The difference between belief and reality for Viktorin's method of inter-pupillary distance measurement.

The variation in the measurement of an inter-pupillary distance and monocular centration distance by 54 registered practitioners using a 'pd rule' was investigated. Their estimate of the repeatability of the measurement was also recorded, as was any correction for their own inter-pupillary distance. There was found to be considerable variation between practitioners in all measurements, and the practitioners' estimated repeatability of the measurement was rather better than the reality.

The effect of pH, temperature and plaque thickness on the hydrolysis of monofluorophosphate in experimental dental plaque.

Monofluorophosphate (MFP), an anti-caries agent commonly used in toothpaste, is known to be degraded to fluoride and orthophosphate by bacterial phosphatases in dental plaque. We have examined the effect of pH, temperature, plaque thickness and some ions on this process. Both natural plaque and artificial microcosm plaque incubated with purified MFP at pH 4-10 showed an optimum pH of approximately 8 for hydrolysis. Diffusion and concomitant hydrolysis were examined in an apparatus in which artificial plaque was held between rigid membranes separating two chambers. When MFP diffused through a plaque of 0.51-mm thickness over 4 h it was almost completely hydrolysed at pH 8, but hydrolysis on diffusion decreased as the pH deviated from 8. MFP in toothpaste extract showed a similar pH susceptibility to hydrolysis, according to the inherent pH of the toothpaste. Hydrolysis of MFP in the toothpaste was reduced by no more than 10% when compared with a matched-pH control, suggesting that other toothpaste ingredients had no major influence on hydrolysis. Transport was slower and hydrolysis at pH 6 more complete the thicker the plaque, but hydrolysis was not significantly slower at 23 degrees C than at 37 degrees C. The addition of various potential activating or inhibiting ions at 0.1 and 1.0 mmol/l had small and non-significant effects on hydrolysis. The results suggest that MFP toothpaste should be formulated and used to maximise enzymic hydrolysis of this complex anion, and that plaque pH control is probably the most important factor.

Saturation of human saliva with respect to calcium salts.

It may be assumed that free ionic concentrations of calcium and phosphate in resting saliva tend to equilibrate with those in plaque fluid, and that salivary data can therefore be used to illustrate chemical conditions in both saliva and plaque. In the present study, salivary data collected from the literature or obtained in our laboratory were used to calculate degrees of super- and undersaturation with respect to apatites, brushite, beta-tricalcium phosphate, octacalcium phosphate, calcium carbonate and calcium fluoride in the pH range from 3 to 9. Concentrations of calcium, phosphate, fluoride, carbonate, and background ion strength of resting parotid saliva, resting submandibular saliva, and resting and stimulated whole saliva were entered into a computer program, and curves illustrating saturation in the pH range 3-9 constructed. It was found that oral fluids are supersaturated with respect to apatites above pH 5.3 and with respect to octacalcium phosphate and beta-tricalcium phosphate above pH 6. Parotid saliva was undersaturated with respect to brushite whilst submandibular saliva was supersaturated with respect to that salt in the pH range 6-8. Stimulated whole saliva with 25 mmol/l carbonate became supersaturated with respect to calcium carbonate only above pH 7.3, which may explain the absence of this salt in the human oral cavity. To maintain the saturation of oral fluids with respect to calcium fluoride, i.e. to ensure its survival in the mouth required 6 ppm fluoride in the aqueous phase. Therefore, this salt, the outcome of topical fluoride therapy, will inevitably dissolve in the oral fluids.

Calcium phosphate deposition in human dental plaque microcosm biofilms induced by a ureolytic pH-rise procedure.

The objectives were to develop and characterize a procedure based on a ureolytic pH rise to deposit calcium phosphate into microcosm dental plaque biofilms and to test the importance of the plaque pH range. Plaque biofilms were cultured in a multiplaque culture system ('artificial mouth') with a continuous supply of a simulated oral fluid (basal medium mucin; BMM) with 146 mmol/l (5% w/v) sucrose periodically applied over 6 min every 8h. After initial plaque growth, the biofilms were periodically exposed for up to 16 days to 6-min applications of calcium phosphate monofluorophosphate urea (CPMU) solution containing 20 mmol/l CaCl(2), 12 mmol/l NaH(2)PO(4), 5 mmol/l monofluorophosphate and 500 mmol/l urea (pH 5.0). Three application regimes were examined, one included a sucrose-induced acidic pH fluctuation. Plaque hydrolysis of the urea in CPMU caused the pH to rise to between 8.2 and 8.8, depositing fluoridated and carbonated calcium phosphates, and possibly some calcium carbonate, into the plaque. Calcium, phosphate and fluoride deposition was rapid for about 4 days and then slowed. After 10 days' treatment under standard conditions (BMM containing 1 mmol/l urea and 1 mmol/l arginine), plaque calcium and phosphate concentrations had increased up to 50-fold and 10-fold to approximately 2-4 and 1-2 mmol/g plaque protein, respectively. The calcium, phosphate and fluoride content increased steadily. Calcium phosphate deposition was proportional to the plaque resting pH, increasing over four-fold when the BMM urea concentration was increased from 0 to 20 mmol/l, which raised the resting pH from 6.4 to 7.2 and yielded a mean plaque calcium concentration of 14.3 mmol/g protein, one subsample reaching 20.8 mmol/g protein. Supplementation of BMM with 20% human serum inhibited deposition. These results support the hypothesis that an alkaline pH in plaque is critical in promoting plaque mineralization and that mineral deposition is modulated by serum. These factors are likely to be important in regulating calculus formation.

Defluoridation of drinking water by boiling with brushite and calcite.

Existing methods for defluoridating drinking water involve expensive high technology or are slow, inefficient and/or unhygienic. A new method is now suggested, encompassing brushite and calcite suspension followed by boiling. Our aim was to examine the efficiency of the method and the chemical reactions involved. Brushite, 0.3-0.5 g, and an equal weight of calcite were suspended in 1 litre water containing 5-20 ppm fluoride. The suspensions were boiled in an electric kettle, left to cool and the calcium salts to sediment. Solution ion concentrations were determined and sediments were examined by X-ray diffraction. In distilled water initially containing 5, 10 and 20 ppm fluoride the concentration was reduced to 0.06, 0.4 and 5.9 ppm, respectively. Using Aarhus tap water which contained 2.6 mmol/l calcium the final concentrations were 1.2, 2.5 and 7.7 ppm, respectively, and runs without calcite gave results similar to those with calcite. Without boiling the fluoride concentration remained unaltered, as did the brushite and calcite salts, despite occasional agitation by hand. All solutions were supersaturated with respect to fluorapatite and hydroxyapatite and close to saturation with respect to brushite. Boiling produced well-crystallised apatite and traces of calcite, while boiling of brushite alone left a poorly crystallised apatite. We conclude that boiling a brushite/calcite suspension rapidly converts the two salts to apatite which incorporates fluoride if present in solution, and that this process may be exploited to defluoridate drinking water.

The effect of sucrose application frequency and basal nutrient conditions on the calcium and phosphate content of experimental dental plaque.

A reduced pool of calcium in dental plaque would be expected to increase the ability of plaque fluid to dissolve the underlying enamel when the pH falls during sugar exposure. We have examined the relationship between frequency of sugar application and Ca and P(i) concentrations in artificial mouth plaque microcosm biofilms. Ten plaques were grown simultaneously from a human saliva inoculum using a continuous flow of simulated saliva, DMM, supplemented with either urea or glucose to modulate the resting pH. In addition the plaques received sucrose applications of varying frequency: 12-, 8-, 6-, or 4-hourly, or not at all. After 15 days the plaques were sampled by taking 4 full-thickness specimens of each, and acid-extractable Ca and P(i), and alkali-soluble protein and carbohydrate were determined. Ca and P(i) concentrations were in a range comparable with those in human plaque, except in the DMM + urea plaque receiving no sucrose, when concentrations were higher. Plaque Ca concentration decreased significantly as sucrose application frequency increased. Increasing sucrose application frequency also reduced the protein, i.e. the cell biomass, content of the plaques and, in the case of DMM + urea plaques, increased the water-insoluble hexose content, presumably extracellular polysaccharide. Reduced biomass was partly due to the bulking of plaque with extracellular polysaccharide, but the marked effect of urea on polysaccharide formation is not understood. This study shows that increasing frequency of sugar application alters dental plaque by reducing its mineral protection capacity.

The influence of salivary variables on fluoride retention in dental plaque exposed to a mineral-enriching solution.

This study was carried out to examine interindividual differences in salivary variables related to plaque accumulation and to estimate their influence on the fluoride retention in plaque in vivo by a mineral-enriching solution. Two saliva samples were taken from 10 subjects, once after brushing and once after 24 h without brushing. Calcium, phosphate and monofluorophosphatase (MFPase) activity in the saliva samples were determined. The salivary flow rate and the debris index were also recorded. After plaque had formed over 3 days within in situ plaque-generating devices, subjects were instructed to rinse with a mineral-enriching mouthrinse three times a day on 4 consecutive days. Plaque exposed to distilled water plus flavoring agents served as a control. Fluoride-free dentifrice was used during the experimental period. Twenty-four hours after the last rinsing, the samples were removed from the mouth, and fluoride and mineral distributions in plaque analyzed using a method previously reported by the authors. Salivary flow, MFPase activity and calcium concentration in saliva were significantly higher after 24 h of plaque accumulation. Rinsing with the mineral-enriching solution produced retention of fluoride and phosphate in the outer and middle layers of plaque. Salivary calcium concentration had a direct effect on fluoride uptake in plaque, but no obvious relationship was found between other salivary variables and the plaque fluoride retention. The salivary calcium effect may be due to enhanced bacterial cell wall binding of fluoride via calcium bridging.

Inhibitory effect of ZnCl(2) on glycolysis in human oral microbes.

Although the inhibition of bacterial glycolysis by zinc ions might be expected to moderate dental caries, there has not been a comparison of the effect of Zn on different organisms under both fixed pH and free-fall conditions. Here, the effect of ZnCl(2) on Streptococcus salivarius, Strep. mutans, Strep. sobrinus, Actinomyces naeslundii and Lactobacillus casei, as well as on mixtures of oral organisms outgrown from human dental plaque and saliva, was surveyed. pH-stat experiments were performed at pH 7, 6 or 5 in a solution containing 5% glucose and a suspension of the test organism; pH-fall experiments started at pH 7. In both cases, acid production was monitored for 60 min, when samples were taken for Zn and lactate determinations. Under pH-stat conditions, acid production was inhibited by Zn most strongly in Strep. sobrinus and Strep. salivarius. In terms of total acid production averted, however, the effect of Zn under both pH-stat and pH-fall conditions was clearly greatest with Strep. salivarius. A. naeslundii was inhibited the least strongly under pH-stat conditions. Cultured oral organism mixtures were more sensitive to moderate concentrations of zinc (0.2-0.3mM initial concentration) than were the single species to higher concentrations (1mM). Packed cell layers responded to Zn quite differently from suspensions, the pH often falling in the presence of 1mM Zn at a rate similar to the no Zn control. As streptococci had the highest acidogenesis rates in both pH-stat and pH-fall experiments, it seems likely that inhibition of acid production with these organisms would be of more value in moderating caries than the inhibition of less acidogenic organisms such as A. naeslundii.

What does the phenol red thread test actually measure?

PURPOSE: This study attempts to resolve whether the phenol red thread test (PRT) is a test of tear volume or tear production through comparisons with other techniques. METHODS: Twenty asymptomatic subjects (10 men and 10 women; average age 30.6 +/- 10.8 years) had PRT (Zone Quick, Menicon) results compared with tear turnover rate (by fluorophotometry; Fluorotron Master, OcuMetrics) and tear volumes (from tear meniscus height and back extrapolation from fluorometric data). RESULTS: PRT wetting was not correlated with either tear turnover or volume (by fluorophotometry or tear meniscus height) on a Pearson product moment correlation test (p > 0.05). CONCLUSIONS: No clear experimental evidence in favor of the PRT being a measure of tear production or volume was found. It is probable that the PRT measures uptake of a (small) amount of fluid residing in the eye, stimulates a low degree of reflex tearing, and reflects the absorption characteristics of the thread dependent on the biophysics or composition of tears.

Plaque mineral induction and inhibition properties in the formation of supragingival calculus.

Several individual species of dental plaque bacteria have the ability to initiate the precipitation of calcium phosphate minerals in vitro; other plaque components have been shown to inhibit mineralisation. We have examined subjects' overall plaque mineralisation promoter and inhibitor properties, and have attempted to correlate them with supragingival calculus development over 6 months. Three-day-old plaque was collected from 22 adult subjects at the start and end of the study. To detect promoter activity, the plaque was placed in a suspension of brushite, the liquid phase of which was supersaturated with respect to hydroxyapatite. The extent of mineralisation was determined by the rise in phosphate concentration over 4 days. To detect inhibitor activity, plaque was placed in a similar suspension, which also contained hydroxyapatite. Promoter activity was compared with that hydroxyapatite, and inhibitor activity was compared with polyaspartate. The subjects' teeth were scaled at the start of the study, and calculus deposition was measured at the end using the Volpe Manhold method. Most plaque samples showed some promoter or inhibitor activity, or both, but no significant correlation existed between these activities and a subject's development of calculus. A significant inverse correlation existed between plaque mineralisation promoter activity and its inhibitor activity at the start of the study. Our results suggest that the nucleating and mineralisation inhibitory properties of young plaque will probably not be a useful target for a practical preventive methodology for supragingival calculus.

An improved fluorophotometric method for tear turnover assessment.

PURPOSE: Scanning fluorophotometry is considered the "gold standard" to determine tear turnover rates (TTR). In this study, we attempted to improve the precision of basal TTR by fluorophotometry through a number of methodological changes. METHODS: By means of a timer program that produces audio cues, a new methodology that ensures confluence and a constant thickness of the tear film and minimization of reflex lacrimation as a result of the inhibition of the blink reflex was developed. This was compared with the standard protocol, both in a paired study and by a review of unpaired data. The minimum length of time required to monitor TTR measurements was also investigated. RESULTS: A significant improvement in correlation coefficient (r) of the log decay curve was observed as a result of changes in the protocol in both the paired (p = 0.016) and unpaired study (p < 0.0001). We determined that it was sufficient to monitor TTR for 10 min to obtain accurate results. CONCLUSIONS: We advocate that the changes be adopted as widely as possible for the precise and rapid measurement of basal tear turnover rates.

Effect of oral contraceptives on tear physiology.

Variations in sex hormones due to the menstrual cycle (and oral contraceptives) were evaluated for their effect on tear physiology. Subjects were 18 females taking oral contraceptives or their age matched controls (no medication), aged 21-33 years. Symptoms of ocular discomfort (visual analog scale), tear film structure (TearScope), non-invasive tear thinning time (HirCal grid), evaporation rate (ServoMed evaporimeter), osmolality (Nanolitre Osmometer), tear turnover rate, tear volume (Fluorotron Master), and tear protein levels (HPLC) were measured around day 2 (D2), actually day 1-4 and day 19 (D19) actually day 18-21 of the cycle. No significant differences were found for any tear parameters between D2 and D19 for either pill users or controls alone, or in the comparison of pill users with controls. No effect on tear physiology was found for serum hormone changes induced by oral contraceptive use or by normal cyclic variations in healthy young females.

Spatial location studies on the chemical composition of human tear ferns.

Tear ferns are the patterns formed when tears are allowed to dry on a clean smooth surface. The patterns have been categorised according to confluence of pattern, with less confluent ferns indicative of tear film abnormalities. Little is known about the mechanism that gives rise to this phenomenon. Previous work has concentrated on correlation of alterations in tear film composition on the ferning phenomenon. The present study takes a new approach by analysing the locations of chemical elements in the fern to understand how the ferns form. By means of SEM coupled with EDXA, Na, K and Cl were detected within the fern. Sulphur, indicative of the presence of macromolecules such as mucins and proteins, was only found at the very periphery of the dry tear drop. The results suggest that macromolecules do not play a direct role in the formation of the ferns. A model is put forward to explain the observation whereby proteins and mucins are precipitated selectively by increasing salt concentration at the drop margin.

Plaque pH and associated parameters in relation to caries.

Intensified plaque acidogenicity in caries-prone subjects was reported many years ago, but emerging evidence has suggested that the relationship may not be as strong as once thought. We have now determined a range of acidogenicity variables in subjects having both caries prevalence and incidence data, and have included plaque mineral data in the analysis. pH measurements were made in 20 randomly selected subjects from a high-caries group (mean DMFS = 8. 95) and 20 from a caries-free group of Beijing children aged 12 years participating in a caries prediction study. Subgroups with a 12-month DMFS increment >/=2 or = 0 were also formed from the two groups, respectively. Measurements were made with an iridium oxide electrode inserted between teeth 13/14, 23/24, 34/35 and 44/45, before and every 5 min for 30 min after rinsing with 10% sucrose, and the 4 resulting 'Stephan curves' averaged using a plaque pH analysis program. Supragingival plaque was collected from buccal and lingual smooth surfaces of posterior and upper anterior teeth and its acid extract analysed for Ca, P and F. Caries-free subjects (based on past experience) had a significantly higher maximum plaque pH and pH value after 30 min (reflecting a faster return to resting pH), a lower minimum enamel dissolution capacity of plaque and recorded less time below pH 7.0 than did high-caries subjects. No other differences were significant, including those of the principal acidogenic parameters 'minimum pH attained after a sugar rinse', 'curve area below the critical pH of 5.5' and 'time below the critical pH'. Selection of the caries groups on the basis of both experience and incidence did not reveal significant differences in more parameters. Upper arch plaque was significantly more acidogenic than lower arch plaque, and there was a consistently strong association between upper and lower arch values in individuals. Ca, P and F in the subjects' plaque had little or no influence on the principal acidogenic parameters. Our failure to find a relationship between caries prevalence or activity and these principal acidogenicity parameters may be related to differences between fissure and smooth surface plaque, temporal variations in acidogenicity and/or to use of F toothpaste during the 1-year observation period. These results support the view that factors such as the frequency of acidogenic episodes may be more important in caries progression than the degree of acidogenicity during any one episode.

Effect of in situ plaque mineral supplementation on the state of saturation of plaque fluid during sugar-induced acidogenesis.

Dental plaque fluid is normally supersaturated with respect to enamel mineral but this may change to a state of undersaturation when plaque pH falls following sugar exposure, placing the adjacent enamel at risk of caries. We have determined the saturation status of the fluid in both resting and fermenting plaque following mineral supplementation. Eleven subjects abstained from oral hygiene and rinsed their mouth 3 times/d for 3 d with a placebo solution or with test solutions designed to enrich plaque with hydroxyapatite or fluorhydroxyapatite. On the morning of day 4, plaque samples were collected before and after exposure to 10% sucrose. Compared to the placebo, use of the test rinses resulted in significantly higher concentrations of Ca, P and F in plaque residue. In plaque fluid, higher post-sucrose Ca2+ free concentrations and saturation levels with respect to enamel mineral and fluorapatite were found after use of the hydroxyapatite rinse compared to the placebo, effects that probably resulted from the release of cell-bound Ca2+ as well as from the dissolution of apatite. Thus, some evidence was obtained that the test mouthrinses can counteract the fall in saturation level found when plaque is exposed briefly to sucrose. Potential long-term benefits of the test mouthrinses deserve further study.

Individual variations of pH, buffer capacity, and concentrations of calcium and phosphate in unstimulated whole saliva.

In order to evaluate the risk of development of dental caries and/or of formation of dental calculus, salivary variables have often been used, but not with particular success. A reason for the apparent lack of association could be that the individual temporal variation of a characteristic was so substantial relative to the overall variation that it is not possible to characterize an individual by a single salivary measurement. The aim here was to examine the individual variation of pH, buffer capacity, and concentrations of calcium and phosphate and to compare it with the overall variation of the characteristics in order to shed light on the above problem. Eight weekly samples of up to 4 ml of unstimulated whole saliva were collected from 11 dental students before tooth brushing on their arrival at 8 a.m. in the dental school. Calcium was determined by atomic absorption spectroscopy, phosphate colorimetrically, and pH electrometrically. The buffer capacity was assessed by titration of the saliva sample from the pH initially observed to pH 3. It was found that within each individual the concentration of calcium and of phosphate, pH, the hydroxyapatite ion product and the buffer capacity varied considerably over the 7 weeks. The individual range frequently covered more than a third of the total range. Further, within each of the variables, single individuals could be found whose samples covered 60% or more of the overall range, whilst others covered less than 10% of the range. It was therefore concluded that, although collected at the same time of the day, pH, buffer capacity and concentrations of calcium and phosphate in unstimulated whole saliva in the single individual vary so much that characterization of individuals and of their saliva based on a single salivary analysis is unreliable and hazardous.

Prophylactic mineral enrichment of approximal dental plaque.

Most research on the chemical modification of dental plaque has used material from buccal and lingual smooth surfaces of teeth--plaque that is readily accessible to treatment solutions. The aim of this study was to test, on the more sheltered approximal plaque, the effect of a mouthrinse previously found to be effective in raising calcium, phosphate, and fluoride concentrations in accessible plaque. Five young adults participated in a blind, cross-over trial to compare a Ca-P-MFP-urea test rinse and a NaCl placebo rinse. For 6 days, subjects rinsed twice per day after their normal toothbrushing, using 10 ml of rinse solution for 30 seconds and a fresh 10 ml for a further 30 seconds. A non-fluoride toothpaste was used and normal flossing was withheld. Plaque was collected before rinsing and again 16 hours after the final rinse from all approximal sites except those between the lower anterior teeth. Pre- and post-rinse calcium values were 291 +/- 100 and 511 +/- 107 nmol/mg protein (mean +/- SD) respectively for the test rinse, and 368 +/- 223 and 306 +/- 125 nmol/mg protein respectively for the placebo. Analysis of variance showed that the test rinse had a significant effect on plaque calcium but not on phosphate or fluoride. Mean calcium values of about 2,400 nmol/mg protein have been obtained previously in smooth-surface plaque after use of the test rinse. The reduced effect here is probably due to restricted access of the rinse solution.

Fluoride distribution in sound and carious root tissues of human teeth.

Proton probe analysis has been used to provide for the first time quantitative F concentration data in carious root tissues from subjects consuming water containing 1 ppm F. In small lesions at the neck of the tooth with minimal tissue loss the F concentration was significantly higher at the outer lesion edge than at the outer edge of adjacent sound root tissue. In one sample with high F values the lesion edge had 19,000 ppm F and the adjacent sound root surface 5,400 ppm F microg F/g apatite). In large lesions with extensive cavitation F was again concentrated in the outer edge of the lesion and was significantly higher (1,800-4,100 ppm) than in adjacent sound inner dentine (190-290 ppm). Fluoride concentrations varied markedly along the outer edge of both normal and carious root tissues. Fluoride increase at the lesion edge is not an effect of tissue shrinkage but probably a result of remineralisation events during caries. This additional F may be expected to increase tissue resistance to further acid attacks.