaF) inhibited dissolution of native HA by 12%, while potassium and sodium dodecyl phosphates (PDP, SDP), at 0.1% or higher, inhibited dissolution by 26-34%. MAPs, but not fluoride, also showed persistence of action. MAPs at 0.5% and fluoride at 300 mg/l were then tested separately against HA pre-treated with human saliva for 2 or 18 h. Agents were applied with brushing to half the specimens, and without brushing to the other half. In control (water-treated) specimens, pre-treatment of HA with human saliva reduced dissolution rate on average by 41% (2 h) and 63% (18 h). Brushing did not have a statistically significant effect on dissolution rate of saliva-coated specimens. In brushed specimens, fluoride significantly increased the inhibition due to 2- or 18-hour saliva pre-treatment. It is hypothesised that brushing partially removes the salivary film and allows KOH-soluble calcium fluoride formation at the surfaces of HA particles. Inhibition was reduced by PDP in 2-hour/non-brushed specimens and in 18-hour/brushed specimens. PDP did not affect dissolution rates in the remaining groups and SDP did not affect dissolution rate in any group. Possible reasons for these variable results are discussed. The experiments show that pre-treatment with saliva can significantly modify results of tests on potential anti-erosive agents and it is recommended that saliva pre-treatment should be a routine part of testing such agents.

Inhibition of erosive dissolution by sodium fluoride: evidence for a dose-response.

OBJECTIVES: The aim of this study was to investigate the protective effects of sodium fluoride solutions and commercial mouthrinses on hydroxyapatite (HA) dissolution in citric acid in vitro, with and without a salivary pellicle. METHODS: A rapid-throughput HA solubility-reduction model was employed in which HA dissolution was quantified using ion chromatography. Two HA substrates were selected, a high-resolution powder and 80 µm diameter beads, and studied in the presence and absence of a salivary pellicle (pooled human saliva, 2 h). Immediately prior to acid exposure, substrates were exposed to one of a number of pre-treatments that included aqueous fluoride (F(-)) solutions and commercially available mouthrinses with F(-) concentrations of 0-450 µg/g (as NaF). Dissolution reduction was calculated relative to a deionised water negative control. RESULTS: For aqueous solutions and mouthrinses, a fluoride dose-response was observed with a plateau around 100 µg/g F(-) for both HA substrates, with or without pellicle. Concentrations as low as 10 µg/g F(-) significantly reduced HA dissolution. The HA substrate had little impact on the fluoride dose-response, and the fluoride was equally effective in the presence of a pellicle as in its absence. CONCLUSIONS: Fluoride significantly reduced HA dissolution at concentrations of 10 µg/g and higher. A fluoride dose-response was seen at low concentrations. This study illustrates the use of a powerful rapid-throughput HA solubility-reduction model for investigating HA dissolution in citric acid in the presence of dissolution inhibitors. CLINICAL SIGNIFICANCE: A single exposure to fluoride solutions with fluoride concentrations and exposure time representative of brushing or rinsing with mainstream oral care products was shown to significantly inhibit HA dissolution under conditions relevant to dental erosion. A similar efficacy was observed in the presence and absence of salivary pellicle.

Screening and prediction of erosive potential.

The literature on the erosive potential of drinks and other products is summarised, and aspects of the conduct of screening tests as well as possible correlations of the erosive potential with various solution parameters are discussed. The solution parameters that have been suggested as important include pH, acid concentration (with respect to buffer capacity and concentration of undissociated acid), degree of saturation, calcium and phosphate concentrations, and inhibitors of erosion. Based on the available data, it is concluded that the dominant factor in erosion is pH. The effect of buffer capacity seems to be pH dependent. The degree of saturation probably has a non-linear relationship with erosion. While calcium at elevated concentrations is known to reduce erosion effectively, it is not known whether it is important at naturally occurring concentrations. Fluoride at naturally occurring concentrations is inversely correlated with erosive potential, but phosphate is probably not. Natural plant gums, notably pectin, do not inhibit erosion, so they are unlikely to interfere with the prediction of erosive potential. The non-linearity of some solution factors and interactions with pH need to be taken into account when developing multivariate models for predicting the erosive potential of different solutions. Finally, the erosive potential of solutions towards enamel and dentine might differ.

Inhibition of dental erosion by casein and casein-derived proteins.

The application of milk-derived proteins such as casein as anti-erosion agents in oral healthcare products is of current interest. The aim of this study was to investigate the potential of 3 commercially available, milk-derived proteins as agents to inhibit enamel erosion. Aqueous solutions of 0.5% w/v casein, casein phosphopeptide (CPP) or glycomacropeptide (GMP) with and without 300 ppm fluoride (F, as NaF) were investigated with regard to enamel softening and tissue loss, in comparison with a deionised water (DIW) negative control and 300 ppm F positive control. Casein and F reduced enamel surface softening compared to DIW, but CPP and GMP did not (DIW: 58.2% reduction in hardness; F: 13.3%; casein: 21.8%; CPP: 50.8%; GMP: 62.4%). Similar results were obtained with solutions containing protein and F, and the effects were statistically indistinguishable from protein alone (casein + F: 19.1%; CPP + F: 48.2%; GMP + F: 66.1%). By contrast, all protein solutions and F significantly reduced tissue loss (p < 0.050; DIW: 25.8 µm tissue loss; F: 21.6 µm; casein: 20.3 µm; CPP: 20.5 µm; GMP: 20.0 µm). Solutions containing protein and F reduced erosion more than protein alone, but this difference was only significant from protein alone for casein (casein + F: 12.2 µm; CPP + F: 17.3 µm; GMP + F: 18.2 µm). Casein and casein-derived proteins may therefore have the potential to act as agents to reduce or prevent enamel erosion. Furthermore, the erosion-reducing efficacy is not reduced by F, and is in some cases enhanced.

Enamel erosion in dietary acids: inhibition by food proteins in vitro.

The aim of this study was to investigate the effects of two common food proteins on human enamel erosion in vitro. Erosion was measured by non-contact profilometry in citric, malic and lactic acids at pH 2.8, 3.2 and 3.8 and five commercially available soft drinks, in the presence of a salivary pellicle. Whole milk casein or hen egg ovalbumin was added to the acid solutions and drinks at 0.2% w/v, and the effect on erosion was determined by comparison with the corresponding solution without protein. Casein significantly reduced erosion in all but two solutions. The effects of the individual subfractions of casein in citric acid at pH 3.2 were similar to that of whole casein. Ovalbumin reduced erosion in some solutions, but the magnitude of the reduction was less than that with casein. A greater proportional reduction in erosion was seen in citric acid than in malic or lactic acids. We postulate that the mechanism involves adsorption of proteins to the pellicle or the enamel surface, forming a protein film with enhanced erosion-inhibiting properties. The citrate ion may play an active stabilising role, since erosion reduction was less in the other acids. In conclusion, casein and, to a lesser extent, ovalbumin show promise as potential anti-erosive additives to drinks.

Effects of pH and acid concentration on erosive dissolution of enamel, dentine, and compressed hydroxyapatite.

The aims of this study were to determine the effects of pH and acid concentration on the dissolution of enamel, dentine, and compressed hydroxyapatite (HA) in citric acid solutions (15.6 and 52.1 mmol l(-1) ; pH 2.45, 3.2, and 3.9), using a pH-stat system. After an initial adjustment period, the dissolution rates of enamel and HA were constant, while that of dentine decreased with time. The dissolution rate increased as the pH decreased, and this was most marked for enamel. To compare substrates, the rate of mineral dissolution was normalized to the area occupied by mineral at the specimen surface. For a given acid concentration, the normalized dissolution rate of HA was always less than that for either dentine or enamel. The dissolution rate for dentine mineral was similar to that for enamel at pH 2.45 and greater at pH 3.2 and pH 3.9. The concentration of acid significantly affected the enamel dissolution rate at pH 2.45 and pH 3.2, but not at pH 3.9, and did not significantly affect the dissolution rates of dentine or HA at any pH. The variation in response of the dissolution rate to acid concentration/buffer capacity with respect to pH and tissue type might complicate attempts to predict erosive potential from solution composition.

A single-substrate model to interpret intra-annual stable isotope signals in tree-ring cellulose.

The carbon and oxygen stable isotope composition of wood cellulose (delta(13)C(cellulose) and delta(18)O(cellulose), respectively) reveal well-defined seasonal variations that contain valuable records of past climate, leaf gas exchange and carbon allocation dynamics within the trees. Here, we present a single-substrate model for wood growth to interpret seasonal isotopic signals collected in an even-aged maritime pine plantation growing in South-west France, where climate, soil and flux variables were also monitored. Observed seasonal patterns in delta(13)C(cellulose) and delta(18)O(cellulose) were different between years and individuals, and mostly captured by the model, suggesting that the single-substrate hypothesis is a good approximation for tree ring studies on Pinus pinaster, at least for the environmental conditions covered by this study. A sensitivity analysis revealed that the model was mostly affected by five isotopic discrimination factors and two leaf gas-exchange parameters. Modelled early wood signals were also very sensitive to the date when cell wall thickening begins (t(wt)). Our model could therefore be used to reconstruct t(wt) time series and improve our understanding of how climate influences this key parameter of xylogenesis.

3D surface imaging in dentistry - what we are looking at.

3D imaging has been widely used within various fields of dentistry to aid diagnosis, in treatment planning and appliance construction. Whereas traditionally this has involved the use of impression materials together with plaster or stone models, modern techniques are continually evolving which use virtual 3D images. These electronic virtual images are created using either contact or non-contact optical scanning techniques, but there are limitations, the most important of which is that any new virtual surface image is created from a series of discrete data points. It is not created from a continuous stream of data relating to the original object. This means that computer software has to be used to recreate a possible best fit, virtual surface from the data obtained. This paper describes the principles behind 3D scanning technology, the limitations of 3D imaging as well as current and possible uses of such imaging in clinical dentistry.

Inhibition of hydroxyapatite dissolution by ovalbumin as a function of pH, calcium concentration, protein concentration and acid type.

Hydroxyapatite is the main constituent of the dental hard tissues, and in vivo its dissolution in acids leads to the pathological condition of dental erosion. Food proteins which inhibit hydroxyapatite dissolution may find application as erosion-reducing agents in food and drink products. The aim of this study was to investigate the egg protein ovalbumin as a potential inhibitor of hydroxyapatite dissolution in acidic solutions, with conditions representative of dental erosion. The dissolution rate of hydroxyapatite discs was measured in an acidic solution as a function of pH, calcium concentration, ovalbumin concentration and acid type. All experiments were performed in triplicate. 0.2% w/v ovalbumin significantly reduced the dissolution rate in citric acid by 50-75% over the pH range 2.80-4.00, and by 45-60% in solutions with calcium concentrations of up to 20 mM (p < 0.05). The effect was persistent for several rinses after the initial exposure to the protein. 0.02% w/v ovalbumin significantly reduced the dissolution of hydroxyapatite in citric acid by 30-55%. Ovalbumin did not, however, statistically significantly reduce the hydroxyapatite dissolution rate in malic or lactic acids. The effect is ascribed to adsorption and partial, reversible denaturation of ovalbumin on the hydroxyapatite surface. There may be some interaction between ovalbumin and the citrate ion which promotes the adsorption of protein in the presence of citric acid. Ovalbumin shows promise as a potential erosion-reducing additive to citrus-based drinks.

Erosion of enamel by non-carbonated soft drinks with and without toothbrushing abrasion.

OBJECTIVE: To investigate how enamel loss due to erosion, and due to cycling of erosion and abrasion, depends on compositional parameters of soft drinks, and particularly whether the thickness of the erosive softened layer is a function of drink composition. SETTING: University dental hospital research laboratory in the UK, 2004. MATERIALS AND METHODS: Six drinks were chosen based on their popularity and composition: apple juice, orange juice, apple drink, orange drink, cranberry drink and 'ToothKind' blackcurrant drink. Group A samples (n = 36) were exposed to soft drinks at 36 degrees C for six consecutive 10 minute periods. Group B samples (n = 36) were subjected to alternating erosion and toothbrushing, repeated six times. Enamel loss was measured using optical profilometry. RESULTS: Group A: significant enamel loss was seen for all drinks (p < 0.001). Erosion was correlated with pH and calcium concentration but not phosphate concentration or titratable acidity. Group B: significant additional material loss due to toothbrush abrasion occurred with all drinks. Abrasive enamel loss differed between the drinks and was positively correlated with drink erosive potential. CONCLUSION: Enamel loss by erosion is exacerbated by subsequent abrasion. The amount of softened enamel removed by toothbrushing is a function of the chemical composition of the erosive medium.

An in vitro investigation of the effect of some analgesics on human enamel.

The sale of over-the-counter pain relief medication has increased dramatically in recent years, and typically amounts to several hundred thousands of pounds per year in the UK. Many soluble analgesic preparations contain citric acid, and it has been suggested that these formulations may cause dental erosion. The aim of this study was to investigate the effect of some over-the-counter analgesics on tooth surface loss from human enamel. Six commonly available analgesics were chosen for this study and the effect of immersing unerupted human enamel was examined using non-contact optical profilometry. Two of the six analgesics investigated caused no detectable erosion (Boots soluble aspirin and Anadin Extra). Three caused statistically significant enamel erosion, but this was very slight and is thought to be clinically insignificant (Alka Seltzer, Panadol and Solpadeine). Only one analgesic caused possible potentially clinical significant enamel erosion. Further studies are needed to determine whether Aspro causes clinically significant enamel erosion.

Incidence of enterohemorrhagic Escherichia coli, Escherichia coli O157, Salmonella, and Listeria monocytogenes in retail fresh ground beef, sprouts, and mushrooms.

The objective of this study was to determine the prevalence of enterohemorrhagic Escherichia coli (EHEC), E. coli O157, Salmonella, and Listeria monocytogenes in retail food samples from Seattle, Wash. A total of 2,050 samples of ground beef (1,750 samples), mushrooms (100 samples), and sprouts (200 samples) were collected over a 12-month period and analyzed for the presence of these pathogens. PCR assays, followed by culture confirmation were used to determine the presence or absence of each organism. Of the 1,750 ground beef samples analyzed, 61 (3.5%) were positive for EHEC, and 20 (1.1%) of these were positive for E. coli O157. Salmonella was present in 67 (3.8%) of the 1,750 ground beef samples. Of 512 ground beef samples analyzed, 18 (3.5%) were positive for L. monocytogenes. EHEC was found in 12 (6.0%) of the 200 sprout samples, and 3 (1.5%) of these yielded E. coli O157. Of the 200 total sprout samples, 14 (7.0%) were positive for Salmonella and none were positive for L. monocytogenes. Among the 100 mushroom samples, 4 (4.0%) were positive for EHEC but none of these 4 samples were positive for E. coli O157. Salmonella was detected in 5 (5.0%) of the mushroom samples, and L. monocytogenes was found in 1 (1.0%) of the samples.

The relationship between enamel softening and erosion caused by soft drinks at a range of temperatures.

OBJECTIVES: Investigations of the erosive potential of soft drinks are usually performed at room or body temperature, but drinks are more frequently served chilled, with ice, or hot. Since the rate of chemical reactions usually increases with temperature, it is predicted that erosion is more severe at high temperatures and reduced at low temperatures. The aim of this study was to investigate the correlation between enamel softening, enamel erosion, and temperature. METHODS: Atomic force microscopy nanoindentation and non-contact optical profilometry were used to assess changes in enamel nanomechanical properties after 5 min and erosive material loss after 30 min exposure to two different non-carbonated soft drinks at 4, 25, 50 and 75 degrees C. RESULTS: For one drink (Robinson's Original Juice Drink), there was a statistically significant difference between nanomechanical properties and erosion depth at all temperatures, with softening and erosion increasing with temperature. For another drink (Ribena ToothKind Juice Drink), there was a slight softening and virtually no material loss, and temperature had no statistically significant impact on erosion. There was a good linear correlation (R2 = 0.94) between nanomechanical properties and material loss. CONCLUSIONS: The difference between the drinks can be explained by their composition. For the erosive drink, material loss increased, and nanohardness decreased, approximately linearly with temperature. The correlation between softening and erosion demonstrated that nanomechanical properties after very short erosion times can be considered a good predictor of bulk material loss after considerably longer erosion times.

Components of ecosystem evaporation in a temperate coniferous rainforest, with canopy transpiration scaled using sapwood density.

Here we develop and test a method to scale sap velocity measurements from individual trees to canopy transpiration (E(c)) in a low-productivity, old-growth rainforest dominated by the conifer Dacrydium cupressinum. Further, E(c) as a component of the ecosystem water balance is quantified in relation to forest floor evaporation rates and measurements of ecosystem evaporation using eddy covariance (E(eco)) in conditions when the canopy was dry and partly wet. Thermal dissipation probes were used to measure sap velocity of individual trees, and scaled to transpiration at the canopy level by dividing trees into classes based on sapwood density and canopy position (sheltered or exposed). When compared with ecosystem eddy covariance measurements, E(c) accounted for 51% of E(eco) on dry days, and 22% of E(eco) on wet days. Low transpiration rates, and significant contributions to E(eco) from wet canopy evaporation and understorey transpiration (35%) and forest floor evaporation (25%), were attributable to the unique characteristics of the forest: in particular, high rainfall, low leaf area index, low stomatal conductance and low productivity associated with severe nutrient limitation.

Human enamel erosion in constant composition citric acid solutions as a function of degree of saturation with respect to hydroxyapatite.

The objective of this study was to investigate human enamel erosion under constant composition conditions, as a function of solution degree of saturation (DS) with respect to hydroxyapatite. The experimental conditions were relevant to the initial stages of enamel erosion by soft drinks. Nanoindentation was used to compare enamel surface softening caused by a control mineral water and two citric acid solutions with DS = 0.000 and DS = 0.032, both having pH 3.30. Enamel hardness and reduced elastic modulus were measured after 0, 30, 60, 120, 300 and 600 s exposure. A statistically significant change in enamel hardness was detected after 30 s exposure to both citric acid solutions, indicating that nanoindentation is extremely sensitive to the initial stages of erosion. There was a statistically significant difference between the mechanical properties of enamel exposed to the two citric acid solutions after 30, 60 and 120 s. At these times, the solution with DS = 0.000 caused twice as much enamel softening as that with DS = 0.032. This demonstrates that it may be possible to design a soft drink with a low erosive potential and a good taste by a small change in DS, at a typical drink pH.